Artículos de revistas sobre el tema "New York Free Circulating Library"

In 1996, its centenary year, the New York Public Library opened its Science, Industry and Business Library (SIBL) in a former department store in mid-town Manhattan, occupying 160,000 square feet of usable floor space. The building, which has received six awards, is designed to be both attractive and highly functional. The $100 million project was funded by a combination of private and government funds. The concept is of a specialized high technology research centre with unparallelled older and current print collections (1.2 million books and serials) and access to electronic resources, which also incorporates a 50,000-item circulating library of popular print, audiovisual and multimedia materials. All of the resources are available to the public at no charge. Much of the collection is on open access. There are several professionally staffed information service points. The provision of extensive training sessions is proving to be an outstanding success, more than 20,000 people having registered since SIBL opened. A three-year grant from the W.K. Kellogg Foundation has enabled SIBL to train information professionals in the three crucial areas of technological competence, customer service and professional development.

This article discusses commuter students’ experiences with the academic library, drawn from a qualitative study at the City University of New York. Undergraduates at six community and baccalaureate colleges were interviewed to explore how they fit schoolwork into their days, and the challenges and opportunities they encountered. Students identified physical and environmental features that informed their ability to successfully engage in academic work in the library. They valued the library as a distraction-free place for academic work, in contrast to the constraints they experienced in other places—including in their homes and on the commute.

The New York Times has been a leader in metadata since the birth of The New York Times Index in 1913. Today, a team of taxonomists uses natural-language processing and a human-in-the-loop model to organize, analyze, and push content to readers. Looking to the future, the taxonomy team hopes to free up resources with the help of new technologies so that the metadata practices already in place can be expanded to other areas beyond core news content.

Circulating free fatty acids (FFAs) may worsen heart failure (HF) due to myocardial lipotoxicity and impaired energy generation. We studied cardiac and whole body effects of 28 days of suppression of circulating FFAs with acipimox in patients with chronic HF. In a randomized double-blind crossover design, 24 HF patients with ischemic heart disease [left ventricular ejection fraction: 26 ± 2%; New York Heart Association classes II ( n = 13) and III ( n = 5)] received 28 days of acipimox treatment (250 mg, 4 times/day) and placebo. Left ventricular ejection fraction, diastolic function, tissue-Doppler regional myocardial function, exercise capacity, noninvasive cardiac index, NH2-terminal pro-brain natriuretic peptide (NT-pro-BNP), and whole body metabolic parameters were measured. Eighteen patients were included for analysis. FFAs were reduced by 27% in the acipimox-treated group [acipimox vs. placebo ( day 28 − day 0): −0.10 ± 0.03 vs. +0.01 ± 0.03 mmol/l, P < 0.01]. Glucose and insulin levels did not change. Acipimox tended to increase glucose and decrease lipid utilization rates at the whole body level and significantly changed the effect of insulin on substrate utilization. The hyperinsulinemic euglycemic clamp M value did not differ. Global and regional myocardial function did not differ. Exercise capacity, cardiac index, systemic vascular resistance, and NT-pro-BNP were not affected by treatment. In conclusion, acipimox caused minor changes in whole body metabolism and decreased the FFA supply, but a long-term reduction in circulating FFAs with acipimox did not change systolic or diastolic cardiac function or exercise capacity in patients with HF.

Reindeer (Rangifer tarandus tarandus) are important livestock for arctic and subarctic herders, including those in North America, but as climate change affects traditional herding practices, alternative methods of rearing (such as captive rearing) will likely become common. Proper nutrition is critical in livestock production, but there is minimal information available on circulating nutrient concentrations in reindeer, who are adapted to a unique climate. This study looks at 2 important antioxidants. Blood and serum were taken from female reindeer from three herds: a free-ranging herd from the Seward Peninsula, Alaska (AK), during the summer, and two captive herds (one in Fairbanks, AK and one in Upstate New York (NY) during the summer and winter. Selenium (Se) and vitamin E concentrations were described stratified on season (when possible), location, and management practices (captive or free range). Herd mean values across seasons for Se ranged from 2.42 to 4.88 µmol/L. Herd mean values across seasons for vitamin E ranged from 5.27 to 6.89 µmol/L.

Purpose – The purpose of this paper is to highlight the challenges to born-digital institutional archiving using a New York Archive Museum (NYAM) as a case. Design/methodology/approach – The digital record-keeping practices at NYAM were studied using three data sources: focus groups with staff, totaling 81 individuals, or approximately one-third of all staff; analysis of network file storage; and analysis of digital records in archival storage, or specifically removable media in acid-free archive boxes. Findings – This case study indicates that the greatest challenges to born-digital institutional archiving are not necessarily technological but social and cultural. Or rather, the challenge is getting individuals to transfer material to a digital archive so that it can undergo the technological transformations needed to ensure its long-term availability. However, transfer is impeded by a variety of factors which can be addressed through education, infrastructure development and proactive appraisal for permanent retention. Practical implications – This paper highlights the challenges to born-digital institutional archiving, yet notes that these challenges can be overcome by following a multi-pronged approach. Originality/Value – This paper outlines the challenges to born-digital institutional archiving, which is not often discussed in the literature outside of the context of higher education.

Introduction: The analysis of cell-free DNA (cfDNA) for genetic abnormalities is a promising new approach for the diagnosis and prognosis of pancreatic cancer patients. Insights into the molecular characteristics of pancreatic cancer may provide valuable information, leading to its earlier detection and the development of targeted therapies. Material and Methods: We conducted a systematic review and a meta-analysis of studies that reported cfDNA in pancreatic ductal adenocarcinoma (PDAC). The studies were considered eligible if they included patients with PDAC, if they had blood tests for cfDNA/ctDNA, and if they analyzed the prognostic value of cfDNA/ctDNA for patients’ survival. The studies published before 22 October 2020 were identified through the PubMED, EMBASE, Web of Science and Cochrane Library databases. The assessed outcomes were the overall (OS) and progression-free survival (PFS), expressed as the log hazard ratio (HR) and standard error (SE). The summary of the HR effect size was estimated by pooling the individual trial results using the Review Manager, version 5.3, Cochrane Collaboration. The heterogeneity was assessed using the Cochran Q test and I2 statistic. Results: In total, 48 studies were included in the qualitative review, while 44 were assessed in the quantitative synthesis, with the total number of patients included being 3524. Overall negative impacts of cfDNA and KRAS mutations on OS and PFS in PDAC (HR = 2.42, 95% CI: 1.95–2.99 and HR = 2.46, 95% CI: 2.01–3.00, respectively) were found. The subgroup analysis of the locally advanced and metastatic disease presented similar results (HR = 2.51, 95% CI: 1.90–3.31). In the studies assessing the pre-treatment presence of KRAS, there was a moderate to high degree of heterogeneity (I2 = 87% and I2 = 48%, for OS and PFS, respectively), which was remarkably decreased in the analysis of the studies measuring post-treatment KRAS (I2 = 24% and I2 = 0%, for OS and PFS, respectively). The patients who were KRAS positive before but KRAS negative after treatment had a better prognosis than the persistently KRAS-positive patients (HR = 5.30, 95% CI: 1.02–27.63). Conclusion: The assessment of KRAS mutation by liquid biopsy can be considered as an additional tool for the estimation of the disease course and outcome in PDAC patients.

ObjectiveStudies have confirmed that patients with circulating tumor cells (CTCs) in their peripheral blood (PB) or disseminated tumor cells (DTCs) in bone marrow (BM) might have bad prognosis. In this paper, we discuss whether CTCs/DTCs would be an appropriate biomarker to predict the prognosis of ovarian cancer.MethodsWe systematically searched PubMed, EMBASE, Cochrane library, and Chinese National Knowledge Infrastructure to collect relevant studies published from the time the database were created to February 2014. Studies quality was assessed by Newcastle-Ottawa Scale. The effect size was estimated by hazard ratio (HR) and corresponding 95% confidence interval (95% CI). Meta-analysis was conducted with STATA Version 12.0.ResultsEight studies of 1184 patients were included in the final analysis. In the PB group, it showed that patients with positive CTCs had significantly shorter overall survival and disease-free survival than patients with negative CTCs (HR, 2.09; CI, 1.13–3.88 and HR, 1.72; CI, 1.32–2.25, respectively). The same result was shown with DTCs in the BM group (HR, 1.61; CI, 1.27–2.04 and HR, 1.44; CI, 1.15–1.80, respectively). We also discussed the influence of CTCs/DTCs on International Federation of Gynecology and Obstetrics stage, pathological grade with odds ratio and 95% CI. However, it did not show any statistical significance.ConclusionsThe CTCs/DTCs might be a new biomarker to predict the prognosis of ovarian cancer. Future studies are needed to confirm this consequence.

AbstractARt Image Exploration Space (ARIES) is a free, cloud-based dynamic environment offering art historians and others an extensive array of practical tools for analysing images. It is the product of a successful collaboration between art historians, librarians, computer scientists, and engineers from the Frick Art Reference Library, New York University's Tandon School of Engineering, and Brazil's Universidade Federal Fluminense. ARIES is a powerful tool for art historians, both replicating and augmenting traditional methods they have long-used to study images.1 With the advent of the prevalent use of digital photos, art historians lacked the technology capable of replacing what they had previously been able to accomplish in the analogue world. Wood Ruby and Deutch realized that art historians needed an out-of-the-box solution that didn't require extensive knowledge of other disciplines (computer science and engineering). The result of successful collaborations and a generous donation, ARIES is now available in BETA form at www.artimageexplorationspace.com.

5046 Background: Methylation profiling of circulating cell-free DNA (cfDNA) is a promising approach for non-invasive tumor detection due to the presence of tissue-specific epigenetic signatures that are detectable in cfDNA. Cell-free methylated DNA immunoprecipitation and high-throughput sequencing (cfMedDIP-seq) is a sensitive, low-input, cost-effective, bisulfite-free approach to profiling cfDNA methylomes, capable of detecting and classifying various tumor types. We tested the feasibility of cfMeDIP-seq to detect urothelial carcinoma (UC) in plasma samples. Methods: We performed cfMeDIP-seq on plasma samples from 43 patients (pts): 18 metastatic UC (UC) pts, 12 pre-cystectomy non-metastatic UC pts, and 13 cancer-free controls. Six (50%) of pre-cystectomy cases were non-muscle invasive UC. cfDNA was immunoprecipitated and enriched using an antibody targeting 5-methylcytosine and PCR-amplified to create a sequence-ready library. The top differentially methylated regions (DMRs) between UC and control samples were used to train a regularized binomial generalized linear model using 80% of the samples as a training set. The 20% of withheld test samples were then assigned a probability of being UC or control. This process was repeated 100 times. Results: The average amount (standard deviation) of cfDNA isolated from 1 ml of UC plasma samples was 29.2 (27.4) ng/µL and 8.02 (3.58) ng/µL in cancer-free controls. We identified 9,826 DMRs in plasma samples at an adjusted p-value of < 0.01, which partitioned UC and control samples. Iterative training and classification of held out samples using the top 300 DMRs resulted in a mean AUROC of 0.987. Conclusions: cfMeDIP-seq is an interesting new approach for non-invasive detection of UC. cfMeDIP-seq demonstrates high sensitivity to detect UC across all stages of UC, including non-muscle invasive disease.

Cardiovascular inflammation and vascular endothelial dysfunction are involved in chronic heart failure (CHF), and cellular adhesion molecules are considered to play a key role in these mechanisms. We evaluated temporal patterns of 12 blood biomarkers of cell adhesion in patients with CHF. In 263 ambulant patients, serial, tri-monthly blood samples were collected during a median follow-up of 2.2 (1.4–2.5) years. The primary endpoint (PE) was a composite of cardiovascular mortality, HF hospitalization, heart transplantation and implantation of a left ventricular assist device and was reached in 70 patients. We selected the baseline blood samples in all patients, the two samples closest to a PE, or, for event-free patients, the last sample available. In these 567 samples, associations between biomarkers and PE were investigated by joint modelling. The median age was 68 (59–76) years, with 72% men and 74% New York Heart Association class I–II. Repeatedly measured levels of Complement component C1q receptor (C1qR), Cadherin 5 (CDH5), Chitinase-3-like protein 1 (CHI3L1), Ephrin type-B receptor 4 (EPHB4), Intercellular adhesion molecule-2 (ICAM-2) and Junctional adhesion molecule A (JAM-A) were independently associated with the PE. Their rates of change also predicted clinical outcome. Level of CHI3L1 was numerically the strongest predictor with a hazard ratio (HR) (95% confidence interval) of 2.27 (1.66–3.16) per SD difference in level, followed by JAM-A (2.10, 1.42–3.23) and C1qR (1.90, 1.36–2.72), adjusted for clinical characteristics. In conclusion, temporal patterns of C1qR, CDH5, CHI3L1, EPHB4, ICAM2 and JAM-A are strongly and independently associated with clinical outcome in CHF patients.

Abstract Due to its noninvasive nature and being readily available in peripheral blood and other bodily fluids, analysis of cell-free DNA (cfDNA) has become a promising application in cancer diagnosis, prognosis and treatment monitoring. However, disease-relevant cfDNA is present in a very limited amount in the huge background of normal cfDNA. This remains a challenge for any detection technology. Many currently available target enrichment and library preparation methods use regular DNA polymerase and amplification processes that introduce substantial bias and artifacts. This results in artifactual errors that greatly limit the detection of true low-frequency variants below 0.5% in heterogeneous samples, such as cfDNA. Here, we present a new targeted cfDNA workflow that overcomes challenges such as biases and artifacts. The workflow uses a highly optimized, high-fidelity reaction chemistry and incorporates UMIs into a single gene-specific, primer-based targeted enrichment process. Compared to regular DNA polymerase, this high-fidelity chemistry resulted in a five- to ten-fold decrease in base substitution error. An individual cancer panel was designed to specifically cover cancer-relevant hotspots and copy number genes with a dense primer design to accommodate the short length of cfDNA. Due to its high-fidelity chemistry and optimal panel design, our streamlined workflow can be completed in a single day. We also report consistent panel performance across different samples. Detection sensitivity and specificity were evaluated on a reference cfDNA sample and a simulated cfDNA sample by mixing enzyme-digested Genome in a Bottle samples, NA12878 and NA24385. We achieved close to 90% detection sensitivity and above 99.9% specificity for 0.1% variant. In addition, copy number variation could be successfully detected with a 1.5-fold difference over normal control samples. These results demonstrate an efficient targeted cfDNA panel workflow that enables cancer-relevant mutation detection with high sensitivity and accuracy. The applications presented here are for research use only. Not for use in diagnostic procedures. Citation Format: Michelle Baird, Mariam Ashraf, Emil Chistensen, Christa Haldrup, Zhong Wu, Eric Lader. Ultra-sensitive targeted DNA panel for very low-frequency mutation detection in circulating cell-free DNA [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB004.

