bluuebunny/crossref_metadata_2025 huggingface dataset:
COVID-19 Incidence in Asymptomatic Cancer Patients Undergoing Chemotherapy
Authors: Monteiro Claudia , Gomes Cecília , Marani Leticia , Colli Leandro , Calado Rodrigo T. , Figueiredo-Pontes Lorena
Published: November 2021

COVID-19 is an infectious disease caused by the virus SARS-CoV-2, which was first described at the end of 2019. Since then, it has affected a growing portion of the world's population because of its high transmissibility. Most patients are asymptomatic or present with mild symptoms, but approximately 5-10% of cases can develop more serious manifestations, such as severe acute respiratory syndrome, acute kidney injury, shock, myocardial injury and even death. These features seem to occur more commonly in patients with essential hypertension, diabetes mellitus, obesity and chronic pulmonary disease. However, there are few studies that clarify the natural history of the disease and its broad clinical spectrum owing to the fact that it is a new entity. Since individuals with malignancies tend to present some degree of immunological deficiency and are more prone to opportunistic infections, especially those being treated with immunosuppressive drugs, it is possible that this group has a higher incidence of COVID-19. The current recommendations of oncology specialists advise to postpone treatments and to use less toxic drugs when possible. However, we still do not know how much these measures will affect in cancer mortality. Also, the incidence of COVID-19 in this population remains undetermined. We do not know if infectious symptoms are a good parameter to motivate these therapeutic changes or if there is benefit to test asymptomatic patients. In this context, this research submitted 100 patients with hematological malignancies or solid tumors on chemotherapy at the Ribeirão Preto Medical School of the University of São Paulo's Hospital, asymptomatic for COVID-19, to RT PCR to determine the SARS-CoV-2 infection incidence in this population. Only two patients were diagnosed with COVID-19. Both had gastrointestinal cancer. One of them developed symptoms, but none presented severe manifestations. Both had their treatment postponed initially and reinitiated after the appropriate period of isolation. Hence, we believe that it's reasonable not to test every asymptomatic patient when the resource for that is scarce, prioritizing those at greater risk of infection and those more prone to severe outcomes as long as the appropriate preventive measures are being taken.

Disclosures

Calado: Team Telomere, Inc.: Membership on an entity's Board of Directors or advisory committees; Agios: Membership on an entity's Board of Directors or advisory committees; Instituto Butantan: Consultancy; Alexion Brasil: Consultancy; AA&MDS International Foundation: Research Funding; Novartis Brasil: Honoraria.
The Synergistic Anticancer Effect of a Combination of Chidamide and Etoposide Against NK/T Cell Lymphoma
Authors: Song Wenting , Chen Zhan , Shi Cunzhen , Gao Yuyang , Feng Xiaoyan , Li Hongwen , Li Zhaoming , Zhang Mingzhi
Published: November 2021

Natural killer/T cell lymphoma (NKTCL) is a highly aggressive hematological malignancy. However, there is currently no consensus on first-line therapies for refractory/relapsed patients. Chidamide is a self-researched and developed HDACs inhibitor, and when combined with DNA-damaging agents, exhibited a clinical synergistic effect for the treatment of some solid tumors and hematological malignancies. Thus in this study, a series of in vitro and in vivo experiments were conducted to explore the efficacy and potential mechanisms of combined chidamide and etoposide treatment in NKTCL. We demonstrated that chidamide or etoposide alone dose- and time-dependently inhibited the cell viability of NKTCL cell lines, YT, NKYS and KHYG-1. Functional experiments suggested that combined chidamide and etoposide treatment exerted synergistic antiproliferation effect and enhanced cell apoptotic death both in vitro and in vivo. Furthermore, the expression of DNA damage related proteins was detected and we also examined the alternations in histone acetylation, cell cycle progression, and mitochondrial membrane potential (MMP). The results suggested that increased histone acetylation, cell cycle arrest at the G2/M phase and loss of MMP, converging to greater DNA damage, might account for the synergism of the combination of chidamide and etoposide in NKTCL. Taken together, our study supplements the clinical application of combining HDACs inhibitors and DNA-damaging agents on treating hematological malignancies but also provide an experimental basis for improved therapeutic efficacy and decreased complications for patients with NKTCL.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Hypomethylating Agents As Maintenance Treatment Following Allogeneic Hematopoietic Transplant in Children's Patients with Acute Myeloid Leukemia or Myelodysplastic Syndrome: A Single Center Study
Authors: Ruan Yongsheng , Xie Danfeng , Liu Xuan , Liu Qiujun , Li Chunfu , Wu Xuedong , He Yuelin , Feng Xiaoqin
Published: November 2021

