bluuebunny/crossref_metadata_2025 huggingface dataset:
Establishment of a Nomogram for Patients with Primary Mediastinal Large B-Cell Lymphoma Based on the SEER Database and Validation of Real-World Data
Authors: Wang Hua , Chen Guanjun , Fu Bibo
Published: November 2021

Background

Primary mediastinal large B-cell lymphoma (PMBCL) is a rare disease with the majority of patients being rapidly progressive anterior mediastinal large tumors. Due to the rarity of PMBCL, information on the incidence, clinical features, prognostic factors and models of PMBCL is limited. The present study is one of the largest studies on the incidence and prognostic factors of PMBCL and is the first to establish a nomogram model of PMBCL and validate it with real-world data. We also compared the newly established Nomogram with the existing IPI prognostic model. We believe that our findings can help clinicians to quickly and accurately assess the predicted survival of patients and help them to perform individualized risk stratification of patients.

Methods

Based on data from the Surveillance, Epidemiology and End Results (SEER) database, 797 patients diagnosed with PMBCL were enrolled in this study. The 797 patients were randomly divided into training and internal validation groups in a 7:3 ratio, and 116 patients diagnosed with PMBCL were included in the external validation group based on data from the Sun Yat-sen University Cancer Center and the First Affiliated Hospital of Guangzhou Medical University. Independent prognostic factors were screened by Cox regression analysis. R-coding was used to construct nomograms predicting overall survival (OS). Discriminations and corrections of the new model were assessed using the consistency index (C-index), subject operating characteristic curves (ROC) and calibration curves, and compared with the conventional international prognostic index (IPI) using decision curve analysis (DCA) to assess its accuracy and benefit.

Results

From 2001 to 2016, the incidence of primary mediastinal large B-cell lymphoma showed a relatively stable increasing trend with an APC of 11.8% (95% confidence interval 8.8-14.0, P<0.05), and this trend was more pronounced in the female population. Multivariate models showed that age and Ann arbor staging were significantly associated with OS, while the variable of extra-nodal invasion was included in the modeling based on clinical experience. In the training cohort, the C-index of the nomogram for OS was 0.712. the C-index for the internal and external validation cohorts was 0.667 and 0.690, respectively. The calibration curve also showed high predictive accuracy. The C-index and ROC curves of nomogram showed better results compared to IPI scores, and also yielded better net gains in decision curve analysis.

Conclusion

In summary, we successfully established a validated nomogram for predicting OS in patients with PMBCL, which can help clinicians to select appropriate individualized treatment for their patients.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Use of Emicizumab in Patients with Hemophilia a with and without Inhibitors: A Single Center Experience
Authors: Giuffrida Gaetano , Nicolosi Daniela , Condorelli Annalisa , Markovic Uros , Di Raimondo Francesco
Published: November 2021

Emicizumab is a humanized bispecific recombinant monoclonal antibody that binds the enzyme factor IXa and the substrate factor X and mimics the function of FVIII. It solved some unmet needs of haemophilia A (HA) treatment, such as regimen (once weekly up to once monthly infusion) and route of administration (subcutaneous). Most importantly, emicizumab reduce bleeding episodes and improved the quality of life of these patients. Emicizumab received FDA approval for prophylaxis in HA patients with inhibitors in November 2017 and for patients without inhibitors in October 2018.The aim of this study is to provide information on treatment with emicizumab in patients with severe haemophilia A with and without inhibitors in terms of efficacy, safety and quality of life. Here we report our experience with 9 patients with severe haemophilia A (5 with inhibitors and 4 without inhibitors) that were switched from replacement therapy to emicizumab prophylaxis. Median age at initiation of treatment was 26.8 years for patients with inhibitors and 18.7 years for those without inhibitors. Median duration of therapy was 12 months for patients with inhibitors and 8 months for those without inhibitors. All patients were started on emicizumab with a loading dose of 3 mg/kg once weekly for 4 weeks followed by a maintenance dose of 1.5 mg/kg once weekly or every two weeks. The mean annual bleeding rates (ABR) in patients without inhibitor prior to emicizumab was 1.5 compared to 0 while on emicizumab prophylaxis. In subjects with inhibitors ABR was 1.8 compared to 0.4 while on emicizumab prophylaxis. Furthermore, no adverse events including thrombotic events occurred in course of emicizumab prophylaxis. Bleeding events in course of emicizumab prophylaxis did not request any treatment with replacement therapy or bypassing agents. The questionnaire evaluating quality of life (HAL v 2.0 - 2015 ITA; pedHAL v 2.0 - 2015 ITA) was administered to the patients before switch to emicizumab and after a follow up of six months. The scores showed a significant improvement in terms of quality of life in course of emicizumab prophylaxis. In conclusion, our experience suggests that emicizumab prophylaxis is safe and improved protection against bleeding and quality of life compared to replacement therapy in patients with severe A haemophilia.

Disclosures

Di Raimondo: Amgen: Honoraria; Jazz Pharmaceutical: Honoraria; Pfizer: Honoraria; Janssen Pharmaceuticals: Honoraria; Bristol Myers Squibb: Honoraria; AbbVie: Honoraria.
Neutrophil Extracellular Traps (NETs) Contribute to the Formation of Microvascular Thrombosis in Immune Thrombotic Thrombocytopenic Purpura
Authors: Yada Noritaka , Sui Jingrui , Zheng Liang , Zheng X. Long
Published: November 2021

Introduction. Immune thrombotic thrombocytopenic purpura (iTTP), a potentially fatal blood disorder, is primarily caused by severe deficiency of plasma ADAMTS13 activity resulting from immunoglobulin (Ig) G-mediated inhibition of plasma ADAMTS13 activity. However, severe ADAMTS13 deficiency is necessary but not sufficient to cause acute iTTP. An environmental factor such as infection or acute inflammation may be necessary to trigger the acute onset of the disease. We and others have previously reported that plasma markers of neutrophil activation and neutrophil extracellular traps (NETs) formation are significantly elevated in patients with acute iTTP, which returns to normal during remission. However, the pathogenetic role of NETs in acute iTTP is not fully understood.