Introdution Wikipedia is one of the most visited knowledge resources in the world. The Alexa traffic rankings put it at number 7, well above the New York Times (104), the BBC (106), the Library of Congress (1,175), and the venerable Encyclopedia Britannica (3711) (Alexa, 2016). As historian Roy Rosenzweig puts it, Wikipedia has become "perhaps the largest work of online historical writing, the most widely read work of digital history, and the most important free historical resource on the World Wide Web" (Rosenzweig 2006, p. 52). Wikipedia has become so ingrained in our everyday search for information that users rarely give thought to the mechanisms and agency underneath its production of knowledge: who produces its content? And what visible and invisible structures govern this production? Indeed, we have come to take its presence for granted to a degree that editorial contributions to many Wikipedia pages are in fact stagnating (Wikipedia, n.p.; Ford, 2011).

PurposeTo update the recommendations for the use of tumor marker tests in the prevention, screening, treatment, and surveillance of breast cancer.MethodsFor the 2007 update, an Update Committee composed of members from the full Panel was formed to complete the review and analysis of data published since 1999. Computerized literature searches of MEDLINE and the Cochrane Collaboration Library were performed. The Update Committee's literature review focused attention on available systematic reviews and meta-analyses of published tumor marker studies. In general, significant health outcomes (overall survival, disease-free survival, quality of life, lesser toxicity, and cost-effectiveness) were used for making recommendations.Recommendations and ConclusionsThirteen categories of breast tumor markers were considered, six of which were new for the guideline. The following categories showed evidence of clinical utility and were recommended for use in practice: CA 15-3, CA 27.29, carcinoembryonic antigen, estrogen receptor, progesterone receptor, human epidermal growth factor receptor 2, urokinase plasminogen activator, plasminogen activator inhibitor 1, and certain multiparameter gene expression assays. Not all applications for these markers were supported, however. The following categories demonstrated insufficient evidence to support routine use in clinical practice: DNA/ploidy by flow cytometry, p53, cathepsin D, cyclin E, proteomics, certain multiparameter assays, detection of bone marrow micrometastases, and circulating tumor cells.

Abstract BACKGROUND Accurate molecular diagnosis of infiltrating brainstem tumors, including diffuse midline gliomas and IDH mutant astrocytomas, is essential for prognostication and optimal therapy. Surgical biopsy of the brainstem carries risk of permanent neurological deficits and may yield insufficient or non-diagnostic tissue. We hypothesized that minimally invasive “liquid biopsy” analyzing circulating tumor cell-free DNA (cfDNA) in cerebrospinal fluid (CSF) could represent an alternative molecular diagnostic modality. METHODS We performed MSK-IMPACT, a New York State Department of Health authorized hybridization capture-based next-generation panel DNA sequencing clinical assay on 44 CSF samples from 37 unique patients with brainstem tumors. Ages at time of CSF collection ranged from 1–47 years (median: 29 years). For samples in which diagnostic alterations could not be detected, we performed a series of secondary analyses for known hotspot mutations such as H3F3A, IDH1/2, and BRAF, including manual review for mutant reads below the official clinical reporting threshold, as well as droplet digital PCR (ddPCR). RESULTS Among 44 samples included in this study, 10/44 (23%) samples had mutations detected by MSK-IMPACT using standard clinical calling criteria and 14 of the remaining 34 samples (41%) had supporting evidence of a diagnostically relevant mutation based on manual review. Testing by ddPCR was performed on 11 samples (based on DNA availability), uncovering 4 additional mutations. Overall positive diagnostic yield was 64% (28/44). In biopsied patients, CSF confirmed disease-defining mutations in 80% (16/20) of CSF samples, including H3F3A K28M (9/10; 90%) and IDH1/2 (5/6; 83%). Among 24 unbiopsied patients, a disease-defining mutation was identified in 60% (15/25) of CSF samples. CONCLUSIONS Analysis of CSF cfDNA in patients with brainstem tumors has a high diagnostic yield and obviates the need for tissue biopsy in a majority of patients. A tiered testing approach using next-generation sequencing and ddPCR assays helps maximize diagnostic information and sensitivity.

Poliomyelitis, or polio, is a highly infectious disease and can result in permanent flaccid paralysis of the limbs. Singapore was certified polio-free by the World Health Organization (WHO) on 29 October 2000, together with 36 other countries in the Western Pacific Region. The last imported case of polio in Singapore was in 2006. Fortunately, polio is vaccine-preventable—the world saw the global eradication of wild poliovirus types 2 and 3 achieved in 2015 and 2019, respectively. However, in late 2022, a resurgence of paralytic polio cases from vaccine-derived poliovirus (VDPV) was detected in countries like Israel and the US (specifically, New York); VDPV was also detected during routine sewage water surveillance with no paralysis cases in London, UK. Without global eradication, there is a risk of re-infection from importation and spread of wild poliovirus or VDPV, or new emergence and circulation of VDPV. During the COVID-19 pandemic, worldwide routine childhood vaccination coverage fell by 5% to 81% in 2020–2021. Fortunately, Singapore has maintained a constantly high vaccination coverage of 96% among 1-year-old children as recorded in 2021. All countries must ensure high poliovirus vaccination coverage in their population to eradicate poliovirus globally, and appropriate interventions must be taken to rectify this if the coverage falters. In 2020, WHO approved the emergency use listing of a novel oral polio vaccine type 2 for countries experiencing circulating VDPV type 2 outbreaks. Environmental and wastewater surveillance should be implemented to allow early detection of “silent” poliovirus transmission in the population, instead of relying on clinical surveillance of acute flaccid paralysis based on case definition alone. Keywords: Acute flaccid paralysis, infectious diseases, polio vaccine, poliovirus, surveillance

e23065 Background: Advances in non-invasive tumor biomarker research have shown that tumor cells release fragments of DNA called circulating tumor DNA (ctDNA) into peripheral blood. Somatic mutations representing the tumors could be successfully detected from isolated ctDNA, providing new potential for tumor sample assessment in addition to traditional tissue biopsy methods. However, the low amount of ctDNA in the blood, which can be less than 1% allelic frequency, presents significant challenges for reliable variant detection with NGS assays. Improvement of sequencing accuracy at low allelic frequency is a critical factor in the implementation of NGS in ctDNA liquid biopsy research. Methods: We demonstrate the technical feasibility for a sample-to-variant NGS workflow that utilizes a broad multi-gene panel to survey a comprehensive list of variants relevant to multiple tumor types for liquid biopsy research. The method includes novel library preparation and analysis reporting for Ion Torrent™ sequencing platforms. 20ng of input cell-free DNA was subjected to the library generation protocol. Prepared libraries were templated on Ion Chef™ and sequenced on Ion S5™. Results: We successfully optimized an NGS workflow that enables the simultaneous examination of more than 360 driver and resistance hotspot mutations in a single-pool assay panel, achieving high sensitivity and specificity with limit of detection at 0.1% allelic frequency. The targeted regions span genes and variants relevant to multiple tumor types for comprehensive variant detection across high-value content reviewed by industry experts and researchers. Sequencing on the Ion S5™ delivered > 95% on-target reads and uniform amplification across targeted regions with deep sequencing depth ( > 40,000x). The workflow is compatible with single or multiple pooled samples on Ion Torrent™ sequencing chips. Conclusions: We demonstrate the ability to accurately detect high-value variants implicated in multiple tumors at 0.1% allelic frequency on Ion Torrent™ NGS. (For Research Use Only. Not for use in diagnostic procedures.)

The study focuses on the adaptation of a politically engaged dramaturgical work from Bolshevik Russia — Vladimir Kirshon’s and Andrey Uspensky’s play Konstantin Terekhin (Rust) — to the specific requirements of Broadway, the commercial theater of the USA, and the textual changes of the Soviet original associated with it. The basic principles of the Broadway theater creative and organizational model, drastically different from the repertory theater of post-revolutionary Russia, are defined — the primacy of commerce over artistry, the absence of state support and censorship, a respectable audience that does not accept radical political ideas. On the American stage the Soviet play was produced in 1929, with the changed title (Red Rust), and the text subjected to changes and distortions. The paper considers these changes in the context of American socio–economic life of the Red Thirties. The discrepancy between the original dramaturgical material and the specific requirements of the American commercial theater is analyzed. The free handling of the text from Bolshevik Russia in the US theater is due to the absence of copyright regulations between the two countries. The process of exporting the play to the United States — via Paris and London — is being reconstructed. Three sources that have carried out the textual transformation of the Soviet original are characterized: the authors of the French-language adaptation from Russian (Fernand Nozière and Vladimir Bienstock), British translators from French into English (Virginia and Frank Vernon) and Broadway stage director who previously visited Moscow and sought to introduce into the text what, in his opinion, Soviet censorship would not allow (Herbert Biberman). The study is based on the materials from the Beinecke Library of Rare Books and Manuscripts collections (Yale University), as well as documents from the Houghton Library (Harvard University), the New York Public Library for Performing Arts, the Russian State Archive of Literature and Art (Moscow), and the museum of the Mossovet State Academic Theater (Moscow). The study is aimed at expanding the understanding of the stage history of the Russian drama in America.

In the present study, we evaluated circulating pro-inflammatory mediators and markers of oxidative stress in patients with decompensated CHF (congestive heart failure) and assessed whether clinical recompensation by short-term inotropic therapy influences these parameters. Patients with worsening CHF (n=29, aged 61.9±2.7 years), NYHA (New York Heart Association) class III–IV, and left ventricular ejection fraction of 23.7±1.8% were studied. Controls comprised age-matched healthy volunteers (n=15; 54.1±3.2 years). Plasma levels of cytokines [IL (interleukin)-6 and IL-18], chemokines [MCP-1 (monocyte chemotactic protein-1)], adhesion molecules [sICAM (soluble intercellular adhesion molecule), sE-selectin (soluble E-selectin)], systemic markers of oxidation [TBARS (thiobarbituric acid-reactive substances), 8-isoprostaglandin F2α and nitrotyrosine] and hs-CRP (high-sensitivity C-reactive protein) were measured by ELISA and colorimetric assays at admission and 30 days following 72-h milrinone (n=15) or dobutamine (n=14) infusion. Plasma IL-6, IL-18, sICAM, E-selectin, hs-CRP and oxidative markers were significantly higher in patients on admission before inotropic treatment compared with controls (P<0.05). Short-term inotropic support improved clinical status as assessed by NYHA classification and by the 6-min walk test and significantly decreased plasma levels of IL-6, IL-18, sICAM, hs-CRP and markers of oxidation (P<0.05) at 30 days. The effects of milrinone and dobutamine were similar. In conclusion, our results demonstrate that patients with decompensated CHF have marked systemic inflammation and increased production of oxygen free radicals. Short-term inotropic support improves functional status and reduces indices of inflammation and oxidative stress in patients with decompensated CHF.

Abstract Introduction The introduction of new treatment regimens has significantly increased the progression free survival (PFS) of newly diagnosed multiple myeloma (MM) patients. However, even with these novel treatments, for some the disease remains refractory, highlighting the need to identify the pathobiology of high-risk MM. In MM patients, high levels of circulating tumor cells (CTCs) is associated with an inferior prognosis independent of high-risk cytogenetics (Chakraborty et al., 2016), suggesting that CTC numbers are a relevant reflection of tumor cell biology. We hypothesized that high levels of CTCs in MM patients are either the result of a transcriptionally distinct tumor clone with enhanced migration capacities, or driven by transcriptional differences present in the bone marrow (BM) tumor cells. To test these hypotheses, we 1) compared MM cells from paired blood and BM samples, and 2) compared BM tumor cells of patients with high and low CTC levels, using single cell RNA-sequencing. Results We isolated plasma cell (PCs) from viably frozen mononuclear cells of paired peripheral blood (PB) and BM aspirates from five newly diagnosed MM patients (0.5%-8% CTCs) to determine the presence of a distinct CTC subclone. We generated single cell transcriptomes from 44,779 CTCs and 35,697 BM PCs. In the total 9 clusters common to BM PCs and CTCs were identified upon single cell data integration, but no cluster specific for either source was detected. Only 25 genes were significantly differential expressed between CTCs and BM PCs. The absence of transcriptional clusters unique to either CTCs or BM PCs, and the transcriptional similarity between these two anatomical sites makes it highly unlikely that CTC levels are driven by the presence of a transcriptionally-primed migratory clone. We next set out to identify possible transcriptional differences in BM PCs from eight patients with high (2-22%) versus thirteen patients with low (0.004%-0.08%) percentages of CTCs. Recurrent high-risk mutations were present in both groups. Single cell transcriptomes were generated from 74,830 BM PCs. Single cell data integration across all patients led to the identification of 8 distinct PC clusters, one of which was characterized by enhanced proliferation as defined by STMN1 and MKI67 transcription. Interestingly, this proliferative cluster was increased in patients with a high percentage of CTCs. Furthermore, cell cycle analyses based on canonical G2M and S phase markers revealed that actively cycling PCs were more frequent in the BM of patients with a high percentage of CTCs (64% versus 30%, p&lt;0.001), irrespective of the transcriptional cluster of origin. We hypothesized that plasma cell-extrinsic cues from the bone marrow micro-environment might be driving tumor proliferation. In order to substantiate this, we isolated BM immune cells from the same 21 patients and generated a library of 301,045 single immune cell transcriptomes. This library contained all major immune cell subsets, including CD4 + and CD8 + T cells, NK cells, B cells and monocytes. Comparative analyses of these cell populations in patients with either high or low levels of CTC are ongoing. Conclusion Through single cell transcriptomic analyses, we demonstrate that CTCs and BM PCs are transcriptionally similar. Importantly, we identify increased BM PC proliferation as a significant difference between patients with high and low levels of CTCs, implicating an increased tumor proliferation as one of the potential mechanisms driving CTC levels and MM disease pathobiology. The relation of the BM immune micro-environment to this altered proliferative state is currently under investigation. Disclosures van der Velden: Janssen: Other: Service Level Agreement; BD Biosciences: Other: Service Level Agreement; Navigate: Other: Service Level Agreement; Agilent: Research Funding; EuroFlow: Other: Service Level Agreement, Patents & Royalties: for network, not personally. Sonneveld: SkylineDx: Honoraria, Research Funding; Karyopharm: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding; Celgene/BMS: Consultancy, Honoraria, Research Funding; Janssen: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding. Broyl: Sanofi: Honoraria; Janssen Pharmaceuticals: Honoraria; Celgene: Honoraria; Bristol-Meyer Squibb: Honoraria; Amgen: Honoraria.