Regarding children's patients undergoing allogeneic hematopoietic stem cell transplant (HSCT) for acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS), disease relapse remains the most common reason for transplant failure and patient death. We implemented a single center study to investigate hypomethylating agents (HMA) as maintenance treatment after HSCT for children AML or MDS. We retrospectively analyzed 30 patients with AML or MDS including 63 courses of azacytidine (AZA) and 69 courses of decitabine (DEC) from January 1, 2018 to June 30, 2021 (Figure 1A). Either AZA (37.5mg/m 2 to 75 mg/m 2 for consecutive 5-7 days) or DEC (5 mg/m 2 for consecutive 5 days) was administrated 90 days post-transplantation as first course. Furthermore, the interval of HMA treatment was 1.5-month hereafter for in total of a year (Figure 1B). Patients that received at least one course of HMA treatment were included in this study. The patient characteristics was shown in Table 1. We found that the disease-free survival (DFS) and overall survival (OS) were 82.96±6.95% and 85.51±6.72%, respectively (Figure 1C, D). Based on Common Terminology Criteria for Adverse Events (CTCAE) criteria, only Grade 1 to 2 leukopenia (P<0.001), anemia (P=0.043), and thrombocytopenia (P=0.033) were found between complete blood count (CBC) of pre-treatment and post-treatment. Subgroup analysis indicated that the difference was mainly come from DEC group [leukopenia (P<0.001), anemia (P=0.001), and thrombocytopenia (P=0.190)] instead of AZA group [leukopenia (P=0.476), anemia (P=0.443), and thrombocytopenia (P=0.095)]. No nausea/vomiting, or elevated ALT, or elevated creatinine, or rash, or de novo/exacerbated GVHD were observed. Moreover, there were no difference of activation T cells populations (CD25 and HLA-DR) among 7 courses except CD3+HLADR+/CD3+ and CD8+HLADR+/CD8+ in course 1 vs course 4 (P= 0.0354 and P=0.0163, respectively, Figure 1E). Furthermore, a trend of increased percentage of CD3+HLADR+/CD3+, or CD4+HLADR+/CD4+, or CD8+HLADR+/CD8+ and decreased percentage of CD3+CD25+/CD3+, or CD4+CD25+/CD4+, or CD8+CD25+/CD8+ was found in relapsed group, despite there was no significant difference. Interestingly, DEC significantly reduced population of CD3+HLADR+/CD3+, or CD4+HLADR+/CD4+, or CD8+HLADR+/CD8+ compared to AZA (P<0.001, P<0.001, or P<0.001, respectively). In conclusion, both AZA and DEC as HMA maintenance treatment following HSCT in children AML or MDS was safe, and the outcome was encouraging. A further large cohort of randomization of AZA and DEC study is required.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Spatial Transcriptomics Reveals DPP4 As Novel Marker of a More Proliferative Phenotype in Early AML Progression
Authors: Haase Christa , Gustafsson Karin , Mei Shenglin , Milosevic Jelena , Yeh Shu-Chi , Sykes David , Kharchenko Peter , Scadden David T. , Lin Charles
Published: November 2021

Acute myeloid leukemia (AML) is a hematologic malignancy with poor prognosis for which the standard-of-care chemotherapy treatment regimen has remained virtually unchanged over the past 40 years. We have employed "Image-Seq", a new technology that was developed in our laboratory, to study spatial variations in early leukemia progression in a mouse model of HoxA9-Meis1 AML. We visualized leukemia cells with differing proliferative phenotype using intravital microscopy, captured these cells under image guidance from individual bone marrow microenvironments and studied their differential expression by single-cell RNA sequencing. This analysis identified DPP4 as a key upregulated gene in AML cells from more proliferative bone marrow compartments and associated DPP4 expression with a cell cluster enriched in progenitor cell markers for HoxA9-Meis1 AML, including Flt3, Itgb7 and Ddx4. Strikingly, DPP4 is not expressed in vitro, and its expression in vivo (as quantitated by FACS analysis) correlated with disease progression and marked a more proliferative phenotype both at the 1-week and 2-week time-points during disease progression.

Disclosures

Sykes: Clear Creek Bio: Current equity holder in publicly-traded company; SAFI Biosolutions: Consultancy, Current equity holder in publicly-traded company; Keros Therapeutics: Consultancy. Scadden: Magenta Therapeutics: Current holder of individual stocks in a privately-held company, Membership on an entity's Board of Directors or advisory committees; VCanBio: Consultancy; LifeVaultBio: Current holder of individual stocks in a privately-held company, Membership on an entity's Board of Directors or advisory committees; Inzen Therapeutics: Membership on an entity's Board of Directors or advisory committees; Garuda Therapeutics: Current holder of individual stocks in a privately-held company, Membership on an entity's Board of Directors or advisory committees; FOG Pharma: Consultancy; Fate Therapeutics: Current holder of individual stocks in a privately-held company; Editas Medicines: Current holder of individual stocks in a privately-held company, Membership on an entity's Board of Directors or advisory committees; Dainippon Sumitomo Pharma: Other: sponsored research; Clear Creek Bio: Current holder of individual stocks in a privately-held company, Membership on an entity's Board of Directors or advisory committees; Agios Pharmaceuticals: Current holder of individual stocks in a privately-held company, Membership on an entity's Board of Directors or advisory committees.
Rural-Urban Disparities in Place of Death in Hematologic Malignancies in the US from 2003 to 2019
Authors: Hussaini Syed M. Qasim , Gupta Arjun
Published: November 2021

Background: more than 60,000 people die annually from hematologic malignancies in the united states (us). Patients with hematologic malignancies more frequently receive aggressive care toward the end-of-life and are more likely to die in a hospital compared to those with a solid tumor. Appropriate care of such patients is very dependent on an existing healthcare infrastructure. There are notable challenges to rural healthcare in the united states which contains less than 1/5th of all hospices in the us. In this study, we sought to investigate rural-urban disparities in place of death the us in individuals that died from hematologic malignancies.

Methods: we utilized the us centers for disease control and prevention wide-ranging online data for epidemiologic research database to analyze all deaths from hematologic malignancies in the us from 2003 to 2019. A population classification utilizing the 2013 us census was made using the national center for health statistics urban-rural classification scheme. These classifications included: large metropolitan area (1 million), small- or medium-sized metropolitan area (50 000-999 999), and rural area (<50 000). We estimated deaths in a medical facility, hospice, home, or nursing care facility. We stratified the results by age, sex, and race/ethnicity. The annual percentage change (apc) in deaths was estimated. All data was publicly available and de-identified.

