bluuebunny/crossref_metadata_2025 huggingface dataset:
An improved sliding-mode observer-based equivalent-input-disturbance approach for permanent magnet synchronous motor drives with faults in current measurement circuits
Authors: Huang Gang , Huang Wei , Li Zhengtan , Li Jiajun , He Jing , Zhang Changfan , Zhao Kaihui
Published: April 2021

The reliability of permanent magnet synchronous motor (PMSM) systems is very important in high-precision industrial drives. However, disturbance or sensor fault may cause the performance degradation of the system. This paper presents an improved sliding-mode-observer (SMO)-based equivalent-input-disturbance (EID) approach for the rejection of faults in current measurement circuits of a PMSM drive. A system model, which contains faults in current measurement circuits, is first constructed by using EIDs in control input circuits. Then, an improved SMO is designed to estimate the equivalent-input-faults. The effect of the faults on the system is rejected based on the EID theory. Moreover, the global stability and convergence analysis is also provided. Experiments and comparisons demonstrate the effectiveness of the method.
Predictive extended state observer-based robust control for uncertain linear systems with experimental validation
Authors: Pawar Sushant N , Chile Rajan H , Patre Balasaheb M
Published: April 2021

This paper describes a predictive extended state observer-based robust control for uncertain process control applications. The technique discussed in the article uses the extended state observer (ESO) that can estimate the dynamics of the system as well as total disturbance encountered in the system. The disturbances, parametric uncertainties associated with the processes are treated as an extended state variable to be estimated in real-time using ESO. With the implementation of a predictive algorithm with an ESO, the proposed control structure extends its applicability to time-delayed higher-order processes. The proposed control technique utilizes the simple first-order modified predictive ESO even in the case of higher-order processes. The novel predictive ESO is able to obtain a delay less estimation of total disturbance as compared with existing normal ESO. Also, novel predictive ESO maintains its stability margin in presence of time delay as well provides better response as compared with normal ESO. Numerical simulations show that the proposed scheme provides a significant improvement in transient response as compared with internal model control-based proportional-integral-derivative (IMC-PID) control. The proposed scheme requires less knowledge of the process as compared with the IMC-PID structure. The implementation of the proposed control is tested on a real-life single tank level control system. Because of its merit, the suggested technique can be used as automatic for online tuning, as it is less reliant on the process model.
Control and Surveillance in Work Practice: Cultivating Paradox in ‘New’ Modes of Organizing
Authors: de Vaujany François-Xavier , Leclercq-Vandelannoitte Aurélie , Munro Iain , Nama Yesh , Holt Robin
Published: April 2021

The new world of work is being characterized by the emergence of what are, apparently, increasingly autonomous ways of working and living. Mobile work, coworking, flex office, platform-based entrepreneurship, virtual collaborations, Do It Yourself (DIT), remote work, digital nomads, among other trends, epitomize ways of organizing work practice that purportedly align productivity with freedom. But most ethnographical research already reveals many paradoxical experiences associated with these new practices and processes. Indeed, it appears that with autonomy comes surveillance and control, to a point where, as Foucault observed way back, subjectivity and subject become synonyms, and the current pandemic both strengthens and makes visible this situation. In this introduction to the special issue we make a foray into this situation, using four open and related themes developed in the five papers we selected: managerial control and technology; surveillance and platform capitalism; time and space; and new organizational forms and autonomy. Paradoxical movements are identified for each of them, before we conclude by reflecting on a grounding paradox which appears at the centre of this special issue and the themes it covers.
A systematic review of randomized controlled trials examining workplace wellness interventions
Authors: Murphy Bridget , Parekh Niyati , Vieira Dorice L. , O’Connor Joyce A.
Published: April 2021

Background:

One of the influencing factors associated with weight gain is overeating as a maladaptive coping strategy to process or avoid the emotional impact of psychological stress. Psychological stress is chronically and pervasively associated with stress stemming from the workplace environment. Workplace wellness interventions have a unique opportunity to change environmental factors impacting psychological stress, which can improve individual food choice and weight management efforts.

Aim:

To synthesize evidence from randomized controlled trials on workplace wellness interventions that impact employee psychological stress and food choice or weight management.

Methods:

A systematic review was completed according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Selected studies were limited to English-language articles exploring randomized interventions at workplaces among adult employees and included measurements of psychometric stress and food choice (qualitative or quantitative) or biometric weight management. From the search, 10 studies were included in the final review.

Results:

Results were inconsistent across studies. There was no observable association between psychological stress reduction and food choice or weight management. Mid-length interventions (ranging from 6 to 9 months) had more consistent associations between intervention program implementation, reduced psychological stress, and improved food choice or weight management.

Conclusions:

The studies examining employee food choices and weight management efforts remained very heterogeneous, indicating that more research is needed in this specific area of employee wellness program planning and measurement. Consistent research methodology and assessment tools are needed to measure dietary intake.
Biomechanical Effects of Torquing on Upper Central Incisor with Thermoplastic Aligner: A Comparative Three-Dimensional Finite Element Study with and Without Auxillaries
Authors: Sandhya V , Arun AV , Reddy Vinay P , Mahendra S , Chandrashekar BS , Aravind
Published: April 2021

Background and Objectives:

This study was conducted to determine the effective method to torque the incisor with thermoplastic aligner using a three-dimensional (3D) finite element method.

Materials and Methods:

Three finite element models of maxilla and maxillary dentition were developed. In the first model, thermoplastic aligner without any auxiliaries was used. In the second and third models, thermoplastic aligner with horizontal ellipsoid composite attachment and power ridge were used, respectively. The software used for the study was ANSYS 14.5 FE. A force of 100 g was applied to torque the upper right central incisor. The resultant force transfer, stress distribution, and tooth displacement were evaluated.

