Artykuły w czasopismach na temat „Broken cell serving”

The current standard for investigating tumors is surgical biopsy, which is costly, invasive, and difficult to perform serially. As an adjunct, circulating tumor cells (CTCs)—cells that have broken away from the primary tumor or metastatic sites—can be obtained from a blood draw and offer the potential for obtaining serial genetic information and serving as biomarkers. Here, we detail the potential for melanoma CTCs to serve as biomarkers and discuss a clinically viable methodology for single-cell CTC isolation and analysis that overcomes previous limitations. We explore the use of melanoma CTC biomarkers by isolating and performing single-cell RNA sequencing on CTCs from melanoma patients. We then compared transcriptional profiles of single melanoma CTCs against A375 cells and peripheral blood mononuclear cells to identify unique genes differentially regulated in circulating melanoma tumor cells. The information that can be obtained via analysis of these CTCs has significant potential in disease tracking.

The poly (ADP-ribose) polymerase 1 (PARP1) enzyme is one of the promising molecular targets for the discovery of antitumor drugs. PARP1 is a common nuclear protein (1-2 million molecules per cell) serving as a sensor for DNA strand breaks. Increased PARP1 expression is sometimes observed in melanomas, breast cancer, lung cancer, and other neoplastic diseases. The PARP1 expression level is a prognostic indicator and is associated with a poor survival prognosis. There is evidence that high PARP1 expression and treatment-resistance of tumors are correlated. PARP1 inhibitors are promising antitumor agents, since they act as chemo- and radiosensitizers in the conventional therapy of malignant tumors. Furthermore, PARP1 inhibitors can be used as independent, effective drugs against tumors with broken DNA repair mechanisms. Currently, third-generation PARP1 inhibitors are being developed, many of which are undergoing Phase II clinical trials. In this review, we focus on the properties and features of the PARP1 inhibitors identified in preclinical and clinical trials. We also describe some problems associated with the application of PARP1 inhibitors. The possibility of developing new PARP1 inhibitors aimed at DNA binding and transcriptional activity rather than the catalytic domain of the protein is discussed.

Sleep appears to play an important role in the lives of honey bees, but to understand how and why, it is essential to accurately identify sleep, and to know when and where it occurs. Viewing normally obscured honey bees in their nests would be necessary to calculate the total quantity and quality of sleep and sleep’s relevance to the health and dynamics of a honey bee and its colony. Western honey bees (Apis mellifera) spend much of their time inside cells, and are visible only by the tips of their abdomens when viewed through the walls of an observation hive, or on frames pulled from a typical beehive. Prior studies have suggested that honey bees spend some of their time inside cells resting or sleeping, with ventilatory movements of the abdomen serving as a telltale sign distinguishing sleep from other behaviors. Bouts of abdominal pulses broken by extended pauses (discontinuous ventilation) in an otherwise relatively immobile bee appears to indicate sleep. Can viewing the tips of abdomens consistently and predictably indicate what is happening with the rest of a bee’s body when inserted deep inside a honeycomb cell? To distinguish a sleeping bee from a bee maintaining cells, eating, or heating developing brood, we used a miniature observation hive with slices of honeycomb turned in cross-section, and filmed the exposed cells with an infrared-sensitive video camera and a thermal camera. Thermal imaging helped us identify heating bees, but simply observing ventilatory movements, as well as larger motions of the posterior tip of a bee’s abdomen was sufficient to noninvasively and predictably distinguish heating and sleeping inside comb cells. Neither behavior is associated with large motions of the abdomen, but heating demands continuous (vs. discontinuous) ventilatory pulsing. Among the four behaviors observed inside cells, sleeping constituted 16.9% of observations. Accuracy of identifying sleep when restricted to viewing only the tip of an abdomen was 86.6%, and heating was 73.0%. Monitoring abdominal movements of honey bees offers anyone with a view of honeycomb the ability to more fully monitor when and where behaviors of interest are exhibited in a bustling nest.

Tribbles (TRIB) proteins are pseudokinase mediators of eukaryotic signalling that have evolved important roles in lipoprotein metabolism, immune function and cellular differentiation and proliferation. In addition, an evolutionary-conserved modulation of PI3K/AKT signalling pathways highlights them as novel and rather unusual pharmaceutical targets. The three human TRIB family members are uniquely defined by an acidic pseudokinase domain containing a ‘broken’ α C-helix and a MEK (MAPK/ERK)-binding site at the end of the putative C-lobe and a distinct C-terminal peptide motif that interacts directly with a small subset of cellular E3 ubiquitin ligases. This latter interaction drives proteasomal-dependent degradation of networks of transcription factors, whose rate of turnover determines the biological attributes of individual TRIB family members. Defining the function of individual Tribs has been made possible through evaluation of individual TRIB knockout mice, siRNA/overexpression approaches and genetic screening in flies, where the single TRIB gene was originally described 15 years ago. The rapidly maturing TRIB field is primed to exploit chemical biology approaches to evaluate endogenous TRIB signalling events in intact cells. This will help define how TRIB-driven protein–protein interactions and the atypical TRIB ATP-binding site, fit into cellular signalling modules in experimental scenarios where TRIB-signalling complexes remain unperturbed. In this mini-review, we discuss how small molecules can reveal rate-limiting signalling outputs and functions of Tribs in cells and intact organisms, perhaps serving as guides for the development of new drugs. We predict that appropriate small molecule TRIB ligands will further accelerate the transition of TRIB pseudokinase analysis into the mainstream of cell signalling.

Abstract Introduction: Terminally ill patients are often faced with the decision to forgo potentially life-prolonging treatment or to accept hospice care leading to a peaceful death. The decision process in such situations is heavily affected by emotions, chief among them is regret. Modern cognitive science increasingly accepts a dual processing approach to human cognition which takes into account both emotion-based (type 1) and analytical-based (type 2) cognitive processing. Because regret is a human emotion (type 1), which involves counterfactual deliberations (type 2), we have previously proposed that it can activate both cognitive domains by serving as a link between type 1 and type 2 processes and therefore help with end of life decisions more precisely than other decision making methodologies. Here, we report the application of a regret-based model built to facilitate referral to hospice while helping patients clarify their preferences related to how they wish to spend the remaining days of their lives. Methods: Between March 2013 to December 2015, we conducted a prospective cohort study at the Tampa General Hospital and the Moffitt Cancer Center that enrolled 178 consecutive adult patients aware of the terminal nature of their disease. Eligible patients were those who were at the point in their care where they had to decide between continuing potentially "curative/life-prolonging" treatment (Rx) or accepting hospice care. The study was broken down into 4 steps. First, we computed the patient's probability of survival at 6 months using a validated Palliative Performance Score (PPS-based) predictive model. This probability was communicated to patients as i. percentage, ii. pictorial, and iii. life expectancy in days. Then, we used the Dual Visual Analog Scale technique (DVAS) to elicit patient preferences towards continuing current treatment vs. accepting hospice care. The first scale in DVAS measured the levels of regret of omission (RGO) (e.g. failure to reap hospice benefits and incurring treatment harms) while the second scale measured regret of commission (RGC) (e.g. incurring harms from hospice and failing to provide potential benefits of treatment). The ratio RGO/RGC was used to compute the threshold probability at which a patient is indifferent between accepting hospice care or continuing current treatment. Each patient's threshold was contrasted against the previously estimated survival probability to suggest a patient specific management plan, which was later compared with the patient's actual choice. The final step of the study involved asking each patient a series of qualitative questions to evaluate the usefulness of the regret model in the hospice referral process. Results: 96% (171/178) of the patients found the information provided by the model helpful; 90% (160/178) stated that it will influence their care decision. 85% (151/178) of the patients agreed with the model's recommendations to either accept hospice care or continue with current treatment [p<0.000001]. The regret model predicted the actual choices for 72% (128/178) of patients [p <0.00001]. Logistic regression analysis showed that people who were initially inclined to be referred to hospice and were predicted to choose hospice over disease-directed treatment by the regret model had close to 98% probability of choosing hospice care at the end of their lives. No other factors (age, gender, race, educational status and pain level) affected the patient actual choice. Conclusions: To our knowledge, this is the first formal study in which helping patient clarify their preferences enabled them to make actual choices with high level of satisfaction. The regret model was well received by patients and its recommendations were largely accepted. We found that people suffering from a terminal disease who are initially inclined to choose hospice and do not regret such a choice will select hospice care with high level of certainty. We conclude that using the regret model to elicit patient choices is both descriptively and prescriptively valid and can be easily implemented in the actual practice. Disclosures No relevant conflicts of interest to declare.

Abstract Objectives The cotyledon cell wall of the common bean can be a factor in Fe bioavailability. This study evaluated the iron bioavailability of two bean varieties (white or black) either boiled (intact cell walls) or extruded into pasta formulated from heat treated bean flour as the major ingredient (100% bean flour; broken cell walls). Methods In vitro Fe bioavailability was determined via the Caco-2 Cell Bioassay. In vivo Fe bioavailability was measured by the capacity of a bean-based or bean pasta-based diet to generate and maintain total body hemoglobin iron (Hb-Fe) during a 6 week poultry feeding trial. Results The iron and phytate concentrations of the bean-based and bean pasta-based diets consisting of tomato paste, carrot, cabbage, milk, potato and corn oil were not significantly different. The Caco-2 cell bioassay predicted that the white bean pasta diet would have the highest Fe bioavailability, closely followed by the white bean diet. The bioassay predicted the black bean diet and the black bean pasta diet would have significantly lower Fe bioavailability. For the in vivo studies, animals fed the white bean pasta diet (broken cell walls) had significantly (p ≤ 0.05) higher hemoglobin, Hb-Fe and hemoglobin maintenance efficiencies than animals fed a white bean, black bean or black bean pasta-based diet. Although cotyledon cells were broken, the iron bioavailability of the black bean pasta was not improved after processing into pasta. The low iron bioavailability of black beans was associated with their high concentrations of seed coat procyanidin, cinnamtannin and anthocyanidin compounds that have a negative impact on the absorption of iron. Conclusions This study shows that white colored dry bean possesses a combination of traits that result in improved iron bioavailability after processing into pasta. The enhanced Fe bioavailability from the white bean pasta is likely due to the breakage of the cotyledon cell wall during processing, thus allowing enhanced bioaccessibility of the intracellular Fe. In the black bean pasta diet, this enhancement was not observed due to the presence of the seed coat polyphenols which interacted with the released intracellular Fe and prevented Fe absorption. Funding Sources Funded by the United States Department of Agriculture, Agricultural Research Service.

MAR (Mobile Augmented Reality) is delegated an innovation that gives similar capacities as AR (Augmented Reality), yet without the actual limitations of the area of an exploration office or testing region. A Location-Based Service (LBS) is an application for portable figuring which gives clients administrations dependent on their topographical area. Area based administrations are getting progressively famous with the ascent of cell phones with an ever increasing number of highlights (particularly Apple's iPhone and Android-based gadgets). Increasingly more substance is improved with geo-information and can subsequently be seen in a virtual climate, however in real, portable conditions and in a setting delicate way fit to the requirements of the client. The definition and advantages of versatile increased reality and area based administrations and the mix of portable enlarged reality and area based administrations are broke down in this article. The issues are examined alongside the upsides and downsides.

22 Background: A call for development of a new model for funding Stem Cell Transplant (SCT) services in Ontario led to a costing initiative described below. The new model needed to be in alignment with current costs and clinical practices, and to avoid over or under-funding the required components of care for all types of transplant (autologous, allogeneic-related, allogeneic un-related). Methods: The patient care path was broken down by transplant type into the following phases: Pre-Transplant, Graft Harvesting or Procurement, Transplant, and Follow-Up. The following Canadian and Ontario data sources were used for fiscal year 2013/14 and 2014/15 to identify costs of care: -Case Costing data submitted by the six hospitals providing SCT services, -Discharge Abstract Database (DAD), -National Ambulatory Care Reporting System (NACRS), -Other data sources, as needed (e.g. Specialized Services Oversight Information System, Health Indicator Tool). Additionally, the costs for high cost drugs, infrastructure, psychosocial support, and pathology services components were specifically identified. Broad consultation with clinical, administrative, and costing experts ensured the data was complete and appropriate to patient care needs. Once the costs for each component and phase were estimated, the funding model was developed using the bundling approach where the costs of specific activities and/or phases were bundled together for the purposes of funding. Results: The SCT Funding Model developed based on this costing initiative consists of the following four bundles covering the continuum of a patient’s care from pre-transplant to post-transplant care follow-up: 1.) Pre-Transplant, 2.)Graft Acquisition, 3.)Transplant (includes transplant and follow-up), and 4.) High Cost Drugs. Conclusions: The new SCT Funding Model reflects up-to-date costs and addresses the gaps of the previous approach. It has been implemented and is widely supported due to the extensive stakeholder engagement throughout the process. It is an important component of provincial-level planning for SCT service delivery.

In this article, the authors propose a system to gather patient information continuously, perform proper non-intrusive checking, and propose restorative or potentially way of life engagements, at whatever point required and proper. The structure, which depends on administration arranged service as application(SOAs) and the Cloud, permits a consistent combination of various innovations, applications, also, administrations. It additionally integrates mobile advances to easily gather, convey imperative information from a patient's wearable biosensors while considering a cell phones' restricted abilities and control seepage, notwithstanding discontinuous system detachments. At that point, information is put away in the Cloud and made accessible by means of SOA to permit access by doctors, paramedics, or other approved authorities. A contextual investigation has been created to assess the convenience of the structure, and the preparatory outcomes that have been broken down are demonstrating exceptionally encouraging outcomes. To secure the data transmission, they have applied identity-based data encryption scheme. It enhances the security in the authors' framework.