Abstract Introduction: Liquid biopsies, which interrogate body fluids in search of analytes released by cancer cells, are becoming increasingly common in clinical oncology. Foremost among the technologies used are those that employ nucleic acids, such as cell-free DNA (cfDNA) and cell-free RNA (cfRNA). Currently, many NGS-based tests perform near the limit of assay detection, making the identification of critical mutations, fusions, or expression patterns like finding a needle in a haystack. Further hindering successful assay development is the variability in analyte availability and detectability that can occur prior to analysis due to blood collection, transport, and storage. Here, we demonstrate that Nucleic Acid BCT stabilizes draw-time levels of plasma nucleic acids for downstream use with NGS-based assays. Methods: Blood from self-proclaimed healthy donors was drawn into EDTA and Nucleic Acid BCT and stored at ambient temperature for up to 7 days before plasma isolation with a general double-spin protocol and storage at -80 °C. Plasma nucleic acids were isolated using the MAGicBead cfDNA Kit (Zymo, cfDNA) or the QIAamp Circulating Nucleic Acid Kit (QIAGEN, cfRNA). Library prep for cfDNA was carried out with added synthetic mutant spike-in oligonucleotides using the Illumina TruSight Tumor 15 kit (Illumina). cfRNA library prep was done with the NEBNext Ultra II RNA Library Prep Kit for Illumina with upfront rRNA depletion (New England Biolabs). Sequencing was performed using an Illumina NextSeq 500 system, and all data were saved in and analyzed with Illumina BaseSpace applications. Results: When blood samples were stored in EDTA tubes for up to 7 days, spiked mutant alleles (EGFRL858R, EGFRdel19, KRASG12D, and PIK3CAE545K) were no longer detected, likely due to a dilution of the spiked DNA concentration following the release of genomic DNA containing wildtype alleles during white blood cell degradation. In contrast, equivalent blood samples stored in Nucleic Acid BCTs displayed mutant allele frequencies reflective of draw-time levels. Interrogation of plasma RNA expression patterns in samples stored in Nucleic Acid BCT versus EDTA revealed similar results, where differentially expressed plasma transcripts in blood samples stored in EDTA were found at high levels (&gt;1,000 transcripts) versus donor-matched blood samples stored in Nucleic Acid BCT (&lt;50 transcripts). Conclusions: Our data demonstrate that Nucleic Acid BCT maintains mutant allele frequencies and the plasma transcriptome profile of blood samples during ambient temperature storage. This provides precious sample integrity during whole blood storage and transport, offering laboratories and assay developers reduced preanalytical variability for NGS-based analysis of gene mutations, fusions, indels, and the plasma transcriptome. Nucleic Acid BCT is for Research Use Only. Not for use in diagnostic procedures. Citation Format: Nicholas M. George, Lisa Bartron. Use of Streck nucleic acid BCT with plasma nucleic acid next-generation sequencing workflows [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5035.

The primary cause of atherosclerotic cardiovascular disease (ASCVD) is elevated levels of low-density lipoprotein cholesterol (LDL-C). Proprotein convertase subtilisin/kexin type 9 (PCSK9) plays a crucial role in this process by binding to the LDL receptor (LDL-R) domain, leading to reduced influx of LDL-C and decreased LDL-R cell surface presentation on hepatocytes, resulting higher circulating levels of LDL-C. As a consequence, PCSK9 has been identified as a crucial target for drug development against dyslipidemia and hypercholesterolemia, aiming to lower plasma LDL-C levels. This research endeavors to identify promising inhibitory candidates that target the allosteric site of PCSK9 through an in silico approach. To start with, the FDA-approved Drug Library from Selleckchem was selected and virtually screened by docking studies using Glide extra-precision (XP) docking mode and Smina software (Version 1.1.2). Subsequently, rescoring of 100 drug compounds showing good average docking scores were performed using Gnina software (Version 1.0) to generate CNN Score and CNN binding affinity. Among the drug compounds, amikacin, bestatin, and natamycin were found to exhibit higher docking scores and CNN affinities against the PCSK9 enzyme. Molecular dynamics simulations further confirmed that these drug molecules established the stable protein–ligand complexes when compared to the apo structure of PCSK9 and the complex with the co-crystallized ligand structure. Moreover, the MM-GBSA calculations revealed binding free energy values ranging from −84.22 to −76.39 kcal/mol, which were found comparable to those obtained for the co-crystallized ligand structure. In conclusion, these identified drug molecules have the potential to serve as inhibitors PCSK9 enzyme and these finding could pave the way for the development of new PCSK9 inhibitory drugs in future in vitro research.

Abstract Whole genome sequencing (WGS) is widely used for cancer diagnostic and therapeutic applications in clinical practice and clinical trials. Currently, the Truseq PCR-free library preparation methods are routinely used in genomics laboratories. However, microgram amounts of DNA input is a limitation due to limited materials often from clinically-derived specimens. Here, we evaluated a novel low input (100-300 ng) PCR-free tagmentation (TAG) based library preparation for WGS. Replicates of TAG- and ligation-based (Illumina TruSeq) sequencing libraries were prepared from 3 pairs of breast cancer-derived tumors (HCC1195, HCC1143 and HCC1187) and matched B-lymphocyte-derived normal (HCC1195BL, HCC1143BL and HCC1187BL) cell lines. Libraries were sequenced on Illumina Novaseq 6000 platforms to target 30x and 90x mean coverage for normal and tumor samples respectively. Raw sequencing data were aligned to the hg38 by Isaac Aligner before variant analysis by Strelka2. Technical sequencing metrics demonstrated high similarity between TAG- and ligation-based workflows, including passing filter reads, Q30%, aligned reads, mean coverage. Germline variants of HCC1395BL showed greater than 98% precision and sensitivity using ligation-based variants as a reference. For somatic mutation calling from TAG-based libraries, HCC1395, HCC1143 and HCC1187, 88%, 85% and 86% precision and 81%, 70%, 68% sensitivity was observed, respectively, compared to the ligation-based method. Furthermore, using the SEQC2 Consortium high-confidence mutation set, TAG- and ligation- based variant calls of HCC1395 had 90% and 93% precision and 74% and 73% sensitivity, respectively with this high confidence reference. Further investigation showed that a high fraction of false negative calls was associated with low variant allele frequency (&lt;10%). Compared to an available reference dataset for benchmarking somatic calling pipelines from New York Genome Center, TAG- and ligation-based variant calls of HCC1143 yielded 85% and 83% precision and 63% and 63% sensitivity, respectively; TAG- and ligation-based variant calls of HCC1187 resulted in 91% and 92% precision and 81% and 79% sensitivity, respectively. In summary, the low input TAG-based WGS protocol produced highly reproducible variant calls with highly concordant somatic variant calls compared to the commonly-used PCR-free ligation-based methods and to reference callset. Further evaluation is warranted to broaden clinical application from minimal starting material.The views expressed in this abstract are solely of the authors and do not reflect the official policy of the Departments of Army/Navy/Air Force, Department of Defense, USUHS, HJF, or U.S. Government. Citation Format: Liqun Jiang, Xijun Zhang, Camille Alba, Gauthaman Sukumar, Elizabeth A. Rice, Matthew D. Wilkerson, Clifton L. Dalgard. The performance characteristic of the low input tagmentation-based whole genome sequencing in high quality somatic variant calling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 211.

Abstract. At the 78th UN General Assembly Session in New York, the Indonesian Minister of Foreign Affairs expressed Indonesia's concern about the condition of the Palestinian people in the Israeli-Palestinian conflict. This speech also emphasized that Indonesia would not stop providing support for the Palestinian people to gain independence. During the conflict, both parties have violated human rights, namely the right to live in safety and other rights. This phenomenon is in stark contrast to the foundation of the Indonesian state, Pancasila. Pancasila, as a national ideology, aspires to justice and prosperity for every individual. In the 1945 Constitution, Indonesia stated that it rejected colonialism and was ready to participate in world order. This commitment is proven by Indonesia's diplomatic activities for world peace. The free and active political system and the Non-Aligned Movement practiced by Indonesia really support Indonesia to become a neutral mediator in regional and international conflicts. This article examines the implementation of Pancasila in world peace, especially in efforts to mediate the Israeli-Palestinian conflict. The research was carried out using qualitative methods with analytical descriptions of data collected using library study techniques. World peace diplomacy based on Pancasila is non-interventionist and respects the sovereignty of other countries. Apart from that, diplomacy based on Pancasila upholds humanity, social justice and the welfare of all people. This is practiced in efforts to mediate the Israeli-Palestinian conflict. Pancasila, with its principles, provides a strong basis for supporting world peace. Indonesia, as a country that upholds Pancasila, has an important role in promoting these values at the international level and contributing to joint efforts to achieve a more peaceful, just and civilized world.

Background: Circulating cell-free DNA (ccfDNA) is highly fragmented DNA in plasma that is released by normal or tumor cells when they undergo apoptosis or necrosis. ccfDNA allows for non-invasive sampling of somatic genomic alterations and is informative in various solid tumors, including as a marker of measurable residual disease (MRD). We sought to assess the utility of baseline assessment and tracking of leukemia-associated mutations through peripheral blood sampling of ccfDNA in patients (pts) with acute leukemias. Methods: Plasma ccfDNA was isolated and analyzed using a next-generation sequencing (NGS) assay of 275 genes. This NGS analysis is based on Single Primer Extension library preparation with unique molecular identifier (Qiagen, Germantown, MD); a sequence coverage ≥ 100X (after removing duplicates) was required. Amplicon-based NGS was also performed on DNA extracted from the bone marrow (BM) in a CLIA-certified molecular diagnostics laboratory. This BM panel detects mutations in the coding sequence of 28 leukemia-associated genes, with an analytic sensitivity of 5-10%. The ccfDNA panel included all 28 genes evaluated on the BM NGS panel (ABL1, ASXL1, BRAF, DNMT3A, EGFR, EZH2, FLT3, GATA1, GATA2, HRAS, IDH1, IDH2, IKZF1, JAK2, KIT, KRAS, MDM2, MLL, MPL, MYD88, NOTCH1, NPM1, NRAS, PTPN11, RUNX1, TET2, TP53, WT1). Established bioinformatics pipelines were used to identify somatic variants. Results: Twenty-four pts (AML, n=22; ALL, n=2) underwent paired ccfDNA and BM sequencing at diagnosis prior to receiving frontline intensive chemotherapy. For baseline samples, ccfDNA was collected a median of 6 days after BM collection (range, 0-27 days) and a median of 0.5 days after start of induction chemotherapy (range, -7 to 7 days). Eleven pts (46%) also had ccfDNA collected at ≥1 time point during remission. Among the 28 genes of interest, the median number of mutations per pt detected in BM and in ccfDNA was 1 (range, 0-4) for both assays (P=0.39). A total of 40 mutations were detected: 18 mutations (45%) were detected by both methods, 7 (18%) were detected only in ccfDNA, and 15 (38%) were detected only in BM. Time from start of chemotherapy until ccfDNA collection did not appear to impact the concordance of ccfDNA and BM mutation analysis (P=0.87). Among mutations detected by ccfDNA in baseline samples, the median variant allelic frequency (VAF) was 33.7% (range, 2.7-90.8%). Among the 18 overlapping mutations, the concordance of VAF assessment by both methods was high (R2 = 0.849). Mutations detected by only one of the two methods were generally of lower VAF than those detected by both methods, suggesting that either method may miss small subclonal populations. The median VAF of mutations (as measured in ccfDNA) that were detected by both methods was higher than those detected only in ccfDNA (39.8% vs 25.2%, respectively; P=0.04); similarly, the median VAF of mutations (as measured in BM) that were detected by both methods was higher than those detected only in BM (40.2% vs 6.6%; P=0.001). Among the 7 mutations detected only by ccfDNA, ASXL1 was detected in 2 pts, WT1 in 1 pt, IDH1 in 1 pt, and BRAF and two EGFR mutations in 1 pt. Among the 5 pts in whom mutations were detected in ccfDNA but not BM, 2 eventually relapsed. In both pts, the discordant mutation (IDH1 and ASXL1) was detected in the relapse BM, suggesting that these were true mutations that were missed by NGS of the baseline BM. ccfDNA detected leukemia-associated mutations during remission that appeared to herald overt relapse (Figure 1). Two pts with t(8;21) AML developed new RUNX1 mutations detected by ccfDNA while in remission and subsequently relapsed 3 months and 14 months later. In both of these pts, the new RUNX1 mutation was confirmed in the BM at the time of morphological relapse. Another pt with AML had persistent TP53 and TET2 mutations detected by ccfDNA 1 month after allogeneic stem cell transplant and subsequently relapsed 1 month later. Conclusions: This study demonstrates that sequencing of ccfDNA can identify prognostic or targetable mutations not detected by BM NGS. However, true mutations were missed by both ccfDNA and BM analysis, suggesting that these methodologies may be complementary in the assessment and monitoring of pts with leukemia. The use of ccfDNA as a non-invasive method to detect mutations and track MRD in AML and other leukemias should be evaluated in larger, prospective cohorts. Disclosures Short: Takeda Oncology: Consultancy, Research Funding; AstraZeneca: Consultancy; Amgen: Honoraria. Jabbour:Amgen: Consultancy, Research Funding; BMS: Consultancy, Research Funding; Pfizer: Consultancy, Research Funding; Cyclacel LTD: Research Funding; AbbVie: Consultancy, Research Funding; Adaptive: Consultancy, Research Funding; Takeda: Consultancy, Research Funding. Garcia-Manero:Amphivena: Consultancy, Research Funding; Helsinn: Research Funding; Novartis: Research Funding; AbbVie: Research Funding; Celgene: Consultancy, Research Funding; Astex: Consultancy, Research Funding; Onconova: Research Funding; H3 Biomedicine: Research Funding; Merck: Research Funding. Kantarjian:BMS: Research Funding; Amgen: Honoraria, Research Funding; Agios: Honoraria, Research Funding; Immunogen: Research Funding; Takeda: Honoraria; Novartis: Research Funding; Ariad: Research Funding; Astex: Research Funding; Pfizer: Honoraria, Research Funding; Actinium: Honoraria, Membership on an entity's Board of Directors or advisory committees; Daiichi-Sankyo: Research Funding; Jazz Pharma: Research Funding; Cyclacel: Research Funding; AbbVie: Honoraria, Research Funding. Ravandi:Macrogenix: Consultancy, Research Funding; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Xencor: Consultancy, Research Funding; Menarini Ricerche: Research Funding; Cyclacel LTD: Research Funding; Selvita: Research Funding.