Methods and results. Using flow cytometry, microfluidic shear-based assay, and confocal imaging analysis, we determined the in vivo NETosis in blood samples obtained from patients with acute episode of iTTP and ex vivo NETs formation, as well as the therapeutic efficacy of DNase I on thrombus formation under flow. We showed that by flow cytometry that only very few CitH3+/MPO+ positive neutrophils were present in the healthy donor blood. This population of cells dramatically increased after being stimulated with a bacterial toxin (i.e., Shigatoxin-2) at ~100 ng/mL for 15 min. Importantly, the number of CitH3+/MPO+ positive neutrophils in the sample obtained from a patient with acute iTTP was ~1,000 times higher than that in the healthy controls (Fig. 1), suggesting a massive NETosis in patients with acute iTTP. Microfluidic shear-based assay and confocal imaging analysis further confirmed a dramatic increase in adhesion and aggregation of murine platelets (stained with Alexa647 anti-CD41) and neutrophil (stained with Hoechst), as well as formation of NETs (stained with Syto green) following a perfusion of an Adamts13 -/- murine whole blood (anti-coagulated with thrombin inhibitor, PPACK) under arterial shear (15 dyne/cm 2) over a stimulated murine endothelial surface. Interestingly, an addition of DNase I (100 U/mL) significantly reduced the overall surface coverage of platelets and neutrophils on the murine endothelial surface under the same conditions (Fig. 2).

Conclusions. These results demonstrate for the first time NETosis and NETs formation are common in patients with acute iTTP and in Adamts13 -/- mice after being stimulated with shigatoxin; DNase I appears to be highly efficacious eliminating the NETs and platelet/neutrophil-dominant thrombosis under arterial flow. Our findings support the pathogenetic role of NETs in the onset and progression of iTTP, and the therapeutic potential of DNase I in such a fatal disease.

Figure 1 Figure 1.

Disclosures

Zheng: Alexion: Speakers Bureau; Sanofi-Genzyme: Honoraria, Speakers Bureau; Takeda: Consultancy, Honoraria; Clotsolution: Other: Co-founder; AJMC: Honoraria.
SATB1 Regulates Chromatin Organization and HSP70 Expression in Early Erythropoiesis and Is Downregulated in Models of Diamond Blackfan Anemia
Authors: Wilkes Mark C , Shibuya Aya , Scanlon Vanessa M , Chae Hee-Don , Narla Anupama , Sakamoto Kathleen M.
Published: November 2021

Diamond Blackfan Anemia (DBA) is a rare genetic disease predominantly caused by mutations carried within one of at least 20 ribosomal genes. DBA is characterized by red blood cell aplasia and normal myeloid and megakaryocyte progenitors, indicating that early uncommitted progenitors are relatively unaffected by the mutations. In DBA, the formation of BFU-E colonies and subsequent erythroblasts are severely restricted and indicate a defect in one of the earliest stages of erythroid expansion.

To identify critical molecular mechanisms that may regulate early erythropoiesis, we used shRNAs against the ribosomal protein RPS19 (the most commonly mutated gene in DBA) in cord blood derived CD34+ hematopoietic stem and progenitor cells (HSPCs) and performed bulk RNA-seq. After 3 days in an erythroid culture media, the transcriptomes in CD71+ erythroid progenitors were examined. We found that the special AT binding protein 1 (SATB1) was downregulated in RPS19-insufficient HSPCs compared to healthy cord blood HSPCs.

SATB1 is modestly expressed in hematopoietic stem cells but is induced during lymphoid expansion and has been previously reported to suppress myeloid/erythroid progenitor (MEP) expansion. Our results showed that maintaining SATB1 expression is required for optimal expansion of MEP progenitors and that the premature loss of SATB1 in DBA contributes to the anemia phenotype.

SATB1 binds to 3 specific regions upstream of the 5'UTR of the HSP70 genes and induces the formation of 2 chromatin loops. An enhancer element associates with the proximal promoters of the two HSP70 genes and facilitates the induction of HSP70. In DBA, HSP70 is not induced and contributes to DBA pathogenesis.

HSPA1A is induced 4.3-fold while HSPA1B is induced 3.1-fold. Increased expression of the master erythroid transcription factor GATA1 during erythropoiesis occurs in two phases. The first induction precedes a more dramatic induction that accompanies later stages of erythroid differentiation. The absence of SATB1 or HSP70 reduced the earlier GATA1 induction that accompany MEP expansion by 46.1% and 49.3% respectively. The number of MEPs in SATB1 knockdown HSPCs was reduced, resulting in a 24.5% reduction in CD235+ erythroid and 20.8% reduction in CD41+ megakaryocytes.

While SATB1-independent effects of RPS19-insufficiency contribute more significantly to erythroid defects in DBA, we have uncovered that SATB1 contributes to regulation of the earliest stages of erythropoiesis by facilitating the induction of HSP70 and subsequent stabilization of an early induction of GATA1.