Results:

The overall tooth displacement and stress distribution appeared high in the model with power ridge, whereas the root movement was more in the horizontal ellipsoid composite attachment model. The model without any auxillaries produced least root movement and stress distribution.

Conclusion:

Horizontal ellipsoid composite attachment achieved better torque of central incisor than the model with power ridge and model without any auxillaries.
Plateformes digitales et concurrence par la rapidité. Le cas des vêtements d’occasion
Authors: Juge Elodie , Pomiès Anissa , Collin-Lachaud Isabelle
Published: April 2021

Des recherches récentes explorent ce que les plateformes digitales font et font faire aux utilisateurs. Dans leur prolongement, cet article étudie comment les plateformes digitales P2P accélèrent les activités marchandes, et comment cette accélération modèle les relations entre utilisateurs. Une étude qualitative à caractère ethnographique a été menée au sein du marché des vêtements d’occasion de 2013 à 2020. En mobilisant la théorie de l’accélération sociale, nous montrons que les affordances des plateformes digitales encouragent une accélération du rythme de vie, une accélération du renouvellement des objets possédés, et une accélération technique multi-domaines. Ces formes d’accélération aboutissent à une concurrence par la rapidité entre utilisateurs : chacun d’eux se doit d’être plus réactif, plus à la pointe, et plus averti que les autres. De plus, les plateformes constituent un espace digital particulier qui invite à repenser l’aliénation sous plusieurs formes, et notamment l’aliénation par rapport à l’espace et par rapport au temps. Outre ses contributions théoriques, cette recherche offre des contributions sociétales visant à faire prendre conscience aux utilisateurs de l’ampleur de leurs activités marchandes accélérées sur les plateformes digitales.
Guest editorial: EU agencies in transnational criminal enforcement: From a coordinated approach to an integrated EU criminal justice
Authors: Öberg Jacob
Published: April 2021

The articles in this special issue consider the institutional foundations of the Union’s criminal policy – a highly critical question for the future development of the Area of Freedom, Security and Justice. The ratification of the Lisbon Treaty and the subsequent legal and political developments have entailed an unprecedented reinforcement of the powers of the EU’s criminal justice agencies Europol, Eurojust and, recently, the establishment of a novel criminal justice body – the European Public Prosecutor’s Office. On the basis of the Treaty mandate, the EU legislator has adopted important reforms such as the EPPO Regulation, and new Europol and Eurojust regulations. In light of these developments, this special issue explores via a multi-disciplinary investigation the extent to which the increased competences of the EU and the stronger presence of EU criminal justice agencies have transformed EU criminal law from an ‘intergovernmental’ regime to a ‘supranational’ and ‘integrated’ framework. We expect that this special issue will enhance further debate on EU criminal justice agencies, encourage novel paths to bridge the boundaries between disciplinary epistemic communities in the study of EU criminal justice and more broadly contribute to an advanced understanding of the role of law in social and political integration.
Abandonment/decommissioning under Nigerian legal regimes: A comparative analysis
Authors: Anyatang Brian F.I. , Kooffreh Bassey E.
Published: April 2021

The overall objective of this article is to analyse critically the legal instruments that regulate decommissioning of onshore and offshore platforms in the Nigerian Petroleum Industry vis-à-vis other jurisdictions such as The United States of America (USA) and United Kingdom (UK) with a view to making requisite recommendations for improvement in the Nigerian Petroleum Industry. This article in its introductory part will be devoted to conceptual clarifications of decommissioning and abandonment and differences (if any). Part two is the critical analysis of legislations on decommissioning in Nigeria. Part three is also a critical analysis on legal regimes and practices on decommissioning in other jurisdictions such as USA and UK. Part four is a comparative analysis of legislations and decommissioning practices in Nigeria and other jurisdiction. Part five consists of findings/recommendations made during the research. In the end, conclusions are drawn, part of which is a call for proactive actions by megacorporation and the Nigerian government in the sphere of timeous decommissioning of obsolete and failed platforms, enactment and, or, review of obsolete legislations regulating decommissioning as well as fulfilment of obligations under multilateral environmental treaties that regulate decommissioning and sustainable environmental management and protection.
Who should fight domination? Individual responsibility and structural injustice
Authors: Gädeke Dorothea
Published: April 2021

Who is responsible for fighting domination? Answering this question, I argue, requires taking the structural dimension of domination seriously to avoid unwillingly reproducing domination in the name of justice. Having cast domination as a structural injustice that refers to structurally constituted positions of power and disempowerment, I show that the outcome-based, the capacity-based and the social connection model suggested in literature on responsibility, fail to fully meet this challenge. Drawing on insights from all of them, I propose an account that proves more sensitive towards the power dynamics at play in fighting domination. It is based on a fundamental duty of justice, which gives rise to two kinds of responsibility. Dominators, dominated and peripheral agents share political responsibility for domination in virtue of reproducing domination by occupying a position within structures of dominating power; they are required to acknowledge and undermine their position of power or disempowerment rather than simply using and thus tacitly reaffirming it. Political responsibility for domination is distinct from moral responsibility for acting within contexts of domination; in fact, ignoring this difference risks reproducing rather than transforming relations of domination. Bystanders who are not implicated in reproducing domination bear limited remedial responsibility to support this struggle.
Game Exposure, Player Characteristics, and Neuromuscular Performance Influence Injury Risk in Professional and Youth Field Hockey Players
Authors: Mason Joel , Wellmann Kai , Groll Andreas , Braumann Klaus-Michael , Junge Astrid , Hollander Karsten , Zech Astrid
Published: April 2021

Background:

Identifying factors that contribute to the occurrence of injury is an important step in designing training programs to minimize the risk of injury. However, despite high injury rates, variables contributing to injury in field hockey players remain relatively unexplored.