Many people use social networks to spend their free time. News, especially at the time of great world changes, began to gain considerable popularity. Washington Post, New York Times, Time, Reuters, Forbes are among the most famous global newspaper publications. An average analyst can spend up to 40 hours a week collecting information about competitors and researching the most popular posts. According to the conducted research, an average of 40 new posts with news per day. The data processing process can be automated using modern information tools to facilitate the routine work of analysts. To analyze the target audience and reach, it is worth considering the text of the message, the number of likes, comments and links. This information was obtained using the Selenium automated web page testing tool using the Java programming language. The time spent on collecting data in the described way from four newspaper editions amounts to approximately 12 hours. The Tensorflow library using the JavaScript programming language is applied to the collected information. Based on information about the number of shares, comments, likes, frequency of news posts, an analysis was carried out using machine learning algorithms. Based on the clustering data, we can observe such a tendency that posts with a large number of likes receive a large number of comments and vice versa. An analysis of the most active hours of users in the network based on news posts is performed. As a result, the highest activity is observed at least three times a day, namely: in the morning hours from 9:00 to 11:00, in the lunch time of the day from 12:00 to 15:00 and in the evening time period from 18:00 to 20:00. This trend is due to the work schedule of most employees during the working week. The resulting statistical information in the work can be used for other content or user behavior in social networks.

Abstract Background: Circulating tumor DNA (ctDNA), a new class of tumor marker, has been demonstrated the clinical validity of: 1) early relapse prediction, 2) therapeutic efficacy evaluation, and 3) relapse-free corroboration in the context of cancer treatment. Variant allele frequencies (VAFs) of ctDNA were under 0.1% even in most advanced cancer patients. Since the detection limit of next-generation sequencing (NGS) is above 1%, higher sensitivity is needed for sensitive ctDNA monitoring. Digital PCR (dPCR) offers excellent sensitivity at an affordable cost compared with NGS. A mutation-specific primer/probe (P/P) is required for the dPCR assay, but a limited number of P/Ps are commercially available. Materials and Methods: To provide essential probes for the rapid monitoring of sensitive ctDNA, we developed an off-the-shelf (OTS) dPCR P/P library (OTS-Probes) that includes probes that can detect &gt;1,000 mutations found in human cancer. The majority of the mutations to be detected by these P/P sets have been selected using an originally developed statistical scoring from hot spot mutations reported in the Catalogue of Somatic Mutations in Cancer (COSMIC) database. To evaluate the coverage of OTS-Probes in human cancer specimens, the Cancer Genome Atlas (TCGA) and the Center for Cancer Genomics and Advanced Therapeutics (C-CAT) datasets were used. TCGA is an international landmark cancer genomic program in the U.S., and C-CAT is a Japanese cancer genome profiling database. Results: At least one mutation was matched to the OTS-Probes for ctDNA monitoring in lung, colorectum, stomach, liver, pancreas, breast, uterus, and prostate from TCGA and C-CAT cancer specimens, reflecting 63% and 77% of corresponding tumor types, respectively. The mutation coverage of the OTS-Probes tends to be lower in TCGA than in C-CAT, partially due to the fact that information about the TERT promoter mutation is absent in TCGA. We found that some ARID1A deletion mutations that have a high count in COSMIC were not observed in either the TCGA or the C-CAT database. We also analyzed the coverage of the OTS-Probes in different organs and driver genes. Currently, more than 250 P/P sets have been designed, synthesized, and validated with human tumor DNA or synthesized DNA fragments. Importantly, nearly 98% of P/P sets work with the same PCR condition. With our proprietary oligo synthesis method with modified bases, only a small percentage of P/P sets required optimization of the annealing temperature. Conclusion: We developed a dPCR P/P library called OTS-Probes. Our analysis suggests that 60-80% of cancer patients could immediately apply at least one P/P set for sensitive ctDNA monitoring by dPCR. A quarter of the P/P library has already been validated by tumor DNA or synthesized DNA fragments. OTS-Probes is a great tool for ctDNA- related studies and for clinical use when a higher sensitivity than NGS can offer is required. Citation Format: Hayato Hiraki, Akiko Yashima-Abo, Takeshi Iwaya, Satoshi S. Nishizuka. A digital-PCR primer/probe library for pan-cancer ctDNA monitoring established from public databases for somatic mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6704.

Abstract Introduction: Multiple Myeloma (MM) is an incurable hematologic malignancy characterized by the abnormal growth of clonal plasma cells in the bone marrow (BM). In most cases MM develops from early, asymptomatic disease stages known as Monoclonal Gammopathy of Undetermined Significance (MGUS) and Smoldering Multiple Myeloma (SMM). Despite effective new therapies, most MM patients inevitably relapse and require further treatment, highlighting the need for better early detection methods for precursor patients and targeted interventions to prevent early disease from progressing. The initial diagnosis of MGUS/SMM remains an incidental process following the identification of increased clonal immunoglobulin in the blood. BM biopsy is the gold standard for diagnosis and monitoring of MM progression, but is intrusive, painful, and comes with possible secondary complications for patients. Consequently, repeated assessment is not a feasible option for MGUS and SMM patients who are asymptomatic. Here we tested the utility of circulating multiple myeloma cells (CMMCs) from non-invasive blood biopsy to accompany BM as a method to monitor disease development, by enumerating CMMCs from MGUS/SMM patients. Methods: Peripheral blood from 185 precursor patients (75 MGUS and 110 SMM) from the Dana-Farber Cancer Institute observational PCROWD study (IRB #14-174) was collected in CellRescue TM Preservative Tubes and processed on the CellSearch CellTracks Autoprep system using the CMMC assay kit using 4mL of blood. This assay employs the enrichment of CMMCs through the immunophenotype of CD138 +CD45 -19 -, and leukocyte exclusion based on CD45 +CD19 +. Nucleated cells were identified using DAPI staining. The CellTracks Analyzer II fluorescence microscope system was subsequently used to scan captured CMMC cartridges, with software allowing the automated scoring and enumeration of CMMCs. Additional molecular analyses were carried out on SMM patients. Briefly, minipools of CMMCs were sorted by DEPArray and underwent whole genome amplification using Ampli1 kit, PCR-free library construction, quantification and low pass whole genome sequencing (~0.5x) on the Illumina HiSeq2500. To assess whether molecular analyses can be performed to detect hyperdiploidy as a genomic biomarker of MM disease, ichorCNA analyses was performed to determine copy number variant (CNV) events and infer tumour fraction. Results: CMMCs were detected in 27% of MGUS patients collected, with a median count of 2 CMMCs (range 0 to 1328). Comparably, CMMCs were detected in 57% of SMM patients, with a median enumeration of 13 CMMCs (range 0 to 43836). Enumeration of CMMCs illustrated a correlation with clinical measure of disease including the International Myeloma Working Group 2/20/20 risk stratification model. A higher CMMC count was associated with increasing risk group based on the 3-risk factor model, with a median of 5, 29 and 59 CMMCs detected at low, intermediate, and high-risk SMM groups, respectively. CMMC counts were significantly increased at intermediate (P = 5.0 x 10 -4) and high-risk stages (P = 3.7 x 10 -3) compared to low-risk. While enumeration provides a correlative measure of CMMCs that may be of tumor origin, downstream molecular characterization can confirm MM-associated genetic alterations. At the precursor stages, a low tumour burden is evident clinically, thus both normal and malignant plasma cells are present. Therefore, to determine the concordance between bone marrow and peripheral blood CMMCs, we performed genomic analyses to identify arm level gain or loss events. Molecular analyses of CMMCs was carried out in patients who had matched BM and clinical fluorescent in situ hybridization (FISH) results. We showed that CMMCs can capture 100% of clinically annotated BM FISH CNV events. Furthermore, CMMC samples identified additional yield, with further CNVs identified that were not observed by FISH. In cases that did not have BM biopsy results, sequencing of CMMCs revealed the existence of genetic aberrations. Conclusion: Our results demonstrate clinical correlation and molecular characterization of CMMCs from MGUS/SMM patients. This study provides a foundation for non-invasive detection, enumeration and genomic interrogation of rare CMMCs from the peripheral blood of MGUS/SMM, illustrating the clinical potential of using liquid biopsies for monitoring and managing disease in the precursor setting of MM. Disclosures Getz: IBM, Pharmacyclics: Research Funding; Scorpion Therapeutics: Consultancy, Current holder of individual stocks in a privately-held company, Membership on an entity's Board of Directors or advisory committees. Ghobrial: AbbVie, Adaptive, Aptitude Health, BMS, Cellectar, Curio Science, Genetch, Janssen, Janssen Central American and Caribbean, Karyopharm, Medscape, Oncopeptides, Sanofi, Takeda, The Binding Site, GNS, GSK: Consultancy.

Background: Smoldering multiple myeloma (SMM) was stratified into risk classes based on several models including Mayo clinic and Spanish myeloma working group models. After the revision of diagnostic criteria for multiple myeloma (MM) in 2014, the ultra-high risk SMM patients (>80% clonal plasma cells at two years) were re-classified as active MM patients. Thus, predictors of progression in patients currently diagnosed as SMM are unknown and reassessment of existing models is required. We aim to identify the risk factors associated with progression in SMM patients classified according to updated guidelines. Methods We performed a literature search following PRISMA guidelines and used following bibliographic databases: MEDLINE (Ovid and PubMed), EMBASE, The Cochrane Library and Cochrane Central Register of Controlled Trials (CENTRAL), as well as annual meetings abstracts from inception till 1st,August 2019. We used MeSH and Emtree terms as well as performed open search for "smoldering multiple myeloma", "smoldering myeloma", and "asymptomatic multiple myeloma". Two independent reviewers screened the literature. We used snowballing technique to screen abstracts and reference within articles to include titles. Cochrane collaboration tool was used to asses risk of bias among included studies Results Our search retrieved 419 titles. After going through the titles and abstracts 38 articles were selected for full text review. Final review led to inclusion of 11 articles. Levels of serum M proteins, percentage of bone marrow plasma cells (BMPCs), serum free light chain ratio (FLCr) and PET/CT scan findings of whole body were most consistently and reliably indicated the progression of SMM to MM (Table 1). New studies are suggesting that B-cell maturation levels (BCMA), evolving M-proteins (eMP) and evolving hemoglobin levels (eHb) are also an accurate measure of SMM progression and should be incorporated in the risk stratification models. A study by Gonsalves WI et al. also suggested that levels of circulating clonal plasma cells with a cutoff of 150 was an important prognostic marker in their study. Immunoparesis status and role of Bence Jones proteins in reliably predicting the progression of SMM was debatable because they were significant in univariate analysis but were not significant in multivariate analysis (Table 1). Conclusion Serum M protein levels (2 g/dL), percentage of BMPCs (20%), serum FLCr (20) and PET/CT scan were reliable in predicting the prognosis of smoldering MM. New techniques like B-cell maturation levels(74.4 ng/mL), evolving M-proteins and evolving hemoglobin levels can play a significant role in proposing future risk predictive models of SMM. Role of immunoparesis and Bence Jones proteins is debatable. Table 1 Disclosures Anwer: Seattle Genetics: Membership on an entity's Board of Directors or advisory committees; In-Cyte: Speakers Bureau.

Abstract Background: Cell free miRNA has been widely used for disease diagnosis, particularly in cancers. The latest research has also demonstrated that isomiRs (isoform miRNAs) can serve as superior markers, displaying enhanced diagnostic performance in cancer diagnosis. However, the low detection sensitivities of miRNA/isomiRs in previous methods in plasma have limited their broad implementations for screening purpose. In this study, we propose a new isomiRs feature of isomiR relative proportion (IRP), which was validated with high performance in Colorectal Cancer (CRC) early diagnosis. Methods: A total of 1036 participants were enrolled from the National Colorectal Cancer Cohort (NCRCC), and 836 participants were eligible for plasma collection, including 238 controls, 373 CRC patients, 41 inflammatory bowel disease (IBD), 5 intramucosal carcinoma (Tis) and 179 advanced precancerous lesions (APL). Total circulating small RNA was extracted, followed by library preparation using UMI-based approach. Subsequently, small RNA sequencing was conducted using DNB-seq, and bioinformatic analysis was performed thereafter. The samples were randomly assigned into training (49%), test (30%) and validation (21%) sets. Models based on isomiR relative proportion (IRP), isomiR relative expression (IRE) or both features were constructed by beam search and random forest algorithms in training and test sets, and results were verified in validation set. Results: Differential IRP or IRE features were identified between cancer and controls. IRP identified 132 specific differential expressed isomiRs. Models constructed using either IRP or IRE features showed high performance (both AUC &gt; 0.9). However, there was improvement in IRP feature model in the validation set (sensitivity of IRP: 0.923, sensitivity of IRE: 0.859) compared with IRE model. We then found that there may be complementarities between IRP and IRE features, and the combination model (34 IRP+IRE features) improved cancer detection performance (AUC: 0.953). Finally, the risk scores predicted by this combination model increased gradually across CRC progression (R=0.921), and the sensitivity of APL was 0.626. Conclusion: We improved the small-RNA sequencing method and developed an IRP method, and present that IRP as a new feature not only demonstrate the ability of CRC early detection, but also revealed the CRC progressing process. Besides, the combination of IRP and IRE improved the performance of diagnosis. These results highlight the potential value of the IRP feature in early cancer detection. Citation Format: Yeting Hu, Wei Lu, Chengcheng Liu, Xiaozhuan Dai, Jiasheng Xu, Xiangxing Kong, Yunfeng Zhu, Yao Ye, Lihao Chen, Xiaolong An, Fan Wu, Shiqing Ma, Nana Wang, Qian Xiao, Jun Liu, Kefeng Ding. IsomiR relative proportion (IRP) within miRNA family as an efficient biomarker for noninvasive colorectal cancer detection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1063.