Disclosures

No relevant conflicts of interest to declare.
Clinical Outcomes of COVID-19 Infection Among Children with Cancer: A Single-Center Experience
Authors: Akkoyun Esra , Sotelo Jesus , Cherry Giselle , Bisceglia Maria , Slone Tamra , Aquino Victor , Sue Paul
Published: November 2021

Background

Outcomes of COVID-19 infection among children with underlying malignancy remain unclear, with limited data available to date. Here we report clinical characteristics and outcomes following COVID-19 infection among children with underlying malignancy or a history of hematopoietic stem cell transplant (HSCT) during the first year of the global COVID-19 pandemic, in Dallas, TX.

Methods:

We retrospectively reviewed children with a history of malignancy or prior HSCT and positive SARS-CoV-2 polymerase chain reaction (PCR) result between March 1, 2020 - March 31, 2021. Demographic, clinical, and outcome data were reviewed, and analyzed according to underlying condition and disease severity.

Results:

Forty-six patients with underlying malignancy or HSCT met inclusion criteria. Median age was 9 years, with twenty (43%) female, and twenty-seven (59%) of Hispanic ethnicity. Underlying conditions included Acute Lymphoblastic Leukemia (67%), Acute Myelogenous Leukemia (9%), HSCT (11%), sarcoma (9%), solid tumors (9%), and Hodgkin's lymphoma (2%). Fifteen subjects (33%) were asymptomatic, with twenty-four children (52%) classified as having mild, five (11%) with severe, and two (4%) with critical COVID-19 disease. The most common symptoms at presentation were fever (46%), cough (39%), and gastroenteritis (19%).

Thirty-four children (74%) were hospitalized, with seven (15%) requiring intensive care unit (ICU) care. 17% of patients were admitted for treatment of COVID-19 alone, with other indications including neutropenic fever, relapse, and chemotherapy. Six (13%) patients required non-invasive ventilation, and two patients (4%) required invasive mechanical ventilation. Median length of stay was 9 days (IQR 4.5-24.5), with median ICU stay of 3.5 days (IQR 2-26.7).

Seven children (15%) received COVID-19 targeted therapy including steroids, remdesivir or convalescent plasma. Among severe and critically ill children, six (86%) had underlying hematologic malignancies (5 HR ALL, 1 AML), and two (28%) a prior history of HSCT. Two children (4%) died.

Thirty-six patients (78%) achieved documented clearance of SARS-CoV-2 by PCR, at median of 43.5 days (IQR 28.7-65.2).

Conclusion:

Among children with underlying malignancy or a history of HSCT, COVID-19 results in a spectrum of illness ranging from asymptomatic disease to death. Rates of hospitalization are high compared to the general pediatric population, and illness may be complicated by additional factors including co-infection, neutropenic fever, relapse of malignancy and need for chemotherapy. Children with hematologic malignancy or a history of HSCT may be at increased risk for severe disease. Additional studies are urgently needed to elucidate risk factors for severe/critical COVID-19 disease among children with underlying malignancy.

Disclosures

No relevant conflicts of interest to declare.
Engineering Human Invariant Natural Killer T (iNKT) Cells to Overexpress Immunomodulatory Cytokines
Authors: Mavers Melissa , Hollingsworth Derek , Boonchalermvichian Chaiyaporn , Liu Jeffrey , Baker Jeanette , Lopes Ramos Teresa , Lohmeyer Juliane K. , Lin Po-Yu , Roncarolo Maria Grazia , Negrin Robert S.
Published: November 2021

Many studies have shown the important role that invariant natural killer T (iNKT) cells play in suppression of graft-versus-host disease (GVHD). Murine studies have also demonstrated multiple subsets of iNKT cells, including those with pro-inflammatory and immunosuppressive properties. The prevention of murine acute GVHD by the adoptive transfer of immunomodulatory iNKT cells suggests that this approach would be ideal for clinical translation. However, the heterogeneity of human iNKT cells remains poorly understood, and the uncertain ability to identify immunosuppressive human iNKT cells and expand them to clinically relevant numbers remains a limitation in clinical translation. Therefore, engineering bulk human iNKT cells to enhance immunosuppressive capacity represents a novel approach to ensuring sufficient cell numbers of immunoregulatory cells for adoptive transfer. Herein, we describe the lentiviral transduction of human iNKT cells to overexpress immunomodulatory cytokines. We focused on IL-4, because murine iNKT cell-mediated GVHD suppression is IL-4 dependent, and IL-10, because an immunosuppressive regulatory T cell-like subset of murine iNKT cells highly produces IL-10. Human iNKT cells were transduced with a lentiviral construct expressing IL-4 or IL-10 (or green fluorescent protein (GFP) as a control) and an inert truncated nerve growth factor receptor (deltaNGFR) to facilitate sorting of transduced cells. Enzyme linked immunosorbent assay revealed significantly increased expression of IL-4 or IL-10 by cells transduced with the corresponding lentivirus, as compared to untransduced, mock transduced, or GFP transduced cells, replicated with 3 iNKT donors (p<0.03) (Figure 1). This proof-of-concept data demonstrates the ability to effectively transduce human iNKT cells to overexpress immunomodulatory cytokines. Future studies will focus on evaluating the immunosuppressive capacity of these engineered cells both in vitro and in vivo (using the xenograft GVHD model). Successful suppression of GVHD by engineered iNKT cells will provide a critical step forward in identifying novel approaches to bring these promising cells to the clinic.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Real World Experience of the BTK Inhibitors. Comparing Cardiac, Vascular and Infectious Toxicities
Authors: Master Samip R , Ramadas Poornima , Mansour Richard Preston
Published: November 2021

Background: Cardiac and vascular toxicities are known complications on Bruton Tyrosine kinase inhibitor (BTKi)drugs. Ibrutinib, acalibrutinib and zanubrutinib are all approved BTKi drugs. We did a retrospective analysis on adverse effects (AE) of BTKi's that has been made available to public by the FDA.