Findings: from 2003-2019, there were a total 1,088,589 deaths form hematologic malignancies in the united states, predominantly in large metropolitan areas (50.2%), followed by small or medium sized metropolitan areas (31.7%) and rural areas (18.2%). All regions noted decreases in medical facility and nursing facility related deaths, and increase in hospice and home deaths. While rural areas demonstrated the quickest uptake of hospice care (apc 61.5), they had the lowest overall presence of hospice care (8.3% of all rural deaths in 2019 vs. 14.9% for small or medium metropolitan vs. 12% for large metropolitan) and larger share of nursing facility related deaths (15.8% of all rural deaths in 2019 vs 12.3% for small or medium metropolitan vs 10.6% for large metropolitan).

Discussion: we demonstrate end-of-life disparities in hematologic malignancies based on where an individual resides in the us with rural areas having notably lower share of deaths in hospice facilities. Older infrastructure, inadequate access to care, and financial barriers add to the medical complexity of care for all patients, and especially hematologic patients with high needs and complex treatment planning. These have been aggravated by rural hospital closures in the previous 18 months. The us senate is currently debating a bipartisan infrastructure that may add billions in building rural healthcare infrastructure to state budgets. Our findings are timely in helping inform congressional policy.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Testing for Heparin Induced Thrombocytopenia in Hospitalized Patients: In Line with Evidence?
Authors: Grewal Udhayvir Singh , Gaddam Shiva Jashwanth , Thotamgari Sahith Reddy , Brown Tyiesha , Beedupalli Kavitha , Mills Glenn M.
Published: November 2021

Background: Heparin-induced thrombocytopenia (HIT) is an immune complication of heparin therapy caused by antibodies to complexes of platelet factor 4 (PF4) and heparin. Both clinical probability and laboratory testing are needed for establishing a diagnosis of HIT. The 4Ts clinical scoring system, due to a very high negative predictive value when low, offers a robust means to exclude a diagnosis of HIT. However, these strategies are under-employed in clinical practice and limited evidence indicates a high prevalence of over-testing for HIT.

Methods: This retrospective analysis was conducted to identify patients who underwent heparin/PF4 antibody testing over a period of 12 months. The testing was performed using an ELISA-based IgG anti-heparin/PF4 antibody assay and an optical density (OD) of 0.4 was used as a cut-off for a positive value. Electronic medical records were reviewed for 4T score documentation, anti-PF4 results, SRA testing and 4T scores were retrospectively calculated for all the patients. SAS v9.4 (Cary, NC) was used for statistical analysis.

Results: A total of 105 patients who underwent anti-PF4 antibody testing were included for analysis. Majority of the patients in our cohort were admitted in an intensive care unit setting (75/105,71.4%). On chart review, only 17 patients (16.2%) were noted to have documentation of 4T score. Based on the retrospectively calculated 4T scores, 60 patients (57.1%) had low pre-test probability, 41 (39%) had intermediate pre-test probability and 4 (3.8%) patients were noted to have high pre-test probability. Anti-PF4/heparin antibodies were positive in 9 patients, of which 5 (55.5%) patients did not undergo concomitant SRA testing. Out of 9, 4 (44.4%) had weakly positive (0.4-1.0 OD units), 2 (21.1%) had strongly positive (1.0-2.0 OD units) and 2 (21.1%) patients had very strongly positive (>2 OD units) anti-PF4 antibody titers. Out of 105 patients, SRA was tested in 11 patients (10.5%) and was noted to be positive in 1 (0.95%). Overall, 2 patients were diagnosed and treated for HIT, out of which the diagnosis was not confirmed with SRA in 1 patient (due to high pre-test probability and very strong anti-PF4 titers). In the remaining patients, sepsis (48, 46.6%) and drug-induced thrombocytopenia (29, 28.2%) emerged as the most common possible causes of thrombocytopenia.

Conclusion: Among hospitalized patients, over-testing for HIT is common. Practices to promote 4T score documentation and evidence-based anti-PF4 testing may help prevent unnecessary costs associated with serological testing and costly alternate anticoagulants. To improve overall outcomes, clinicians should also attempt to identify and treat other more likely causes of thrombocytopenia, especially in patients with low pre-test probability for HIT.

Disclosures

No relevant conflicts of interest to declare.
CD200 Expression on Multiple Myeloma Cells Induces Attenuation of T Cell-Mediated Cytotoxicity Via DOK2
Authors: Shah Pooja , Stuehmer Thorsten , Bruennert Daniela , Munawar Umair , Leich Ellen , Kraus Sabrina , Hudecek Michael , Chatterjee Manik , Schlosser Andreas , Bargou Ralf C , Einsele Hermann , Berberich-Siebelt Friederike , Steinbrunn Torsten
Published: November 2021

Introduction

The CD200/CD200 receptor (CD200R) axis is known to exert immunoregulatory effects in myeloid-derived cells and constitutes a putative immune checkpoint in hematological malignancies, in which CD200 expression is associated with poor prognosis. In multiple myeloma (MM), CD200 is expressed in the majority of patient-derived primary cells. However, its functional importance as well as the related downstream mechanism upon CD200 ligand binding to its CD200R on T cells are not well understood. In this study, we analyze the functional role of CD200 as a potential immune checkpoint in MM and decipher the mechanism of CD200-mediated immune escape.