OBJECTIVES/SPECIFIC AIMS: This study aims to understand the potential immunomudualtory effect of APOL1 variants in auto-antigen activated myeloid cells by assessing lysosomal integrity in activated cells expressing APOL1. The primary stimuli were: 1. ssRNA hY3 as a proxy for the Ro immune complex; 2. in an bulk RNA seq model, interferon-response gene, Siglec 1, as a read out of interferon activity. The primary outcomes were: 1. Myeloid cell APOL1 expression both in primary macrophage cultures and ex-vivo patient derived macrophages; 2. Lysosome integrity as measured by fluorescence intensity of lysotracker dye on light microscopy. METHODS/STUDY POPULATION: All recruited subjects provided written informed consent as per the NEW YORK UNIVERSITY Division of Rheumatology-wide Specimen and Matched Phenotype Linked Examination (SAMPLE) protocol. Subjects were African American; SLE subjects met 4 American College of Rheumatology criteria for SLE. Healthy donor monocytes representing each genotype in duplicate (reference allele: G0/G0; heterozygote variant: RV/G0; and homozygote variant RV/RV) were cultured with GM-CSF to yield macrophages which were incubated in serum free media or with hY3 ssRNA (TLR 7/8 agonist) to yield inflammatory M1 macrophages. Fold increase of APOL1 in untreated vs hY3 treated macrophages was measured using qPCR. Live cells were then cultured on glass chamber slides with DNA dye, DAPI, and Lysotracker red, a fluorescent dye that stains acidic lysosomes. As a proof of concept, interferon response gene, Signlec1, and APOL1 transcriptional activity in peripheral blood monocytes (PBMCs) were measured and correlated in 17 SLE patients by RNA seq. RESULTS/ANTICIPATED RESULTS: Regardless of genotype, hY3 increased APOL1 expression by 29 (+/−18.4) fold (P = 0.007 vs no treatment). Genotyping of the qPCR product showed concordance with the chromosomal DNA with the RV heterozygotes expressing both alleles. To examine lysosomal membrane integrity, live hY3-treated macrophages were stained with lysosotracker dye and fluorescence intensity was measured. Compared to reference allele carrying macrophages, each additional variant allele corresponded with a lesser degree of lysosome compartment staining. In SLE PBMCs, we found that APOL1 was highly expressed, and significantly correlated with Siglec1 (F=10.5; P = 0.005) supporting an association between circulating interferons and APOL1 accumulation in monocytes. DISCUSSION/SIGNIFICANCE OF IMPACT: Given that the “cytokine milieu” in SLE elicits APOL1 expression, induces inflammatory cell metabolic rewiring, and stimulates autophagy thereby exposing defects in autophagic flux, this gene-environment interaction may underpin the relationship between chronic inflammation and heightened APOL1 polymorphism-attributed cardiovascular risk. These data support further inquiry into the intersection between chronic autoimmunity and APOL1’s functional role in the vascular microenvironment. The in vitro studies herein extend our prior work by demonstrating a mechanistic link between SLE-associated inflammation, APOL1 risk variant status and CVD via a lysosomal defect which converges on common autophagic and metabolic pathways in mononuclear cells.

Abstract A blood biomarker for early detection of renal cell carcinoma (RCC) would help decrease the burden of 15,000 deaths annually in the U.S. (1). Antibodies against tumor (neo)antigens are thought to arise early in cancer development and might serve as a more sensitive biomarker for RCC than tumor-derived biomolecules. Serum Epitope Repertoire Analysis (SERA) of human serum/plasma uses a random bacterial peptide display library and next generation sequencing to allow for unbiased highly multiplexed profiling of B cell epitopes. In combination with bioinformatic methods and machine learning, a classifier was trained to predict the presence of RCC using a cohort of 163 treatment-naïve patients (median age 62, stage: I 35%, II 5%, III 20%, IV 40%) with clear cell RCC (ccRCC) and 438 age-matched healthy controls (median age 61). Classifier performance was evaluated by sensitivity, specificity, and receiver operator curve (ROC) area under the curve (AUC) analysis in an independent validation cohort of 43 treatment-naïve patients with ccRCC (age 22-86, median 63, stage: I 33%, II 2%, III 26%, IV 39%) and 1,759 healthy controls (age 18-89, median 40). A total of 1,465 individual B cell epitope features were discovered and enriched in ccRCC vs. controls and included in a random forest model. In the validation cohort, the ROC-AUC was 0.65, and sensitivities of 9%, 16%, 30%, and 36% were achieved at specificities of 99%, 98%, 95%, and 90%, respectively. Performance was better in both early stage ccRCC and low tumor grade (grade 1 or 2). For stage I, the ROC-AUC was 0.88, and sensitivities of 7%, 14%, 36%, and 57% were achieved at specificities of 99%, 98%, 95%, and 90%, respectively. For low tumor grade, the ROC-AUC was 0.86, and sensitivities of 10%, 20%, 50%, and 60% of were achieved at specificities of 99%, 98%, 95%, and 90%, respectively. The classifier did not miscategorize benign renal lesions as ccRCC (Mann-Whitney p-value 0.52, ROC-AUC 0.54) in a separate cohort of 23 patients with benign renal tumors vs. the 1,759 healthy controls. This preliminary performance for a B cell epitope-based classifier is promising: the better performance in early stage ccRCC suggests that B cell epitope biomarkers may complement the lower sensitivity of cell-free DNA for detecting early-stage cancer (2). Future work will further validate these results in larger multi-institutional cohorts to improve classification performance and explore if B cell epitope biomarkers can be combined with existing assays to develop an early detection composite biomarker for RCC and tumor grade. (1)Siegel RL, Miller KD, Wagle NS, Jemal A. Cancer statistics, 2023. CA Cancer J Clin. 2023 Jan;73(1):17-48.(2)Francini, E.; Fanelli, G.N.; Pederzoli, F.; Spisak, S.; Minonne, E.; Raffo, M.; Pakula, H.; Tisza, V.; Scatena, C.; Naccarato, A.G.; et al. Circulating Cell-Free DNA in Renal Cell Carcinoma: The New Era of Precision Medicine. Cancers 2022, 14, 4359. Citation Format: Thomas W. Campbell, Christian R. Hoerner, Elizabeth Alli, John T. Leppert, John C. Shon, Alice C. Fan. Development of a B-cell epitope classifier for early detection of renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1056.

Abstract Introduction: Leptomeningeal metastases (LM) are diagnosed in up to 10% of metastatic cancer patients, the most frequent cancers being carcinomas of breast and lung or melanoma. Due to a dismal prognosis and an overall median survival of only four months new approaches for molecular analysis could help to identify individual treatment options. Material and Methods: In this study, we collected 28 cerebrospinal fluid (CSF) samples from patients with different primary tumors but suspected LM. CSF was divided in two parts for standard neuropathologic cytomorphology and detection of circulating tumor cells (CTCs) by using the CellSearch® CTC Kit, respectively. Subsequently, single Cytokeratin-positive CTCs were isolated for whole genome amplification by Ampli1TM and molecular analysis. Additionally, we were able to isolate cell-free DNA (cfDNA) from CSF in a subset of patients which we amplified by an adapted Ampli1TM library protocol. In a small subset of patients, we collected slices of formalin-fixed paraffin embedded (FFPE) primary tumors and systemic metastases for dissociation and isolation of pure subpopulations by DEPArrayTM technology. Isolated CTCs, cfDNA and subpopulations of tumor and stromal cells were analyzed for copy number variations (CNV) applying the Ampli1TM LowPass protocol on the Illumina MiSeqTM platform. Results: By comparing results from classical cytomorphology and CellSearch® workflow, we could find high concordance (24/28 samples) with a slightly higher tumor cell detection rate by CellSearch® (11 positive cases vs. 9 by cytomorphology). Importantly, we could confirm the malignant origin of isolated cells by CNV analysis in all patients positive in CellSearch® assay. Moreover, by comparing single cell data with cfDNA analysis of a subset of 20 samples, we could also detect a higher sensitivity for tumor detection in CTC-based analysis (8/20 cases positive) in comparison to cfDNA-based analysis (5/20 cases positive). In a comprehensive analysis of multiple samples of an individual patient, including CTCs and cfDNA from CSF at different time points, CTCs from blood and biopsies from multiple visceral metastasis, we could identify divergent genomic evolution between tumor cells from the central nervous syste Conclusion: In this proof-of-concept study we show that a liquid biopsy-based approach for single cell analysis from CSF holds potential for innovative diagnostic assays for patients with suspected LM. Combining CTC quantification with molecular single cell analysis could enable us to define predictive tests to select novel treatment options for patients with LM in the future. Citation Format: Caecilia Koestler, Giancarlo Feliciello, Florian Lueke, Kathrin Weidele, Saida Zoubaa, Peter Hau, Tobias Pukrop, Markus J. Riemenschneider, Christoph A. Klein, Bernhard Polzer. Single cell analysis from CSF of patients with suspected leptomeningeal metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3709.

Background: Infections caused by microorganisms are the major cause of death today. The tremendous and improper use of antimicrobial agents leads to antimicrobial resistance. Aim and Objective: Various currently available antimicrobial drugs are inadequate to control the infections and lead to various adverse drug reactions. Efforts based on computer-aided drug design (CADD) can excavate a large number of databases to generate new, potent hits and minimize the requirement of time as well as money for the discovery of newer antimicrobials. Pharmaceutical sciences also have made development with advances in drug designing concepts. The current research article focuses on the study of various G-6-P synthase inhibitors from literature cited molecular database. Docking analysis was conducted and ADMET data of various molecules was evaluated by Schrodinger Glide and PreADMET software, respectively. Here, the results presented efficacy of various inhibitors towards enzyme G-6-P synthase. Docking scores, binding energy and ADMET data of various molecules showed good inhibitory potential toward G-6-P synthase as compared to standard antibiotics. This novel antimicrobial drug target G-6-P synthase has not so extensively been explored for its application in antimicrobial therapy, so the work done so far proved highly essential. This article has helped the drug researchers and scientists to intensively explore about this wonderful antimicrobial drug target. Materials and Methods: The Schrodinger, Inc. (New York, USA) software was utilized to carry out the computational calculations and docking studies. The hardware configuration was Intel® core (TM) i5-4210U CPU @ 2.40GHz, RAM memory 4.0 GB under 64-bit window operating system. The ADMET data was calculated by using the PreADMET tool (PreADMET ver. 2.0). All the computational work was completed in the Laboratory for Enzyme Inhibition Studies, Department of Pharmaceutical Sciences, M.D. University, Rohtak, INDIA. Results: Molecular docking studies were carried out to identify the binding affinities and interaction between the inhibitors and the target proteins (G-6-P synthase) by using Glide software (Schrodinger Inc. U.S.A.-Maestro version 10.2). Grid-based Ligand Docking with Energetic (Glide) is one of the most accurate docking softwares available for ligand-protein, protein-protein binding studies. A library of hundreds of available ligands was docked against targeted proteins G-6-P synthase having PDB ID 1moq. Results of docking are shown in Table 1 and Table 2. Results of G-6-P synthase docking showed that some compounds were found to have comparable docking score and binding energy (kj/mol) as compared to standard antibiotics. Many of the ligands showed hydrogen bond interaction, hydrophobic interactions, electrostatic interactions, ionic interactions and π- π stacking with the various amino acid residues in the binding pockets of G-6-P synthase. Conclusion: The docking study estimated free energy of binding, binding pose andglide score and all these parameters provide a promising tool for the discovery of new potent natural inhibitors of G-6-P synthase. These G-6-P synthase inhibitors could further be used as antimicrobials. Here, a detailed binding analysis and new insights of inhibitors from various classes of molecules were docked in binding cavity of G-6-P synthase. ADME and toxicity prediction of these compounds will further accentuate us to study these compounds in vivo. This information will possibly present further expansion of effective antimicrobials against several microbial infections.

BackgroundAtrial fibrillation (AF) can be treated using a maze procedure during planned cardiac surgery, but the effect on clinical patient outcomes, and the cost-effectiveness compared with surgery alone, are uncertain.ObjectivesTo determine whether or not the maze procedure is safe, improves clinical and patient outcomes and is cost-effective for the NHS in patients with AF.DesignMulticentre, Phase III, pragmatic, double-blind, parallel-arm randomised controlled trial. Patients were randomised on a 1 : 1 basis using random permuted blocks, stratified for surgeon and planned procedure.SettingEleven acute NHS specialist cardiac surgical centres.ParticipantsPatients aged ≥ 18 years, scheduled for elective or in-house urgent cardiac surgery, with a documented history (> 3 months) of AF.InterventionsRoutine cardiac surgery with or without an adjunct maze procedure administered by an AF ablation device.Main outcome measuresThe primary outcomes were return to sinus rhythm (SR) at 12 months and quality-adjusted life-years (QALYs) over 2 years after randomisation. Secondary outcomes included return to SR at 2 years, overall and stroke-free survival, drug use, quality of life (QoL), cost-effectiveness and safety.ResultsBetween 25 February 2009 and 6 March 2014, 352 patients were randomised to the control (n = 176) or experimental (n = 176) arms. The odds ratio (OR) for return to SR at 12 months was 2.06 [95% confidence interval (CI) 1.20 to 3.54;p = 0.0091]. The mean difference (95% CI) in QALYs at 2 years between the two trial arms (maze/control) was –0.025 (95% CI 0.129 to 0.078;p = 0.6319). The OR for SR at 2 years was 3.24 (95% CI 1.76 to 5.96). The number of patients requiring anticoagulant drug use was significantly lower in the maze arm from 6 months after the procedure. There were no significant differences between the two arms in operative or overall survival, stroke-free survival, need for cardioversion or permanent pacemaker implants, New York Heart Association Functional Classification (for heart failure), EuroQol-5 Dimensions, three-level version score and Short Form questionnaire-36 items score at any time point. Sixty per cent of patients in each trial arm had a serious adverse event (p = 1.000); most events were mild, but 71 patients (42.5%) in the maze arm and 84 patients (45.5%) in the control arm had moderately severe events; 31 patients (18.6%) in the maze arm and 38 patients (20.5%) in the control arm had severe events. The mean additional cost of the maze procedure was £3533 (95% CI £1321 to £5746); the mean difference in QALYs was –0.022 (95% CI –0.1231 to 0.0791). The maze procedure was not cost-effective at £30,000 per QALY over 2 years in any analysis. In a small substudy, the active left atrial ejection fraction was smaller than that of the control patients (mean difference of –8.03, 95% CI –12.43 to –3.62), but within the predefined clinically equivalent range.LimitationsLow recruitment, early release of trial summaries and intermittent resource-use collection may have introduced bias and imprecise estimates.ConclusionsAblation can be practised safely in routine NHS cardiac surgical settings and increases return to SR rates, but not survival or QoL up to 2 years after surgery. Lower anticoagulant drug use and recovery of left atrial function support anticoagulant drug withdrawal provided that good atrial function is confirmed.Further workContinued follow-up and long-term clinical effectiveness and cost-effectiveness analysis. Comparison of ablation methods.Trial registrationCurrent Controlled Trials ISRCTN82731440.FundingThis project was funded by the NIHR Health Technology Assessment programme and will be published in full inHealth Technology Assessment; Vol. 22, No. 19. See the NIHR Journals Library website for further project information.

