bluuebunny/crossref_metadata_2025_split huggingface dataset:
Modifications to the Framework Regions Eliminate Chimeric Antigen Receptor Tonic Signaling
Authors: Landoni Elisa , Fucá Giovanni , Wang Jian , Chirasani Venkat R. , Yao Zhiyuan , Dukhovlinova Elena , Ferrone Soldano , Savoldo Barbara , Hong Lee K. , Shou Peishun , Musio Silvia , Padelli Francesco , Finocchiaro Gaetano , Droste Miriam , Kuhlman Brian , Shamshiev Abdijapar , Pellegatta Serena , Dokholyan Nikolay V. , Dotti Gianpietro 
Published: February 2021

AbstractChimeric antigen receptor (CAR) tonic signaling, defined as spontaneous activation and release of proinflammatory cytokines by CAR-T cells, is considered a negative attribute because it leads to impaired antitumor effects. Here, we report that CAR tonic signaling is caused by the intrinsic instability of the mAb single-chain variable fragment (scFv) to promote self-aggregation and signaling via the CD3ζ chain incorporated into the CAR construct. This phenomenon was detected in a CAR encoding either CD28 or 4-1BB costimulatory endodomains. Instability of the scFv was caused by specific amino acids within the framework regions (FWR) that can be identified by computational modeling. Substitutions of the amino acids causing instability, or humanization of the FWRs, corrected tonic signaling of the CAR, without modifying antigen specificity, and enhanced the antitumor effects of CAR-T cells. Overall, we demonstrated that tonic signaling of CAR-T cells is determined by the molecular instability of the scFv and that computational analyses of the scFv can be implemented to correct the scFv instability in CAR-T cells with either CD28 or 4-1BB costimulation.
Performance analysis of real-time PSO tuned PI controller for regulating voltage and frequency in an AC microgrid
Authors: Sultana Umbrin , Qazi Sajid Hussain , Rasheed Nadia , Mustafa M. W. 
Published: February 2021

<p>In this study, a control strategy based on the self-tuning method and synchronous reference frame (SRF) with PI regulator is proposed to achieve optimum quality of power in an autonomous micro grid (MG). The MGS is based on multiple distributed generation (DG) connected with 120 kV power grid. The proposed system is first simulated with fixed gain values for PI controller which are not optimal for sudden changes in the system i.e. transition of MG to islanding mode, load variations. So, the particle swarm optimization (PSO) has been utilized for tuning of PI controller parameters which ensure flexible performance and superior quality of power. The principal parameters considered in this study are, regulation of voltage and frequency, steady-state and dynamic response and harmonic distortion, mainly when microgrid is islanded. The performance of PI and PI-PSO is compared in this study by simulating AC microgrid in the MATLAB/Simulink environment. Summarized results of the system are provided to authenticate viability of proposed arrangement. The proposed controller performs intelligently while regulating voltage and frequency of the MGS and utility system.</p><div> </div>
“He Needed Just About Everything”: Caring for Aging Adults Postincarceration
Authors: Jimenez Laylla B. , Cross Sarah H. , Boucher Nathan A. 
Published: February 2021

Little is known about the experiences of caregivers for older adults reentering communities following incarceration. We sought to explore the challenges faced by such caregivers in Central North Carolina. Methods: We conducted a thematic analysis on semi-structured interviews with 11 caregivers. Results: Caregivers discussed how prior incarceration coupled with aging present barriers to housing, employment, and safety-net benefits—making caregiving more difficult. Caregivers assist their older care recipients to develop essential life skills (e.g., scheduling tasks, applying for services) and navigate the dynamic aging process (e.g., loneliness, illness). Caregivers struggle to gain care recipients’ trust, often due to their past trauma. Discussion: Caregivers play essential roles supporting older adults postincarceration, yet they experience significant challenges with limited resources. This study informed the development of a resource guide to assist caregivers. Dedicated programs and policies are needed to address these needs at the local level.
Physiological Pacing: A New Road to Future
Authors: Arora Vanita , Suri Pawan 
Published: February 2021