Methods

Primary MM cells and MM cell lines were analyzed for CD200 surface expression by flow cytometry. To overexpress CD200 on non-expressing MM cell lines we used a Sleeping Beauty transposon vector system. CD200+/- MM cell lines L363, U266 and MM.1s were co-cultured with CD3/CD28-activated healthy donor T cells. T cell-mediated cytotoxicity in these co-cultures was assessed with flow cytometry and/or luciferase assay. Moreover, to analyze the effects of CD200R activation on downstream signaling pathways, activated T cells were treated with recombinant human CD200 (rhCD200) and/or anti-CD200 blocking antibody and subsequently, Western blotting was performed.

Results

Of n=120 primary MM samples (n=120) analyzed, CD200 protein expression could be detected in ca. 75 % of the cases. In contrast, all n=9 MM cell lines tested neither displayed surface nor cytoplasmic CD200 expression. Therefore, using a Sleeping Beauty transposon vector system we stably expressed CD200 on MM cell lines for further analyses. In the presence of CD200-expressing MM cells up to 50% decrease in CD3+ T cell-mediated cytotoxicity against MM cells was observed in flow cytometry and luciferase assay. Proliferation rates of MM cell lines remained unchanged regardless of the level of CD200 overexpression as determined by Alamar blue assays. In myeloid-derived cells, CD200R directly interacts with docking protein-2 (DOK2). In activated T cells, we observed DOK2 phosphorylation upon CD200 binding when treated with rhCD200 in a time- and concentration-dependent manner. Applying an anti-CD200 blocking antibody, this effect could be reversed, thus revealing a possible mechanism for the observed attenuation of T cell function.

Conclusion

Our study shows that anti-MM cytotoxicity from primary healthy donor CD3+ T cells is attenuated by CD200 expression on MM cells. We also demonstrate that this inhibitory mechanism in CD3+ T cells is mediated via DOK2, providing a potential target for immunotherapeutic approaches in MM.

Disclosures

Einsele: Janssen, Celgene/BMS, Amgen, GSK, Sanofi: Consultancy, Honoraria, Research Funding.
Carfilzomib-Lenalidomide-Dexamethasone in the Management of Lenalidomide-Refractory Multiple Myeloma
Authors: Cerchione Claudio , Nappi Davide , Pareto Anna Emanuele , Pane Fabrizio , Catalano Lucio , Martinelli Giovanni
Published: November 2021

Carfilzomib is an epoxyketone proteasome inhibitor of second generation, proved to be effective and safe in relapsed and refractory Multiple Myeloma (rrMM), in combination with dexamethasone or lenalidomide and dexamethasone.

In this retrospective observational trial, it has been evaluated efficacy and safety of carfilzomib, in combination with lenalidomide-dexamethasone (KRD) as salvage regimen in patients with rrMM, refractory to lenalidomide, where lenalidomide-based regimens have no proven efficacy.

41 patients (23 M/18 F), with rrMM, median age at diagnosis 63.7 years (r. 43-82), median age at start of treatment 67 years (r. 48-84) previously treated with several lines of treatments (median 3, r. 2-11), underwent to KRD regimen (ASPIRE trial schedule) for a median treatment cycles of 8 (r 2-18). ISS was equally distributed, and all patients had previously been treated with bortezomib and IMIDs, and were refractory to this agents. 61% (19/31) of them had undergone at least to a single ASCT.

According to IMWG criteria, after a median follow-up of 9 months (r. 2-18), ORR was 68,2% (28/41: 9 CR, 12 VGPR, 7 PR) with 5 progressive diseases (PD) and 8 patients in stable disease (SD): this can be considered as an impressive result in this subset of rrMM patients, refractory to lenalidomide. In particular, for 11 patients, KRD was, after having achieved at least a PR, a bridge to second/third autologous SCT. Median time to response was 1.3 months (r.1-4), median OS from diagnosis was 62 months (r. 9-170), median OS from start of Carfilzomib was 11 months (r. 2-18). Carfilzomib was well tolerated, with grade 2 anemia in 39%(16/41) of patients, successfully managed by ESAs, without necessity of blood transfusions; 29% (12/41) grade 3-4 neutropenia (pegfilgrastim in primary prophylaxis was given, no ospedalization was required, no septic shocks were observed); 34% (14/41) grade 2, 21% (9/41) grade 3 and 12% (5/41) grade 4 thrombocytopenia, without hemorrhagic events and transfusion-dependency. Moreover, it was observed pneumonia in 39% (16/41) of patients, treated by common antibiotic drugs and always solved. A cardiac monitoring was performed for all patients: hypertension (grade 2-3) in 34% (14/41) of patients; fatigue in 39% (16/31) of patients.

Carfilzomib-Lenalidomide-Dexamethasone has shown significant efficacy in a particularly severe setting of patients, relapsed and refractory to all available therapeutic resources, also lenalidomide, and it could be considered as a bridge to a second autologous or allogenic SCT.