BackgroundImmune checkpoint inhibitors have improved clinical responses and overall survival for patients with non-small cell lung cancer (NSCLC). However, the response is not equal and known NSCLC biomarkers are not sufficient in predicting therapy outcome. Deep proteomic analysis of NSCLC patient‘s plasma treated with anti-PD-1-blockade using a state-of-the-art data independent acquisition mass spectrometry (DIA-MS) is a powerful and unbiased way of identifying protein signatures associated with disease stage or response to treatment. However, to unravel these associations large-scale omics data should be analyzed with respect to available clinical information. To achieve this goal, we have used an approach previously applied by Uhlen et al., 20171 for transcriptomic datasets. In this approach survival data is used to set the most optimal thresholds for candidate biomarkers.Methods125 plasma samples were analyzed by capillary flow liquid chromatography coupled to DIA-MS. Data were extracted with latest SpectronautTM and proteins were quantified. Each recorded protein intensity was used as a threshold for two groups of samples for which Kaplan-Meier estimates were generated using ‘survival’2 package in R. Benjamini-Hochberg correction was applied and p-values with corresponding intensity cut-offs were extracted to generate panels of potential biomarkers.Results125 plasma samples (in total 75 baseline and 50 after 8-weeks treatment) from advanced NSCLC patients treated with an anti-PD-1 inhibitor following at least 1 prior line of treatment were analyzed. 727 unique proteins were quantified across all samples. Data analysis was performed separately for each line of treatment and treatment status resulting in more than 100’000 p-values. For each group, panels of proteins with best performance in separating progression free survivals were defined at FDR of 0.10, giving 64 unique proteins which were mapped to acute phase response, platelet degranulation and complement activation. Several of these proteins were listed in the Early Detection Research Network database of the National Cancer Institute, and one of them – LYPD3, was a potential therapeutic target in a preclinical study for NSCL treatment.3 Selected proteins were then used to cluster patients into cohorts that showed association with the response to therapy.ConclusionsDeep proteomic profiling of plasma samples using DIA-MS in conjunction with clinical outcome enables a holistic and stringent analysis of potential circulating biomarkers. Such analysis generates functional insights into the plasma proteome that enable deeper understanding and comprehensive integration of clinical data with proteomics markers at different disease stages and treatment phases.ReferencesUhlen M, Zhang C, Lee S, Sjöstedt E, Fagerberg L, Bidkhori G, Benfeitas R, Arif M, Liu Z, Edfors F, Sanli K, von Feilitzen K, Oksvold P, Lundberg E, Hober S, Nilsson P, Mattsson J, Schwenk J.Therneau TM, Grambsch PM. Modeling Survival Data: Extending the Cox Model. Springer. 2000, New York, ISBN 0-387-98784-3.Willuda J, Linden L, Lerchen H, Kopitz C, Stelte-Ludwig B, Pena C, Lange C, Golfier S, Kneip C, Carrigan P E, Mclean K, Schuhmacher J, von Ahsen O, Müller J, Dittmer F, Beier R, El Sheikh S, Tebbe J, Leder G, Apeler H, Jautelat R, Ziegelbauer K, Kreft B, Preclinical Antitumor Efficacy of BAY 1129980-a Novel Auristatin-Based Anti-C4.4A (LYPD3) Antibody-Drug Conjugate for the Treatment of Non-Small Cell Lung Cancer. Mol Caner Ther 2017;16(5):893–904

Introduction Cancer-associated venous thromboembolism (VTE) is a major source of oncologic cost, morbidity and mortality. Identifying an optimal population for prophylactic anticoagulation is challenging and adds to clinician overhead. Circulating tumor (ct)DNA sequencing assays (“liquid biopsies”) are increasingly deployed in clinic, with multiple US FDA approvals for matching to molecularly targeted therapy. Preliminary data suggest that cell-free (cf)DNA, which is also measured as part of liquid biopsies and may consist of tumor or wild-type DNA, is thrombogenic due to its association with neutrophil extracellular traps. However, clinical validation of cfDNA for VTE prediction is lacking. ctDNA detection is associated with aggressive tumor biology and worse survival; however, whether it is associated with VTE is unknown. To address these gaps, we studied the clinical utility of ctDNA and cfDNA from liquid biopsies for predicting VTE. Methods We conducted an observational study in two cohorts: 1) Patients with any cancer type at Memorial Sloan Kettering Cancer Center (MSK, New York USA), an academic medical center, with plasma ctDNA sequencing using MSK-ACCESS, a NY State-approved, 129-gene assay (N=4,141). 2) A nonoverlapping cohort of patients at MSK or GenesisCare (Sydney, Australia), a community oncology setting, with stage IV or recurrent non-small cell lung cancer and ctDNA sequencing using a separate commercial assay (Agilent ctDx Lung, N=463). We studied the independent contribution of ctDNA, cfDNA and related variables for predicting VTE using Cox proportional hazards. We studied the potential utility of DNA liquid biopsies for predicting VTE within six months using machine learning models, comparing their dynamic area under the receiver operating curve, precision, and recall to those of the Khorana score, the most widely implemented risk metric. We tested the impact of ongoing anticoagulation for non-VTE indications on future VTE in patients with and without detectable ctDNA using nonrandomized, real-world evidence. Results ctDNA detection was associated with an increased risk of VTE in a dose-dependent manner (Figure 1). This association was observed across assays and treatment sites and regardless of genomic subtype. ctDNA was associated with VTE risk in non-small cell lung, breast, pancreatic, prostate, and less represented cancers as well as melanoma but not in bladder, hepatobiliary, or colorectal cancer. By contrast, cfDNA concentration was associated with VTE risk in all cancer types. In multivariate analysis, ctDNA detection, cfDNA concentration, Khorana score, chemotherapy treatment within 30 days, and number of organ sites of metastasis were all independently associated with VTE. In a cohort of patients with treatment naive lung adenocarcinoma with segmented positron emission tomography scans, ctDNA was an independent predictor of VTE after controlling for metabolic tumor volume. A random survival forest machine learning model to predict VTE within six months of plasma draw using liquid biopsy data (i.e. cfDNA and ctDNA levels as well as genomic content), cancer type, and chemotherapy receipt (“LB+”) outperformed the Khorana score in 5-fold cross-validation and was similarly superior in the external lung cohort (Table 1). The superiority of the model was observed in patients treated or not treated with chemotherapy. Additional clinical and radiologic data, including demographics, metastatic sites of disease, and Khorana score components (“All”), did not improve the ctDNA assay-based LB+ model. Patients with anticoagulation had lower VTE rates if ctDNA was detectable (adjusted HR 0.54, 95%CI 0.33-0.87) but not if ctDNA was undetectable. Conclusion In patients with cancer, ctDNA and cfDNA are independent biomarkers for VTE. Risk models incorporating liquid biopsy data are feasible in academic or community settings and, if prospectively validated, would provide a means of VTE risk stratification at scale.

<p><strong>Abstract.</strong> It seems that the aim of conventional studies on tactile maps for visually impaired people have been to improve their utility in terms of instrumentalism. However, given the recent progress of disability studies in social science and post-representational approach in recent map studies, it is necessary to examine how tactile maps work in the society and how they relate to “disability” as a social phenomenon. These are also important issues to think about how geospatial information technology for visually impaired people embed in the society. Hence, regarding the map as more-than-representation, this paper analysed various documents (e.g. newspaper articles, essays, instruction manuals for orientation and mobility specialist), and considered the social position of tactile maps in Japan. The results showed the following.</p><p>The tactile map has a long history as a teaching material of geography. It was used for education for the blind in Western countries in the late 18th century. This situation was introduced to Japan by overseas memoir. For example, in the late Edo era, Namura Gohachirou who was a member of Japanese Embassy to the United States witnessed geography education using tactile maps at the blind school in New York. In his diary named <i>Akou Nikki</i>, Namura said that the education for blinds using tactile maps seemed to “translation” from the words of those who can see both eyes well to the words of visually impaired people. His statement clearly shows the material difference of tactile maps. Many tactile maps have acquired social status as “translated objects”. The material form of tactile maps is different from the “map” (visual map) known to the general public, so it makes awareness of physical differences between the body of visually impaired people and sighted people. In 1880, the first blind school in Japan, Kyoto Moua In had exhibited teaching materials and other instruments to the exposition, including a tactile map representing the city of Kyoto. Such a social event also had a role to attract the attention of sighted people to the material heterogeneity of the tactile map <i>and</i> visually impaired people.</p><p>The 1960s-80s was the period of the situation of tactile maps changed significantly in Japan. In the 1960s, rehabilitation techniques for visual impairments was introduced to Japan from USA, and the concept of “orientation” which was lacking in conventional walking training diffused. In accordance with these movement, the teaching manual of orientation and mobility (O&amp;M) training became write the methodologies to make and use the tactile map for the training. In 1964, Kazuo Honma, the founder of Japan Brail Library, visited all over the world and bought lots of tools for visually impaired people, including tactile maps, at the blind libraries in various places and brought them back to Japan. He held an exhibition to show those materials in the following year. Furthermore, as the International Year of Disabled Persons 1981 and the subsequent enforcement of various laws related to people with disability, it has been emphasized to create cities where physically impaired people can go out of their homes. Since that time the tactile maps began to be installed in public facilities, and were introduced in the assistive technology catalogs. In this way, the tactile map was incorporated into the context of “outdoor behavior” and “walking”. Not only the body of visually impaired people but also the tactile maps as material objects have increasingly been exposed to the “outside” spaces (e.g. city, road). As a result, the tactile map became understood from the viewpoint of “safety”, and it became involved with human and non-human actors (e.g. government offices, volunteer organizations, barrier-free laws, traffic guidelines) different from those of geography education.</p>

BackgroundHome oxygen therapy (HOT) is commonly used for patients with severe chronic heart failure (CHF) who have intractable breathlessness. There is no trial evidence to support its use.ObjectivesTo detect whether or not there was a quality-of-life benefit from HOT given as long-term oxygen therapy (LTOT) for at least 15 hours per day in the home, including overnight hours, compared with best medical therapy (BMT) in patients with severely symptomatic CHF.DesignA pragmatic, two-arm, randomised controlled trial recruiting patients with severe CHF. It included a linked qualitative substudy to assess the views of patients using home oxygen, and a free-standing substudy to assess the haemodynamic effects of acute oxygen administration.SettingHeart failure outpatient clinics in hospital or the community, in a range of urban and rural settings.ParticipantsPatients had to have heart failure from any aetiology, New York Heart Association (NYHA) class III/IV symptoms, at least moderate left ventricular systolic dysfunction, and be receiving maximally tolerated medical management. Patients were excluded if they had had a cardiac resynchronisation therapy device implanted within the past 3 months, chronic obstructive pulmonary disease fulfilling the criteria for LTOT or malignant disease that would impair survival or were using a device or medication that would impede their ability to use LTOT.InterventionsPatients received BMT and were randomised (unblinded) to open-label LTOT, prescribed for 15 hours per day including overnight hours, or no oxygen therapy.Main outcome measuresThe primary end point was quality of life as measured by the Minnesota Living with Heart Failure (MLwHF) questionnaire score at 6 months. Secondary outcomes included assessing the effect of LTOT on patient symptoms and disease severity, and assessing its acceptability to patients and carers.ResultsBetween April 2012 and February 2014, 114 patients were randomised to receive either LTOT or BMT. The mean age was 72.3 years [standard deviation (SD) 11.3 years] and 70% were male. Ischaemic heart disease was the cause of heart failure in 84%; 95% were in NYHA class III; the mean left ventricular ejection fraction was 27.8%; and the median N-terminal pro-B-type natriuretic hormone was 2203 ng/l. The primary analysis used a covariance pattern mixed model which included patients only if they provided data for all baseline covariates adjusted for in the model and outcome data for at least one post-randomisation time point (n = 102: intervention,n = 51; control,n = 51). There was no difference in the MLwHF questionnaire score at 6 months between the two arms [at baseline the mean score was 54.0 (SD 18.4) for LTOT and 54.0 (SD 17.9) for BMT; at 6 months the mean score was 48.1 (SD 18.5) for LTOT and 49.0 (SD 20.2) for BMT; adjusted mean difference –0.10, 95% confidence interval (CI) –6.88 to 6.69;p = 0.98]. At 3 months, the adjusted mean MLwHF questionnaire score was lower in the LTOT group (–5.47, 95% CI –10.54 to –0.41;p = 0.03) and breathlessness scores improved, although the effect did not persist to 6 months. There was no effect of LTOT on any secondary measure. There was a greater number of deaths in the BMT arm (n = 12 vs.n = 6). Adherence was poor, with only 11% of patients reporting using the oxygen as prescribed.ConclusionsAlthough the study was significantly underpowered, HOT prescribed for 15 hours per day and subsequently used for a mean of 5.4 hours per day has no impact on quality of life as measured by the MLwHF questionnaire score at 6 months. Suggestions for future research include (1) a trial of patients with severe heart failure randomised to have emergency oxygen supply in the house, supplied by cylinders rather than an oxygen concentrator, powered to detect a reduction in admissions to hospital, and (2) a study of bed-bound patients with heart failure who are in the last few weeks of life, powered to detect changes in symptom severity.Trial registrationCurrent Controlled Trials ISRCTN60260702.FundingThis project was funded by the NIHR Health Technology Assessment programme and will be published in full inHealth Technology Assessment; Vol. 19, No. 75. See the NIHR Journals Library website for further project information.

In December 2004, Google Inc. announced its plans to digitize millions of books from prestigious libraries such as Harvard, Stanford, and the New York Public Library. Most of the books are in the public domain and will be available for free on the Internet. The Google initiative is one among many, including the American Memory Project of the Library of Congress, the San Francisco-based Internet Archive, and Gallica, the digital library of the Bibliothèque nationale de France. All of these programs offer free access to good-quality digital materials. Another common feature is that they are heavily funded.

The Junior Scholars Program (JSP) is a tuition-free youth learning program at the Schomburg Center for Research and Black Culture in Harlem, New York. At JSP, Damaris, a former JSP instructor, was introduced to Kadiatou, the Schomburg Center's former education and outreach programs manager. They write collaboratively to (re)member the dialectical relationship between Black women's labor, Black Space, and Radical Black Joy.