Purpose:

To identify factors that may predict injury in professional and youth field hockey players.

Study Design:

Cohort study; Level of evidence, 2.

Method:

Professional and youth hockey players completed preseason neuromuscular performance testing and were monitored for injuries, training, and game exposure throughout the subsequent 12-month indoor and outdoor hockey season. Least absolute shrinkage and selection operator analysis was conducted to identify injury risk factors. Receiver operating characteristic curves were then calculated to determine the individual predictive accuracy of the identified variables.

Results:

A total of 83 players (mean ± SD age, 20.7 ± 4.9 years; 34.9% female) with complete performance, injury, and exposure data were included for data analysis. Almost half of players (44.6%) sustained a time-loss injury during the season, and 73% of these injuries occurred in the lower limb. Playing more games and having an older age, asymmetrical and poor dynamic postural control, and better explosive performance were identified as jointly influencing the risk of injury. When considered individually, number of games played throughout the season was the most accurate predictor of injury risk (area under the curve [AUC] = 0.74; P < .001), while asymmetrical and poor dynamic postural control (AUCs = 0.61-0.65; P = .01-.04) and better explosive athletic performance (AUCs = 0.65-0.67; P < .01) were identified as moderate individual predictors of sustaining a general or lower limb injury.

Conclusion:

A number of modifiable factors were individually and jointly associated with an increased injury risk in field hockey players, providing initial evidence for the design of targeted and sport-specific training programs to mitigate the risk of injury.
Exploring the Experiences and Information Needs of Parents Caring for a Child With a Urinary Tract Infection: A Qualitative Study
Authors: Campbell Alyson , Hartling Lisa , Louie-Poon Samantha , Scott Shannon D.
Published: April 2021

Urinary tract infections (UTIs) are common in children, however, clinical practice variations can leave parents unsure how to care for their child. We aim to develop knowledge tools that provide evidence-based information about pediatric UTIs. To inform tool development, we asked parents to share their experiences and information needs in caring for a child with a UTI. Using qualitative description methods, 16 semistructured interviews were conducted with 18 parents. Parents were recruited through the emergency department (ED) of a major Canadian urban pediatric hospital. Five major themes emerged: (1) parent descriptions of their child’s symptoms and behaviors; (2) UTIs have an effect on the entire family; (3) reasons for going to the ED; (4) parent experiences with UTI treatment, management, and follow-up strategies for their child; and (5) parent information needs and preferred information sources for UTIs. Our findings highlight the diversity of UTI symptoms children experience, which causes uncertainty and confusion for parents. This diversity suggests the development of knowledge tools for parents about UTIs is needed.
The Firm as an Engine of Imagination: Organizational prospection and the making of economic futures
Authors: Beckert Jens
Published: April 2021

What do organizations do, and why? An important but only selectively scrutinized aspect of the doings of organizations consists in their creation of imaginaries of economic futures. Under conditions of uncertainty, it is through ‘imagined futures’ that organizations motivate and find the rationale for their decisions, coordinate activities, manage stakeholders and compete with one another. This article suggests making the construction of imagined futures a vantage point for the study of organizations and processes of organizing. It focuses on ‘instruments of imagination’ used by firms to create ‘fictional expectations’ which are used to come to terms with an uncertain future – and to proactively shape this future. Instruments discussed here include strategic planning, technological projections, economic forecasting, and business plans among others. The article argues that a fruitful general analytical perspective can be developed by bringing the constitution, contestation and effects of imagined futures to the forefront of organizational analysis.
Tumor-Intrinsic RIG-I Signaling Promotes Anti-CTLA-4 Checkpoint Inhibitor-Mediated Anticancer Immunity
Authors: Heidegger Simon , Wintges Alexander , Bek Sarah , Schmickl Martina , Ruland Jürgen , van den Brink Marcel R.M. , Peschel Christian , Haas Tobias , Poeck Hendrik
Published: April 2021

Strong inter-individual variation in clinical response to checkpoint inhibitors such as anti-CTLA-4 remains a major challenge. In a retrospective analysis, expression of viral defense genes in human melanomas including the cytosolic RNA receptor RIG-I (DDx58) has been associated with clinical benefit to CTLA-4 blockade. However, possible interconnections of these two distinct pathways remain unknown. Using CRISPR/Cas9 technology to generate B16 melanoma cells lines that lack nucleic acid receptors or downstream signaling molecules (RIG-I, STING, IRF3/7) together with available genetically deficient mouse models, we addressed the importance of nucleic acid receptor signaling in both tumor and host cells for the efficacy of anti-CTLA-4 immunotherapy. We here provide experimental proof that anti-CTLA-4 immunotherapy relies on tumor cell-intrinsic RIG-I but not STING signaling. Following anti-CTLA-4 treatment, tumor-intrinsic RIG-I signaling critically impacts on cross-presentation of tumor-associated antigen by CD103+ dendritic cells, the expansion of tumor antigen-specific CD8+ T cells and ultimately the accumulation of CD8+ T cells within tumor tissue. Consistently, therapeutic targeting of RIG-I with 5'-phosphorylated-RNA in both tumor and non-malignant cells potently augmented the efficacy of CTLA-4 checkpoint blockade. These processes were additionally dependent on host STING, MAVS and type I interferon signaling and were closely linked to RIG-I-mediated tumor cell death. In summary, our study identifies both tumor- and host-intrinsic RIG-I signaling as fundamental requirements for anti-CTLA-4-mediated antitumor immunity. Our data predict that targeting RIG-I may serve as a basis for the development of new combination strategies to increase the response rate of checkpoint inhibitor-based immunotherapy of malignancy including lymphoma, particularly in individuals that do not have a sufficient spontaneous antitumor T-cell immune response.