Anatomy and physiology are the basis of human body functioning and as we have progressed in management of various diseases, we have understood that physiological intervention is always better than an anatomical one. For more than 50 years, a standard approach to permanent cardiac pacing has been an anatomical placement of transvenous pacing lead at the right ventricular apex with a proven benefit of restoring the rhythm. However, the resultant ventricular dyssynchrony on the long-term follow-up in patients requiring more than 40% ventricular pacing led to untoward side effects in the form of heart failure and arrhythmias. To counter such adverse side effects, a need for physiological cardiac pacing wherein the electrical impulse be transmitted directly through the normal conduction system was sought. His bundle pacing (HBP) with an intriguing alternative of left bundle branch pacing (LBBP) is aimed at restoring such physiological activation of ventricles. HBP is safe, efficacious, and feasible; however, localization and placement of a pacing lead at the His bundle is challenging with existing transvenous systems due to its small anatomic size, surrounding fibrous tissue, long-learning curve, and the concern remains about lead dislodgement and progressive electrical block distal to the HBP lead. In this article, we aim to take the reader through the challenging journey of HBP with focus upon the hardware and technique, selective versus nonselective HBP, indications and potential disadvantages, and finally the future prospects.
Lyn Kinase Activity Is Required for Akt Mediated Erythroleukemia Cell Differentiation
Authors: Plani-LAM Janice H. C. , Maxwell Mhairi , Slavova-Azmanova Neli , Kucera Nicole , Louw Alison , Satiaputra Jiulia , Wu Susan , Bastow Edward , Le Cindy , Quilici Cathy , Hibbs Margaret L , Ingley Evan 
Published: February 2021

Erythroleukemia (M6 subtype of Acute Myeloid Leukaemia) is uncommon but has a poor prognosis, with reports of successful differentiation therapy using erythropoietin (Epo). Signaling through the Epo-receptor, which involves JAK2 and Lyn tyrosine kinases, controls red blood cell progenitor development. We have highlighted the importance of Lyn for regulating downstream Akt, and feed-back inhibitory signaling of the Epo-receptor through analysis of Lyn-/-, Lynup/up (hyperactive Lyn) and Cbp-/- (Csk-binding protein, a negative regulator of Lyn) erythroid cells. However, the importance of maintaining Lyn activity as opposed to Lyn protein for erythroid cell development and signaling, has not been delineated. To address this, we generated LynKD/KD mice (expressing a kinase dead K275M mutant Lyn), and analysed their erythroid compartment and signaling in immortalized erythroid progenitors. We show that LynKD/KD mice display splenic extramedullary erythropoiesis and have evidence of elevate bone marrow erythropoiesis, similar to Lyn-/- mice but with a less severe phenotype. Immortalized erythroid progenitors from LynKD/KD mice show impaired Epo-induced differentiation and a greater dependence on Epo for viability, but unaltered proliferation, compared to wild-type cells. Epo-induced signaling of LynKD/KD cells showed enhanced pJAK2/pSTAT5, reduced pAkt/pGAB2, and substantially reduced ALAS-e levels, compared to wild-type cells. Importantly, elevating Akt signaling in LynKD/KD cells by addition of phosphatase inhibitors (okadaic acid or Calyculin A), or via expression of active Akt, restored their differentiation capacity (and ALAS-e levels) and reduced their dependence on Epo for viability. We have unveiled that Lyn kinase activity, and not just its expression, is required for correct signaling of Akt to GATA-1 to maintain ALAS-e expression in erythroid cells, enabling hemoglobin production and viability during differentiation.

Disclosures
No relevant conflicts of interest to declare.
An Engineered RNA Platform to Neutralize DNMT1 Function and Control DNA Methylation for Myelodysplastic Syndrome
Authors: Esposito Carla Lucia , Autiero Ida , Bassal Mahmoud Adel , Annamaria Sandomenico , Ummarino Simone , Borchiellini Marta , Ruvo Menotti , Catuogno Silvia , Ebralidze Alexander , De Franciscis Vittorio , Di Ruscio Annalisa 
Published: February 2021