Disclosures

Martinelli: Jazz Pharmaceuticals: Consultancy; Stemline Therapeutics: Consultancy; Astellas: Consultancy, Speakers Bureau; Celgene /BMS: Consultancy, Speakers Bureau; Abbvie: Consultancy; Daichii Sankyo: Consultancy; Roche: Consultancy; Pfizer: Consultancy, Speakers Bureau; Incyte: Consultancy.
Spontaneous Calvarial Infarction with Epidural Hematoma and Subgaleal Hemorrhage in a Teenager with Sickle Cell SS Disease and Glucose-6-Phosphate Dehydrogenase Deficiency
Authors: Nestor Jennifer , Sanders Danielle , Mysore Sukrita , Ahmed Rafat , Milano Aubri , Ostrowicki Renata , Nwankwor Odiraa , Presenza Thomas , Scattergood Emily
Published: November 2021

Introduction:

Sequelae of sickle cell disease (SCD) are often the result of vaso-occlusion (VOC) related to the dysmorphic shape of sickled red blood cells (RBC). Bone infarction occurs commonly in patients with sickle cell, most notably in long bones and axial skeleton. Infarct occurs much less frequently in the bones of the skull. Calvarial bone infarct results in MRI findings of T2 hyperintensity in areas of bone marrow edema with associated swelling or hemorrhage. Symptomatic headaches in children with SCD are more likely to be due to severe underlying cerebral pathologies than in those children without SCD. When evaluating a child with SCD presenting with headache, it is important to consider calvarial infarct.

Case Description:

A 15 year old male with a history of sickle cell disease (type SS) and G6PD deficiency presented with chief complaints of headache and swelling over the left parietal region of the skull for two days with associated left sided visual disturbance. He denied head trauma or seizure-like activity. Upon presentation he was febrile with tachycardia and a normal neurological exam. CT scan of his head was significant for a left subgaleal hemorrhage and bilateral epidural midline and left parietal fluid collections consistent with bone marrow infarcts and associated subacute epidural hematomas, without evidence of brain parenchymal injury or sinus venous thrombosis. A follow up MRI obtained during hospitalization indicated that the left subgaleal hematoma remained stable with breakdown of subgaleal blood products. The patient was treated primarily with supportive care including hydration, packed red blood cell transfusions and pain management. He did not require neurosurgical intervention. His hospital course was complicated by VOC, sickle cell hepatopathy and acute chest syndrome.

Discussion:

Rarely, cranial VOC has resulted in epidural hematoma. In this case, calvarial infarct with unilateral headache, acute vision loss and acute cranial swelling resulted in subgaleal hemorrhage. This clinically significant finding results from bleeding between the periosteum and scalp galea aponeurosis, which may lead to hypovolemic shock. Calvarial infarcts should be considered on the differential in sickle cell disease patients presenting with a headache as prompt diagnosis can improve morbidity outcomes.

Disclosures

No relevant conflicts of interest to declare.
p53 Is a Crucial Regulator of Hemogenic Endothelial Cell Differentiation from Human Induced Pluripotent Stem Cells
Authors: Kanaujiya Jitendra K. , Lingenheld Elizabeth G. , Skarnes William C. , Oguro Hideyuki
Published: November 2021

De novo generation of hematopoietic stem cells (HSCs) from human induced pluripotent stem cells (hiPSCs) could provide a virtually unlimited supply of autologous HSCs for clinical transplantation, and offer various approaches that enable gene therapy, drug discovery, disease modeling, and in vitro modeling of human hematopoietic development. However, the derivation of long-term self-renewing HSCs from hiPSCs in culture remains elusive. The tumor suppressor protein p53 plays important roles in normal and malignant hematopoiesis, and Trp53-deficient mice exhibit increased number of HSCs. Although activation of p53 is known to promote differentiation of hPSCs and hPSCs recurrently acquire TP53 dominant negative mutations, its role in hematopoietic differentiation of hiPSCs has not been explored.

To differentiate hiPSCs into hematopoietic stem and progenitor cells (HSPCs), we used embryoid body (EB) formation method to first differentiate hiPSCs into hemogenic endothelial (HE) cells that express the CD34 highCD144 +CD73 -CD184 -CD43 -CD235a - cell-surface markers. HE cells were then transferred onto a Matrigel-coated plate to undergo endothelial-to-hematopoietic transition (EHT) to generate HSPCs that express the CD34 midCD45 mid cell-surface markers. Developed HSPCs were functionally evaluated by colony forming assay.

We observed that the expression of CDKN1A, a p53 target gene, was upregulated in hiPSC-derived EBs and HSPCs over the course of differentiation. To investigate the role of p53 in the generation of HSPCs from hiPSCs, we genetically deleted TP53 in hiPSCs followed by hematopoietic differentiation. While TP53 deletion increased the growth of EBs, it resulted in severe impairment of differentiation into HE cells and overall production of HSPCs that can form colonies. During HE differentiation from hiPSCs, TP53-deficient EBs showed significant reduction of endothelial-lineage gene expression, such as ETV2, CDH5, and PECAM1, as well as expression of RUNX1, a master transcription factor required for HE specification. These results indicate the indispensable role of p53 in HE differentiation from hiPSCs.

We then examined the effect of p53 activation on HE differentiation from hiPSCs by pharmacological activation of p53 in hiPSC-derived cells. Transient activation of p53 by Nutlin-3, a small molecule that inhibits the p53-HDM2 interaction and protects p53 from proteasomal degradation, only during HE differentiation but not during EHT significantly promoted HSPC generation as compared to the vehicle treated control.

Our findings shed light on the importance of selecting hiPSC lines that retain normal p53 activity for HE differentiation, and provide an approach to promote hematopoietic differentiation of hiPSCs by transiently activating p53 during HE differentiation.