Disclosures

van den Brink: Jazz Pharmaceuticals: Consultancy; Seres: Research Funding; PureTech Health: Consultancy; Therakos Institute: Other: Speaking engagement.
Single Cell-Resolution Analysis of HSC Dysfunction in Mir-146a knockout Mice
Authors: Grants Jennifer , Wegrzyn Joanna , Knapp David , Hui Tony , O'Neill Kieran , Parker Jeremy , Deng Deborah , Boldin Mark , Hirst Martin , Eaves Connie J. , Karsan Aly
Published: April 2021

MicroRNA miR-146a is frequently depleted in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Loss of miR-146a may be an initiating event in tumorigenesis, as miR-146a loss in mouse models is sufficient to cause features of MDS and eventual progression to AML. To define how miR-146a loss initiates tumorigenesis, we analyzed hematopoietic stem cell (HSC) function from miR-146a knockout (KO) mice prior to onset of an overt malignant phenotype. Tracking cell division kinetics, proliferation, and differentiation of single long-term HSC (LT-HSC; EPCR+CD45+CD48-CD150+) in culture, we found evidence that miR-146a KOreduces HSC quiescence and promotes differentiating cell divisions. Our data show that miR-146a KO HSC dysfunction may stem from loss of a CD150-bright EPCR-bright sub-population, which has previously been associated with robust HSC activity. In line with this, single cell DNA methylation profiling revealed a reduction in a primitive sub-population of LT-HSCs in miR-146a KO animals. In addition, single cell LT-HSC transplants revealed a myeloid repopulation bias. As reduced HSC cell cycle quiescence has been linked to impaired HSC self-renewal upon hematopoietic stress, such as serial transplantation, we assessed the frequency of serially transplantable HSCs by performing secondary transplants with limiting dilution. Serially transplantable HSC frequency was reduced in miR-146a KO compared to wild type, suggesting impaired HSC self-renewal. Transcriptome profiling of miR-146a KO hematopoietic stem and progenitor cells identified tumor necrosis factor (TNF) signaling activation as a potential driver of HSC dysfunction. LT-HSC cell cycle quiescence and the CD150-bright EPCR-bright LT-HSC sub-population were restored in miR-146a/TNF double KO mice, suggesting that aberrant TNF signaling activation drives HSC dysfunction upon loss of miR-146a. Gene expression levels in the TNF signaling network are inversely correlated with miR-146a levels in human AML, implying that TNF signaling may similarly disrupt HSC function in miR-146a- depleted myeloid malignancies. Overall, our findings suggest that miR-146a promotes HSC cell cycle quiescence and inhibits differentiation by antagonizing TNF signaling, in order to maintain a primitive sub-population of long-term self-renewing HSCs.

Disclosures

Eaves: Experimental Hematology: Other: Editor of journal; StemCell Technologies Inc: Other: Wife of owner.
Deficiency of Cux1, Encoded on Human Chromosome 7q, Causes Aberrant Hematopoietic Stem Cell Function and Spontaneous Myeloproliferative Disease in Mice
Authors: An Ningfei , Khan Saira , Imgruet Molly , Gurbuxani Sandeep , Burgess Michael R. , McNerney Megan E.
Published: April 2021

Loss of all or part of chromosome 7 [-7/del(7q)] is a common, high-risk cytogenetic abnormality identified in a variety myeloid malignancies, including myelodysplastic syndrome (MDS), acute myeloid leukemia, therapy-related myeloid neoplasms, juvenile myelomonocytic leukemia (JMML) and chronic myelomonocytic leukemia (CMML). CUX1 is a non-clustered, homeodomain-containing transcription factor encoded on the commonly deleted region of 7q22. CUX1 inactivating mutations occur across solid tumors and myeloid neoplasms, and carry a poor prognosis in myeloid malignancies independently of 7q status. Previously, we reported that CUX1 is a conserved, haploinsufficient, myeloid tumor suppressor. To determine the mechanism by which CUX1 regulates leukemogenesis, we generated an inducible, shRNA-transgenic, Cux1 -knockdown mouse model. Insufficient Cux1 levels caused early mortality in mice, due to a spontaneous myeloproliferative disorder with features of CMML/JMML and MDS. Cux1 deficient mice had a significant increase in myelomonocytic cell numbers in the peripheral blood, bone marrow, and spleen. Cux1 deficiency led to extramedullary hematopoiesis, splenomegaly, and an increase in megakaryocyte numbers and megakaryocyte dysplasia. In addition, Cux1 is required for normal red cell development, as Cux1 knockdown led to a normocytic anemia due to an erythroblast maturation arrest. To identify CUX1 transcriptional targets, we performed gene expression profiling after CUX1 knockdown in CD34+ human hematopoietic stem and progenitor cells (HSPCs). Differentially expressed genes were enriched for genes deregulated in JMML and -7/del(7q) MDS, in addition to altered expression of quiescent, proliferative, and myeloid differentiation pathways. Consistent with these gene signatures, Cux1 maintains hematopoietic stem cell quiescence and represses HSPC proliferation, self-renewal, and granulocyte/monocyte differentiation. Thus Cux1 has pleotropic roles in multiple myeloid lineages. Deletion of CUX1, in the context of -7/del(7q), may facilitate myeloid transformation through aberrant transcriptional regulation of HSPC differentiation and proliferation.