AbstractThis article reviews the factors that determine the impact of explosive weapons on urban services in space and time, with a focus on drinking water services. The evidence comes from published and unpublished research and records, as well as experience restoring or maintaining such services. Urban services are seen as interconnected, and each composed of interdependent components of people, consumables and hardware. Elements that make up the components are labelled “upstream”, “midstream” and “downstream”, to reflect their location and hierarchy in the production and delivery of any urban service. The impact of explosive weapons is broken into the direct effects on any of the components of a service, and the reverberating effects on up- and or downstream components of the same service, or on other services. The effects are most commonly observed in service infrastructure, and determined chiefly by the extent of the damage to the functionality of any component. The spatial extent of the impact is found to be determined primarily by the hierarchy of the component suffering the direct impact, with attacks on upstream components being the furthest-reaching. The duration of the impact is determined primarily by the pre-explosion “baseline resilience” of the service, itself a function of system redundancies and emergency preparedness and response. The analysis suggests that the impact is more reasonably foreseeable than may commonly be thought, in the sense that the direct effects of explosives are well known and that the most important infrastructure is generally identifiable. It follows that proportionality assessments which involve urban services would benefit from (i) the direct and consistent engagement of specialized engineers within the targeting cell, and (ii) greater familiarity of the weapons controller with services, infrastructure and systems in urban areas.

Photoactivated adenylyl cyclase (PAC) and guanylyl cyclase rhodopsin increase the concentrations of intracellular cyclic nucleotides upon illumination, serving as promising second-generation tools in optogenetics. To broaden the arsenal of such tools, it is desirable to have light-activatable enzymes that can decrease cyclic nucleotide concentrations in cells. Here, we report on an unusual microbial rhodopsin that may be able to meet the demand. It is found in the choanoflagellate Salpingoeca rosetta and contains a C-terminal cyclic nucleotide phosphodiesterase (PDE) domain. We examined the enzymatic activity of the protein (named Rh-PDE) both in HEK293 membranes and whole cells. Although Rh-PDE was constitutively active in the dark, illumination increased its hydrolytic activity 1.4-fold toward cGMP and 1.6-fold toward cAMP, as measured in isolated crude membranes. Purified full-length Rh-PDE displayed maximal light absorption at 492 nm and formed the M intermediate with the deprotonated Schiff base upon illumination. The M state decayed to the parent spectral state in 7 s, producing long-lasting activation of the enzyme domain with increased activity. We discuss a possible mechanism of the Rh-PDE activation by light. Furthermore, Rh-PDE decreased cAMP concentration in HEK293 cells in a light-dependent manner and could do so repeatedly without losing activity. Thus, Rh-PDE may hold promise as a potential optogenetic tool for light control of intracellular cyclic nucleotides (e.g. to study cyclic nucleotide-associated signal transduction cascades).

The ovary is an endocrine and exocrine gland that plays primary role in female development and reproductive activities. The seamen are collecting a lot of heavy metals by serving as filters in the seas. Since the last decades of the nineteenth century and until today, the study of the consequences of human exposure to heavy metals has risen as a central research area in the toxicological field. Among the group of metals with proven human toxicity aluminium (Al) and lead (Pb) are known to be highly neurotoxic. The Mediterranean mussel (Mytilus galloprovincialis) is a species of bivalve, a marine mollusc in the family Mytilidae. In our previous researches, we found Al, zinc (Zn) and iron (Fe) values were higher in mussels taken from Çamburnu region of the Dardanelles. In many tumors, inducible nitric oxide synthase (iNOS) expression is high, however, the role of iNOS during tumor development is very complex and quite perplexing, with both promoting and inhibiting actions having been described. The purpose of the study is to demonstrate the iNOS immunoreactivity in the ovarian tissues of rats which are fed with mussels that are collected from the Çamburnu region of the Dardanelles. The mussels given as food to the rats were removed from the Çamburnu region of the Dardanelles. Average 100±10 g weight were selected. After the beaks were overcooked, the meat broke off and the meat at 100 degrees was dried. Two groups of rats are included in the study, group 1 (n=6), control group fed with standard rat food, group 2 (n=6), 90% mussels and 10% standard rat food daily. To detect iNOS localization in the tissues, the LAB-SA Detection System was used. iNOS immunoreactivity was detected in the interstitial cell cytoplasm of the ovaries of rats fed with mussel. iNOS immunoreactivity was also observed in germ cell cytoplasm in some primordial follicles. There was statistically significant difference between the iNOS immunoreactivity of the interstitial cells in the ovarian parenchyma of the rats in the experimental and control groups (p> 0.05).

Spectrum inefficiency is a significant issue because of the increasingly more solicitation of transmission capacity by the end clients. Accomplishing high transmission rates and elevated levels of Quality of Service (QoS) speaks to in any case an open issue. Long haul Evolution (LTE) has been proposed as the reason for the fourth era versatile cell systems (4G) that points of the LTE standard are higher client bit rates, lower delays, expanded range proficiency, diminished expense, and operational effortlessness. In any case, this innovation is as yet being worked on and a few open issues must be still explored, for example, obstruction coordination, and power utilization, assets the board and handover procedures. The point of this work is to ensure the decrease of intensity utilization utilizing another handover calculation dependent on green arrangement. Also, the proposed conspire ensures the minimization of superfluous handovers. In any case, the issue in this technique is doesn't mulls over of the impact of green strategies in high force hubs. Along these lines, in the proposed framework the impact of high force hubs are thought of. This situation is expanded more than one full scale cell and various kinds of low force hubs, for example, pico-cell and microcells. In this situation, the force sparing is broke down during the handover strategies. Think about these issues; another strategy is proposed called Dynamic base Station arranging (DBSP) for accomplishing vitality productivity. The fundamental idea is to kill a BS individually that will insignificantly influence the system by utilizing a recently presented thought of system sway, which considers the extra burden increases brought to its neighboring BSs. So as to additionally diminish the flagging and execution overhead over the air and back take, utilize the estimated estimations of system sway as their choice measurements. A trial result shows that the proposed strategy accomplishes high vitality proficiency under different situations. In this work the presentation of an UMTS organize situation is assessed by utilizing different estimations of the priority bits of the CBR application.

Abstract The melanomacrophage center (MMC), aggregates of highly pigmented melanomacrophages found within the lymphoid tissues of ectotherm vertebrates, is thought to be the evolutionary precursor to the mammalian germinal center (GC). Studies of cold-blooded vertebrates suggest that the MMC is the site of the humoral adaptive immune response, where B cells proliferate, generate high affinity antibody, and undergo affinity maturation. MMCs transiently increase in size/number following immunization or bacterial infection, supporting the hypothesis that the MMC response is likely linked to the adaptive immune response. However, while morphological studies and shared expression of a GC specific gene further support the idea that the MMC is a proto-GC, melanomacrophages perform many functions not associated with the humoral adaptive immune response. Therefore, to determine if the MMC is serving in a GC-like capacity or is a more generalist feature of ectotherm biology, we turned to the fish model system, three spine stickleback. Fish MMCs increase in size in response to immunization with NP-CGG in alum, but not control injection with adjuvant alone. Fish MMCs are also found in close proximity to lymphocytes and cells expressing the B/T cell activation marker GL7. These preliminary studies not only give further support to the notion of the MMC as a GC homolog, but also provide important tools to broaden the study of adaptive immunity in other fish, reptilian, and amphibian systems.

Introduction: Multiple case reports have described lymphomas and plasma cell neoplasms developing after trauma 1-4. It has been hypothesized that trauma-induced inflammation may contribute to the neoplasia, and that the site of trauma may serve as the nidus (locus minoris resistentiae) 1, 5. However, to our knowledge there are no studies demonstrating causality. Traumatic injuries sustained during combat have been linked with higher rates of hypertension, coronary artery disease, diabetes mellitus and chronic kidney disease 6. Whether severe injury also increases the risk of hematologic malignancies is unknown. We hypothesized that combat injured veterans have higher rates of lymphoma and plasma cell dyscrasias than un-injured combat veterans. Methods: This was a retrospective cohort study of US military personnel injured during combat operations in Iraq or Afghanistan from 2002-2015 extracted from the Department of Defense (DoD) Trauma Registry. Patients were excluded if they died in theater, had multiple battle injuries, had pre-existing cancer diagnoses or missing data. A comparator arm of deployed and un-injured Iraq and Afghanistan combat veterans was obtained from the Military Health System Data Repository (MDR) matched for year of birth, branch of service, and sex. Cancer diagnoses were defined using International Classification of Diseases, Ninth Revision (ICD-9) and Tenth Revision (ICD-10) Clinical Modification codes obtained from both the DoD and the Veterans Health Administration through the MDR and the Veterans Informatics and Computing Infrastructure, respectively. Malignancy classifications by ICD codes are demonstrated in Table 1 and broken down into aggressive B-cell lymphomas, indolent B-cell lymphomas, Hodgkin lymphoma, T-cell lymphomas and plasma cell dyscrasias. Statistical analyses were performed with the chi-square test and fisher's exact test. This study was approved by the IRB at David Grant USAF Medical Center. Results: There were 9,654 subjects each in the injured and un-injured cohorts. Baseline demographics are demonstrated in Table 2. The mean age was 26 and most patients were junior enlisted, members of the Army, males, and of non-Hispanic white ethnicity/race. The average post-trauma follow-up time was 5.3 years. Overall rates of aggressive B-cell lymphomas, indolent B-cell lymphomas, Hodgkin lymphoma, T-cell lymphomas and plasma cell dyscrasias were low, as demonstrated in Table 3, and there were no statistically significant differences in incidence rates between the two cohorts. Discussion: Traumatic injuries can activate innate immune responses involving inflammatory cytokines, complement, coagulation and dendritic cells, resulting in a pro-inflammatory state 7. Preclinical studies reveal that such inflammation may hamper T-cell immune-surveillance needed to eradicate malignancy 8. However, in our cohort of severely injured combat veterans, rates of lymphomas and plasma cell dyscrasias were not increased over un-injured combat veterans. Strengths of this study include a well-characterized cohort with a matched comparator arm enabling vigorous examination of the impacts of traumatic injuries on cancer. Limitations include retrospective design and relatively short follow up. It is possible that differences will emerge with longer follow up. Conclusions: Despite critical combat trauma injuries being associated with a heightened risk of multiple chronic comorbidities, they do not appear to heighten the risk of lymphoid or plasma cell neoplasms within the first 5-years. The opinions and assertions expressed herein are those of the authors and do not necessarily reflect the official policy or position of the U.S. Air Force, David Grant USAF Medical Center, the Uniformed Services University or the Department of Defense. References: 1. Kriwalsky MS, Oral Surg, 2010. 2. Huang J, Medicine, 2019. 3. Stemberga V, Hematol Oncol, 2003. 4. Erdogan B, Eur Spine J, 2005. 5. Lo Schiavo A, Clin Dermatol, 2014. 6. Stewart IJ, Circulation, 2015. 7. Huber-Lang M, Nature Immunology, 2018. 8. Krall J, Science, 2018. Disclosures No relevant conflicts of interest to declare.

Background The COVID-19 pandemic has placed strains on communities. During this public health crisis, health systems have created remote methods of monitoring symptom progression and delivering care virtually. Objective Using an SMS text message-based system, we sought to build and test a remote model to explore community needs, connect individuals to curated resources, and facilitate community health worker intervention when needed during the pandemic. The primary aims of this pilot study were to establish the feasibility (ie, engagement with the text line) and acceptability (ie, participant ratings of resources and service) of delivering automated well-being resources via smartphone technology. Methods Eligible patients (aged 18 years or older, having a cell phone with SMS text messaging capability, and recently visited the emergency department) were identified using the electronic health record. The patients were consented to enroll and begin receiving COVID-19–related information and links to community resources. We collected open-ended and close-ended resource and mood ratings. We calculated the frequencies and conducted a thematic review of the open-ended responses. Results In 7 weeks, 356 participants were enrolled; 13,917 messages were exchanged including 333 resource ratings (mean 4) and 673 well-being scores (mean 6.8). We received and coded 386 open-ended responses, most of which elaborated upon their self-reported mood score (29%). Overall, 77% (n=274) of our participants rated the platform as a service they would highly recommend to a family member or friend. Conclusions This approach is designed to broaden the reach of health systems, tailor to community needs in real time, and connect at-risk individuals with robust community health support.

Background:Immediately following the first wave of the COVID-19 pandemic, the number of giant cell arteritis (GCA) diagnoses noticeably increased at the Royal National Hospital for Rheumatic Diseases in Bath, UK. Furthermore, there was an increase in the proportion of patients with visual complications [1]. The finding supports the viral hypothesis of GCA aetiopathogenesis as previously described [2]. This not only has ramifications for understanding the underlying disease mechanisms in GCA but also has implications for the provision of local GCA services which may have already be affected by the pandemic.Objectives:The objective of the study was to estimate the incidence of giant cell arteritis during the COVID-19 pandemic years of 2020 – 2021 and compare it to 2019 data. Given that there have now been two distinct peaks of COVID-19 as reflected by hospital admissions of COVID-19-positive patients this has allowed us to investigate if there is a temporal relationship between the prevalence of COVID-19 and the incidence of GCA.Methods:The incidence of GCA was calculated by assessing emailed referrals to the GCA service and the hospital electronic medical records to identity positive cases from 2019 to the current date. Local COVID-19 prevalence was estimated by measuring the number of hospital beds taken up by COVID-19 positive patients, available publicly in a UK Government COVID-19 dataset [3].Results:There were 61 (95% Poisson distribution confidence interval [CI] 47 - 78) probable or definite GCA diagnoses made in 2020 compared to 28 (CI 19 – 40) in 2019 (Figure 1). This is an excess of 33 cases in 2020, or an increase in 118%. Given that 41% of the hospital’s catchment population is over 50, this equates to an annual incidence rate of 13.7 per 100,000 in 2019 and 29.8 per 100,000 in 2020. This compares to a previously estimated regional incidence rate of 21.6 per 100,000 for the South West of the UK [4].Figure 1. Prevalence of hospital COVID-19 and incidence of GCA (2019 – 2021). Graph showing the number of hospital beds occupied by COVID-19-positive patients in 2020 – 2021 (blue circles), number of daily GCA diagnoses in 2020 – 2021 (red circles), and previous GCA diagnoses in 2019 (green circles). The broken lines represent moving averages with a period of 7 days for COVID-19 cases and 28 days for GCA diagnoses.A peak in COVID-19-positive inpatients was seen on 10th April 2020 with a corresponding peak of GCA diagnoses on 29th May 2020, giving a lag period of approximately 6 weeks between these peaks (Figure 1).Conclusion:The incidence of GCA in Bath was significantly increased in 2020 compared to 2019. This may be the result of the widespread infection of the local population with the COVID-19 virus as a precipitating factor. Possible mechanisms include, but are not limited to, endothelial disruption by the virus, immune system priming towards T helper cell type 1 (Th1) cellular immunity and/or activation of the monocyte-macrophage system. More work is currently underway to assess the causal relationship between the two diseases.There was a lag period of 6 weeks between the peak during the first wave of the pandemic and the rise in GCA cases. We shall be closely monitoring the number of referrals that follow the current wave of the pandemic.References:[1]Luther R, Skeoch S, Pauling JD, et al. Increased number of cases of giant cell arteritis and higher rates of ophthalmic involvement during the era of COVID-19. Rheumatol Adv Pract 2020;4:1–4. doi:10.1093/rap/rkaa067[2]Russo MG, Waxman J, Abdoh AA, et al. Correlation between infection and the onset of the giant cell (temporal) arteritis syndrome. Arthritis Rheum 1995;38:374–80. doi:10.1002/art.1780380312[3]England PH. GOV.UK Coronavirus (COVID-19) in the UK. 2021.https://coronavirus.data.gov.uk/details/download (accessed 25 Jan 2021).[4]Smeeth L, Cook C, Hall AJ. Incidence of diagnosed polymyalgia rheumatica and temporal arteritis in the United Kingdom, 1990-2001. Ann Rheum Dis 2006;65:1093–8. doi:10.1136/ard.2005.046912Disclosure of Interests:Ben Mulhearn Speakers bureau: Novartis UK, 2019, Grant/research support from: Chugai, 2019, Jessica Ellis: None declared, Sarah Skeoch: None declared, John Pauling: None declared, Sarah Tansley: None declared