DNA methylation is a major signature involved in the regulation of gene expression. Numerous studies have established a link between aberrant DNA methylation and cancer (Herman and Baylin 2003, Baylin and Jones 2011, Feinberg 2018). Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematopoietic malignancies, characterized by ineffective hematopoiesis, cytopenia and risk of progression to acute myeloid leukemia (AML) in approximately 30% of the cases (Khan, Vale et al. 2013, Arber, Orazi et al. 2016). Abnormal DNA methylation is considered the molecular lesion leading to tumor suppressor gene silencing and clonal variation in MDS and evolution to AML (Figueroa, Skrabanek et al. 2009, Jiang, Dunbar et al. 2009, Feinberg 2018).
In the past decades two nucleoside-based compounds, 5-azacytidine and 5-aza 2'-deoxycytidine have been extensively tested to reduce global DNA methylation levels and received approval by the U.S. Food and Drug Administration (FDA) for the treatment of MDS. Both drugs have been used as the frontline therapy for the management of higher-risk-to-transform MDS, that are ineligible for more aggressive treatment such as: allogenic transplants and standard chemotherapy. Unfortunately, cytotoxic and global non-specific demethylation effects, have limited their clinical application, revealing the need for smarter and safer epigenetic drugs.
Nucleic-acid aptamers are a new class of specific targeting molecules and represent high affinity ligands and potential antagonists of disease-associated proteins.
Herein, we present an innovative RNA aptamer-based approach to target DNMT1 function by translating the inherent neutralizing properties of RNA (Di Ruscio, Ebralidze et al. Nature, 2013, 503(7476):371-6) into an aptamer platform. Through a combinatorial chemistry strategy named SELEX (Systematic Evolution of Ligands by EXponential enrichment) (Mercier, Dontenwill et al. 2017), we shortlisted the most specific DNMT1-neutralizing RNA aptamers. These molecules display high specificity and serum stability and reveal strong structural affinity in targeting DNMT1, by molecular modelling and dynamic simulations. Foremost, we found that the selected aptamers specifically inhibit DNMT1 enzymatic activity both in vitro and in cells, thus impairing cell viability and leading to global demethylation.
Collectively, this study provides a proof-of-concept for the generation of the first RNA-based epigenetic therapy as well as an attractive and scalable tool of precision medicine. Such a tool will overcome the toxicity of in-use demethylating protocols and greatly improve care and treatment of patients with MDS and other disease triggered by aberrant DNA methylation.
Figure

Disclosures
No relevant conflicts of interest to declare.
Factors Predictive of Oral Abstract Being Published: Is Gender Disparity Playing a Role?
Authors: Kapoor Ankita , Patel Mehul P. , Khunger Arjun , Jizzini Mazen , Ammad Ud Din Mohammad , Gill Simrat , Nasir Samar , Manogna Dharmini , Gandhi Roopali Goyal , Agrawal Nikhil , Ather Hussain Syed , Gandhi Pulkit , Jamshed Saad 
Published: February 2021

Introduction: It remains unclear what percentage of abstracts proceed to manuscript publication and the characteristics that predict successful publication. This study aimed to determine factors associated with successful peer-reviewed publications following oral presentation at the American Society of Hematology (ASH) annual meeting.
Methods: All oral abstract presentations (n=621) in the hematological malignancy category from 2016 ASH annual meeting were included in the study. Abstract publication was confirmed by searching for the publicly listed abstract on PubMed by title, first, and last author names, and institutional matching. We recorded time to online publication, US versus foreign journal publication, and journal impact factor by 3.5 years from 2016 ASH annual meeting. Abstracts characteristics that were analyzed also included number of authors, gender of first author, gender of last author, and single vs multi-institution studies. Gender of the first and last author was confirmed by looking at their biography details on their institutional website. Descriptive analysis was performed and an association between presenter's or last author's gender and publication matrix was analyzed using Chi-square tests.
Results: Of the 621 abstracts, 350 (56%) were published in full text by three and a half years since the 2016 ASH annual meeting. The abstracts' average time to journal publication was 17.46 months (SD +/- 11.32) (Table 1). Of the published articles, 64% (223/350) were published in U.S. journals; mean impact factor for all publications was 14.46 (SD+/- 11.47).The median number of authors for published and unpublished abstracts were similar. Females presented 37% (228/621) of the abstracts and 35% (123/350) of the journal publications had female first author and 22% (77/350) had female last author. A total of 53.9% (123/228) abstracts presented by a female author were published versus 57.7% (227/393) abstracts presented by a male author (p=&lt;0.001). Males were more often involved in multi-institutional trials (p=0.045) and were more likely to have senior authorship (p=0.005). There was no correlation between the gender of the first author to journal impact factor (p=0.109) or time to publication (p=0.091).
Conclusion: More than half of the oral abstracts were successfully published regardless of gender and number of authors. The rate of successful publication is higher for male authors even though there was no correlation between the gender of the first author to journal impact factor or time to publication. Our study highlights gender disparity in senior authorship, however this difference is not as wide in first authorship.