Disclosures

No relevant conflicts of interest to declare.
Cytokine Release Syndrome and Tumor Responses in a First-in-Man Trial of a Novel Affinity-Enhanced TCR-Gene Transduced T Cell Transfer Targeting NY-ESO-1 Antigen
Authors: Kageyama Shinichi , Ishihara Mikiya , Kitano Shigehisa , Miyahara Yoshihiro , Kato Hidefumi , Mishima Hideyuki , Yamamoto Noboru , Hattori Hiroyoshi , Iwase Hiroaki , Funakoshi Takeru , Kojima Takashi , Inoue Naoki , Kidokoro Tomohide , Tomura Daisuke , Okamoto Sachiko , Chono Hideto , Nukaya Ikuei , Mineno Junichi , Ikeda Hiroaki , Watanabe Takashi , Shiku Hiroshi
Published: April 2021

Adoptive cell transfers of receptor gene-engineered T cells include chimeric antigen receptor-gene transduced T (CAR-T) cell therapy and TCR-gene transduced T (TCR-T) cell therapy. In CD19-CAR-T cell therapy, high incidence of cytokine release syndrome (CRS) is associated with in vivo CAR-T cell proliferation and its clinical efficacy. In human TCR-T cell therapies, there have not been well known about CRS and its association with in vivo T cell kinetics or tumor responses.

We have been developing a novel-type affinity-enhanced NY-ESO-1-specific TCR, and an original retrovirus vector that encodes siRNA to silence endogenous TCR creation. The NY-ESO-1 TCR is mutated for high affinity with replacements of G50A and A51E in CDR2 region, which is restricted with HLA-A*02:01 and A*02:06. We extensively examined potential cross-reactivities to different antigen-peptides in preclinical studies, and the high-affinity NY-ESO-1 TCR did not recognize analogous peptides. The new generation retroviral TCR-vector provides enhanced expression of transduced tumor-specific TCRs and an inhibition effect of formations of self-reactive TCRs.

This is a first-in-man clinical trial of the novel NY-ESO-1-specfic TCR-T cell transfer to evaluate the safety, in vivo cell kinetics and clinical responses. It is designed as a cell-dose escalation from 5 x108 to 5 x109 cells. NY-ESO-1-expressing refractory cancer patients were enrolled, with 3+3 cohort design. Cyclophosphamide with/without fludarabine were administered prior to the TCR-T cell transfer as pre-conditioning.

Six patients were treated with the NY-ESO-1 TCR-T cell transfer, and evaluated for the safety and in vivo cell kinetics. The TCR-T cells appeared in peripheral blood with a dose-dependent manner, associated with in vivo proliferation in an early phase. In three patients given 5x108 cells, no toxicities were seen. Two patients receiving 5x109 cells developed early-phase CRS (G2), with elevations of serum IL-6 and IFN-gamma. They were managed the treatment of anti-IL-6 receptor monoclonal antibody, tocilizumab. In a patient who developed CRS, an event of lung injury (G3) occurred, which was associated with marked infiltration of the NY-ESO-1 TCR-T cells. It was successfully treated with steroid.

Two synovial sarcoma patients exhibited tumor responses of PRs. In one patient, progression-free survival lasted more than 8 months.

In summary, the affinity-enhanced NY-ESO-1 TCR-T cell transfer exhibited CRSs in association with in vivo cell proliferation and sequential tumor responses.

Disclosures

No relevant conflicts of interest to declare.
Bortezomib, Lenalidomide and Low-Dose Dexamethasone (VRD) Versus Lenalidomide and Low-Dose Dexamethasone (Ld) for Newly-Diagnosed Multiple Myeloma- a Randomized Phase III Study
Authors: Mookerjee Anjali , Gupta Ritu , Jasrotia Shivali , Sahoo Ranjit , Kumar Rakesh , Thulkar Sanjay , Sharma Om Dutt , Kumar Lalit
Published: April 2021

Background: In this prospective study, we compared VRD with Ld as induction therapy for newly-diagnosed Multiple myeloma patients. The primary objective of this study is to compare the progression-free survival in the 2 arms.

Methods: Between September 2014 and Oct 2016, 144 patients have been recruited and randomly assigned to receive 4 cycles of either Bortezomib 1.3 mg/m2 SC on days 1, 8, 15 and 22 with Lenalidomide 15mg/day from day 1 to 14 (Arm A) or Lenalidomide 25 mg/day from day 1 to 21 (Arm B). Patients in both arms received oral dexamethasone 40 mg on days 1,8,15 and 22. Both treatment regimens were 28-day cycles. All patients received 75 mg aspirin daily, acyclovir prophylaxis and monthly zoledronic acid. Response assessment was done at the end of the 4th cycle using the International Myeloma Working Group (IMWG) uniform response criteria. The study was approved by the Institute Ethics Committee (Ref IEC/NP-264/01-08-2014, RP-7/2014).