Abstract: Rather than attempting to prevent all overdoses, the Prescription Pill Dispenser Vial (RPPDV) aims to prevent accidental overdoses by controlling how many pills a user can access within a twenty-four hour period. The RPPDV is much like a traditional pill vial. The body is similar to a conventional pill vial; however, it is modified to prevent the cap's opening without a special screw. The cap is heavily modified and uses a CR2032 or CR2025 coin cell battery to power a gate and the unlocking of the pill cap. The RPPDV works first when the user clicks the red button on the cap of the RPPDV, which opens the pill gate and opens space into a pill container section. This section is large enough for only one pill (or two if multiple pills are required simultaneously). Once a tablet has entered the pill container section, the button must be pressed again to close the pill gate. The user cannot open the lid to the pill container section until the pill gate is closed (see Fig 1.4.1). The RPPDV is similar to an escrow service, with the pill holder section acting as a middleman. The size of the pill holder container prevents access to multiple pills at the same time. The number of times the user presses the button is recorded and timed with a 555 timer set for twenty-four hours. For example, if a drug has a maximum dose of three pills per day, the button could be capped at four cycles or eight total pushes (to allow a grace cycle if a tablet is broken or stuck). Similarly, the gate cannot be opened unless the lid to the pill container section is closed. The production price of the RPPDV is significantly higher than a traditional pill vial and can get higher depending on the additional features added to the RPPDV. The necessary components to function include CR2032 or CR2025 lithium batteries, a 555 timer, and a coin battery holder, along with plastic and metal molds. The base model alone produces an estimated average cost per unit of $1.33 USD (see Fig 4.2.1). Additional features include adding a Bluetooth Low Energy (Bluetooth LE) component to customize the number of unlocks per day and RFIDs for storage and tracking. Prices vary depending on the specifications of the RPPDV.

ABSTRACTA defining feature of mycobacterial redox metabolism is the use of an unusual deazaflavin cofactor, F420. This cofactor enhances the persistence of environmental and pathogenic mycobacteria, including after antimicrobial treatment, although the molecular basis for this remains to be understood. In this work, we explored our hypothesis that F420enhances persistence by serving as a cofactor in antimicrobial-detoxifying enzymes. To test this, we performed a series of phenotypic, biochemical, and analytical chemistry studies in relation to the model soil bacteriumMycobacterium smegmatis. Mutant strains unable to synthesize or reduce F420were found to be more susceptible to a wide range of antibiotic and xenobiotic compounds. Compounds from three classes of antimicrobial compounds traditionally resisted by mycobacteria inhibited the growth of F420mutant strains at subnanomolar concentrations, namely, furanocoumarins (e.g., methoxsalen), arylmethanes (e.g., malachite green), and quinone analogues (e.g., menadione). We demonstrated that promiscuous F420H2-dependent reductases directly reduce these compounds by a mechanism consistent with hydride transfer. Moreover,M. smegmatisstrains unable to make F420H2lost the capacity to reduce and detoxify representatives of the furanocoumarin and arylmethane compound classes in whole-cell assays. In contrast, mutant strains were only slightly more susceptible to clinical antimycobacterials, and this appeared to be due to indirect effects of F420loss of function (e.g., redox imbalance) rather than loss of a detoxification system. Together, these data show that F420enhances antimicrobial resistance in mycobacteria and suggest that one function of the F420H2-dependent reductases is to broaden the range of natural products that mycobacteria and possibly other environmental actinobacteria can reductively detoxify.IMPORTANCEThis study reveals that a unique microbial cofactor, F420, is critical for antimicrobial resistance in the environmental actinobacteriumMycobacterium smegmatis. We show that a superfamily of redox enzymes, the F420H2-dependent reductases, can reduce diverse antimicrobialsin vitroandin vivo.M. smegmatisstrains unable to make or reduce F420become sensitive to inhibition by these antimicrobial compounds. This suggests that mycobacteria have harnessed the unique properties of F420to reduce structurally diverse antimicrobials as part of the antibiotic arms race. The F420H2-dependent reductases that facilitate this process represent a new class of antimicrobial-detoxifying enzymes with potential applications in bioremediation and biocatalysis.

Background Bronchiolitis obliterans syndrome (BOS), also known as respiratory chronic Graft versus Host Disease (cGvHD), is an obstructive airway disease of the lungs following alloHSCT that has significant morbidity and mortality. BOS is characterized by T-cell mediated inflammation and fibrosis of bronchiolar walls that reduces the diameter of the bronchioles resulting in progressive and irreversible airflow obstruction. BOS is a well described complication following lung transplant and can also be a complication following alloHSCT, with similar histopathology and clinical symptoms. There is currently no approved therapy for the treatment of BOS. While data on alloHSCT costs are available (Milliman 2017; Broder et al., 2017), little is known about the impact of BOS on healthcare resource use (HRU) and costs in alloHSCT. The aim of this study is to quantify the economic burden of BOS in alloHSCT patients. Methods We performed a retrospective analysis of commercial claims data from the IQVIA PharMetrics Plus™ database for patients aged 0-64 who were treated with alloHSCT from 1/1/2007 to 9/30/2017. Patients were observable 12 months before and after index alloHSCT. Those who developed BOS were identified using International Classification of Disease (ICD) diagnosis codes (ICD-9: 491.8, 491.9, 515, 516.34, 561.8; ICD-10: J41.8, J42, J84.09, J84.89, J84.115) and propensity score matched to identify patients who did not develop BOS after alloHSCT. We calculated mean annual per patient rates of hospitalizations, tests and treatments for BOS, as well as costs in the follow-up year for inpatient admissions, Emergency Department (ED) and non-ED outpatient visits, and prescription medications. Costs for the index HSCT were included in the first follow-up year costs. All analyses were repeated for the subset of patients observable in the second post-alloHSCT year. The effect of BOS was estimated as the difference in HRU and costs between the matched BOS and non-BOS alloHSCT patients. Results We identified 161 alloHSCT patients with commercial insurance coverage who developed BOS. A majority were male (60%). Mean age was 51.0 yrs (+/- SD: 12.13); a small proportion (2%) were under age 18. We also identified 161 matched alloHSCT patients who did not develop BOS. More BOS patients were diagnosed with leukemias or lymphomas compared with non-BOS patients (94% vs. 84%) in the pre-alloHSCT year, and more had pulmonary disease (asthma or COPD; 24% vs. 16%). No other differences were statistically significant at p&gt;0.05. In the year following alloHSCT, BOS patients had higher mean per patient annual numbers of inpatient admissions (3.9 [+/- SD: 3.3] vs. 2.6 [+/- SD: 2.5]). More BOS patients received lung function testing, including spirometry (60% vs. 42%), and more were treated with ventilation, oxygen therapy or pulmonary rehabilitation (61% vs. 39%). Costs were higher for BOS patients ($560,048 vs. $408,764), with inpatient costs responsible for most of this difference ($446,622 vs. $300,146). Lung function test costs were over 3 times as high for BOS patients ($35,744 vs. $10,192) and lung function treatment costs 2.5 times as high ($31,761 vs. $7,994). Costs for patients observable in the second post-alloHSCT year were considerably lower, as the costs for the initial alloHSCT costs were reflected in the first year expenditures. Nonetheless, costs remained higher for BOS patients ($72,829 vs. $43,665), with higher inpatient, outpatient and prescription drug costs and HRU (Table, Figure). Conclusion AlloHSCT patients who develop BOS in the U.S. have higher rates of hospitalization and have more lung function tests and treatments, compared with alloHSCT patients with no diagnosis of BOS in the first 1-2 years post-alloHSCT. These higher rates of healthcare service use are accompanied by mean annual per patient costs that are $151,000 higher in the first post-alloHSCT year. Disclosures Sacks: Breath Therapeutics Inc.: Other: Employee of Precision Health Economics (PHE). PHE received funding from Breath Therapeutics for this research. . Healey:Breath Therapeutics Inc.: Other: Employee of Precision Health Economics (PHE). PHE received funding from Breath Therapeutics for this research. . Cyr:Breath Therapeutics Inc.: Other: Employee of Precision Health Economics (PHE). PHE received funding from Breath Therapeutics for this research. . Batt:Breath Therapeutics Inc.: Other: Scientific Adviser to Precision Health Economics (PHE). PHE receieved funding from Breath Therapeutics for this research.. Henig:Breath Therapeutics Inc.: Employment.