1991 An afternoon in late 1991 found me on a Sydney bus reading Brett Easton Ellis’ American Psycho (1991). A disembarking passenger paused at my side and, as I glanced up, hissed, ‘I don’t know how you can read that filth’. As she continued to make her way to the front of the vehicle, I was as stunned as if she had struck me physically. There was real vehemence in both her words and how they were delivered, and I can still see her eyes squeezing into slits as she hesitated while curling her mouth around that final angry word: ‘filth’. Now, almost fifteen years later, the memory is remarkably vivid. As the event is also still remarkable; this comment remaining the only remark ever made to me by a stranger about anything I have been reading during three decades of travelling on public transport. That inflamed commuter summed up much of the furore that greeted the publication of American Psycho. More than this, and unusually, condemnation of the work both actually preceded, and affected, its publication. Although Ellis had been paid a substantial U.S. $300,000 advance by Simon & Schuster, pre-publication stories based on circulating galley proofs were so negative—offering assessments of the book as: ‘moronic … pointless … themeless … worthless (Rosenblatt 3), ‘superficial’, ‘a tapeworm narrative’ (Sheppard 100) and ‘vile … pornography, not literature … immoral, but also artless’ (Miner 43)—that the publisher cancelled the contract (forfeiting the advance) only months before the scheduled release date. CEO of Simon & Schuster, Richard E. Snyder, explained: ‘it was an error of judgement to put our name on a book of such questionable taste’ (quoted in McDowell, “Vintage” 13). American Psycho was, instead, published by Random House/Knopf in March 1991 under its prestige paperback imprint, Vintage Contemporary (Zaller; Freccero 48) – Sonny Mehta having signed the book to Random House some two days after Simon & Schuster withdrew from its agreement with Ellis. While many commented on the fact that Ellis was paid two substantial advances, it was rarely noted that Random House was a more prestigious publisher than Simon & Schuster (Iannone 52). After its release, American Psycho was almost universally vilified and denigrated by the American critical establishment. The work was criticised on both moral and aesthetic/literary/artistic grounds; that is, in terms of both what Ellis wrote and how he wrote it. Critics found it ‘meaningless’ (Lehmann-Haupt C18), ‘abysmally written … schlock’ (Kennedy 427), ‘repulsive, a bloodbath serving no purpose save that of morbidity, titillation and sensation … pure trash, as scummy and mean as anything it depicts, a dirty book by a dirty writer’ (Yardley B1) and ‘garbage’ (Gurley Brown 21). Mark Archer found that ‘the attempt to confuse style with content is callow’ (31), while Naomi Wolf wrote that: ‘overall, reading American Psycho holds the same fascination as watching a maladjusted 11-year-old draw on his desk’ (34). John Leo’s assessment sums up the passionate intensity of those critical of the work: ‘totally hateful … violent junk … no discernible plot, no believable characterization, no sensibility at work that comes anywhere close to making art out of all the blood and torture … Ellis displays little feel for narration, words, grammar or the rhythm of language’ (23). These reviews, as those printed pre-publication, were titled in similarly unequivocal language: ‘A Revolting Development’ (Sheppard 100), ‘Marketing Cynicism and Vulgarity’ (Leo 23), ‘Designer Porn’ (Manguel 46) and ‘Essence of Trash’ (Yardley B1). Perhaps the most unambiguous in its message was Roger Rosenblatt’s ‘Snuff this Book!’ (3). Of all works published in the U.S.A. at that time, including those clearly carrying X ratings, the Los Angeles chapter of the National Organization for Women (NOW) selected American Psycho for special notice, stating that the book ‘legitimizes inhuman and savage violence masquerading as sexuality’ (NOW 114). Judging the book ‘the most misogynistic communication’ the organisation had ever encountered (NOW L.A. chapter president, Tammy Bruce, quoted in Kennedy 427) and, on the grounds that ‘violence against women in any form is no longer socially acceptable’ (McDowell, “NOW” C17), NOW called for a boycott of the entire Random House catalogue for the remainder of 1991. Naomi Wolf agreed, calling the novel ‘a violation not of obscenity standards, but of women’s civil rights, insofar as it results in conditioning male sexual response to female suffering or degradation’ (34). Later, the boycott was narrowed to Knopf and Vintage titles (Love 46), but also extended to all of the many products, companies, corporations, firms and brand names that are a feature of Ellis’s novel (Kauffman, “American” 41). There were other unexpected responses such as the Walt Disney Corporation barring Ellis from the opening of Euro Disney (Tyrnauer 101), although Ellis had already been driven from public view after receiving a number of death threats and did not undertake a book tour (Kennedy 427). Despite this, the book received significant publicity courtesy of the controversy and, although several national bookstore chains and numerous booksellers around the world refused to sell the book, more than 100,000 copies were sold in the U.S.A. in the fortnight after publication (Dwyer 55). Even this success had an unprecedented effect: when American Psycho became a bestseller, The New York Times announced that it would be removing the title from its bestseller lists because of the book’s content. In the days following publication in the U.S.A., Canadian customs announced that it was considering whether to allow the local arm of Random House to, first, import American Psycho for sale in Canada and, then, publish it in Canada (Kirchhoff, “Psycho” C1). Two weeks later, when the book was passed for sale (Kirchhoff, “Customs” C1), demonstrators protested the entrance of a shipment of the book. In May, the Canadian Defence Force made headlines when it withdrew copies of the book from the library shelves of a navy base in Halifax (Canadian Press C1). Also in May 1991, the Australian Office of Film and Literature Classification (OFLC), the federal agency that administers the classification scheme for all films, computer games and ‘submittable’ publications (including books) that are sold, hired or exhibited in Australia, announced that it had classified American Psycho as ‘Category 1 Restricted’ (W. Fraser, “Book” 5), to be sold sealed, to only those over 18 years of age. This was the first such classification of a mainstream literary work since the rating scheme was introduced (Graham), and the first time a work of literature had been restricted for sale since Philip Roth’s Portnoy’s Complaint in 1969. The chief censor, John Dickie, said the OFLC could not justify refusing the book classification (and essentially banning the work), and while ‘as a satire on yuppies it has a lot going for it’, personally he found the book ‘distasteful’ (quoted in W. Fraser, “Sensitive” 5). Moreover, while this ‘R’ classification was, and remains, a national classification, Australian States and Territories have their own sale and distribution regulation systems. Under this regime, American Psycho remains banned from sale in Queensland, as are all other books in this classification category (Vnuk). These various reactions led to a flood of articles published in the U.S.A., Canada, Australia and the U.K., voicing passionate opinions on a range of issues including free speech and censorship, the corporate control of artistic thought and practice, and cynicism on the part of authors and their publishers about what works might attract publicity and (therefore) sell in large numbers (see, for instance, Hitchens 7; Irving 1). The relationship between violence in society and its representation in the media was a common theme, with only a few commentators (including Norman Mailer in a high profile Vanity Fair article) suggesting that, instead of inciting violence, the media largely reflected, and commented upon, societal violence. Elayne Rapping, an academic in the field of Communications, proposed that the media did actively glorify violence, but only because there was a market for such representations: ‘We, as a society love violence, thrive on violence as the very basis of our social stability, our ideological belief system … The problem, after all, is not media violence but real violence’ (36, 38). Many more commentators, however, agreed with NOW, Wolf and others and charged Ellis’s work with encouraging, and even instigating, violent acts, and especially those against women, calling American Psycho ‘a kind of advertising for violence against women’ (anthropologist Elliot Leyton quoted in Dwyer 55) and, even, a ‘how-to manual on the torture and dismemberment of women’ (Leo 23). Support for the book was difficult to find in the flood of vitriol directed against it, but a small number wrote in Ellis’s defence. Sonny Mehta, himself the target of death threats for acquiring the book for Random House, stood by this assessment, and was widely quoted in his belief that American Psycho was ‘a serious book by a serious writer’ and that Ellis was ‘remarkably talented’ (Knight-Ridder L10). Publishing director of Pan Macmillan Australia, James Fraser, defended his decision to release American Psycho on the grounds that the book told important truths about society, arguing: ‘A publisher’s office is a clearing house for ideas … the real issue for community debate [is] – to what extent does it want to hear the truth about itself, about individuals within the community and about the governments the community elects. If we care about the preservation of standards, there is none higher than this. Gore Vidal was among the very few who stated outright that he liked the book, finding it ‘really rather inspired … a wonderfully comic novel’ (quoted in Tyrnauer 73). Fay Weldon agreed, judging the book as ‘brilliant’, and focusing on the importance of Ellis’s message: ‘Bret Easton Ellis is a very good writer. He gets us to a ‘T’. And we can’t stand it. It’s our problem, not his. American Psycho is a beautifully controlled, careful, important novel that revolves around its own nasty bits’ (C1). Since 1991 As unlikely as this now seems, I first read American Psycho without any awareness of the controversy raging around its publication. I had read Ellis’s earlier works, Less than Zero (1985) and The Rules of Attraction (1987) and, with my energies fully engaged elsewhere, cannot now even remember how I acquired the book. Since that angry remark on the bus, however, I have followed American Psycho’s infamy and how it has remained in the public eye over the last decade and a half. Australian OFLC decisions can be reviewed and reversed – as when Pasolini’s final film Salo (1975), which was banned in Australia from the time of its release in 1975 until it was un-banned in 1993, was then banned again in 1998 – however, American Psycho’s initial classification has remained unchanged. In July 2006, I purchased a new paperback copy in rural New South Wales. It was shrink-wrapped in plastic and labelled: ‘R. Category One. Not available to persons under 18 years. Restricted’. While exact sales figures are difficult to ascertain, by working with U.S.A., U.K. and Australian figures, this copy was, I estimate, one of some 1.5 to 1.6 million sold since publication. In the U.S.A., backlist sales remain very strong, with some 22,000 copies sold annually (Holt and Abbott), while lifetime sales in the U.K. are just under 720,000 over five paperback editions. Sales in Australia are currently estimated by Pan MacMillan to total some 100,000, with a new printing of 5,000 copies recently ordered in Australia on the strength of the book being featured on the inaugural Australian Broadcasting Commission’s First Tuesday Book Club national television program (2006). Predictably, the controversy around the publication of American Psycho is regularly revisited by those reviewing Ellis’s subsequent works. A major article in Vanity Fair on Ellis’s next book, The Informers (1994), opened with a graphic description of the death threats Ellis received upon the publication of American Psycho (Tyrnauer 70) and then outlined the controversy in detail (70-71). Those writing about Ellis’s two most recent novels, Glamorama (1999) and Lunar Park (2005), have shared this narrative strategy, which also forms at least part of the frame of every interview article. American Psycho also, again predictably, became a major topic of discussion in relation to the contracting, making and then release of the eponymous film in 2000 as, for example, in Linda S. Kauffman’s extensive and considered review of the film, which spent the first third discussing the history of the book’s publication (“American” 41-45). Playing with this interest, Ellis continues his practice of reusing characters in subsequent works. Thus, American Psycho’s Patrick Bateman, who first appeared in The Rules of Attraction as the elder brother of the main character, Sean – who, in turn, makes a brief appearance in American Psycho – also turns up in Glamorama with ‘strange stains’ on his Armani suit lapels, and again in Lunar Park. The book also continues to be regularly cited in discussions of censorship (see, for example, Dubin; Freccero) and has been included in a number of university-level courses about banned books. In these varied contexts, literary, cultural and other critics have also continued to disagree about the book’s impact upon readers, with some persisting in reading the novel as a pornographic incitement to violence. When Wade Frankum killed seven people in Sydney, many suggested a link between these murders and his consumption of X-rated videos, pornographic magazines and American Psycho (see, for example, Manne 11), although others argued against this (Wark 11). Prosecutors in the trial of Canadian murderer Paul Bernardo argued that American Psycho provided a ‘blueprint’ for Bernardo’s crimes (Canadian Press A5). Others have read Ellis’s work more positively, as for instance when Sonia Baelo Allué compares American Psycho favourably with Thomas Harris’s The Silence of the Lambs (1988) – arguing that Harris not only depicts more degrading treatment of women, but also makes Hannibal Lecter, his antihero monster, sexily attractive (7-24). Linda S. Kauffman posits that American Psycho is part of an ‘anti-aesthetic’ movement in art, whereby works that are revoltingly ugly and/or grotesque function to confront the repressed fears and desires of the audience and explore issues of identity and subjectivity (Bad Girls), while Patrick W. Shaw includes American Psycho in his work, The Modern American Novel of Violence because, in his opinion, the violence Ellis depicts is not gratuitous. Lost, however, in much of this often-impassioned debate and dialogue is the book itself – and what Ellis actually wrote. 