Results: These are the results from an analysis of 143 patients (arm A-74, arm B-69). Baseline characteristics of patients were similar in both arms with respect to age, gender, ISS and DS stage, immunoglobulin subtype and serum LDH. Patients' median age is 56 years (range 31-70) in arm A and 52 years (range 28-69) in arm B. Gender M/F: Arm A 54/20 and 43/26 in arm B. ISS stage III 51 (68.9%) arm A vs 44 (63.8%) arm B. Serum LDH raised to >250 u/L was observed in 25 (44.6%) vs 31 (52.5%) in arms A and B, p=0.4. Revised staging including ISS and serum LDH at baseline: stage III 47 (81%) and 37 (65%) in arms A and B respectively. 14 (18.9%) and 19 (27.5%) of patients had light chain myeloma in arms A and B respectively. Overall response rates (sCR+CR+VGPR+PR) is 78.4% vs 73.9% in arms A and B respectively, p=0.6; sCR + CR 21 (28.4%) and 21 (30.4%) respectively, p=0.86. Median follow-up 17.1 months (range 1 to 33). Median overall survival (OS) is 30.2 months (95% CI 28.2 to 32.2) and 28.6 months (95%CI 26 to 31.3) in arms A and B respectively, p=0.3. Median progression-free survival (PFS) was 27.8 months (95%CI 25.4 to 30.2) and 28 months (95%CI 24.6 to 31.4) respectively, p=0.3. Estimated one-year OS is 88% vs 85% in arms A and B, and PFS 83% vs 72%, respectively. Grade 3 anemia occurred in one patient in arm B, and grade 3 deep vein thromboses in one patient in arm A. One patient in arm A developed grade 4 myelosuppression leading to therapy change at the end of the first cycle.

Conclusion: In this analysis - response rates and median progression-free survival are similar in both arms.

Disclosures

No relevant conflicts of interest to declare.
Bok Promotes Erythropoiesis in a Mouse Model of Myelodysplastic Syndrome
Authors: Kang Seong-Ho , Perales Oscar , Michuad Michael , Katz Samuel G.
Published: April 2021

BCL-2 Ovarian Killer (BOK) is a pro-apoptotic member of the BCL-2 family of proteins best characterized for its putative ability to induce apoptosis in response to Endoplasmic Reticulum (ER) stress, when stabilized from ER-associated degradation (ERAD). Although ER stress appropriately activates the unfolded protein response (UPR) in BOK-disrupted cells, as measured by PERK and eIF2-alpha phosphorylation, downstream effector signaling, including ATF4 and CHOP, is defective. A functional role for BOK as a tumor suppressor is suggested by its genetic location in one of the 20 most frequent, focally deleted chromosomal regions across all human cancers. To evaluate the consequences of BOK loss in the pathogenesis of myelodysplasia (MDS) and Acute Myeloid Leukemia (AML), we used the Nup98-HoxD13 (NHD13) transgenic mouse model of MDS/AML. In this model, both overexpression of anti-apoptotic BCL-2 and deletion of pro-apoptotic PUMA rescue cytopenias, but surprisingly delay progression to AML. In contrast, AML developed in 36.7% of NHD13 mice lacking BOK between the age of 8 and 13 months with a similar overall survival to the NHD13 mice. However, the loss of BOK exacerbated the anemia of the NHD13 mice, which raised a potential connection between BOK and the regulation of erythropoiesis in cells experiencing stress from the NHD13 translocation. NHD13 mice deficient for BOK exhibited significantly lower hemoglobin (Hb), lower mean cell hemoglobin concentration (MCHC) and higher mean cell volume (MCV) than NHD13 mice, whereas other lineages were unaffected. Mouse colony forming unit assays revealed there is a decreased amount of erythroid progenitor stem cells (BFU-E) in the bone marrow of NHD13-transgenic/BOK-deficient mice, which hinted at a diminished ability to produce RBCs in the absence of BOK. Isolation of various stages of erythroid progenitors in the bone marrow by CD44/TER119 FACS sorting revealed that both NHD13 and NHD13-transgenic/BOK-deficient mice have an increase in proerythroblasts relative to more mature red blood cells. Preliminary RT-QPCR analysis shows decreased expression of UPR components in the RBC progenitors of both BOK-deficient and NHD13-transgenic/BOK-deficient mice. Interestingly, CHOP is not only a component of the UPR, but also an erythropoietin target gene necessary for erythroid differentiation. These results suggest that in addition to its pro-apoptotic function, BOK may have other regulatory roles within the cell, and specifically a role in regulating erythropoiesis when certain RBC progenitors experience ER stress.

Disclosures

Katz: Gene-in-Cell: Equity Ownership.
The Flow Cytometric Osmotic Fragility Test Is an Effective Screening Method for Dehydrated Hereditary Stomatocytosis
Authors: Utsugisawa Taiju , Iwasaki Takuya , Aoki Takako , Okamoto Yoshio , Kawakami Takahiro , Ogura Hiromi , Kanno Hitoshi
Published: April 2021

Introduction: Dehydrated hereditary stomatocytosis (DHSt) or hereditary xerocytosis (HX) is a form of congenital hemolytic anemia characterized by red blood cell (RBC) dehydration. Heterozygous mutations in PIEZO1, a mechanically-activated ion channel, cause DHSt. Recently, KCNN4, which encodes the Gardos channel, has been found to be the second pathogenic gene for DHSt. DHSt is characterized by an alteration in the RBC morphology in target cells, stomatocytes, and/or echinocytes, and RBC deformability assessments by ektacytometry as well as RBC ion flux measurements are currently the standard laboratory tests for DHSt, but their use in laboratories is limited. The flow cytometric osmotic fragility (FCM-OF) test is a useful diagnostic test for hereditary spherocytosis (HS) and also for hereditary elliptocytosis (HE). In this study, we showed that the FCM-OF test could also successfully diagnose DHSt.

Subjects: A total of 46 cases of RBC membrane disorders were examined, and tentative diagnoses were made based on the RBC morphology, acid glycerol lysis time, and eosin 5'-maleimide binding tests, resulting in HS (n=31), HE (n=6), and DHSt (n=9).