Abstract Introduction : IGM-2323 is the first engineered high-affinity, high-avidity bispecific IgM monoclonal antibody TCE to be tested in the clinic. It has 10 binding domains for CD20 and a single binding domain for CD3 and is designed to bind irreversibly to CD20-high and low-expressing cells with more physiologic stimulation to T cells, which may mitigate cytokine release syndrome (CRS)-related toxicity and broaden the therapeutic window. IGM-2323 may act by multiple mechanisms: T-cell dependent cytotoxicity, complement dependent cytotoxicity, and enhanced immune modulation via IFNγ-dominant cytokine stimulation. This phase 1 study is exploring the safety and activity of IGM-2323 using a dose titration schedule intended to optimize repeatable immune stimulation while minimizing toxicity. Methods : This first-in-human Phase 1 study is a global, multicenter, open-label, dose escalation evaluating safety, tolerability, PK, and preliminary efficacy (NCT04082936). Adults with relapsed or refractory CD20 + B-cell NHL with ≥ 2 prior systemic therapies, adequate organ function, and ECOG 0-1 are eligible. IGM-2323 is given IV on Days 1, 8, and 15 of 21-day cycles until disease progression or unacceptable toxicity. Treatment can continue beyond progression if the patient (pt) has benefitted from treatment and intra-patient dose escalation is allowed. This study also utilizes a dose titration scheme where a starting dose is given on Day 1, then higher subsequent doses are given up to a plateau dose. If a pt has symptoms of CRS, the same dose may be repeated, and a higher dose is not given until the dose is well tolerated. Results : As of April 30, 2021, 29 pts have been enrolled: 12 at 5 fixed dose levels (0.5, 2.5, 10, 30, 100 mg) and 17 at 5 dose titration levels (50/100, 50/200, 50/300, 50/600, 50/1000 mg)*. NHL subtypes include: 13 follicular (FL), 11 diffuse large B-cell (DLBCL), 3 mantle cell (MCL), and 2 marginal zone (MZL). Median age is 66 (range, 36-84) and median prior therapies is 3 (range, 2-7). 2 pts had prior autologous transplant and 7 had prior CAR-T. All 29 pts received at least one dose and are safety evaluable. There were no DLTs and no neurotoxicity AEs. No pts discontinued due to AE. There were 3 drug-related SAEs (1 each Gr1/2/3 CRS by ASTCT). 16 pts discontinued treatment: 2 pt/investigator decision and 14 PD. The 13 pts still on treatment have received a median of 9 doses (range, 1-42). At fixed dose levels, 5 of 12 pts had a CRS event, primarily grade 1 (fever). Only 1 of 17 titration pts had CRS (Gr3 in a pt with prior experimental CAR-T and baseline circulating B-cells). The most common drug-related AEs in titration pts were hypophosphatemia (29%), IRR (29%; the distinction between IRR and CRS was determined by the investigator), and nausea (24%). 23 pts were evaluable for efficacy; there were 8 responses (5 CR, 3 PR). 5 responses were in FL/MZL and 3 in DLBCL/MCL. Response kinetics varied, with some CRs at the first scan (Week 6) and others after biopsy-confirmed pseudo-progression or prolonged SD (up to 69 weeks). 7 of 8 responses were ongoing as of the data cut. 5 of 11 evaluable patients in dose titration cohorts responded (3 CR, 2 PR), with a median time to response of 6 weeks. In the titration cohort with the longest follow-up (50/100), there were 2 CR and 1 PR in 4 evaluable pts. Repeatable IFNγ stimulation is detectable in nearly all pts with measurable cytokine elevations, while polycytokine response was observed in CRS cases (Table). Preliminary IHC data in matched baseline and on-treatment biopsies (n=3) shows decrease of CD20 + tumor cells and increase of CD3 + T cells in tumor tissue upon treatment, regardless of clinical response. Conclusions : Preliminary results from this first-in-human study of IGM-2323 show an excellent safety and tolerability profile at up to 1000 mg, with reduced CRS when IGM-2323 was given using a dose titration scheme. Response patterns included rapid responses, deepening responses and pseudo-progression. IGM-2323 also has evidence of an IFNγ-dominant repeatable T-cell activation and preservation of T cell function over time. The MTD of IGM-2323 has not been reached. Clinical activity was observed across multiple histologies. Updated safety, PK, biomarker, and efficacy data, including complete dose escalation data, will be presented at the meeting. * starting and plateau dose referenced Figure 1 Figure 1. Disclosures Budde: Mustang Bio, Inc: Research Funding; Roche: Consultancy; BeiGene: Consultancy; Novartis: Consultancy; AstraZeneca: Research Funding; IGM Biosciences: Research Funding; Merck, Inc: Research Funding; Amgen: Research Funding; Gilead: Consultancy. Gopal: Agios: Research Funding; MorphoSys: Honoraria; Bristol Meyers Squibb: Research Funding; Incyte: Honoraria; Acrotech: Consultancy, Honoraria; Servier: Consultancy, Honoraria; IGM Biosciences: Research Funding; Nurix Inc: Consultancy, Honoraria; Beigene: Consultancy, Honoraria; Astra-Zeneca: Research Funding; Takeda: Research Funding; Teva: Research Funding; ADC Therapeutics: Consultancy, Honoraria; Kite: Consultancy, Honoraria; Cellectar: Consultancy, Honoraria; Genetech: Consultancy, Honoraria, Research Funding; Gilead: Consultancy, Honoraria, Research Funding; I-Mab bio: Consultancy, Honoraria, Research Funding; Epizyme: Consultancy, Honoraria; SeaGen: Consultancy, Honoraria, Research Funding; Janssen: Consultancy, Honoraria, Research Funding; Karyopharm: Consultancy, Honoraria; Merck: Consultancy, Honoraria, Research Funding. Kim: Dong-A Pharmaceutical: Research Funding; Eisai: Research Funding; Celltrion: Research Funding; Johnson & Johnson: Research Funding; Kyowa Kirin: Research Funding; Roche: Research Funding; Sanofi: Research Funding; IGM Biosciences: Research Funding. Flinn: Novartis: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Seagen: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Incyte: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Pharmacyclics LLC, an AbbVie Company: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Forma Therapeutics: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Curis: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Forty Seven: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Infinity Pharmaceuticals: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; MorphoSys: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Constellation Pharmaceuticals: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Yingli Pharmaceuticals: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Verastem: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Calithera Biosciences: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; TG Therapeutics: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Celgene: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Takeda: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Juno Therapeutics: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Kite, a Gilead Company: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; IGM Biosciences: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Janssen: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Roche: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; AbbVie: Consultancy, Other: All Consultancy and Research Funding payments made to Sarah Cannon Research Institute, Research Funding; Gilead Sciences: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Trillium Therapeutics: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Acerta Pharma: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Karyopharm Therapeutics: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Loxo: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Merck: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Rhizen Pharmaceuticals: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Teva: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Agios: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Unum Therapeutics: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Nurix Therapeutics: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Pfizer: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Great Point Partners: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Portola Pharmaceuticals: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; ArQule: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Iksuda Therapeutics: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Genentech: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; BeiGene: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; AstraZeneca: Consultancy, Other: All consultancy and research funding payments made to Sarah Cannon Research Institute, Research Funding; Triphase Research & Development Corp.: Other: All research funding payments made to Sarah Cannon Research Institute, Research Funding; Century Therapeutics: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Hutchison MediPharma: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Vincerx Pharma: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Sarah Cannon Research Institute: Current Employment; Servier Pharmaceuticals: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Yingli Pharmaceuticals: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Seagen: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Servier Pharmaceuticals: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute; Unum Therapeutics: Consultancy, Other: All consultancy payments made to Sarah Cannon Research Institute, Research Funding; Johnson & Johnson: Current holder of individual stocks in a privately-held company; Seattle Genetics: Research Funding. Cheah: Ascentage Pharma: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; AstraZeneca: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; BMS: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Roche: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel, Accommodations, Expenses, Research Funding; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Gilead: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Lilly: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; TG therapeutics: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; MSD: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Beigene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Abbvie: Research Funding. Nastoupil: Bayer: Honoraria; Denovo Pharma: Other: DSMC; Novartis: Honoraria, Research Funding; Genentech: Honoraria, Research Funding; Pfizer: Honoraria, Research Funding; TG Therapeutics: Honoraria, Research Funding; Takeda: Honoraria, Other: DSMC, Research Funding; IGM Biosciences: Research Funding; Caribou Biosciences: Research Funding; Gilead/Kite: Honoraria, Research Funding; Bristol Myers Squibb/Celgene: Honoraria, Research Funding; Epizyme: Honoraria, Research Funding; ADC Therapeutics: Honoraria; Janssen: Honoraria, Research Funding; MorphoSys: Honoraria. Matasar: Daiichi Sankyo: Consultancy; TG Therapeutics: Consultancy, Honoraria; Merck Sharp & Dohme: Current holder of individual stocks in a privately-held company; Juno Therapeutics: Consultancy; Rocket Medical: Consultancy, Research Funding; Genentech, Inc.: Consultancy, Honoraria, Research Funding; Teva: Consultancy; F. Hoffmann-La Roche Ltd: Consultancy, Honoraria, Research Funding; Memorial Sloan Kettering Cancer Center: Current Employment; Bayer: Consultancy, Honoraria, Research Funding; GlaxoSmithKline: Honoraria, Research Funding; Merck: Consultancy; IGM Biosciences: Research Funding; Pharmacyclics: Honoraria, Research Funding; ImmunoVaccine Technologies: Consultancy, Honoraria, Research Funding; Janssen: Honoraria, Research Funding; Seattle Genetics: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria. Diefenbach: Janssen: Consultancy, Honoraria, Research Funding; Morphosys: Consultancy, Honoraria, Research Funding; Merck Sharp & Dohme: Consultancy, Honoraria, Research Funding; Gilead: Current equity holder in publicly-traded company; Trillium: Research Funding; IGM Biosciences: Research Funding; Celgene: Research Funding; AbbVie: Research Funding; Perlmutter Cancer Center at NYU Langone Health: Current Employment; Incyte: Research Funding; MEI: Consultancy, Research Funding; Genentech, Inc./ F. Hoffmann-La Roche Ltd: Consultancy, Honoraria, Research Funding; Seattle Genetics: Consultancy, Honoraria, Research Funding; Bristol-Myers Squibb: Consultancy, Honoraria, Research Funding; IMab: Research Funding. Gregory: Roche: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel fees, Speakers Bureau; Janssen: Consultancy; Novartis: Consultancy. Qazi: IGM Biosciences: Current Employment. Pang: IGM Biosciences: Consultancy. Leabman: IGM Biosciences: Current Employment. Hernandez: IGM Biosciences: Current Employment. Sison: IGM Biosciences: Current Employment. Keyt: IGM Biosciences: Current Employment. Chen: IGM Biosciences: Ended employment in the past 24 months. Armand: Enterome: Consultancy; C4: Consultancy; GenMab: Consultancy; Infinity: Consultancy; Otsuka: Research Funding; Regeneron: Consultancy; Pfizer: Consultancy; Morphosys: Consultancy; Daiichi Sankyo: Consultancy; Epizyme: Consultancy; Affimed: Consultancy, Research Funding; Adaptive: Consultancy, Research Funding; AstraZeneca: Consultancy; Genentech: Consultancy, Research Funding; Roche: Research Funding; Tensha: Research Funding; IGM: Research Funding; Kite: Research Funding; ADC Therapeutics: Consultancy; BMS: Consultancy, Honoraria, Research Funding; Sigma Tau: Research Funding; Merck: Consultancy, Honoraria, Research Funding; Celgene: Consultancy; Miltenyi: Consultancy; Tessa Therapeutics: Consultancy.

e20513 Background: Lung cancer is the leading cause of cancer death in Taiwan. The poor prognosis of lung cancer patients with advanced disease and recent development of precision medicine motivates the investigation of molecular targets. The purpose of the study is to estimate the prevalence of KRAS gene mutations and 15 other gene targets among non-small cell lung cancer (NSCLC) patients in Taiwan. Methods: This retrospective study included 933 NSCLC patients diagnosed between January 1992 and July 2018 at six medical centers in Taiwan, including Taichung Veteran’s General Hospital, Chung San Medical University Hospital, China Medical University Hospital, Kaohsiung Veteran’s General Hospital, Tri-Service General Hospital and National Taiwan University Hospital. Patients ≥18 years, had primary histologically confirmed NSCLC diagnosis at time before any treatment, and availability of archival tumor tissue with >30% tumor cellularity. Mutational Analysis: The Formalin-Fixed Paraffin-Embedded (FFPE) tumor blocks were analyzed for the prevalence of somatic mutations. The entire coding sequence of 16 genes was analyzed by next generation sequencing of macrodissected DNA from FFPE recut slides at a depth of greater than 300x and interrogated for somatic pathogenic variants. RNA was not available; hence this limits sensitivity to detect fusions. Results: The mean age at diagnosis was 62 years, 498/933 (53%) were women, 411/843 (49%) were smokers or former smokers, and 829/933 (89%) of adenocarcinoma histopathology. Stage was available for 723 patients with a distribution of 328/723 (45%) Stage I, 120/723 (17%) Stage II, 210/723 (29%) Stage III, and 65/723 (9%) Stage IV. EGFR mutations were identified in 327/933 (35.1%) and TP53 mutations in 179/933 (19.2%). KRAS mutations were identified in 72/933 (7.7%) patients, with KRAS G12C mutations observed in 23/933 (2.5%), KRAS G12D in 16/933 (1.7%), and KRAS G12V in 14/933 (1.5%) (Table). In addition, co-occurring mutations with KRAS were observed and are highlighted, including 14 TP53, 4 PIK3CA, 2 EGFR, 2 CTNNB1, 1 STK11, 1 BRAF and 1 PTEN. Conclusions: EGFR and TP53 were the most frequent mutations identified among all stage NSCLC in Taiwanese populations, followed by KRAS. KRAS G12C and KRAS G12D, the two most frequent KRAS mutations, were identified in 32% and 22% of the KRAS-mutant tumors. Although the prevalence of any KRAS mutation or KRAS G12C mutation is lower than typically seen in Western countries, it is similar to observations among Asian populations. These findings broaden the understanding of the mutational landscape of NSCLC patients in Taiwan to inform development of new therapeutic approaches. [Table: see text]