Disclosures

Kanaujiya: Synthego: Other: Scientific Advisory; eGenesis: Other: Scientific Advisory.
Fotemustine-Containing Regimens Versus Regimens Based on High-Dose Methotrexate in Newly Diagnosed PCNSL
Authors: Wu Jingjing , Zhang Mingzhi , Wang Wenhua
Published: November 2021

Objective: This study was conducted to compare the effectiveness and safety of fotemustine-containing regimens and high-dose methotrexate-based regimens in newly diagnosed primary central nervous system lymphoma(PCNSL). Methods: From April 2012 to April 2020, 88 patients with PCNSL who received fotemustine-containing regimens and regimens based on high-dose methotrexate were retrospectively assessed in this study.47 patients were treated with fotemustine-containing regimens, of which 25 patients were treated with R-FPD (Rituximab , formustine, pemetrexed, dexamethasone) regimen, and the other 22 patients were treated with FVD (formustine, vincristine,dexamethasone) regimen. 41 patients were treated with high-dose methotrexate-based regimens, of which 26 patients were treated with MA(Methotrexate ,Cytarabine ) regimen, of which 8 patients were treated with R-MT(Rituximab ,methotrexate ,temozolomide)regimen,of which 5 patients were treated with R-MA(Rituximab ,methotrexate ,Cytarabine) regimen, and the other 2 patients were treated with RM(Rituximab ,methotrexate) combined Zebutinib regimen. Results: In the fotemustine-containing regimen groups (n=47), 18 cases were complete response(CR), 19 cases were partial response(PR), 6 cases were

stable disease (SD), 4 cases were disease progression (PD), objective response rate(ORR )was 79%, and diease contral rate(DCR) was 91%.10 cases were CR in the high-dose methotrexate groups (n=41) , 17 cases of PR, 7 cases of SD, 7 cases of PD, ORR was 66%, DCR was 83%.there was no significant difference in ORR (79%vs66%, P=0.176)and DCR(91%vs83% , P=0.226) between the two groups.No significant difference was observed in the 2-year PFS rate (19.4%vs10%, P=0.149) and 3-year OS rate (24%vs15%, P=0.200) between the two groups. The main adverse reactions in the high-dose methotrexate groups were myelosuppression and mucositis. In the fotemustine-containing regimen groups, myelosuppression was the main adverse reaction. The incidence of leukopenia of grade 3 and above in the high-dose methotrexate groups and fotemustine groups was 54% and 23%, respectively. The high-dose methotrexate groups had more severe leukopenia (P= 0.001). The incidence of mucositis in the high-dose methotrexate groups (68%) was significantly higher than that in the fotemustine group (26%). Conclusion: The fotemustine-containing regimens for the treatment of newly diagnosed PCNSL has the similar efficacy to the high-dose methotrexate group, with fewer adverse reactions, better safety and tolerability, and may be a feasible regimen.

Disclosures

No relevant conflicts of interest to declare.
Recommendation on Advanced Molecular Testing in Hematolymphoid Malignancies in the Veterans Population
Authors: Neff Jadee L , Mosse Claudio A. , Wang-Rodriguez Jessica , Ahmed Sara , Kelley Michael
Published: November 2021

The Veterans Health Administration is the largest integrated provider of cancer services in the nation, with approximately 50,000 new cancer cases diagnosed each year. The goal of the VA's National Oncology Program is to establish a System of Excellence in cancer care with the help of tools, resources, programs, and best practices across the enterprise to ensure every veteran, regardless of location, receives the same high level of cancer care and access to precision diagnostics. This is mediated through a variety of programs, including the National TeleOncology Service and the National Precision Oncology Program which includes standardized clinical pathways for molecular testing, a second opinion service on diagnostic consultations, test result interpretation and treatment recommendations, as well as a molecular oncology tumor board.

The VA National Oncology Program and the Pathology and Laboratory Medicine Program offices have collaborated on a recommended guideline for advanced laboratory and molecular testing algorithm in hematologic diagnosis. We aim to offer best practice for all VA providers who must decide the type of ancillary tests at the time of initial clinical presentation in order to gain prognostic information and guide therapy. The recommendations include traditional laboratory tests, such as morphology, flow cytometry, and immunohistochemistry, as well as the use of fluorescent in situ hybridization (FISH), karyotype, polymerase chain reaction (PCR), and next generation sequencing (NGS). An overview of the draft algorithm for acute leukemias and chronic myeloid neoplasms is presented in Table 1; details will be discussed at the annual meeting.

Conclusion: The joint effort from VA Oncology and Pathology presents advanced testing algorithms in a variety of hematolymphoid malignancies as a systematic approach to hematopathology diagnosis, classification, as well as offering prognostic and therapeutic information. The recommendations across VA Healthcare would ensure standardized care for all VA patients with hematologic malignancy to benefit from the latest advances in precision medicine.

Figure 1 Figure 1.

Disclosures

Neff: Spring Discovery: Consultancy, Ended employment in the past 24 months; EUSA Pharma: Speakers Bureau; Enzyvant: Consultancy.
Epidemiology of Aggressive NK-Cell Leukemia in the United States: A SEER Population-Based Study
Authors: Amisha Fnu , Malik Paras , Konda Manojna , Kakadia Sunilkumar , Fugere Tyler , Mukherjee Akash
Published: November 2021

INTRODUCTION

Aggressive NK-cell leukemia ( ANKL) is a systemic lymphoproliferative disorder of natural killer (NK) cells frequently associated with Epstein-Barr virus (EBV) which has been grouped under histiocytic/dendritic neoplasms in the 2008 WHO classification of hematopoietic neoplasms. Due to rarity of diagnosis, the current available literature is limited to case reports and case series. This population-based study using Surveillance Epidemiology and End result program (SEER) is the largest to explore demographic characteristics, survival patterns and long-term outcomes in patients with ANKL in the United States.