Methods: The number of RBCs in isotonic and hypotonic buffers were measured by flow cytometry. The degree of osmotic fragility was expressed as the "percentage residual RBCs (%RRC)". We confirmed the DHSt diagnosis by the massively paralleled sequencing using our custom panels targeting 68 hemolytic anemia-related genes with the next-generation sequencer.

Results: Both HS and HE patients showed a decrease in %RRC; HS (18.0±8.9%, p<0.001) and HE (41.8±15.7%, p<0.001) compared to normal control (66.7±1.5%). DHSt patients showed a significant increase (112.6±34.5%, p<0.001) in FCM-OF. Additionally, next-generation sequencing revealed consistent causative gene mutations for DHSt; PIEZO1 (p.R2488Q and p.E2496ELE) or KCNN4 (p.P204R, p.A279T and p.R352H).

Discussion: We examined 77 patients with congenital hemolytic anemia recently, and 59 cases were confirmed by diagnostic tests (76.6%). The results were as follows: 48 cases of RBC membrane abnormality (62.3%), 6 cases of RBC enzymopathy (7.8%), and 5 cases of hemoglobinopathy (6.5%). Of the cases of RBC membrane disorders, 31 cases of HS, 9 cases of DHSt, and 8 cases of HE were identified. These observations suggest that DHSt is the second-most common RBC membranopathy in Japan, and that the FCM-OF test and targeted sequencing efficiently discriminate DHSt from other RBC membrane disorders.

Disclosures

No relevant conflicts of interest to declare.
Restriction of HIV-1 Infection in Sickle Cell Disease Trait
Authors: Kumari Namita , Khan Javed , Ammosova Tatiana , Ahmad Asrar , Diaz Sharmin , Rana Sohail , Nekhai Sergei
Published: April 2021

We recently showed that in Sickle Cell Disease (SCD), activation of anti-viral restriction factor SAMHD1 and NF-κB inhibitor, IkBα prevented ex vivo HIV-1 infection. SAMHD1 activation was due to its reduced phosphorylation by CDK2, whose activity was inhibited by reduced intracellular iron levels in SCD peripheral blood mononuclear cells (PBMC). We also found that a hazard ratio associated with HIV infection among 11,412 SCD trait subjects was 1.0% comparing to >2% HIV-1 infection among African-Americans in USA. We compared 9 HbAS HIV-1 infected individuals with 107 HbAA or HbAC HIV-1 infected individuals who were enrolled at Howard University clinic. While hemoglobin levels in these two groups were not significantly different, HIV-1 viral load was significantly lower in HbAS group as well the number of HIV-1 associated complications (Hospitalization) were reduced. In the present study, we analyzed ex vivo HIV-1 infection of SCD trait PBMC and determined the expression of iron and hypoxia-regulated host factors including HO-1, NFκB, IKK, IKBα that are implicated in HIV-1 replication. We also analyzed CDK2 activity, SAMHD1 phosphorylation and RNR2 expression in SCD trait PBMCs. One round HIV-1 infection was significantly reduced in in SCD trait PBMC. Expression of HO-1 and IKBα was upregulated in SCD trait whereas IKK and NF-κB expression was downregulated. While CDK2 activity and SAMHD1 phosphorylation were not changed, RNR2 expression was reduced. Expression levels of HIV-1 env and gag mRNA were significantly lower in HIV-1 infected SCD trait subjects. In the same patients, HO-1 and IKBα mRNA levels were upregulated comparing to HIV-1+ HbAA or HbAC subjects. Our findings suggest that HIV-1 infection might be deregulation in SCD trait and that iron metabolism and hypoxia might play a role in this deregulation.

This work was supported by NIH Research Grants 1P50HL118006, 1R01HL125005, and 5G12MD007597. The content is solely the responsibility of the authors and does not necessarily represent the official view of NHLBI, NIMHD or NIH.

Disclosures

No relevant conflicts of interest to declare.
Impairment in the Mesenchymal Bone-Marrow Niche of Beta-Thalassemia Patients and Association with Prolonged Iron Exposure
Authors: Crippa Stefania , Rossella Valeria , Silvestri Laura , Aprile Annamaria , Scaramuzza Samantha , Rivis Silvia , Avanzini maria Antonietta , Zecca Marco , Marktel Sarah , Ciceri Fabio , Aiuti Alessandro , Ferrari Giuliana , Bernardo Maria Ester
Published: April 2021

Bone marrow (BM) contains a population of mesenchymal stromal cells (MSC) that, together with endothelial cells and osteoblasts, provide a specific microenvironment to support hematopoietic stem cell homeostasis. Beta-thalassemia (BT) is an hereditary blood disorder characterized by reduced or absent synthesis of hemoglobin beta-chains. Data on the mesenchymal compartment in BT patients are scarce. We isolated and characterized MSCs from BT and healthy donor (HD) BM samples. BT-MSCs showed a reduced clonogenic capacity, delay in colony formation, lower numbers of CFU-Fs and longer population doubling time. Similarly, we observed an altered differentiation capacity into adipocytes and osteoblasts. Both HD- and BT-MSCs express the canonical mesenchymal markers. On the contrary, the expression of CD146 and CD271 was extremely reduced in BT-MSCs, indicating a pauperization of the most primitive stem cell pool. We analyzed the expression of genes involved in the crosstalk between MSCs and HSCs and found a reduced expression of Cxcl12, SCF and Angp1 in BT-MSCs. Several genes relevant for MSC functionality were also downregulated in BT-MSCs. We demonstrated that MSCs are able to uptake and store iron. ROS levels were increased in BT-MSCs possibly due to altered anti-oxidant response and iron overload associated with blood transfusions. Importantly, higher level of ROS and reduced expression of primitive markers were observed in HD-MSCs cultured in presence of iron, underlying the importance of iron level for normal MSC function. In conclusion, we showed an impairment in the mesenchymal niche of BT-BM possibly associated with prolonged iron exposure.