Background: Thrombopoietin receptor agonists (TPO-RAs) (e.g., romiplostim, eltrombopag, avatrombopag) are used to stimulate platelet production in patients with primary immune thrombocytopenia (ITP). While it was previously thought that patients would need to remain on a TPO-RA indefinitely, case reports and cohort studies have shown that some patients have discontinued TPO-RAs while maintaining a hemostatic platelet count. We convened a panel of experts to develop clinical recommendations on when it is appropriate to consider tapering and how to taper TPO-RAs in children and adults with persistent or chronic primary ITP. Methods: Using a RAND/UCLA modified Delphi panel, we convened 9 hematologists with an average of 25 years of experience and 1 patient representative. Experts were provided with a summary of evidence from 12 case reports, 11 cohort studies, and 2 clinical trial analyses on sustained remission in patients with ITP treated with TPO-RAs. Experts collaboratively developed and then rated (on a 1 to 9 scale) how appropriate it would be to recommend tapering (with the aim of discontinuing) TPO-RA monotherapy in 432 patient scenarios. Each scenario was a simplified patient history which varied by current platelet count, history of bleeding, intensification of treatment, trauma risk, use of anticoagulants or platelet inhibitors, duration of ITP, months on TPO-RA monotherapy, and early platelet response to TPO-RA (Table 1). In addition, the rating form included different ways to taper patients off therapy, how to monitor patients after discontinuation, and how to restart therapy. Ratings were completed independently by each expert before a full-day meeting. Median ratings were grouped into 3 categories (1-3, 4-6, 7-9) and disagreement was defined as ≥2 ratings of 1-3 and ≥2 ratings of 7-9 for a given scenario. During the meeting, discordant ratings were discussed. At the conclusion, experts completed ratings again (final round). Chi-square tests were conducted to determine if each patient characteristic significantly impacted ratings. Final ratings were used to describe the circumstances when it is inappropriate or appropriate to consider tapering TPO-RA monotherapy, how to taper TPO-RAs, how to monitor patients after discontinuation, and how to restart therapy. The panel was double-blinded while work was ongoing: the sponsor did not know the identity of the experts and the experts did not know the identity of the sponsor. The sponsor did not provide input on study design, methods, results, or interpretation of findings. Results: The proportion of items with disagreement decreased from 20% to 10% following the meeting. In the final round, 5 patient characteristics were found to significantly impact ratings and thus the appropriateness of tapering TPO-RA treatment: platelet count (p&lt;0.001), history of bleeding (p=0.001), intensification of treatment (p&lt;0.001), trauma risk (p&lt;0.001), use of anticoagulants or platelet inhibitors (p&lt;0.001). These characteristics were used to describe when it is inappropriate or appropriate to consider tapering TPO-RA monotherapy (Table 1). Experts agreed that it is inappropriate to consider tapering TPO-RA monotherapy in responding patients with low platelet counts, in patients with less than normal but still adequate platelet counts who have a history of major bleeding, or in patients who have a high risk of trauma and are using anticoagulants or platelet inhibitors (regardless of platelet count). It is appropriate to consider tapering TPO-RA monotherapy in patients with normal/above normal platelet counts, no history of major bleeding, and who have not required an intensification of treatment in the past 6 months. Recommendations on how to taper patients off therapy, how to monitor patients after discontinuation, and how to restart therapy were also developed. Conclusion: A validated methodology was used to assist an expert panel in developing clinical recommendations on when it is inappropriate or appropriate to consider tapering TPO-RA monotherapy and how to safely taper patients off therapy. The guidance reflects areas of greatest agreement based on clinical experience and currently available limited evidence. These recommendations could serve as a guide to clinical care and inform the development and design of clinical trials that prospectively test the safety of tapering TPO-RA monotherapy in patients with ITP. Disclosures Cuker: Pfizer: Research Funding; Novartis: Research Funding; Novo Nordisk: Research Funding; Sanofi: Research Funding; Spark: Research Funding; Takeda: Research Funding; Alexion: Research Funding; Bayer: Research Funding; Synergy CRO: Consultancy. Despotovic:Dova: Consultancy; Amgen: Consultancy, Research Funding; Novartis: Consultancy, Honoraria, Research Funding. Grace:Novartis: Research Funding; Dova: Membership on an entity's Board of Directors or advisory committees; Agios: Research Funding; Pfizer: Research Funding. Kruse:UCB: Other: Grant and consultancy fee, all paid to PDSA; Rigel: Other: Grant paid to PDSA; Principia: Other: Grant paid to PDSA; Pfizer: Other: Grant and consultancy fee, all paid to PDSA; Argenx: Other: Grant paid to PDSA; Amgen: Other: Grant and honorarium, all paid to PDSA; Novartis: Other: PDSA received payment for recruiting patients to I-WISh and for promoting I-WISh on the globalitp.org website. Grant and consultancy fee, all paid to PDSA outside the submitted work; CSL Behring: Other: Grant paid to PDSA. Lambert:Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Platelet Disorder Support Association (PDSA): Consultancy; ClinGen: Honoraria; Principia: Consultancy, Membership on an entity's Board of Directors or advisory committees; Argenix: Consultancy; Bayer: Consultancy; ITP Australia: Consultancy; AstraZeneca: Research Funding; Sysmex: Research Funding; Dova: Consultancy, Membership on an entity's Board of Directors or advisory committees; CdLS Foundation: Consultancy; RDMD ITP study: Consultancy; 22qSociety: Consultancy; Octapharma: Consultancy, Research Funding; Educational Concepts in Medicine: Consultancy; Shionogi: Consultancy. Liebman:Janssen: Consultancy; Amgen: Research Funding; Novartis: Honoraria, Research Funding; Kezar: Research Funding; Argenix: Research Funding; Alexion: Other; Genzyme: Consultancy; BMS: Consultancy; Rigel: Consultancy, Research Funding; Portola: Consultancy; Principia Biopharma: Consultancy. Lyons:Novartis: Honoraria; Texas Oncology/US Oncology: Current Employment. McCrae:Dova: Consultancy; Rigel: Consultancy; Momenta Pharmaceuticals: Consultancy; Novartis: Honoraria. Pullarkat:Dova: Consultancy, Honoraria; Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Servier: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Jazz Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Pfizer: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Genetech: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; AbbVie, Inc.: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau. Wasser:Amgen: Consultancy; Biogen: Current equity holder in publicly-traded company, Other: He and his wife have equity ownership; Eli Lilly: Current equity holder in publicly-traded company, Other: He and his wife have equity ownership; Novartis: Honoraria, Speakers Bureau; Pfizer: Current equity holder in publicly-traded company, Other: He and his wife have equity ownership, Research Funding; Incyte: Research Funding; Merck: Current equity holder in publicly-traded company, Other: He and his wife have equity ownership, Research Funding. Beenhouwer:Dr. Beenhouwer reports other from Novartis during the conduct of the study; other from AbbVie, other from Akcea, other from ASPC, other from Amgen, other from AstraZeneca, other from BMS, other from Boston Scientific Corporation, other from Celgene, other: Other: Dr. Beenhouwer reports other from Novartis during the conduct of the study; other from AbbVie, other from Akcea, other from ASPC, other from Amgen, other from AstraZeneca, other from BMS, other from Boston Scientific Corporation, other from Celgene, other. Gibbs:Ms. Gibbs reports other from Novartis during the conduct of the study; other from AbbVie, other from Akcea, other from ASPC, other from Amgen, other from AstraZeneca, other from BMS, other from Boston Scientific Corporation, other from Celgene, other from: Other: SN Gibbs is an employee of the Partnership for Health Analytic Research (PHAR), LLC, which was paid by Novartis to conduct this research.. Yermilov:Dr. Yermilov reports other from Novartis during the conduct of the study; other from AbbVie, other from Akcea, other from ASPC, other from Amgen, other from AstraZeneca, other from BMS, other from Boston Scientific Corporation, other from Celgene, other f: Other: Dr. Yermilov reports other from Novartis during the conduct of the study; other from AbbVie, other from Akcea, other from ASPC, other from Amgen, other from AstraZeneca, other from BMS, other from Boston Scientific Corporation, other from Celgene, other f. Broder:Dr. Broder reports other from Novartis during the conduct of the study; other from AbbVie, other from Akcea, other from ASPC, other from Amgen, other from AstraZeneca, other from BMS, other from Boston Scientific Corporation, other from Celgene, other fro: Other: MS Broder is an employee of the Partnership for Health Analytic Research (PHAR), LLC, which was paid by Novartis to conduct this research..

Background Multiple groups of pts, including elderly/frail pts and those with comorbidities, are typically under-represented in randomized controlled trials (RCTs). A recent study found an average of 16 eligibility criteria per cancer trial, 60% of which were related to comorbidity or performance status (PS; Unger, JNCI 2014). Phase 3 RCTs in MM have similar extensive eligibility criteria, resulting in populations that are not reflective of RW MM pts. Data from CONNECT-MM (Shah, CLML, 2017) and CoMMpass (Fiala, IMW 2017) suggest that 22-40% of RW pts are ineligible for MM RCTs, and an analysis of US RW relapsed/refractory MM (RRMM) pts showed that only 25-47% of pts would have been eligible for the phase 3 ASPIRE, TOURMALINE-MM1, ELOQUENT-2, and POLLUX studies, based on their differing eligibility criteria (Chari, EHA 2018). Further, data from CONNECT MM show that clinical trial ineligibility is associated with poorer long-term outcome (Shah, CLML, 2017). Thus, it is important to characterize RW MM pts and understand the discrepancies vs RCT populations. INSIGHT MM (NCT02761187) is the largest prospective, observational study in MM to date, following ~4,200 pts from 15 countries. Here we analyze RCT eligibility in INSIGHT MM pts, with a focus on the treatment of frail MM pts in the real world. Methods INSIGHT MM is following newly diagnosed (≤3 mos since treatment initiation) MM (NDMM) and RRMM (≤3 prior lines) pts. Demographics and disease characteristics, including medical history, comorbidities, PS, and frailty status (per IMWG Frailty Index criteria, Palumbo, JCO 2015), are collected using electronic case report forms at study baseline visit. For this analysis, pt data were reviewed vs 20 standard RCT eligibility criteria, using a conservative approach of classifying 'not available' data as 'eligible'; laboratory/PS and medical history exclusion criteria are summarized in the Table. Presence of hypertension was reviewed but omitted, as INSIGHT MM only collected data on 'hypertension requiring treatment' vs the standard RCT exclusion criterion of 'uncontrolled hypertension'. Results Data from 3,201 pts (1,761 NDMM, 1,440 RRMM) were analyzed. The proportions of pts who would be ineligible for RCTs based on each individual parameter, and the overall rate of ineligible pts, are shown in the Table. Overall, 39.2% of pts would have been ineligible for RCTs based on not meeting at least one of the 20 standard eligibility criteria included in this analysis, including 38.8% of NDMM and 39.7% of RRMM pts. The most common criteria excluding pts overall were another prior malignancy (7.5%), CrCl ≤30 mL/min (6.4%), cardiac arrhythmias (5.4%), platelets ≤75,000/mm3 (5.1%), and hemoglobin <8.0 g/dL (4.8%). Common criteria were similar in NDMM and RRMM pts, except for higher rates of CrCl ≤30 mL/min (7.3%) and hemoglobin <8.0 g/dL (5.9%) in NDMM pts, and higher rates of platelets ≤75,000/mm3 (7.4%) in RRMM pts. Frail pts are particularly likely to be ineligible for RCTs based on these standard criteria. Per study team evaluation applying the IMWG Frailty Index parameters, 320/1,685 evaluable pts (19%) were determined to be frail in INSIGHT MM. Of these 320 frail pts, applying the 20 standard eligibility criteria used in this analysis, 74.7% would have been ineligible for RCTs. Of 126 evaluable frail 1st line pts, 30 (24%) and 83 (66%) received doublet and triplet therapy, respectively, with median duration of treatment (DoT; Kaplan-Meier analysis) of 5.4 and 6.2 mos. Of 48 evaluable 2nd to 4th line pts, 26 (54%) and 21 (44%) received doublet and triplet therapy, respectively, with median DoT of 4.4 and 5.3 mos. Additional analyses of treatments received, reasons for treatment discontinuation, and DoT by frailty status and RCT eligibility will be presented. Conclusions A high proportion of INSIGHT MM pts (39.2%) were RCT-ineligible, consistent with or higher than previous reports. Common reasons for ineligibility included prior history of malignancy, low CrCl, and cardiac arrhythmias; more RRMM vs NDMM pts were excluded due to low platelet count. Analysis of frail pts suggests that these pts are particularly under-represented in RCTs using standard eligibility criteria. These findings emphasize the discrepancy between RW and RCT pt populations and the importance of RW data for evaluating effectiveness of treatment options. Initiatives are also ongoing to broaden oncology clinical trial eligibility criteria. Disclosures Hungria: Abbvie: Consultancy, Membership on an entity's Board of Directors or advisory committees; BMS: Consultancy, Honoraria, Speakers Bureau; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; BMS: Consultancy, Honoraria, Speakers Bureau; Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Abbvie: Consultancy, Membership on an entity's Board of Directors or advisory committees; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau. Lee:Janssen: Consultancy, Research Funding; Takeda: Consultancy, Research Funding; Daiichi Sankyo: Research Funding; Celgene: Consultancy, Research Funding; GlaxoSmithKline plc: Research Funding; Sanofi: Consultancy; Amgen: Consultancy, Research Funding. Abonour:Takeda: Consultancy, Research Funding; Janssen: Consultancy, Research Funding; Celgene: Consultancy, Research Funding; BMS: Consultancy. Rifkin:Takeda: Membership on an entity's Board of Directors or advisory committees; Amgen: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees. Terpos:Janssen: Honoraria, Other: Travel expenses, Research Funding; Genesis: Honoraria, Other: Travel expenses, Research Funding; Medison: Honoraria; Takeda: Honoraria, Other: Travel expenses, Research Funding; Celgene: Honoraria; Amgen: Honoraria, Research Funding. Leleu:Oncopeptide: Honoraria; Karyopharm: Honoraria; Amgen: Honoraria; Carsgen: Honoraria; Incyte: Honoraria; Novartis: Honoraria; Celgene: Honoraria; Janssen: Honoraria; BMS: Honoraria; Merck: Honoraria; Sanofi: Honoraria; Takeda: Honoraria. Costello:Celgene: Consultancy, Honoraria, Research Funding; Janssen: Research Funding; Takeda: Honoraria, Research Funding. van Rhee:Takeda: Consultancy; Sanofi Genzyme: Consultancy; EUSA: Consultancy; Castleman Disease Collaborative Network: Consultancy; Karyopharm Therapeutics: Consultancy; Kite Pharma: Consultancy; Adicet Bio: Consultancy. Weisel:Juno: Consultancy; Adaptive Biotech: Consultancy, Honoraria; Celgene Corporation: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding; GSK: Honoraria; Takeda: Consultancy, Honoraria; Janssen: Consultancy, Honoraria, Research Funding; Sanofi: Consultancy, Honoraria, Research Funding; Bristol-Myers Squibb: Consultancy, Honoraria. Puig:Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Takeda: Consultancy, Honoraria; Amgen: Consultancy, Honoraria; The Binding Site: Honoraria; Janssen: Consultancy, Honoraria, Research Funding. Berdeja:AbbVie Inc, Amgen Inc, Acetylon Pharmaceuticals Inc, Bluebird Bio, Bristol-Myers Squibb Company, Celgene Corporation, Constellation Pharma, Curis Inc, Genentech, Glenmark Pharmaceuticals, Janssen Biotech Inc, Kesios Therapeutics, Lilly, Novartis, Poseida: Research Funding; Poseida: Research Funding; Amgen Inc, BioClinica, Celgene Corporation, CRISPR Therapeutics, Bristol-Myers Squibb Company, Janssen Biotech Inc, Karyopharm Therapeutics, Kite Pharma Inc, Prothena, Servier, Takeda Oncology: Consultancy. Cook:Janssen: Consultancy, Honoraria, Research Funding, Speakers Bureau; Karyopharm: Consultancy, Honoraria, Speakers Bureau; Takeda: Consultancy, Honoraria, Research Funding, Speakers Bureau; Sanofi: Consultancy, Honoraria, Speakers Bureau; Celgene: Consultancy, Honoraria, Research Funding, Speakers Bureau. Usmani:Bristol-Myers Squibb: Consultancy, Research Funding; Janssen: Consultancy, Patents & Royalties, Research Funding, Speakers Bureau; Pharmacyclics: Patents & Royalties, Research Funding; Takeda: Consultancy, Patents & Royalties, Research Funding, Speakers Bureau; Amgen: Consultancy, Patents & Royalties, Research Funding, Speakers Bureau; Merck: Consultancy, Research Funding; Celgene: Consultancy, Patents & Royalties, Research Funding, Speakers Bureau; Array Biopharma: Patents & Royalties, Research Funding; Sanofi: Patents & Royalties, Research Funding, Speakers Bureau; Skyline DX: Consultancy. Thompson:GSK: Membership on an entity's Board of Directors or advisory committees; Doximity: Equity Ownership, Membership on an entity's Board of Directors or advisory committees; BMS: Research Funding; UpToDate: Patents & Royalties: Myeloma reviewer; Celgene: Membership on an entity's Board of Directors or advisory committees; AbbVie: Research Funding; Adaptive: Membership on an entity's Board of Directors or advisory committees; Takeda: Membership on an entity's Board of Directors or advisory committees, Research Funding; VIA Oncology: Membership on an entity's Board of Directors or advisory committees; Lynx Bio: Research Funding. Boccadoro:Mundipharma: Research Funding; Sanofi: Honoraria, Research Funding; Celgene: Honoraria, Research Funding; Amgen: Honoraria, Research Funding; Janssen: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; Bristol-Myers Squibb: Honoraria, Research Funding; AbbVie: Honoraria. Zonder:Oncopeptides: Consultancy, Membership on an entity's Board of Directors or advisory committees; BMS: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Janssen: Consultancy, Membership on an entity's Board of Directors or advisory committees; Intellia: Consultancy, Membership on an entity's Board of Directors or advisory committees; Caelum: Consultancy, Membership on an entity's Board of Directors or advisory committees; Alnylam: Consultancy, Membership on an entity's Board of Directors or advisory committees; Amgen: Consultancy, Membership on an entity's Board of Directors or advisory committees; Celgene Corporation: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Takeda: Membership on an entity's Board of Directors or advisory committees. Spencer:Servier: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Sanofi: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Specialised Therapeutics Australia: Consultancy, Honoraria; AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Secura Bio: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Janssen Oncology: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Haemalogix: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees. Hajek:Janssen: Honoraria, Other: Consultant or advisory relationship, Research Funding; Amgen: Honoraria, Other: Consultant or advisory relationship, Research Funding; Celgene: Honoraria, Other: Consultant or advisory relationship, Research Funding; AbbVie: Other: Consultant or advisory relationship; Bristol-Myers Squibb: Honoraria, Other: Consultant or advisory relationship, Research Funding; Novartis: Other: Consultant or advisory relationship, Research Funding; PharmaMar: Honoraria, Other: Consultant or advisory relationship; Takeda: Honoraria, Other: Consultant or advisory relationship, Research Funding. Fanning:Celgene: Speakers Bureau; Takeda: Speakers Bureau. Boyd:Novartis: Consultancy, Honoraria; Janssen: Consultancy, Honoraria; Takeda: Consultancy, Honoraria; Amgen: Consultancy, Honoraria; Celgene: Consultancy, Honoraria. Armour:Millennium Pharmaceuticals: Other: Financial relationship; Janssen: Other: Financial relationship; Amgen: Other: Financial relationship; Celgene: Other: Financial relationship. Morgan:Myeloma Patients Europe, a non-profit organisation, which receives unrestricted grants from the following pharmaceutical companies: Amgen, Bristol Myers Squibb, Takeda, Janssen, Karyopharm, Novartis, Celgene and Sanofi.: Employment. Patel:Takeda: Employment. Carlson:Takeda: Employment. Ferrari:Takeda: Employment. Stull:Takeda: Employment. Ren:Takeda: Employment. Cherepanov:Takeda: Employment. Pottala:PPD, Inc., research organization contracted by Millennium Pharmaceuticals, Inc.: Employment. Chari:Sanofi: Membership on an entity's Board of Directors or advisory committees; Seattle Genetics: Membership on an entity's Board of Directors or advisory committees, Research Funding; Pharmacyclics: Research Funding; Oncoceutics: Research Funding; Novartis Pharmaceuticals: Research Funding; GlaxoSmithKline: Research Funding; Array Biopharma: Research Funding; Karyopharm: Consultancy, Membership on an entity's Board of Directors or advisory committees; Janssen: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Millennium/Takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Amgen: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Bristol-Myers Squibb: Consultancy.