Methods: The FDA has made the data on AEs of various treatments available to general public through the FDA Adverse Events Reports System (FAERS) public dashboard. We investigated the infectious, cardiac and vascular AEs of various BTKi for the years 2019-2021.

Results: The percentage of cardiac AE compared to total AEs reported for ibrutinib, acalibrutinib and zanubrutinib were 12%, 7.3% and 6.2% respectively. The most common cardiac toxicity was atrial fibrillation.

The percentage of vascular AE compared to total AEs reported for ibrutinib, acalibrutinib and zanubrutinib were 8.2 %, 5.2 % and 3.9 % respectively. The most common vascular toxicity was hemorrhage.

The percentage of infectious AE compared to total AEs reported for ibrutinib, acalibrutinib and zanubrutinib were 18.3%, 13.6% and 35.5% respectively. The most common vascular toxicity was pneumonia.

Conclusions: Out of the reported cases of AEs to BTKis approved, ibrutinib have most and acalibrutinib have least cardiac AEs. Zanubrutinib has least vascular AEs. Out of the reported cases of AEs to BTKis approved, zanubrutinib have most infectious AEs.

Disclosures

Master: Blue Bird Bio: Current holder of individual stocks in a privately-held company.
The Loss of Liver Kinase B1 Accelerates Maturation Time in the Developing Erythrocyte
Authors: White III Zollie , Freeman Katie , Delehanty Lorrie L , Goldfarb Adam N
Published: November 2021

In the US, iron-restricted anemia contributes greatly to morbidity. The erythroid iron deprivation response, characterized by a pathway in which erythropoiesis is suppressed during iron restriction, underlies this anemia. In preliminary studies, liver kinase B1 (LKB1) was implicated as a potential key component in the erythroid iron deprivation response. Normal human CD34+ hematopoietic progenitor cells cultured for 3 days in erythroid medium with 100% or 10% transferrin saturations underwent immunoblot of whole cell lysates. Reproducibly, the levels of LKB1 did not change based on the transferrin saturation; however, immunofluorescence imaging showed a shift in subcellular localization when cells were subjected to low iron conditions.

To assess the effects of LKB1 loss in the erythroid compartment, we used control and LKB1 conditional knockout (STK11 F/F; EpoR-Cre+) mice. In steady-state conditions, the loss of LKB1 does not confer a change in RBC count, though, there is a baseline increase in the number of reticulocytes, and a large increase in the level of serum Erythropoietin (Epo). In a model to precipitate anemia, these mice were challenged with intraperitoneal injection of phenylhydrazine (PHZ). It was found that LKB1 is dispensable for an appropriate response to this type of stress erythropoiesis.

To assess the impact of LKB1 on maturation, we performed flow cytometric analysis using an ex vivo culture system of splenic erythroblasts. LKB1-deficient erythroid progenitors show increased percentages of more advanced cells as evidenced by the surface markers CD71 and Ter119 (the mouse analogue of human glycophorin A). Current studies are underway to assess if this change is due to signaling through the AMPK pathway. These studies will provide mechanistic detail of LKB1 function and activity on erythropoiesis, improving our understanding of programs involved in maturation of differentiation and lineage selection which may ultimately help to improve health outcomes and advance treatment for various types of anemias.

Disclosures

No relevant conflicts of interest to declare.
Retrospective Analysis of Minimal Residual Disease Testing By High Throughput Immunosequencing Versus High Sensitivity Flow Cytometry in Multiple Myeloma
Authors: Khan Anwar , Pakasticali Nagehan , Fathalla Omar , Nishihori Taiga , Hussaini Mohammad O
Published: November 2021

Introduction:

Detection of minimal residual disease (MRD) is one of the strongest predictors of outcome in multiple myeloma (MM). Until recently, the most commonly available method to detect MRD in clinical practice has been high sensitivity flow cytometry (FC) which can detect MRD with at 10 -5 sensitivity. In recent years, next-generation sequencing (NGS) has become a viable method to assess the MRD in MM patients with a 10 -6 sensitivity. NGS appears to have some advantages over HC-FC by circumventing subjectivity of analysis. However, real-world comparison between these two methodologies in the literature is limited and is important to inform daily hematopathology and oncology ordering practices.

Methods:

We retrospectively identified all cases of MM with NGS MRD data from bone marrow specimens at the Moffitt Cancer Center and collated corresponding flow MRD data and clinical data (OS, patient demographics) electronically and via chart review.

10-color flow cytometry was performed on a Gallios System and analyzed on Kaluza (Beckman Coulter, IN). Two million events were collected on all cells. Validated lower limit of detection was at least 0.01%. Antibodies included CD28, CD81, CD56, CD138, CD319, CD20, CD19, CD117, CD38, CD45, CD27, CD200 (BD, Biolegend, Beckman Coulter).

clonoSEQ ® (Adaptive Biotechnologies, Seattle, WA) testing was performed which uses multiplex polymerase chain reaction (PCR) and NGS to identify, characterize, and monitor clonotypes of immunoglobulin (Ig) IgH (V-J), IgH (D-J), IgK, and IgL receptor gene sequences, and translocated BCL1/IgH (J) and BCL2/IgH (J) sequences

Statistical analysis was performed by Spearman correlation coefficient and Kaplan-Meier analysis.