Disclosures

Marktel: GSK: Other: B-thalassemia gene therapy was developed by Fondazione Telethon and Ospedale San Raffaele and has been inlicenced by GSK that provides funding for the clinical trial, Research Funding. Ciceri: GSK: Other: B-thalassemia gene therapy was developed by Fondazione Telethon and Ospedale San Raffaele and has been inlicenced by GSK that provides funding for the clinical trial, Research Funding.
Pathological Role of Halted Proteasome Machinery in Sickle Cell Disease
Authors: Song Anren , Angelo d'Alessandro , Sun Kaiqi , Liu Hong , Peng Zhangzhe , Kellems Rodney , Juneja Harinder S. , Xia Yang
Published: April 2021

Although proteasome machinery is a conserved cellular component to maintain their normal function, its function in erythrocyte under stress conditions is largely unknown, especially in sickle cell disease (SCD). To determine whether proteasome machinery is altered in SCD erythrocyte, we conducted western blot to detect total ubiquitinated proteins on the erythrocyte membrane in both mice and humans with or without SCD. We found that ubiquitinated proteins were significantly accumulated in SCD mice and humans compared to WT mice and normal controls, indicating that proteasome machinery is halted in SCD. Next, to determine which specific proteins are ubiquitinated and accumulated in SCD, we conducted robust and nonbiased proteomic profiling by immunoprecipitation ubiquitinated proteins followed by proteomics analysis. We found significant accumulation of several categories of ubiquitinated proteins on the erythrocyte membrane in SCD, including cytoskeleton proteins (Spectrin, Actin, Ankryin), glycolytic enzymes (GAPDH, 2,3-BPG mutase, Pyruvate Kinase, G6PD), transporters (Band3, large neutral AA transporter, calcium transporter, ENT1), hemoglobin, components of proteasome machinery [E2, E3 ligases, and valosin-containing protein (p97)]. Finally, to determine the effect of halted proteasome machinery in SCD functionally, we conducted in vitro hypoxia induced red blood cell (RBC) sickling assay. We found that inhibition of RBC proteasome machinery by targeting p97 using CB-5083 or targeting proteasome using MG132 increases SCD RBC sickling. Overall, our findings reveal a novel role of halted proteasome machinery in the pathophysiology of SCD and open up new therapies for the disease.

Disclosures

No relevant conflicts of interest to declare.
Hairy Platelet-Derived Extracellular Vesicles Promote Lung Vaso-Occlusion in Sickle Cell Disease
Authors: Sundd Prithu , Jimenez Maritza Ann , Bennewitz Margaret F. , Brzoska Tomasz , Tutuncuoglu Egemen , Kato Gregory J. , Gladwin Mark T
Published: April 2021

Background: Acute chest syndrome (ACS) is a type of acute lung injury and the leading cause of mortality in Sickle Cell Disease (SCD). Current treatments for ACS are primarily supportive, and there is a critical need for rescue therapies. ACS is often a sequela of acute systemic vaso-occlusive crisis and preceded by thrombocytopenia. However, the role of platelets in the pathogenesis of ACS remains largely unknown.

Methods: We used our validated model of vaso-occlusive crisis in transgenic, humanized SCD mice, which is triggered by intravenous challenge with nanogram levels of the TLR4 ligand, lipopolysaccharide (LPS). Platelet-neutrophil aggregates and blood flow in the lung microcirculation was visualized in real time in vivo, using multi-photon-excitation microscopy of intact lung in live SCD mice. SCD or control human blood was perfused through microfluidic channels in vitro and neutrophil-platelet aggregation was visualized using fluorescence microscopy. Platelet derived extracellular vesicles were characterized using nanoparticle tracking and biochemical approaches.

Results: We have made a novel finding that the arrest of blood flow and injury in the lung is secondary to blockade of pulmonary arterioles by platelet-neutrophil aggregates. Using in vitro microfluidic studies, we confirmed that platelet-neutrophil aggregation is higher in LPS-treated SCD patient blood compared with healthy controls, and this correlates with increased numbers of platelet-derived extracellular vesicles (EVs) that express IL-1β. Our studies also reveal that platelet-neutrophil aggregation in pulmonary arterioles of SCD mice is associated with an increase in peripheral blood levels of platelet-derived EVs containing IL-1β. Remarkably, inhibition of TLR4 or TLR4/NLRP3-inflammasome activated caspase-1, or inhibition of IL-1β signaling, attenuated release of platelet EVs and platelet-neutrophil aggregation in the lung arterioles of SCD mice in vivo and SCD human blood in vitro .

Conclusions: TLR4 and NLRP3-inflammasome-mediated caspase-1 activation in platelets during vaso-occlusive crisis leads to release of IL-1β-containing EVs into the circulation. These circulating platelet EVs promote platelet-neutrophil aggregation in pulmonary arterioles, which results in arrest of blood flow in the lung, leading to ACS. Therapeutic inhibition of TLR4/NLRP3-caspase-1 signaling in platelets or IL-1β signaling is a potential therapy for ACS in SCD patients.

Acknowledgments: This study was supported by 1R01HL128297-01 (P.S.) and VMI startup funds (P.S.).

Disclosures

No relevant conflicts of interest to declare.