Disclosures
Jamshed: Takeda, Amgen and Celgene: Honoraria.
Pembrolizumab in mycosis fungoides with PD-L1 structural variants
Authors: Beygi Sara , Fernandez-Pol Sebastian , Duran George , Wang Erica B. , Stehr Henning , Zehnder James L. , Ramchurren Nirasha , Fling Steven P. , Cheever Martin A. , Weng Wen-Kai , Kim Youn H. , Khodadoust Michael S. 
Published: February 2021

Key Points
PD-L1 structural variants are recurrent in mycosis fungoides with large cell transformation. PD-L1 structural variants in relapsed/refractory mycosis fungoides should prompt consideration of treatment with PD-1 inhibitors.
Constitutive activation of NF-κB during early bone marrow development results in loss of B cells at the pro-B-cell stage
Authors: Paun Andrea , Claudio Estefania , Siebenlist Ulrich K. 
Published: February 2021

AbstractThere is a considerable body of work exploring the role of NF-κB family of transcription factors in the maturation and functions of later stage B cells; however, their role in the earlier bone marrow stages of development is less well understood despite the demonstration that NF-κB activity is present at all early stages of B-cell development. To explore the consequences of early, B cell–targeted constitutive activation of both NF-κB pathways on B-cell development, we generated mice that have either or both. NF-κB pathways constitutively activated beginning in early pro-B cells. In marked contrast to activating a single pathway, we found mice with both pathways constitutively activated displayed a profound loss of B cells, starting with early pro-B cells and peaking at the late pro-B-cell stage, at least in part as a result of increased apoptosis. This effect was found to be cell autonomous and to have striking phenotypic consequences on the secondary lymphoid organs and circulating antibody levels. This effect was also found to be temporal in nature as similar activation under a Cre expressed later in development did not result in generation of a similar phenotype. Taken together, these findings help to shed further light on the need for tight regulation of the NF-κB family of transcription factors during the various stages of B-cell development in the bone marrow.
Measurable residual disease in elderly acute myeloid leukemia: results from the PETHEMA-FLUGAZA phase 3 clinical trial
Authors: Simoes Catia , Paiva Bruno , Martínez-Cuadrón David , Bergua Juan-Miguel , Vives Susana , Algarra Lorenzo , Tormo Mar , Martinez Pilar , Serrano Josefina , Herrera Pilar , Ramos Fernando , Salamero Olga , Lavilla Esperanza , Gil Cristina , Lopez Jose-Luis , Vidriales Maria-Belen , Labrador Jorge , Falantes Jose-Francisco , Sayas María-José , Ayala Rosa , Martinez-Lopez Joaquin , Villar Sara , Calasanz Maria-Jose , Prosper Felipe , San-Miguel Jesús F. , Sanz Miguel Ángel , Montesinos Pau 
Published: February 2021

Abstract
The value of measurable residual disease (MRD) in elderly patients with acute myeloid leukemia (AML) is inconsistent between those treated with intensive vs hypomethylating drugs, and unknown after semi-intensive therapy. We investigated the role of MRD in refining complete remission (CR) and treatment duration in the phase 3 FLUGAZA clinical trial, which randomized 283 elderly AML patients to induction and consolidation with fludarabine plus cytarabine (FLUGA) vs 5-azacitidine. After consolidation, patients continued treatment if MRD was ≥0.01% or stopped if MRD was &lt;0.01%, as assessed by multidimensional flow cytometry (MFC). On multivariate analysis including genetic risk and treatment arm, MRD status in patients achieving CR (N = 72) was the only independent prognostic factor for relapse-free survival (RFS) (HR, 3.45; P = .002). Achieving undetectable MRD significantly improved RFS of patients with adverse genetics (HR, 0.32; P = .013). Longer overall survival was observed in patients with undetectable MRD after induction though not after consolidation. Although leukemic cells from most patients displayed phenotypic aberrancies vs their normal counterpart (N = 259 of 265), CD34 progenitors from cases with undetectable MRD by MFC carried extensive genetic abnormalities identified by whole-exome sequencing. Interestingly, the number of genetic alterations significantly increased from diagnosis to MRD stages in patients treated with FLUGA vs 5-azacitidine (2.2-fold vs 1.1-fold; P = .001). This study supports MRD assessment to refine CR after semi-intensive therapy or hypomethylating agents, but unveils that improved sensitivity is warranted to individualize treatment and prolong survival of elderly AML patients achieving undetectable MRD.
A patient with a germlineGATA2mutation and primary myelofibrosis
Authors: Rütsche Cyrill V. , Haralambieva Eugenia , Lysenko Veronika , Balabanov Stefan , Theocharides Alexandre P. A. 
Published: February 2021