Results:

192 samples from 122 unique patients were identified that had both NGS and FC data performed on the same sample. FC+ values ranged from 1x10 -7 to 0.39. NGS+ values ranged from 2.3 x 10 -7 to 0.15. Spearman correlation coefficient showed moderate concordance between NGS and FC at r=0.67 (p<0.001). Six samples were positive by FC (mean tumor burden (MTB)= 0.0007) but missed by NGS; whereas 59 samples were positive by NGS (MTB= 0.002) but missed by flow cytometry. Two cases by FC were equivocal and these were both definitively designated as MRD+ by NGS. Overall survival was worse for MRD+ (by NGS or FC) vs MRD(-) (Figure 1).

Conclusion:

Our study confirms the importance of MRD detection in MM and shows the robust utility of NGS for MRD detection in routine hematopathology practice. While both FC and NGS are complementary given that each can potentially detect MRD missed by another method, the data supports the increased sensitivity of NGS over FC.

Figure 1 Figure 1.

Disclosures

Nishihori: Novartis: Research Funding; Karyopharm: Research Funding. Hussaini: Stemeline Therapeutics: Honoraria.
Rates of Neutropenic Fever in Patients with Hematologic Malignant Diseases with Central Venous Access
Authors: Abbagoni Sujith , Malik Sarah , Fox Ashley D , Mian Mohammad A. H. , Anwar Tahsin , Bradshaw Danielle , Bryan Locke J. , Jillella Anand P , Kolhe Ravindra , Savage Natasha M. , Padala Sandeep , Kota Vamsi
Published: November 2021

Background: Patients who are diagnosed with hematologic malignant diseases (HMD) almost invariably receive placement of central venous catheters (CVCs), whether temporary or permanent. These catheters are used for administration of various chemotherapies, antibiotics and even nutrition. Although the patients with HMD are immunocompromised, prophylactic use of antibiotics among these patients, especially related to CVC placement, remains controversial. This retrospective analysis evaluates the rates of development of neutropenic fever in patients with HMD with CVCs receiving prophylactic antibiotics.

Methods: Data were collected retrospectively in a cohort of 268 patients with HMD who received treatment at our academic institution between 2002 to 2019. Demographic and other related data was abstracted and descriptive analysis was performed using Epi-Info - 7 software.

Results: Of the 268 patients with HMD, 175 patients (65.3%) received prophylactic antibiotics and among these patients, 180 (67.2%) had a permanent CVC and 109 (40.7%) had a temporary CVC. Comparatively, 93 patients (34.7%) did not receive prophylactic antibiotics and among these individuals, 64 (68.8%) had a permanent CVC and 34 (36.6%) had a temporary CVC. In patients with a permanent central line, 84.3% of patients receiving prophylactic antibiotics developed neutropenic fever while 85% of those who did not receive prophylactic antibiotic also developed neutropenic fever. Among the individuals with a temporary CVC, 79.7% of patients receiving prophylactic antibiotics developed neutropenic fever and 65.4% of patients who were not receiving prophylactic antibiotics also developed neutropenic fever. None of the differences were statistically significant (p=0.8990 and p=0.1547 respectively).

Conclusion: In this cohort of HMD patients, the rate of neutropenic fever development in patients with central venous access receiving prophylactic antibiotic was not significantly different compared to the rate of neutropenic fever development in patients with central venous access who did not receive any prophylactic antibiotic. This data suggests that there does not appear to be a significant clinical utility for prophylactic antibiotics in the prevention of neutropenic fever in patients with HMD who have central venous catheters in place. Further studies analyzing the impact of prophylactic antibiotics on true infections as an etiology for neutropenic fevers in patients with HMD and central venous catheters is warranted.

Disclosures

No relevant conflicts of interest to declare.
A Novel Type V CRISPR System with Potential for Genome Editing in the Liver
Authors: Cost Gregory J. , Temoche-Diaz Morayma , Mei Janet , Butterfield Cristina N. , Brown Christopher T. , Brooks Alan R
Published: November 2021

RNA guided CRISPR genome editing systems can make specific changes to the genomes of mammalian cells and have the potential to treat a range of diseases including those that can be addressed by editing hepatocytes. Attempts to edit the liver in vivo have relied almost exclusively on the Cas9 nucleases derived from the bacteria S treptococcus pyogenes or Staphylococcus aureus to which humans are commonly exposed. Pre-existing immunity to both these proteins has been reported in humans which raises concerns about their in vivo application. In silico analysis of a large metagenomics database followed by testing in mammalian cells in culture identified MG29-1, a novel CRISPR system which is a member of the Type V family but exhibits only 41 % amino acid identity to Francisella tularensis Cas12a/cpf1. MG29-1 is a 1280 amino acid RNA programmable nuclease that utilizes a single guide RNA comprised of a 22 nucleotide (nt) constant region and a 20 to 25 nt spacer, recognizes the PAM KTTN (predicted frequency 1 in 16 bp) and generates staggered cuts. MG29-1 was derived from a sample taken from a hydrothermal vent and it is therefore unlikely that humans will have developed pre-existing immunity to this protein. A screen for sgRNA targeting serum albumin in the mouse liver cell line Hepa1-6 identified 6 guides that generated more than 80% INDELS. The MG29-1 system was optimized for in vivo delivery by screening chemical modifications to the guide that improve stability in mammalian cell lysates while retaining or improving editing activity. Two lead guide chemistries were evaluated in mice using MG29-1 mRNA and sgRNA packaged in lipid nanoparticles (LNP). Three days after a single IV administration on-target editing was evaluated in the liver by Sanger sequencing. The sgRNA that was the most stable in the in vitro assay generated INDELS that ranged from 20 to 25% while a sgRNA with lower in vitro stability failed to generate detectable INDELs. The short sgRNA and small protein size compared to spCas9 makes MG29-1 an attractive alternative to spCas9 for in vivo editing applications. Evaluation of the potential of MG29-1 to perform gene knockouts and gene additions via non-homologous end joining is ongoing.