MATERIALS AND METHODS

We utilized SEER 18 November 2020 submission database to select all patients (> 1 years of age) diagnosed from 2000-2018 with ANKL using International Classification of Diseases for Oncology edition 3 (ICD-O-3) code of 9948/3. Patients were divided into various cohorts based on age (<60 years, 60-79 years, >80 years), sex, race (Caucasians, African American, Asian/Pacific islander and American Indians/Alaska natives) and median household income of county of residence (< $ 50,000 vs > $50,000). We used SEER*stat to calculate age adjusted incidence rate using 2000 US standard population. Kaplan Meier curve was utilized to calculate 5-year overall survival. Cox proportional hazard model was used for multivariate analysis of factors associated with survival and p<0.05 was considered significant.

RESULTS

A total of 140 patients were identified with ANKL. The median age at diagnosis is 58 years. The crude, age-adjusted to 2000 US standard population and age-specific incidence rate of HCD in Unites states is < 1/100, 000 respectively. The incidence in males is 1.9 times that of females- 92 males (65.7%) and 48 females (34.2%). Between 2001-2018, minimum number of cases were diagnosed in 2010 (n=2).Out of the cohort, 107 patients were White (76.4%), 21 patients were Asian or Pacific islander (15 %), 9 patients were African Americans (6.4%) and 3 patients were American Indian/Alaska native (2.1%). The median overall survival was 7 months (95% CI; 0-16). Survival rates at 1year, 2 years and 5 years were found to be 45.3% , 37.1 % and 31.1% respectively [Figure 1]. 5 year overall survival rates are as follows- Whites ( 69.7% ), Blacks ( 63.1%), American Indian (59.9%), Asian or Pacific Islander ( 66.8%). On multivariate analysis, black race was associated with poor outcomes (p 0.008), whereas sex, income and age had no significant effect on cancer outcomes.

CONCLUSIONS

Our study shows that ANKL is a rare hematological malignancy in general population with a poor median survival of less than one year. Males are twice more likely to be affected than females with poor outcomes in africo-americans.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Ophthalmic Rosai–Dorfman disease: a multi-centre comprehensive study
Authors: Alzahem Tariq A. , Cruz Antonio Augusto , Maktabi Azza M. Y. , Chahud Fernando , Alkatan Hind
Published: November 2021

Background

To provide basic demographic information and clinicopathologic features of ophthalmic Rosai–Dorfman disease (RDD) with a literature review.

Methods

A multi-centre retrospective case series reviewing all patients with histopathologically confirmed ophthalmic RDD at three tertiary eye care centres between January 1993 and December 2018.

Results

Eleven eyes of eight patients with histopathologically confirmed ophthalmic RDD were included, with equal numbers of males and females. The median age was 40.25 years (range: 26.6–72.4). Two patients had familial RDD. The orbit was the most commonly involved site (90.9% eyes). One patient (one eye) presented with a scleral nodule, anterior uveitis and cystoid macular oedema. Visual acuity ranged from 20/25 to light perception. Six patients had an extra-nodal ophthalmic disease, and the remaining two had an associated submandibular lymphadenopathy (nodal RDD).

Conclusions

Ophthalmic RDD can be the only manifestation of this systemic disease, with the orbit being the most commonly involved site, exhibiting bone destruction, intracranial and/or sinus involvement and variable degree of visual loss. Ophthalmic familial RDD represent a severe form with a malignant course. Steroid monotherapy may be inadequate to control orbital RDD; thus, combined treatment is usually necessary. A comprehensive approach to assessment and management is recommended.
Metabolome and microbiome of chronic periapical periodontitis in permanent anterior teeth: a pilot study
Authors: Huang Yun , Zhou Peng , Liu Siqi , Duan Wei , Zhang Qinqin , Lu Ying , Wei Xin
Published: November 2021

Background

Periapical periodontitis is a common oral inflammatory disease that affects periapical tissues and is caused by bacteria in the root canal system. The relationship among the local metabolome, the inflammatory grade, and the type and abundance of microorganisms associated with periapical periodontitis is discussed in this study.

Methods

The inflammatory grades of periapical samples from 47 patients with chronic periapical periodontitis in permanent anterior teeth were determined based on the immune cell densities in tissues subjected to haematoxylin and eosin staining. The metabolome was evaluated using ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectrometry, followed by principal component analysis and orthogonal partial least squares discriminant analysis. The microbiome was accessed using 16 S rRNA high-throughput sequencing. The differences in the metabolomes and microbiomes of the periapical periodontitis samples were assessed using Spearman’s correlation analysis.

Result

N-acetyl--glucosamine, -tryptophan, -phenylalanine, and 15 other metabolites were identified by the comparison between samples with severe inflammation and mild or moderate inflammation. Four amino acid metabolism pathways and one sugar metabolism pathway were associated with the inflammatory grade of periapical periodontitis. The abundance of Actinomycetes was negatively correlated with the abundance of glucosamine (GlcN), while the abundance of Tannerella was positively correlated with the abundance of -methionine.

Conclusions

The local metabolome of periapical periodontitis is correlated with the inflammatory grade. The abundance of the local metabolites GlcN and -methionine is correlated with the abundance of the major microorganisms Actinomycetes and Tannerella, respectively.
Factors related to turnover intention among staff of elderly caring social organizations in Anhui Province, China: a cross-sectional study
Authors: Chen Xuefei , Tang Ling , Liu Liu , Bai Zhongliang , Chen Ren
Published: November 2021

Objective

Turnover intention of employees in elderly caring social organizations has a significant impact on elderly care service delivery. This study investigated the associated factors of turnover intention among employees of elderly caring social organizations in Anhui Province, China.