21-years-old when Less than Zero was published, Ellis was still only 26 when American Psycho was released and his youth presented an obvious target. In 1991, Terry Teachout found ‘no moment in American Psycho where Bret Easton Ellis, who claims to be a serious artist, exhibits the workings of an adult moral imagination’ (45, 46), Brad Miner that it was ‘puerile – the very antithesis of good writing’ (43) and Carol Iannone that ‘the inclusion of the now famous offensive scenes reveals a staggering aesthetic and moral immaturity’ (54). Pagan Kennedy also ‘blamed’ the entire work on this immaturity, suggesting that instead of possessing a developed artistic sensibility, Ellis was reacting to (and, ironically, writing for the approval of) critics who had lauded the documentary realism of his violent and nihilistic teenage characters in Less than Zero, but then panned his less sensational story of campus life in The Rules of Attraction (427-428). Yet, in my opinion, there is not only a clear and coherent aesthetic vision driving Ellis’s oeuvre but, moreover, a profoundly moral imagination at work as well. This was my view upon first reading American Psycho, and part of the reason I was so shocked by that charge of filth on the bus. Once familiar with the controversy, I found this view shared by only a minority of commentators. Writing in the New Statesman & Society, Elizabeth J. Young asked: ‘Where have these people been? … Books of pornographic violence are nothing new … American Psycho outrages no contemporary taboos. Psychotic killers are everywhere’ (24). I was similarly aware that such murderers not only existed in reality, but also in many widely accessed works of literature and film – to the point where a few years later Joyce Carol Oates could suggest that the serial killer was an icon of popular culture (233). While a popular topic for writers of crime fiction and true crime narratives in both print and on film, a number of ‘serious’ literary writers – including Truman Capote, Norman Mailer, Kate Millet, Margaret Atwood and Oates herself – have also written about serial killers, and even crossed over into the widely acknowledged as ‘low-brow’ true crime genre. Many of these works (both popular or more literary) are vivid and powerful and have, as American Psycho, taken a strong moral position towards their subject matter. Moreover, many books and films have far more disturbing content than American Psycho, yet have caused no such uproar (Young and Caveney 120). By now, the plot of American Psycho is well known, although the structure of the book, noted by Weldon above (C1), is rarely analysed or even commented upon. First person narrator, Patrick Bateman, a young, handsome stockbroker and stereotypical 1980s yuppie, is also a serial killer. The book is largely, and innovatively, structured around this seeming incompatibility – challenging readers’ expectations that such a depraved criminal can be a wealthy white professional – while vividly contrasting the banal, and meticulously detailed, emptiness of Bateman’s life as a New York über-consumer with the scenes where he humiliates, rapes, tortures, murders, mutilates, dismembers and cannibalises his victims. Although only comprising some 16 out of 399 pages in my Picador edition, these violent scenes are extreme and certainly make the work as a whole disgustingly confronting. But that is the entire point of Ellis’s work. Bateman’s violence is rendered so explicitly because its principal role in the novel is to be inescapably horrific. As noted by Baelo Allué, there is no shift in tone between the most banally described detail and the description of violence (17): ‘I’ve situated the body in front of the new Toshiba television set and in the VCR is an old tape and appearing on the screen is the last girl I filmed. I’m wearing a Joseph Abboud suit, a tie by Paul Stuart, shoes by J. Crew, a vest by someone Italian and I’m kneeling on the floor beside a corpse, eating the girl’s brain, gobbling it down, spreading Grey Poupon over hunks of the pink, fleshy meat’ (Ellis 328). In complete opposition to how pornography functions, Ellis leaves no room for the possible enjoyment of such a scene. Instead of revelling in the ‘spine chilling’ pleasures of classic horror narratives, there is only the real horror of imagining such an act. The effect, as Kauffman has observed is, rather than arousing, often so disgusting as to be emetic (Bad Girls 249). Ellis was surprised that his detractors did not understand that he was trying to be shocking, not offensive (Love 49), or that his overall aim was to symbolise ‘how desensitised our culture has become towards violence’ (quoted in Dwyer 55). Ellis was also understandably frustrated with readings that conflated not only the contents of the book and their meaning, but also the narrator and author: ‘The acts described in the book are truly, indisputably vile. The book itself is not. Patrick Bateman is a monster. I am not’ (quoted in Love 49). Like Fay Weldon, Norman Mailer understood that American Psycho posited ‘that the eighties were spiritually disgusting and the author’s presentation is the crystallization of such horror’ (129). Unlike Weldon, however, Mailer shied away from defending the novel by judging Ellis not accomplished enough a writer to achieve his ‘monstrous’ aims (182), failing because he did not situate Bateman within a moral universe, that is, ‘by having a murderer with enough inner life for us to comprehend him’ (182). Yet, the morality of Ellis’s project is evident. By viewing the world through the lens of a psychotic killer who, in many ways, personifies the American Dream – wealthy, powerful, intelligent, handsome, energetic and successful – and, yet, who gains no pleasure, satisfaction, coherent identity or sense of life’s meaning from his endless, selfish consumption, Ellis exposes the emptiness of both that world and that dream. As Bateman himself explains: ‘Surface, surface, surface was all that anyone found meaning in. This was civilisation as I saw it, colossal and jagged’ (Ellis 375). Ellis thus situates the responsibility for Bateman’s violence not in his individual moral vacuity, but in the barren values of the society that has shaped him – a selfish society that, in Ellis’s opinion, refused to address the most important issues of the day: corporate greed, mindless consumerism, poverty, homelessness and the prevalence of violent crime. Instead of pornographic, therefore, American Psycho is a profoundly political text: Ellis was never attempting to glorify or incite violence against anyone, but rather to expose the effects of apathy to these broad social problems, including the very kinds of violence the most vocal critics feared the book would engender. Fifteen years after the publication of American Psycho, although our societies are apparently growing in overall prosperity, the gap between rich and poor also continues to grow, more are permanently homeless, violence – whether domestic, random or institutionally-sanctioned – escalates, and yet general apathy has intensified to the point where even the ‘ethics’ of torture as government policy can be posited as a subject for rational debate. The real filth of the saga of American Psycho is, thus, how Ellis’s message was wilfully ignored. While critics and public intellectuals discussed the work at length in almost every prominent publication available, few attempted to think in any depth about what Ellis actually wrote about, or to use their powerful positions to raise any serious debate about the concerns he voiced. Some recent critical reappraisals have begun to appreciate how American Psycho is an ‘ethical denunciation, where the reader cannot but face the real horror behind the serial killer phenomenon’ (Baelo Allué 8), but Ellis, I believe, goes further, exposing the truly filthy causes that underlie the existence of such seemingly ‘senseless’ murder. But, Wait, There’s More It is ironic that American Psycho has, itself, generated a mini-industry of products. A decade after publication, a Canadian team – filmmaker Mary Harron, director of I Shot Andy Warhol (1996), working with scriptwriter, Guinevere Turner, and Vancouver-based Lions Gate Entertainment – adapted the book for a major film (Johnson). Starring Christian Bale, Chloë Sevigny, Willem Dafoe and Reese Witherspoon and, with an estimated budget of U.S.$8 million, the film made U.S.$15 million at the American box office. The soundtrack was released for the film’s opening, with video and DVDs to follow and the ‘Killer Collector’s Edition’ DVD – closed-captioned, in widescreen with surround sound – released in June 2005. Amazon.com lists four movie posters (including a Japanese language version) and, most unexpected of all, a series of film tie-in action dolls. The two most popular of these, judging by E-Bay, are the ‘Cult Classics Series 1: Patrick Bateman’ figure which, attired in a smart suit, comes with essential accoutrements of walkman with headphones, briefcase, Wall Street Journal, video tape and recorder, knife, cleaver, axe, nail gun, severed hand and a display base; and the 18” tall ‘motion activated sound’ edition – a larger version of the same doll with fewer accessories, but which plays sound bites from the movie. Thanks to Stephen Harris and Suzie Gibson (UNE) for stimulating conversations about this book, Stephen Harris for information about the recent Australian reprint of American Psycho and Mark Seebeck (Pan Macmillan) for sales information. References Archer, Mark. “The Funeral Baked Meats.” The Spectator 27 April 1991: 31. Australian Broadcasting Corporation. First Tuesday Book Club. First broadcast 1 August 2006. Baelo Allué, Sonia. “The Aesthetics of Serial Killing: Working against Ethics in The Silence of the Lambs (1988) and American Psycho (1991).” Atlantis 24.2 (Dec. 2002): 7-24. Canadian Press. “Navy Yanks American Psycho.” The Globe and Mail 17 May 1991: C1. Canadian Press. “Gruesome Novel Was Bedside Reading.” Kitchener-Waterloo Record 1 Sep. 1995: A5. Dubin, Steven C. “Art’s Enemies: Censors to the Right of Me, Censors to the Left of Me.” Journal of Aesthetic Education 28.4 (Winter 1994): 44-54. Dwyer, Victor. “Literary Firestorm: Canada Customs Scrutinizes a Brutal Novel.” Maclean’s April 1991: 55. Ellis, Bret Easton. American Psycho. London: Macmillan-Picador, 1991. ———. Glamorama. New York: Knopf, 1999. ———. The Informers. New York: Knopf, 1994. ———. Less than Zero. New York: Simon & Schuster, 1985. ———. Lunar Park. New York: Knopf, 2005. ———. The Rules of Attraction. New York: Simon & Schuster, 1987. Fraser, James. :The Case for Publishing.” The Bulletin 18 June 1991. Fraser, William. “Book May Go under Wraps.” The Sydney Morning Herald 23 May 1991: 5. ———. “The Sensitive Censor and the Psycho.” The Sydney Morning Herald 24 May 1991: 5. Freccero, Carla. “Historical Violence, Censorship, and the Serial Killer: The Case of American Psycho.” Diacritics: A Review of Contemporary Criticism 27.2 (Summer 1997): 44-58. Graham, I. “Australian Censorship History.” Libertus.net 9 Dec. 2001. 17 May 2006 http://libertus.net/censor/hist20on.html>. Gurley Brown, Helen. Commentary in “Editorial Judgement or Censorship?: The Case of American Psycho.” The Writer May 1991: 20-23. Harris, Thomas. The Silence of the Lambs. New York: St Martins Press, 1988. Harron, Mary (dir.). American Psycho [film]. Edward R. Pressman Film Corporation, Lions Gate Films, Muse Productions, P.P.S. Films, Quadra Entertainment, Universal Pictures, 2004. Hitchens, Christopher. “Minority Report.” The Nation 7-14 January 1991: 7. Holt, Karen, and Charlotte Abbott. “Lunar Park: The Novel.” Publishers Weekly 11 July 2005. 13 Aug. 2006 http://www.publishersweekly.com/article/CA624404.html? pubdate=7%2F11%2F2005&display=archive>. Iannone, Carol. “PC & the Ellis Affair.” Commentary Magazine July 1991: 52-4. Irving, John. “Pornography and the New Puritans.” The New York Times Book Review 29 March 1992: Section 7, 1. 13 Aug. 2006 http://www.nytimes.com/books/97/06/15/lifetimes/25665.html>. Johnson, Brian D. “Canadian Cool Meets American Psycho.” Maclean’s 10 April 2000. 13 Aug. 2006 http://www.macleans.ca/culture/films/article.jsp?content=33146>. Kauffman, Linda S. “American Psycho [film review].” Film Quarterly 54.2 (Winter 2000-2001): 41-45. ———. Bad Girls and Sick Boys: Fantasies in Contemporary Art and Culture. Berkeley: University of California Press, 1998. Kennedy, Pagan. “Generation Gaffe: American Psycho.” The Nation 1 April 1991: 426-8. Kirchhoff, H. J. “Customs Clears Psycho: Booksellers’ Reaction Mixed.” The Globe and Mail 26 March 1991: C1. ———. “Psycho Sits in Limbo: Publisher Awaits Customs Ruling.” The Globe and Mail 14 March 1991: C1. Knight-Ridder News Service. “Vintage Picks up Ellis’ American Psycho.” Los Angeles Daily News 17 November 1990: L10. Lehmann-Haupt, Christopher. “Psycho: Wither Death without Life?” The New York Times 11 March 1991: C18. Leo, John. “Marketing Cynicism and Vulgarity.” U.S. News & World Report 3 Dec. 1990: 23. Love, Robert. “Psycho Analysis: Interview with Bret Easton Ellis.” Rolling Stone 4 April 1991: 45-46, 49-51. Mailer, Norman. “Children of the Pied Piper: Mailer on American Psycho.” Vanity Fair March 1991: 124-9, 182-3. Manguel, Alberto. “Designer Porn.” Saturday Night 106.6 (July 1991): 46-8. Manne, Robert. “Liberals Deny the Video Link.” The Australian 6 Jan. 1997: 11. McDowell, Edwin. “NOW Chapter Seeks Boycott of ‘Psycho’ Novel.” The New York Times 6 Dec. 1990: C17. ———. “Vintage Buys Violent Book Dropped by Simon & Schuster.” The New York Times 17 Nov. 1990: 13. Miner, Brad. “Random Notes.” National Review 31 Dec. 1990: 43. National Organization for Women. Library Journal 2.91 (1991): 114. Oates, Joyce Carol. “Three American Gothics.” Where I’ve Been, and Where I’m Going: Essays, Reviews and Prose. New York: Plume, 1999. 232-43. Rapping, Elayne. “The Uses of Violence.” Progressive 55 (1991): 36-8. Rosenblatt, Roger. “Snuff this Book!: Will Brett Easton Ellis Get Away with Murder?” New York Times Book Review 16 Dec. 1990: 3, 16. Roth, Philip. Portnoy’s Complaint. New York: Random House, 1969. Shaw, Patrick W. The Modern American Novel of Violence. Troy, NY: Whitson, 2000. Sheppard, R. Z. “A Revolting Development.” Time 29 Oct. 1990: 100. Teachout, Terry. “Applied Deconstruction.” National Review 24 June 1991: 45-6. Tyrnauer, Matthew. “Who’s Afraid of Bret Easton Ellis?” Vanity Fair 57.8 (Aug. 1994): 70-3, 100-1. Vnuk, Helen. “X-rated? Outdated.” The Age 21 Sep. 2003. 17 May 2006 http://www.theage.com.au/articles/2003/09/19/1063625202157.html>. Wark, McKenzie. “Video Link Is a Distorted View.” The Australian 8 Jan. 1997: 11. Weldon, Fay. “Now You’re Squeamish?: In a World as Sick as Ours, It’s Silly to Target American Psycho.” The Washington Post 28 April 1991: C1. Wolf, Naomi. “The Animals Speak.” New Statesman & Society 12 April 1991: 33-4. Yardley, Jonathan. “American Psycho: Essence of Trash.” The Washington Post 27 Feb. 1991: B1. Young, Elizabeth J. “Psycho Killers. Last Lines: How to Shock the English.” New Statesman & Society 5 April 1991: 24. Young, Elizabeth J., and Graham Caveney. Shopping in Space: Essays on American ‘Blank Generation’ Fiction. London: Serpent’s Tail, 1992. Zaller, Robert “American Psycho, American Censorship and the Dahmer Case.” Revue Francaise d’Etudes Americaines 16.56 (1993): 317-25. Citation reference for this article MLA Style Brien, Donna Lee. "The Real Filth in : A Critical Reassessment." M/C Journal 9.5 (2006). echo date('d M. Y'); ?> <http://journal.media-culture.org.au/0610/01-brien.php>. APA Style Brien, D. (Nov. 2006) "The Real Filth in American Psycho: A Critical Reassessment," M/C Journal, 9(5). Retrieved echo date('d M. Y'); ?> from <http://journal.media-culture.org.au/0610/01-brien.php>.

Abstract Background Cell-free DNA (cfDNA), which is extracellular DNA present in the circulating plasma and other body fluids, is currently investigated as a minimally invasive, highly informative biomarker. While nucleosome-sized cfDNA fragments have been investigated intensively, shorter DNA fragments in the plasma have not been studied due to several technical limitations. Results We aimed to investigate the existence of shorter cfDNA fragments in the blood. Using an improved cfDNA purification protocol and a 3′-end-labeling method, we found DNA fragments of approximately 50 nucleotides in length in the human plasma, present at a molar concentration comparable to that of nucleosome-sized fragments. Unfortunately, these short fragments cannot be recovered by widely used cfDNA isolation methods. In addition, they are composed of single-stranded DNA (ssDNA), thus escaping detection in previous studies. Therefore, we established a library-preparation protocol based on our unique ssDNA ligation technique and applied it to the isolated cfDNA. Deep sequencing of these libraries revealed that the short fragments are derived from hundreds of thousands of genomic sites in open chromatin regions and enriched with transcription factor-binding sites. Remarkably, antisense strands of putative G-quadruplex motifs occupy as much as one-third of the peaks by these short fragments. Conclusions We propose a new class of plasma cfDNA composed of short single-stranded fragments that potentially form non-canonical DNA structures.