Disclosures

No relevant conflicts of interest to declare.
Resolvin T-series reduce neutrophil extracellular traps
Authors: Chiang Nan , Sakuma Miyuki , Rodriguez Ana R. , Spur Bernd W. , Irimia Daniel , Serhan Charles N.
Published: November 2021

The newly identified 13-series (T-series) resolvins (RvTs) regulate phagocyte functions and accelerate resolution of infectious inflammation. Because severe acute respiratory syndrome coronavirus 2 elicits uncontrolled inflammation involving neutrophil extracellular traps (NETs), we tested whether stereochemically defined RvTs regulate NET formation. Using microfluidic devices capturing NETs in phorbol 12-myristate 13-acetate–stimulated human whole blood, the RvTs (RvT1-RvT4; 2.5 nM each) potently reduced NETs. With interleukin-1β–stimulated human neutrophils, each RvT dose and time dependently decreased NETosis, conveying ∼50% potencies at 10 nM, compared with a known NETosis inhibitor (10 μM). In a murine Staphylococcus aureus infection, RvTs (50 ng each) limited neutrophil infiltration, bacterial titers, and NETs. In addition, each RvT enhanced NET uptake by human macrophages; RvT2 was the most potent of the four RvTs, giving a >50% increase in NET-phagocytosis. As part of the intracellular signaling mechanism, RvT2 increased cyclic adenosine monophosphate and phospho–AMP-activated protein kinase (AMPK) within human macrophages, and RvT2-stimulated NET uptake was abolished by protein kinase A and AMPK inhibition. RvT2 also stimulated NET clearance by mouse macrophages in vivo. Together, these results provide evidence for novel pro-resolving functions of RvTs, namely reducing NETosis and enhancing macrophage NET clearance via a cyclic adenosine monophosphate–protein kinase A–AMPK axis. Thus, RvTs open opportunities for regulating NET-mediated collateral tissue damage during infection as well as monitoring NETs.
Spliceator: multi-species splice site prediction using convolutional neural networks
Authors: Scalzitti Nicolas , Kress Arnaud , Orhand Romain , Weber Thomas , Moulinier Luc , Jeannin-Girardon Anne , Collet Pierre , Poch Olivier , Thompson Julie D.
Published: November 2021

Background

Ab initio prediction of splice sites is an essential step in eukaryotic genome annotation. Recent predictors have exploited Deep Learning algorithms and reliable gene structures from model organisms. However, Deep Learning methods for non-model organisms are lacking.

Results

We developed Spliceator to predict splice sites in a wide range of species, including model and non-model organisms. Spliceator uses a convolutional neural network and is trained on carefully validated data from over 100 organisms. We show that Spliceator achieves consistently high accuracy (89–92%) compared to existing methods on independent benchmarks from human, fish, fly, worm, plant and protist organisms.

Conclusions

Spliceator is a new Deep Learning method trained on high-quality data, which can be used to predict splice sites in diverse organisms, ranging from human to protists, with consistently high accuracy.
Transcriptome dynamics in developing testes of domestic cats and impact of age on tissue resilience to cryopreservation
Authors: Amelkina Olga , Silva Andreia M. da , Silva Alexandre R. , Comizzoli Pierre
Published: November 2021

Background

Fundamental knowledge of cellular and molecular mechanisms in developing testicular tissues is critical to better understand gonadal biology and responses to non-physiological conditions. The objective of our study was to (1) analyze transcriptome dynamics in developing testis of the domestic cat and (2) characterize age effects on the initial response of the tissue to vitrification. Tissues from adult and juvenile cats were processed for histology, DNA integrity, and RNA sequencing analyses before and after vitrification.

Results

Transcriptomic findings enabled to further characterize juvenile period, distinguishing between early and late juvenile tissues. Changes in gene expression and functional pathways were extensive from early to late juvenile to adult development stages. Additionally, tissues from juvenile animals were more resilient to vitrification compared to adult counterparts, with early juvenile sample responding the least to vitrification and late juvenile sample response being closest to adult tissues.

Conclusions

This is the first study reporting comprehensive datasets on transcriptomic dynamic coupled with structural analysis of the cat testis according to the age and exposure to cryopreservation. It provides a comprehensive network of functional terms and pathways that are affected by age in the domestic cat and are either enriched in adult or juvenile testicular tissues.
Risk of severe COVID-19 and mortality in patients with established chronic liver disease: a nationwide matched cohort study
Authors: Simon Tracey G. , Hagström Hannes , Sharma Rajani , Söderling Jonas , Roelstraete Bjorn , Larsson Emma , Ludvigsson Jonas F.
Published: November 2021

Background and aims

Some, but not all, prior studies have suggested that patients with chronic liver disease are at increased risk of contracting COVID-19 and developing more severe disease. However, nationwide data are lacking from well-phenotyped cohorts with liver histology and comparisons to matched general population controls.