Key PointsFirst description of a patient with a germline GATA2 mutation and diagnosis of primary myelofibrosis. Development of bone marrow failure on a Janus kinase inhibitor.
Prostaglandin-Endoperoxide Synthase 2 (PTGS2) in the Oviduct: Roles in Fertilization and Early Embryo Development
Authors: Anamthathmakula Prashanth , Winuthayanon Wipawee 
Published: February 2021

Abstract
The mammalian oviduct is a dynamic organ where important events such as final maturation of oocytes, transport of gametes, sperm capacitation, fertilization, embryo development, and transport take place. Prostaglandin-endoperoxide synthase 2 (PTGS2), also known as cyclooxygenase 2 (COX-2), is the rate-limiting enzyme in the production of prostaglandins (PGs) and plays an essential role during early pregnancy, including ovulation, fertilization, implantation, and decidualization. Even though the maternal-embryo communication originates in the oviduct, not many studies have systemically investigated PTGS2 signaling during early development. Most of the studies investigating implantation and decidualization processes in Ptgs2-/- mice employed embryo transfer into the uterus, thereby bypassing the mammalian oviduct. Consequently, an understanding of the mechanistic action as well as the regulation of PTGS2 and derived PGs in oviductal functions is far from complete. In this review, we aim to focus on the importance of PTGS2 and associated PGs signaling in the oviduct particularly in humans, farm animals, and laboratory rodents to provide a broad perspective to guide further research in this field. Specifically, we review the role of PTGS2-derived PGs in fertilization, embryo development, and transport. We focus on the actions of ovarian steroid hormones on PTGS2 regulation in the oviduct. Understanding of cellular PTGS2 function during early embryo development and transport in the oviduct will be an important step toward a better understanding of reproduction and may have potential implication in the assisted reproductive technology.
Thyrotropin N-glycosylation and Glycan Composition in Severe Primary Hypothyroidism
Authors: Wide Leif , Eriksson Karin 
Published: February 2021

Abstract

Context
In severe primary hypothyroidism (sPH), the serum thyrotropin (TSH) levels are elevated with an increased degree of sialylation. The circulating TSH comprises 2 different TSH glycoforms: TSHdi with 2 and TSHtri with 3 N-glycans and methods have developed to determine their contents of anionic monosaccharides (AMS), that is, sialic acid (SA) and sulfonated N-acetylglactosamine (SU) residues.

Objective
Characterize N-glycosylation and glycan composition of circulating TSH molecules and determine the effects during levothyroxine treatment in patients with sPH.

Methods
Serum samples were obtained from 25 patients with sPH, from 159 euthyroid individuals, and from 12 women during treatment with levothyroxine for sPH. Degrees of N-glycosylation and concentrations of TSHdi and TSHtri as well as their contents of AMS, SA, and SU residues were determined.

Results
The circulating TSH molecules in sPH patients had lower degrees of N-glycosylation, higher degrees of sialylation, and lower degrees of sulfonation than in euthyroid individuals. Levothyroxin restored sialylation and sulfonation of the glycans already at low free thyroxine (FT4) levels, while degree of N-glycosylation was not restored until the FT4 levels were normal.

Conclusions
The majority of TSH molecules in severe primary hypothyroidism were less N- glycosylated, more sialylated, and less sulfonated compared with euthyroid individuals. This glycan pattern favors a prolonged half-life in the circulation combined with lower in vitro biopotency at the target cells. During levothyroxine treatment of sPH patients, the sialylation and sulfonation of glycans were restored already at low FT4 levels, while N-glycosylation of TSH was not restored until the FT4 levels were normal.