ABSTRACT Microtubules (MTs) regulate numerous cellular processes, but their roles in brain morphogenesis are not well known. Here, we show that CAMSAP3, a non-centrosomal microtubule regulator, is important for shaping the lateral ventricles. In differentiating ependymal cells, CAMSAP3 became concentrated at the apical domains, serving to generate MT networks at these sites. Camsap3-mutated mice showed abnormally narrow lateral ventricles, in which excessive stenosis or fusion was induced, leading to a decrease of neural stem cells at the ventricular and subventricular zones. This defect was ascribed at least in part to a failure of neocortical ependymal cells to broaden their apical domain, a process necessary for expanding the ventricular cavities. mTORC1 was required for ependymal cell growth but its activity was downregulated in mutant cells. Lysosomes, which mediate mTORC1 activation, tended to be reduced at the apical regions of the mutant cells, along with disorganized apical MT networks at the corresponding sites. These findings suggest that CAMSAP3 supports mTORC1 signaling required for ependymal cell growth via MT network regulation, and, in turn, shaping of the lateral ventricles.

The coronavirus pandemic (COVID-19) broke into the lives of people around the world, (WHO, 2020) led to changes in the way of working and studying. For students it represents a challenge. The educational community has made efforts to sustain student learning; This represented the need to use information technologies and depend on their own resources to guarantee the continuity of learning, forcing them to take distance classes through digital means (UNESCO, 2020). This research will allow to know the conditions and how the students of the Administrative Economic area of the Technological University of León face the challenges. For this, a questionnaire was designed in order to collect information, it was validated to measure reliability and it was applied to a sample of 249 students. The data was analyzed and showed how many of them have a computer; if it is for exclusive use or shared; how many have internet and the quality of the service; how many have cell phones; how many hours they use their devices for classes; the platforms they use; if they have what it takes to work at home and if they prefer distance or face-to-face classes.

Spectrum inefficiency is a significant issue because of the increasingly more solicitation of transmission capacity by the end clients. Accomplishing high transmission rates and elevated levels of Quality of Service (QoS) speaks to in any case an open issue. Long haul Evolution (LTE) has been proposed as the reason for the fourth era versatile cell systems (4G) that points of the LTE standard are higher client bit rates, lower delays, expanded range proficiency, diminished expense, and operational effortlessness. In any case, this innovation is as yet being worked on and a few open issues must be still explored, for example, obstruction coordination, and power utilization, assets the board and handover procedures. The point of this work is to ensure the decrease of intensity utilization utilizing another handover calculation dependent on green arrangement. Also, the proposed conspire ensures the minimization of superfluous handovers. In any case, the issue in this technique is doesn't mulls over of the impact of green strategies in high force hubs. Along these lines, in the proposed framework the impact of high force hubs are thought of. This situation is expanded more than one full scale cell and various kinds of low force hubs, for example, pico-cell and microcells. In this situation, the force sparing is broke down during the handover strategies. Think about these issues; another strategy is proposed called Dynamic base Station arranging (DBSP) for accomplishing vitality productivity. The fundamental idea is to kill a BS individually that will insignificantly influence the system by utilizing a recently presented thought of system sway, which considers the extra burden increases brought to its neighboring BSs. So as to additionally diminish the flagging and execution overhead over the air and back take, utilize the estimated estimations of system sway as their choice measurements. A trial result shows that the proposed strategy accomplishes high vitality proficiency under different situations. In this work the presentation of an UMTS organize situation is assessed by utilizing different estimations of the priority bits of the CBR application.