Methods

We conducted a nationwide cohort study of all Swedish adults with chronic liver disease (CLD) confirmed by liver biopsy between 1966 and 2017 (n = 42,320), who were alive on February 1, 2020. CLD cases were matched to ≤ 5 population comparators by age, sex, calendar year and county (n = 182,147). Using Cox regression, we estimated multivariable-adjusted hazard ratios (aHRs) and 95% confidence intervals (CIs) for COVID-19 hospitalization and severe COVID-19 (intensive care admission or death due to COVID-19).

Results

Between February 1 and July 31, 2020, 161 (0.38%) CLD patients and 435 (0.24%) general population controls were hospitalized with COVID-19 (aHR = 1.36, 95% CI = 1.11–1.66), while 65 (0.15%) CLD patients and 191 (0.10%) controls developed severe COVID-19 (aHR = 1.08, 95% CI = 0.79–1.48). Results were similar in patients with CLD due to alcohol use, nonalcoholic fatty liver disease, viral hepatitis, autoimmune hepatitis, and other etiologies. Among patients with cirrhosis (n = 2549), the aHRs for COVID-19 hospitalization and for severe COVID-19 were 1.08 (95% CI 0.48–2.40) and 1.23 (95% CI = 0.37–4.04), respectively, compared to controls. Moreover, among all patients diagnosed with COVID-19, the presence of underlying CLD was not associated with increased mortality (aHR = 0.85, 95% CI = 0.61–1.19).

Conclusions

In this nationwide cohort, patients with CLD had a higher risk of hospitalization for COVID-19 compared to the general population, but they did not have an increased risk of developing severe COVID-19.
Community-level educational attainment and dementia: a 6-year longitudinal multilevel study in Japan
Authors: Takasugi Tomo , Tsuji Taishi , Hanazato Masamichi , Miyaguni Yasuhiro , Ojima Toshiyuki , Kondo Katsunori
Published: November 2021

Background

As the understanding of the association between community-level education and dementia is insufficient, this study examined the contextual association of community-level prevalence of low educational attainment on the risk of dementia incidence. With this study, we further explored the potential differences in the aforementioned associations for urban and non-urban areas.

Methods

We analyzed 6 years of prospective cohort data from the Japan Gerontological Evaluation Study, beginning with the baseline data collected between 2010 and 2012, for 51,186 physically and cognitively independent individuals aged ≥65 years (23,785 men and 27,401 women) from 346 communities in 16 municipalities across 7 prefectures. We assessed dementia incidence using available data from the long-term care insurance system in Japan. We dichotomized education years as ≤9 and ≥ 10 years and aggregated individual-level educational attainment as a community-level independent variable. Model 1 covariates were age and sex. Income, residential years, disease, alcohol, smoking, social isolation, and population density were added in Model 2. We conducted multiple imputation to address the missing data. We performed a two-level (community and individual) survival analysis to calculate hazard ratios (HRs) and 95% confidence intervals (CIs).

Results

The results indicate that the cumulative incidence of dementia during the follow-up period was 10.6%. The mean proportion with educational attainment of ≤9 years was 40.8% (range: 5.1–87.3%). Low community-level educational attainment was significantly associated with higher dementia incidence (HR: 1.04; 95% CI: 1.01–1.07), estimated by 10 percentage points of low educational attainment after adjusting for individual-level educational years and covariates. While the association was significant in non-urban areas (HR: 1.07; 1.02–1.13), there was no association in urban areas (HR: 1.03; 0.99–1.06).

Conclusions

Older people living in communities with low educational attainment among their age demographic develop dementia more often compared with those living in areas with high educational attainment after adjusting for individual-level educational attainment and covariates; the association was pronounced in non-urban areas. Securing education for adolescents as a life course and population approach could thus be crucial in preventing dementia later in life among older people living in non-urban areas.
Spectrum of antibiotic resistance in UTI caused by Escherichia coli among HIV-infected patients in Uganda: a cross-sectional study
Authors: Abongomera George , Koller Maurice , Musaazi Joseph , Lamorde Mohammed , Kaelin Marisa , Tasimwa Hannington B. , Eberhard Nadia , Hongler Jan , Haller Sabine , Kambugu Andrew , Castelnuovo Barbara , Fehr Jan
Published: November 2021

Background

Antimicrobial drug resistance is one of the top ten threats to global health according to the World Health Organization. Urinary tract infections (UTIs) are among the most common bacterial infections and main reason for antibiotic prescription. The incidence of UTIs appears to be high among people living with HIV. We sought to determine the most common UTI pathogens among HIV infected patients and evaluate their susceptibility towards antibiotics.

Methods

We performed a cross-sectional study among HIV-infected patients aged ≥ 18 years presenting at an HIV care specialized clinic with symptoms suggestive of a urethritis. Urine cultures were subjected to antibiotic susceptibility testing according to Clinical Laboratory Standards Institute. The data was analyzed using STATA, we performed Pearson’s Chi-square and Fisher’s exact tests to compare differences between proportions.

Results

Out of the 200 patients, 123 (62%) were female. The median age was 41.9 years (IQR 34.7–49.3). Only 32 (16%) urine cultures showed bacterial growth. Escherichia coli was the most commonly isolated uropathogen (72%), followed by Klebsiella pneumoniae (9%). E. coli was completely resistant to cotrimoxazole and ampicillin; resistance to ciprofloxacin and ceftriaxone was 44% and 35% respectively; 9% to gentamicin; no resistance detected to nitrofurantoin and imipenem.