Methods

A total of 605 participants were selected using a multi-stage stratified random sampling method. A self-administered questionnaire was used to collect information on socio-demographic, social support, and turnover intention from the participants. The data was analyzed through descriptive statistical analysis, one-way variance analysis, Spearman correlation analysis, and multiple linear regression were used to analyze the factors related to turnover intention.

Results

Results of our study showed that the total score of turnover intention, turnover intention I (possibility of quitting a current job),turnover intention II (motivation to find other jobs) and turnover intention III (obtaining the external possibility of work) were 8.84, 2.32, 2.38, and 4.14, respectively. Social support negatively correlated with turnover intention I and turnover intention II. However, it showed positive correlation with turnover intention III and total turnover intention scores; turnover intentionI (coefficient: − 0.082), turnover intention II (coefficient: − 0.071), turnover intention III (coefficient: 0.19), Total score of turnover intention (coefficient: 0.093). Ethnic group, age, education level, and job satisfaction were associated with turnover intention.

Conclusion

Improvement of social support play an important role in reducing the turnover intention of employees in elderly caring social organizations. It is important to increase organizational commitment and strengthen psychological empowerment, combined with decreasing job burnout for stability.
Treatment effect of nafamostat mesylate in patients with COVID-19 pneumonia: study protocol for a randomized controlled trial
Authors: Moon Kyunglan , Hong Kyung-Wook , Bae In-Gyu
Published: November 2021

Background

This study is designed to evaluate the main hypothesis that nafamostat mesilate with standard therapy improves the severity and mortality rate in patients with COVID-19 pneumonia.

Methods

We conduct a randomized, open type, multi-institute/center, 2-group clinical trial with COVID-19 pneumonia patients in Korea. Eighty four patients with COVID-19 pneumonia are randomly assigned to intervention group or control group. Patients in intervention group receive the standard therapy with a dose of 0.1 to 0.2 mg/kg/h (2.4 to 4.8 mg/kg/day) of nafamostat mesilate. Patients in control group receive the standard therapy such as lopinavir/ritonavir, hydroxychloroquine, oxygen therapy, non-invasive and invasive ventilator, antibiotic therapy, renal-replacement therapy, and extracorporeal membrane oxygenation (ECMO). The primary outcome is proportion of patients with clinical improvement as defined by live discharge from hospital or a decline of 2 categories on the seven-category ordinal scale of clinical status, as well as secondary outcome comprised change in National Early Warning Score, duration of hospitalization, incidence of new-non-invasive ventilation or high flow oxygen use or ventilator, mortality at day 28, viral load change, and adverse events.

Discussion

Our study contributes to the establishment of therapeutic strategy in COVID-19 pneumonia by evaluating the therapeutic effect and safety of nafamostat mesilate.

Trial registration

ClinicalTrials.gov NCT04418128. Registered on 5 June 2020.
Unexpected similarity between HIV-1 reverse transcriptase and tumor necrosis factor binding sites revealed by computer vision
Authors: Eguida Merveille , Rognan Didier
Published: November 2021

Rationalizing the identification of hidden similarities across the repertoire of druggable protein cavities remains a major hurdle to a true proteome-wide structure-based discovery of novel drug candidates. We recently described a new computational approach (ProCare), inspired by numerical image processing, to identify local similarities in fragment-based subpockets. During the validation of the method, we unexpectedly identified a possible similarity in the binding pockets of two unrelated targets, human tumor necrosis factor alpha (TNF-α) and HIV-1 reverse transcriptase (HIV-1 RT). Microscale thermophoresis experiments confirmed the ProCare prediction as two of the three tested and FDA-approved HIV-1 RT inhibitors indeed bind to soluble human TNF-α trimer. Interestingly, the herein disclosed similarity could be revealed neither by state-of-the-art binding sites comparison methods nor by ligand-based pairwise similarity searches, suggesting that the point cloud registration approach implemented in ProCare, is uniquely suited to identify local and unobvious similarities among totally unrelated targets.
Seismic Retrofitting of RC Circular Columns Using Carbon Fiber, Glass Fiber, or Ductile PET Fiber
Authors: Choi Donguk , Hong Seongwon , Lim Myung-Kwan , Ha Sang-Su , Vachirapanyakun Sorrasak
Published: November 2021

The effectiveness of seismic retrofitting using three different fibers—carbon fiber (CF), glass fiber (GF), polyethylene terephthalate (PET) fiber—and a fiber combination of aramid fiber (AF) and PET fiber (called hybrid fiber reinforced polymer (HF)) wrapped on reinforced concrete (RC) circular columns was experimentally evaluated. A total of 11 RC circular columns were tested: three control columns and eight retrofitted columns in three different test groups. The purpose of fiber wrapping was flexural strength improvement as well as enhancement of rotational capacity in the plastic hinge region. Mechanical properties of CF, GF, AF, and PET were first defined; that is, CF, GF, and AF exhibited linear stress–strain behavior with limited ultimate strain capacity typically less than 3%, while ductile PET exhibited as much as 15% strain and non-linear stress–strain behavior with a very low elastic modulus. In the RC column tests, all three different fibers and the AF + PET fiber combination were effective in enhancing the strength and ductility but resulted in different structural behaviors and failure modes depending on the fiber type and the fiber amount used. The column sections were then analytically studied by section analysis using the behavior of confined concrete, the non-linear relationship of fiber-reinforced polymer (FRP), and the actual material properties of reinforcement. The analytical and experimental results revealed that ductile PET is beneficial, as it demonstrates more ductile behavior with a degree of strength enhancement similar to that of CF and GF.