In the contemporary urban environment of mass transit, it falls to a small group of public officers to keep large number of travellers safe. The small size of their force and the often limited powers they exert mean that these public safety ‘transit officers’ must project more authority and control than they really have. It is this ambience of authority and control which, in most situations they encounter and seek to influence, is enough to keep the public safe. This paper examines the ambience of a group of transit officers working on the railway lines of an Australian capital city. We seek to show how transit officers are both influenced by, and seek to influence, the ambience of their workplace and the public spaces they inhabit whilst on duty, and here we take ambience to apply to the surrounding atmosphere, the aura, and the emotional environment of a place or situation: the setting, tone, or mood. For these transit officers to keep the public safe, they must themselves remain safe. A transit officer who is disabled in a confrontation with a violent offender is unable to provide protection to his or her passengers. Thus, in the culture of the transit officers, their own workplace safety takes on a higher significance. It affects not just themselves. The ambience exuded by transit officers, and how transit officers see their relationship with the travelling public, their management and other organisational work groups, is an important determinant of their work group’s safety culture. Researching the Working Lives of Transit Officers in Perth Our discussion draws on an ethnographic study of the working lives and communication cultures of transit officers (TOs) employed by the Public Transport Authority (PTA) of Western Australia (WA). Transit officers have argued that to understand fully the challenges of their work it is necessary to spend time with them as they undertake their daily duties: roster in, roster out. To this end, the research team and the employer organisation secured an ARC Linkage Grant in partnership with the PTA to fund doctoral candidate and ethnographer Christine Teague to research the workers’ point of view, and the workers’ experiences within the organisation. The two-hundred TOs are unique in the PTA. Neither of the other groups who ride with them on the trains, the drivers and revenue protection staff (whose sole job is to sell and check tickets), experiences the combination of intense contact with passengers, danger of physical injury or group morale. The TOs of the PTA in Perth operate from a central location at the main train station and the end stations on each line. Here there are change lockers where they can lock up their uniforms and equipment such as handcuffs and batons when not on duty, an equipment room where they sign out their radios, and ticket-checking machines. At the main train station there is also a gym, a canteen and holding cells for offenders they detain. From these end stations and central location, the TOs fan out across the network to all suburbs where they either operate from stations or onboard the trains. The TOs also do ‘delta van’ duty providing rapid, mobile back-up support for their colleagues on stations or trains, and providing transport for arrested persons to the holding cell or police lock up. TOs are on duty whenever the trains are running–but the evenings and nights are when they are mainly rostered on. This is when trouble mostly occurs. The TOs’ work ends only after the final train has completed its run and all offenders who may require detaining and charging have been transferred into police custody. While the public perceive that security is the TOs’ most frequent role, much of the work involves non-confrontational activity such as assisting passengers, checking tickets and providing a reassuring presence. One way to deal with an ambiguous role is to claim an ambience of power and authority regardless. Various aspects of the TO role permit and hinder this, and the paper goes on to consider aspects of ambience in terms of fear and force, order and safety, and role confusion. An Ambience of Fear and Force The TOs are responsible for front-line security in WA’s urban railway network. Their role is to offer a feeling of security for passengers using the rail network after the bustle of the work day finishes, and is replaced by the mainly recreational travels of the after hours public. This is the time when some passengers find the prospect of evening travel on the public transport rail network unsettling–so unsettling that it was a 2001 WA government election promise (WA Legislative Council) that every train leaving the city centre after 7pm would have two TOs riding on it. Interestingly, recruitment levels have never been high enough for this promise to be fully kept. The working conditions of the TOs reflect the perception, and to an extent, the reality that some late night travel on public transport involves negotiating an edgy ambience with an element of risk, rubbing shoulders with people who may be loud, rowdy, travelling in a group, and or drug and alcohol affected. As Fred (all TO names are pseudonyms) comments: You’re not dealing with rational people, you’re not dealing with ‘people’: most of the people you’re dealing with are either drunk or under the influence of drugs, so they’re not rational, they don’t hear you, they don’t understand what you’re saying, they just have no sense of what’s right or wrong, you know? Especially being under the influence, so I mean, you can talk till you’re blue in the face with somebody who’s drunk or on drugs, I mean, all you have to say is one thing. ‘Oh, can I see your ticket please’, ‘oh, why do I need a fucking ticket’, you know? They just don’t get simple everyday messages. Dealing with violence and making arrest is a normal part of this job. Jo described an early experience in her working life as a TO:Within the first week of coming out of course I got smacked on the side of the head, but this lady had actually been certified, like, she was nuts. She was completely mental and we were just standing on the train talking and I’ve turned around to say something to my partner and she was fine, she was as calm as, and I turned around and talked to my partner and the next thing I know I ended up with her fist to the side of my head. And I went ‘what the hell was that’? And she went off, she went absolutely ballistic. I ended up arresting her because it was assault on an officer whether she was mental or not so I ended up arresting her.Although Jo here is describing how she experienced an unprovoked assault in the early days of her career as a TO, one of the most frequent precursors to a TO injury occurs when the TO is required to make an arrest. The injury may occur when the passenger to be arrested resists or flees, and the TO gives chase in dark or treacherous circumstances such as railway reserves and tunnels, or when other passengers, maybe friends or family of the original person of concern, involve themselves in an affray around the precipitating action of the arrest. In circumstances where capsicum spray is the primary way of enforcing compliance, with batons used as a defence tool, group members may feel that they can take on the two TOs with impunity, certainly in the first instance. Even though there are security cameras on trains and in stations, and these can be cued to cover the threatening or difficult situations confronting TOs, the conflict is located in the here-and-now of the exchanges between TOs and the travelling public. This means the longer term consequence of trouble in the future may hold less sway with unruly travellers than the temptation to try to escape from trouble in the present. In discussing the impact of remote communications, Rubert Murdoch commented that these technologies are “a powerful influence for civilised behaviour. If you are arranging a massacre, it will be useless to shoot the cameraman who has so inconveniently appeared on the scene. His picture will already be safe in the studio five thousand miles away and his final image may hang you” (Shawcross 242). Unfortunately, whether public aggression in these circumstances is useless or not, the daily experience of TOs is that the presence of closed circuit television (CCTV) does not prevent attacks upon them: nor is it a guarantee of ‘civilised behaviour’. This is possibly because many of the more argumentative and angry members of the public are dis-inhibited by alcohol or other drugs. Police officers can employ the threat or actual application of stun guns to control situations in which they are outnumbered, but in the case of TOs they can remain outnumbered and vulnerable until reinforcements arrive. Such reinforcements are available, but the situation has to be managed through the communication of authority until the point where the train arrives at a ‘manned’ station, or the staff on the delta vehicle are able to support their colleagues. An Ambience of Order and Safety Some public transport organisations take this responsibility to sustain an ambience of order more seriously than others. The TO ethnographer, Christine Teague, visited public transport organisations in the UK, USA and Canada which are recognised as setting world-class standards for injury rates of their staff. In the USA particularly, there is a commitment to what is called ‘the broken windows’ theory, where a train is withdrawn from service promptly if it is damaged or defaced (Kelling and Coles; Maple and Mitchell). According to Henry (117): The ‘Broken Windows’ theory suggests that there is both a high correlation and a causal link between community disorder and more serious crime: when community disorder is permitted to flourish or when disorderly conditions or problems are left untended, they actually cause more serious crime. ‘Broken windows’ are a metaphor for community disorder which, as Wilson and Kelling (1982) use the term, includes the violation of informal social norms for public behaviour as well as quality of life offenses such as littering, graffiti, playing loud radios, aggressive panhandling, and vandalism.This theory implies that the physical ambience of the train, and by extension the station, may be highly influential in terms of creating a safe working environment. In this case of ‘no broken window’ organisations, the TO role is to maintain a high ‘quality of life’ rather than being a role predominantly about restraining and bringing to justice those whose behaviour is offensive, dangerous or illegal. The TOs in Perth achieve this through personal means such as taking pride in their uniforms, presenting a good-natured demeanour to passengers and assisting in maintaining the high standard of train interiors. Such a priority, and its link to reduced workforce injury, suggests that a perception of order impacts upon safety. It has long been argued that the safety culture of an organisation affects the safety performance of that organisation (Pidgeon; Leplat); but it has been more recently established that different cultural groupings in an organisation conceive and construct their safety culture differently (Leith). The research on ‘safety culture’ raises a problematic which is rarely addressed in practice. That problematic is this: managers frequently engage with safety at the level of instituting systems, while workers engage with safety in terms of behaviour. When Glendon and Litherland comment that, contrary to expectations, they could find no relationship between safety culture and safety performance, they were drawing attention to the fact that much managerial safety culture is premised upon systems involving tick boxes and the filling in of report forms. The broken window approach combines the managerial tick box with managerial behaviour: a dis-ordered train is removed from service. To some extent a general lack of fit between safety culture and safety performance endorses Everett’s view that it is conceptually inadequate to conceive organisations as cultures: “the conceptual inadequacy stems from the failure to distinguish between culture and behavioural features of organizational life” (238). The general focus upon safety culture as a way of promoting improvements in safety performance assumes that compliance with a range of safety systems will guarantee a safe workplace. Such an assumption, however, risks positioning the injured worker as responsible for his or her own predicament and sets up an environment in which some management officials are wont to seek ways in which that injured worker’s behaviour failed to conform with safety rules or safety processes. Yet there are roles which place workers in harm’s way, including military duties, law enforcement and some emergency services. Here, the work becomes dangerous as it becomes disorderly. An Ambience of Roles and Confusion As the research reported here progressed, it became clear that the ambience around the presentation of the self in the role of a TO (Goffman) was an important part of how ‘safety’ was promoted and enacted in their work upon the PTA (WA) trains, face to face with the travelling public. Goffman’s view of all people, not specifically TOs, is that: Regardless of the particular objective which the individual has in mind and of his motive for having this objective, it will be in his interests to control the conduct of the others, especially their responsive treatment of him. This will largely be through influencing the perception and definition that others will come to formulate of him. He will influence them by expressing himself in such a way that the kind of impression given off will lead them to act voluntarily in accordance with his own plan. (3)This ‘influencing of perception’ is an important element of performing the role of a TO. This task of the TOs is made all the more difficult because of confusions about their role in relation to two other officers: police (who have more power to act in situations of public safety) and revenue project officers (who have less), as we now discuss. The aura of the TO role borrows somewhat from those quintessential law and order officers: the police. TOs work in pairs, like many police, to support each other. They have a range of legal powers including the power of arrest, and they carry handcuffs, a baton and capsicum spray as a means of helping ensure their safety and effectiveness in circumstances where they might be outnumbered. The tools of their trade are accessibly displayed on heavy leather belts around their waists and their uniforms have similarities with police uniforms. However, in some ways these similarities are problematic, because TOs are not afforded the same respect as police. This situation underlines of the ambiguities negotiated within the ambience of what it is to be a TO, and how it is to conduct oneself in that role. Notwithstanding the TOs’ law and order responsibilities, public perceptions of the role and some of the public’s responses to the officers can position these workers as “plastic cops” (Teague and Leith). The penultimate deterrent of police officers, the stun gun (Taser), is not available to TOs who are expected to control all incidents arising on duty through the fact that they operate in pairs, with capsicum spray available and, as a last resort, are authorised to use their batons in self defence. Furthermore, although TOs are the key security and enforcement staff in the PTA workforce, and are managed separately from related staff roles, they believe that the clarity of this distinction is compromised because of similarities in the look of Revenue Protection Officers (RPOs). RPOs work on the trains to check that passengers have tickets and have paid the correct fares, and obtain names and addresses to issue infringement notices when required. They are not PTA employees, but contracted staff from an outside company. They also work in pairs. Significantly, the RPO uniform is in many respects identical to that of the TO, and this appears to be a deliberate management choice to make the number of TOs seem greater than it is: extending the TO ambience through to the activities of the RPOs. However, in the event of a disturbance, TOs are required and trained to act, while RPOs are instructed not to get involved; even though the RPOs appear to the travelling public to be operating in the role of a law-and-order-keeper, RPOs are specifically instructed not to get involved in breaches of the peace or disruptive passenger behaviour. From the point of view of the travelling public, who observe the RPO waiting for TOs to arrive, it may seems as if a TO is passively standing by while a chaotic situation unravels. As Angus commented: I’ve spoken to quite a few members of public and received complaints from them about transit officers and talking more about the incident have found out that it was actually [RPOs] that are dealing with it. So it’s creating a bad image for us …. It’s Transits that are copping all the flak for it … It is dangerous for us and it’s a lot of bad publicity for us. It’s hard enough, the job that we do and the lack of respect that we do get from people, we don’t need other people adding to it and making it harder. Indeed, it is not only the travelling public who can mistake the two uniforms. Mike tells of an “incident where an officer [TO] has called for backup on a train and the guys have got off [the train at the next station] and just stood there, and he didn’t realise that they are actually [revenue protection] officers, so he effectively had no backup. He thought he did, but he didn’t.” The RPO uniform may confer an ambience of power borrowed from TOs and communicated visually, but the impact is to compromise the authority of the TO role. Unfortunately, what could be a complementary role to the TOs becomes one which, in the minds of the TO workforce, serves to undermine their presence. This effect of this role confusion is to dilute the aura of authority of the TOs. At one end of a power continuum the TO role is minimised by those who see it as a second-rate ‘Wannabe cop’ (Teague and Leith 2008), while its impact is diluted at the other end by an apparently deliberate confusion between the TO broader ‘law and order’ role, and the more limited RPO revenue collection activities. Postlude To the passengers of the PTA in Perth, the presence and actions of transit officers appear as unremarkable as the daily commute. In this ethnographic study of their workplace culture, however, the transit officers have revealed ways in which they influence the ambience of the workplace and the public spaces they inhabit whilst on duty, and how they are influenced by it. While this ambient inter-relationship is not documented in the organisation’s occupational safety and health management system, the TOs are aware that it is a factor in their level at safety at work, both positively and negatively. Clearly, an ethnography study is conducted at a certain point in time and place, and culture is a living and changing expression of human interaction. The Public Transport Authority of Western Australia is committed to continuous improvement in safety and to the investigation of all ways and means in which to support TOs in their daily activities. This is evident not only in their support of the research and their welcoming of the ethnographer into the workforce and onto the tracks, but also in their robust commitment to change as the findings of the research have progressed. In particular, changes in the ambient TO culture and in the training and daily practices of TOs have already resulted from this research or are under active consideration. Nonetheless, this project is a cogent indicator of the fact that a safety culture is critically dependent upon intangible but nonetheless important factors such as the ambience of the workplace and the way in which officers are able to communicate their authority to others. References Everett, James. “Organizational Culture and Ethnoecology in Public Relations Theory and Practice.” Public Relations Research Annual. Vol. 2. Eds. Larissa Grunig and James Grunig. Hillsdale, NJ, 1990. 235-251. Glendon, Ian, and Debbie Litherland. “Safety Climate Factors, Group Differences and Safety Behaviour in Road Construction.” Safety Science 39.3 (2001): 157-188. Goffman, Erving. The Presentation of the Self in Everyday Life. London: Penguin, 1959. Henry, Vincent. The Comstat Paradigm: Management Accountability in Policing, Business and the Public Sector. New York: Looseleaf Law Publications, 2003. Kelling, George, and Catherine Coles. Fixing Broken Windows: Restoring Order and Reducing Crime in Our Communities. New York: Touchstone, 1996. Leith, David. Workplace Culture and Accidents: How Management Can Communicate to Prevent Injuries. Saarbrücken: VDM Verlag, 2008. Leplat, Jacques. “About Implementation of Safety Rules.” Safety Science 29.3 (1998): 189-204. Maple, Jack, and Chris Mitchell. The Crime Fighter: How You Can Make Your Community Crime-Free. New York: Broadway Books, 1999. Pidgeon, Nick. “Safety Culture and Risk Management in Organizations.” Journal of Cross-Cultural Psychology 22.1 (1991): 129-140. Shawcross, William. Rupert Murdoch. London: Chatto & Windus, 1992. Teague, Christine, and David Leith. “Men of Steel or Plastic Cops? The Use of Ethnography as a Transformative Agent.” Transforming Information and Learning Conference Transformers: People, Technologies and Spaces, Edith Cowan University, Perth, WA, 2008. ‹http://conferences.scis.ecu.edu.au/TILC2008/documents/2008/teague_and_leith-men_of_steel_or_plastic_cops.pdf›. Wilson, James, and George Kelling. “Broken Windows.” The Atlantic Monthly (Mar. 1982): 29-38. WA Legislative Council. “Metropolitan Railway – Transit Guards 273 [Hon Ed Dermer to Minister of Transport Hon. Simon O’Brien].” Hansard 19 Mar. 2009: 2145b.