Conclusions

Our findings are congruent with the Uganda national clinical guidelines which recommends nitrofurantoin as the first line antibiotic for uncomplicated UTI. Significant ciprofloxacin and ceftriaxone resistance was detected. In the era of emerging antibiotic resistance, understanding the local susceptibilities among sub-populations such as HIV infected patients is crucial. Further investigation is needed to address reasons for the low bacterial growth rate observed in the urine cultures.
Stent encrustation or fragmentation? A case report of post stent removal encrustation in postpartum woman and literature review
Authors: Ye Zhaohua , Mi Qiwu , Huang Renzhao
Published: November 2021

Background

Ureteral stents are commonly used in the field of urology to relieve ureteral obstruction. However, complications relating to ureteral stent use, such as encrustation continue to occur, especially with prolonged indwell time.

Case presentation

Here we present a 37-year-old postpartum woman with a foreign body in her bladder after removing a ureteral stent 1 month before. She insisted that the foreign body was the fragment of stent and asked for medical malpractice indemnity payments while the surgeon of her insisted that the stent was intact during the procedure. Finally, the foreign body was confirmed as an encrustation by cystoscopy and the patient received 10,000 yuan ($ 1500) as indemnity payments after encrustation removal.

Conclusion

In the absence of guidelines, stent indwelling time vary with centers’ habits, stent materials and patient’s education. Early detection of stent encrustation and timely removal of the encrusted stent are still the best way to avoid stent retention. Violent stent removal is of danger and not recommended in any case.
Enhancing Cancer care of rural dwellers through telehealth and engagement (ENCORE): protocol to evaluate effectiveness of a multi-level telehealth-based intervention to improve rural cancer care delivery
Authors: Pal Tuya , Hull Pamela C. , Koyama Tatsuki , Lammers Phillip , Martinez Denise , McArthy Jacob , Schremp Emma , Tezak Ann , Washburn Anne , Whisenant Jennifer G. , Friedman Debra L.
Published: November 2021

Background

Despite lower cancer incidence rates, cancer mortality is higher among rural compared to urban dwellers. Patient, provider, and institutional level factors contribute to these disparities. The overarching objective of this study is to leverage the multidisciplinary, multispecialty oncology team from an academic cancer center in order to provide comprehensive cancer care at both the patient and provider levels in rural healthcare centers. Our specific aims are to: 1) evaluate the clinical effectiveness of a multi-level telehealth-based intervention consisting of provider access to molecular tumor board expertise along with patient access to a supportive care intervention to improve cancer care delivery; and 2) identify the facilitators and barriers to future larger scale dissemination and implementation of the multi-level intervention.

Methods

Coordinated by a National Cancer Institute-designated comprehensive cancer center, this study will include providers and patients across several clinics in two large healthcare systems serving rural communities. Using a telehealth-based molecular tumor board, sequencing results are reviewed, predictive and prognostic markers are discussed, and treatment plans are formulated between expert oncologists and rural providers. Simultaneously, the rural patients will be randomized to receive an evidence-based 6-week self-management supportive care program, Cancer Thriving and Surviving, versus an education attention control. Primary outcomes will be provider uptake of the molecular tumor board recommendation and patient treatment adherence. A mixed methods approach guided by the Consolidated Framework for Implementation Research that combines qualitative key informant interviews and quantitative surveys will be collected from both the patient and provider in order to identify facilitators and barriers to implementing the multi-level intervention.

Discussion

The proposed study will leverage information technology-enabled, team-based care delivery models in order to deliver comprehensive, coordinated, and high-quality cancer care to rural and/or underserved populations. Simultaneous attention to institutional, provider, and patient level barriers to quality care will afford the opportunity for us to broadly share oncology expertise and develop dissemination and implementation strategies that will enhance the cancer care delivered to patients residing within underserved rural communities.

Trial registration

Clinicaltrials.gov, NCT04758338. Registered 17 February 2021 – Retrospectively registered, http://www.clinicaltrials.gov/
Therapists’ emotional state after sessions in which suicidality is addressed: need for improved management of suicidal tendencies in patients with borderline personality pathology
Authors: Bühlmann Vera , Schlüter-Müller Susanne , Fürer Lukas , Steppan Martin , Birkhölzer Marc , Schmeck Klaus , Koenig Julian , Kaess Michael , Zimmermann Ronan
Published: November 2021

Introduction

Patient suicidality is a frequently experienced topic for psychotherapists. Especially adolescents with borderline personality pathology (BPP) often exhibit suicidal tendencies. Previous research which examined therapists’ countertransference towards suicidal patients suggested that therapists are negatively affected and distressed by them. We hypothesize that this emotional response of the therapists is related to specific sessions in which suicidality came up as a topic. Accordingly, the objective of this study consists in examining therapists’ emotional state on a session level of analysis.

Methods

The sample consisted of N = 21 adolescents (age 13–19 years) with BPD or subthreshold BPD. Therapists’ emotional states were measured in n = 418 sessions using the Session Evaluation Questionnaire. Principal component analysis was used to reduce dimensionality of the therapist response. The emotional states were compared depending on whether suicidality has been addressed in the session (SS) or not (NSS).

Results

Two components could be identified. Firstly, therapists were more aroused, excited, afraid, angry and uncertain after SS than after NSS. Secondly, therapists were more aroused, excited, definite and pleased after SS than after NSS. Discussion: Suicidality does not always have to be a burden for therapists: Both a “distress” and an “eustress” component occur in this context from which the latter is supposed to help clinicians master a difficult situation. Since countertransference feelings are often not fully conscious, it is necessary to do research on therapists’ emotional states after sessions in which suicidality is addressed. This is crucial to both prevent the therapeutic process from being endangered and preserve clinicians’ mental health. Clinical implications and limitations are discussed.