Like any genre or mode, the Gothic is malleable, changing according to time and place. This is particularly apparent when what is considered Gothic in one era is compared with that of another. The giant helmet that falls from the sky in Horace Walpole’s Castle of Otranto (1764) is a very different threat to the ravenous vampires that stalk the novels of Anne Rice, just as Ann Radcliffe’s animated portraits may not inspire anxiety for a contemporary reader of Stephen King. The mutability of Gothic is also apparent across various versions of national Gothic that have emerged, with the specificities of place lending Gothic narratives from countries such as Ireland, Scotland and Australia a distinctive flavour. In New Zealand, the Gothic is most commonly associated with Pakeha artists exploring extreme psychological states, isolation and violence. Instead of the haunted castles, ruined abbeys and supernatural occurrences of classic Gothics of the eighteenth and nineteenth centuries, such as those produced by writers as diverse as Charles Brockden Brown, Matthew Lewis, Edgar Allen Poe, Radcliffe, Bram Stoker and Walpole, New Zealand Gothic fiction tends to focus on psychological horror, taking its cue, according to Jenny Lawn, from Mary Shelley’s Frankenstein (1818), which ushered in a tendency in the Gothic novel to explore the idea of a divided consciousness. Lawn observes that in New Zealand “Our monsters tend to be interior: they are experiences of intense psychological states, often with sexual undertones within isolated nuclear families” (“Kiwi Gothic”). Kirsty Gunn’s novella Rain (1994), which focuses on a dysfunctional family holidaying in an isolated lakeside community, exemplifies the tendency of New Zealand Gothic to omit the supernatural in favour of the psychological, with its spectres being sexual predation, parental neglect and the death of an innocent. Bronwyn Bannister’s Haunt (2000) is set primarily in a psychiatric hospital, detailing various forms of psychiatric disorder, as well as the acts that spring from them, such as one protagonist’s concealment for several years of her baby in a shed, while Noel Virtue’s The Redemption of Elsdon Bird (1987) is another example, with a young character’s decision to shoot his two younger siblings in the head as they sleep in an attempt to protect them from the religious beliefs of his fundamentalist parents amply illustrating the intense psychological states that characterise New Zealand Gothic. Although there is no reason why Gothic literature ought to include the supernatural, its omission in New Zealand Gothic does point to a confusion that Timothy Jones foregrounds in his suggestion that “In the absence of the trappings of established Gothic traditions – castles populated by fiendish aristocrats, swamps draped with Spanish moss and possessed by terrible spirits” New Zealand is “uncertain how and where it ought to perform its own Gothic” (203). The anxiety that Jones notes is perhaps less to do with where the New Zealand Gothic should occur, since there is an established tradition of Gothic events occurring in the bush and on the beach, while David Ballantyne’s Sydney Bridge Upside Down (1968) uses a derelict slaughterhouse as a version of a haunted castle and Maurice Gee successfully uses a decrepit farmhouse as a Gothic edifice in The Fire-Raiser (1986), but more to do with available ghosts. New Zealand Gothic literature produced in the twentieth and twenty-first centuries certainly tends to focus on the psychological rather than the supernatural, but earlier writing that utilises the Gothic mode is far more focused on spooky events and ghostly presences. There is a tradition of supernatural Gothic in New Zealand, but its representations of Maori ghosts complicates the processes through which contemporary writers might build on that tradition. The stories in D. W. O. Fagen’s collection Tapu and Other Tales of Old New Zealand (1952) illustrate the tendency in colonial New Zealand literature to represent Maori in supernatural terms expressive both of anxieties surrounding Maori agency and indigeneity, as well as Western assumptions regarding Maori culture. In much colonial Gothic, Maori ghosts, burial grounds and the notion of tapu express settler anxieties while also working to contain those anxieties by suggesting the superstitious and hence backward nature of indigenous culture. In Fagan’s story “Tapu”, which first appeared in the Bulletin in 1912, the narrator stumbles into a Maori burial ground where he is confronted by the terrible sight of “two fleshless skeletons” that grin and appear “ghastly in the dim light” (37). The narrator’s desecration of land deemed tapu fills him with “a sort of nameless terror at nothing, a horror of some unknown impending fate against which it was useless to struggle and from which there was no escape” (39). This expresses a sense of the authenticity of Maori culture, but the narrator’s thought “Was there any truth in heathen devilry after all?” is quickly superseded by the relegation of Maori culture as “ancient superstitions” (40). When the narrator is approached by a tohunga following his breach of tapu, his reaction is outrage: "Here was I – a fairly decent Englishman, reared in the Anglican faith and living in the nineteenth century – hindered from going about my business, outcast, excommunicated, shunned as a leper, my servant dying, all on account of some fiendish diablerie of heathen fetish. The affair was preposterous, incredible, ludicrous" (40). Fagan’s story establishes a clear opposition between Western rationalism and “decency”, and the “heathen fetishes” associated with Maori culture, which it uses to infuse the story with the thrills appropriate to Gothic fiction and which it ultimately casts as superstitious and uncivilised. F. E. Maning’s Old New Zealand (1863) includes an episode of Maori women grieving that is represented in terms that would not be out of place in horror. A group of women are described as screaming, wailing, and quivering their hands about in a most extraordinary manner, and cutting themselves dreadfully with sharp flints and shells. One old woman, in the centre of the group, was one clot of blood from head to feet, and large clots of coagulated blood lay on the ground where she stood. The sight was absolutely horrible, I thought at the time. She was singing or howling a dirge-like wail. In her right hand she held a piece of tuhua, or volcanic glass, as sharp as a razor: this she placed deliberately to her left wrist, drawing it slowly upwards to her left shoulder, the spouting blood following as it went; then from the left shoulder downwards, across the breast to the short ribs on the right side; then the rude but keen knife was shifted from the right hand to the left, placed to the right wrist, drawn upwards to the right shoulder, and so down across the breast to the left side, thus making a bloody cross on the breast; and so the operation went on all the time I was there, the old creature all the time howling in time and measure, and keeping time also with the knife, which at every cut was shifted from one hand to the other, as I have described. She had scored her forehead and cheeks before I came; her face and body was a mere clot of blood, and a little stream was dropping from every finger – a more hideous object could scarcely be conceived. (Maning 120–21) The gory quality of this episode positions Maori as barbaric, but Patrick Evans notes that there is an incident in Old New Zealand that grants authenticity to indigenous culture. After being discovered handling human remains, the narrator of Maning’s text is made tapu and rendered untouchable. Although Maning represents the narrator’s adherence to his abjection from Maori society as merely a way to placate a local population, when a tohunga appears to perform cleansing rituals, the narrator’s indulgence of perceived superstition is accompanied by “a curious sensation […] like what I fancied a man must feel who has just sold himself, body and bones, to the devil. For a moment I asked myself the question whether I was not actually being then and there handed over to the powers of darkness” (qtd. in Evans 85). Evans points out that Maning may represent the ritual as solely performative, “but the result is portrayed as real” (85). Maning’s narrator may assert his lack of belief in the tohunga’s power, but he nevertheless experiences that power. Such moments of unease occur throughout colonial writing when assertions of European dominance and rational understanding are undercut or threatened. Evans cites the examples of the painter G. F. Angus whose travels through the native forest of Waikato in the 1840s saw him haunted by the “peculiar odour” of rotting vegetation and Edward Shortland whose efforts to remain skeptical during a sacred Maori ceremony were disturbed by the manifestation of atua rustling in the thatch of the hut in which it was occurring (Evans 85). Even though the mysterious power attributed to Maori in colonial Gothic is frequently represented as threatening, there is also an element of desire at play, which Lydia Wevers highlights in her observation that colonial ghost stories involve a desire to assimilate or be assimilated by what is “other.” Wevers singles out for discussion the story “The Disappearance of Letham Crouch”, which appeared in the New Zealand Illustrated Magazine in 1901. The narrative recounts the experiences of an overzealous missionary who is received by Maori as a new tohunga. In order to learn more about Maori religion (so as to successfully replace it with Christianity), Crouch inhabits a hut that is tapu, resulting in madness and fanaticism. He eventually disappears, only to reappear in the guise of a Maori “stripped for dancing” (qtd. in Wevers 206). Crouch is effectively “turned heathen” (qtd. in Wevers 206), a transformation that is clearly threatening for a Christian European, but there is also an element of desirability in such a transformation for a settler seeking an authentic New Zealand identity. Colonial Gothic frequently figures mysterious experiences with indigenous culture as a way for the European settler to essentially become indigenous by experiencing something perceived as authentically New Zealand. Colonial Gothic frequently includes the supernatural in ways that are complicit in the processes of colonisation that problematizes them as models for contemporary writers. For New Zealanders attempting to produce a Gothic narrative, the most immediately available tropes for a haunting past are Maori, but to use those tropes brings texts uncomfortably close to nineteenth-century obsessions with Maori skeletal remains and a Gothicised New Zealand landscape, which Edmund G. C. King notes is a way of expressing “the sense of bodily and mental displacement that often accompanied the colonial experience” (36). R. H. Chapman’s Mihawhenua (1888) provides an example of tropes particularly Gothic that remain a part of colonial discourse not easily transferable into a bicultural context. Chapman’s band of explorers discover a cave strewn with bones which they interpret to be the remains of gory cannibalistic feasts: Here, we might well imagine, the clear waters of the little stream at our feet had sometime run red with the blood of victims of some horrid carnival, and the pale walls of the cavern had grown more pale in sympathy with the shrieks of the doomed ere a period was put to their tortures. Perchance the owners of some of the bones that lay scattered in careless profusion on the floor, had, when strong with life and being, struggled long and bravely in many a bloody battle, and, being at last overcome, their bodies were brought here to whet the appetites and appease the awful hunger of their victors. (qtd. in King) The assumptions regarding the primitive nature of indigenous culture expressed by reference to the “horrid carnival” of cannibalism complicate the processes through which contemporary writers could meaningfully draw on a tradition of New Zealand Gothic utilising the supernatural. One answer to this dilemma is to use supernatural elements not specifically associated with New Zealand. In Stephen Cain’s anthology Antipodean Tales: Stories from the Dark Side (1996) there are several instances of this, such as in the story “Never Go Tramping Alone” by Alyson Cresswell-Moorcock, which features a creature called a Gravett. As Timothy Jones’s discussion of this anthology demonstrates, there are two problems arising from this unprecedented monster: firstly, the story does not seem to be a “New Zealand Gothic”, which a review in The Evening Post highlights by observing that “there is a distinct ‘Kiwi’ feel to only a few of the stories” (Rendle 5); while secondly, the Gravatt’s appearance in the New Zealand landscape is unconvincing. Jones argues that "When we encounter the wendigo, a not dissimilar spirit to the Gravatt, in Ann Tracy’s Winter Hunger or Stephen King’s Pet Sematary, we have a vague sense that such beings ‘exist’ and belong in the American or Canadian landscapes in which they are located. A Gravatt, however, has no such precedent, no such sense of belonging, and thus loses its authority" (251). Something of this problem is registered in Elizabeth Knox’s vampire novel Daylight (2003), which avoids the problem of making a vampire “fit” with a New Zealand landscape devoid of ancient architecture by setting all the action in Europe. One of the more successful stories in Cain’s collection demonstrates a way of engaging with a specifically New Zealand tradition of supernatural Gothic, while also illustrating some of the potential pitfalls in utilising colonial Gothic tropes of menacing bush, Maori burial caves and skeletal remains. Oliver Nicks’s “The House” focuses on a writer who takes up residence in an isolated “little old colonial cottage in the bush” (8). The strange “odd-angled walls”, floors that seem to slope downwards and the “subterranean silence” of the cottage provokes anxiety in the first-person narrator who admits his thoughts “grew increasingly dark and chaotic” (8). The strangeness of the house is only intensified by the isolation of its surroundings, which are fertile but nevertheless completely uninhabited. Alone and unnerved by the oddness of the house, the narrator listens to the same “inexplicable night screeches and rustlings of the bush” (9) that furnish so much New Zealand Gothic. Yet it is not fear inspired by the menacing bush that troubles the narrator as much as the sense that there was more in this darkness, something from which I felt a greater need to be insulated than the mild horror of mingling with a few wetas, spiders, bats, and other assorted creepy-crawlies. Something was subtlely wrong here – it was not just the oddness of the dimensions and angles. Everything seemed slightly off, not to add up somehow. I could not quite put my finger on whatever it was. (10) When the narrator escapes the claustrophobic house for a walk in the bush, the natural environment is rendered in spectral terms. The narrator is engulfed by the “bare bones of long-dead forest giants” (11) and “crowding tree-corpses”, but the path he follows in order to escape the “Tree-ghosts” is no more comforting since it winds through “a strange grey world with its shrouds of hanging moss, and mist” (12). In the midst of this Gothicised environment the narrator is “transfixed by the intersection of two overpowering irrational forces” when something looms up out of the mist and experiences “irresistible curiosity, balanced by an equal and opposite urge to turn and run like hell” (12). The narrator’s experience of being deep in the threatening bush continues a tradition of colonial writing that renders the natural environment in Gothic terms, such as H. B. Marriot Watson’s The Web of the Spider: A Tale of Adventure (1891), which includes an episode that sees the protagonist Palliser become lost in the forest of Te Tauru and suffer a similar demoralization as Nicks’s narrator: “the horror of the place had gnawed into his soul, and lurked there, mordant. He now saw how it had come to be regarded as the home of the Taniwha, the place of death” (77). Philip Steer points out that it is the Maoriness of Palliser’s surroundings that inspire his existential dread, suggesting a certain amount of settler alienation, but “Palliser’s survival and eventual triumph overwrites this uncertainty with the relegation of Maori to the past” (128). Nicks’s story, although utilising similar tropes to colonial fiction, attempts to puts them to different ends. What strikes such fear in Nicks’s narrator is a mysterious object that inspires the particular dread known as the uncanny: I gave myself a stern talking to and advanced on the shadow. It was about my height, angular, bony and black. It stood as it now stands, as it has stood for centuries, on the edge of a swamp deep in the heart of an ancient forest high in this remote range of hills forming a part of the Southern Alps. As I think of it I cannot help but shudder; it fills me even now with inexplicable awe. It snaked up out of the ground like some malign fern-frond, curving back on itself and curling into a circle at about head height. Extending upwards from the circle were three odd-angled and bent protuberances of unequal length. A strange force flowed from it. It looked alien somehow, but it was man-made. Its power lay, not in its strangeness, but in its unaccountable familiarity; why did I know – have I always known? – how to fear this… thing? (12) This terrible “thing” represents a return of the repressed associated with the crimes of colonisation. After almost being devoured by the malevolent tree-like object the narrator discovers a track leading to a cave decorated with ancient rock paintings that contains a hideous wooden creature that is, in fact, a burial chest. Realising that he has discovered a burial cave, the narrator is shocked to find more chests that have been broken open and bones scattered over the floor. With the discovery of the desecrated burial cave, the hidden crimes of colonisation are brought to light. Unlike colonial Gothic that tends to represent Maori culture as threatening, Nicks’s story represents the forces contained in the cave as a catalyst for a beneficial transformative experience: I do remember the cyclone of malign energy from the abyss gibbering and leering; a flame of terror burning in every cell of my body; a deluge of shrieking unreason threatening to wash away the bare shred that was left of my mind. Yet even as each hellish new dimension yawned before me, defying the limits even of imagination, the fragments of my shattered sanity were being drawn together somehow, and reassembled in novel configurations. To each proposition of demonic impossibility there was a surging, answering wave of kaleidoscopic truth. (19) Although the story replicates colonial writing’s tendency to represent indigenous culture in terms of the irrational and demonic, the authenticity and power of the narrator’s experience is stressed. When he comes to consciousness following an enlightenment that sees him acknowledging that the truth of existence is a limitless space “filled with deep coruscations of beauty and joy” (20) he knows what he must do. Returning to the cottage, the narrator takes several days to search the house and finally finds what he is looking for: a steel box that contains “stolen skulls” (20). The narrator concludes that the “Trophies” (20) buried in the collapsed outhouse are the cause for the “Dark, inexplicable moods, nightmares, hallucinations – spirits, ghosts, demons” that “would have plagued anyone who attempted to remain in this strange, cursed region” (20). Once the narrator returns the remains to the burial cave, the inexplicable events cease and the once-strange house becomes an ideal home for a writer seeking peace in which to work. The colonial Gothic mode in New Zealand utilises the Gothic’s concern with a haunting past in order to associate that past with the primitive and barbaric. By rendering Maori culture in Gothic terms, such as in Maning’s blood-splattered scene of grieving or through the spooky discoveries of bone-strewn caves, colonial writing compares an “uncivilised” indigenous culture with the “civilised” culture of European settlement. For a contemporary writer wishing to produce a New Zealand supernatural horror, the colonial Gothic is a problematic tradition to work from, but Nicks’s story succeeds in utilising tropes associated with colonial writing in order to reverse its ideologies. “The House” represents European settlement in terms of barbarity by representing a brutal desecration of sacred ground, while indigenous culture is represented in positive, if frightening, terms of truth and power. Colonial Gothic’s tendency to associate indigenous culture with violence, barbarism and superstition is certainly replicated in Nicks’s story through the frightening object that attempts to devour the narrator and the macabre burial chests shaped like monsters, but ultimately it is colonial violence that is most overtly condemned, with the power inhabiting the burial cave being represented as ultimately benign, at least towards an intruder who means no harm. More significantly, there is no attempt in the story to explain events that seem outside the understanding of Western rationality. The story accepts as true what the narrator experiences. Nevertheless, in spite of the explicit engagement with the return of repressed crimes associated with colonisation, Nicks’s engagement with the mode of colonial Gothic means there is a replication of some of its underlying notions relating to settlement and belonging. The narrator of Nicks’s story is a contemporary New Zealander who is placed in the position of rectifying colonial crimes in order to take up residence in a site effectively cleansed of the sins of the past. Nicks’s narrator cannot happily inhabit the colonial cottage until the stolen remains are returned to their rightful place and it seems not to occur to him that a greater theft might underlie the smaller one. Returning the stolen skulls is represented as a reasonable action in “The House”, and it is a way for the narrator to establish what Linda Hardy refers to as “natural occupancy,” but the notion of returning a house and land that might also be termed stolen is never entertained, although the story’s final sentence does imply the need for the continuing placation of the powerful indigenous forces that inhabit the land: “To make sure that things stay [peaceful] I think I may just keep this story to myself” (20). The fact that the narrator has not kept the story to himself suggests that his untroubled occupation of the colonial cottage is far more tenuous than he might have hoped. References Ballantyne, David. Sydney Bridge Upside Down. Melbourne: Text, 2010. Bannister, Bronwyn. Haunt. Dunedin: University of Otago Press, 2000. Calder, Alex. “F. E. Maning 1811–1883.” Kotare 7. 2 (2008): 5–18. Chapman, R. H. 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