bluuebunny/crossref_metadata_2025_split huggingface dataset:
Introduction of Two Simultaneous Mutations By Genome Editing Greatly Enhances the Accumulation of the Endogenous Fetal Hemoglobin in Human Normal Erythroid Cells
Authors: Psatha Nikoleta , Li Chang , Wang Hongjie , Dalas Demetri , Stamatoyannopoulos George , Papayannopoulou Thalia , Lieber Andre 
Published: April 2021

Abstract
Thalassemia or sickle cell patients with increased fetal hemoglobin have an ameliorated clinical picture up to transfusion independency. Genome editing with targeted nucleases of the γ-globin silencer BCL11a or of the HBG promoter has been shown to reenact an HPFH-like phenotype by significantly increasing the endogenous fetal globin (HbF) expression. In order to achieve higher efficiency in HbF reactivation we developed an innovative approach, mediated through a highly customizable helper dependent-adenoviral vector (HD-Ad5/35++) with nuclease activity. This vector bares the Cas9 gene and two sequential gRNAs, permitting simultaneous targeting of two different DNA loci, specifically, the erythroid enhancer of BCL11a and the HBG promoter to recreate the -114 to -102 HPFH deletion. This double targeting vector was compared to the respective single gRNA vectors. We observed that disruption of the BCL11a-enhancer increased Gγ globin, whereas disruption of the HBG promoter had a greater impact on the Αγ globin expression. Simultaneous disruption of both loci in human erythroid progenitors increased the overall HbF expression from less than 1% in the control to more than 20%, through a possibly synergistic effect of the two mutations (HbF:2.2% in BCL11a-enhancer only knock-out, 10.8% in HBG-only knock out) by affecting both gamma globin chains. The editing efficiency per locus was similar between the single gRNA and double gRNA vectors. Erythroid cell morphology and phenotypic profile of the double knock-out cells did not differ compared to the single knock-out cells. In addition to our in vitro experiments, we observed that mobilized peripheral blood CD34+ cells transduced and edited by the double gRNA vector, can efficiently engraft in NSG mice. Furthermore, the engrafted edited cells after erythroid differentiation expressed significantly higher levels of gamma globin compared to the control. This strategy has the potential to induce higher levels of HbF reactivation with a clinical benefit in patients with beta globin disorders.

Disclosures
No relevant conflicts of interest to declare.
Engineered Blood Vessels
Authors: Lawson Jeffrey 
Published: April 2021

Abstract
The field of tissue engineering has has made significant progress in the past 20 years. Of the many tissues and organs in development, the area of vascular tissue engineering is now one of the most mature. In that regard, we have developed a novel tissue engineered vascular graft (human acellular vessel [HAV]) that addresses many of the limitations of native vein harvest and the performance of both synthetic ePTFE and autologous vein grafts. The HAV is manufactured in a laboratory by culturing human vascular cells within a biodegradable scaffold that forms a mechanically robust tissue engineered blood vessel. The cells are then completely removed (decellularization), leaving behind a non-immunogenic human collagen-based vascular tissue that can be stored on the shelf for months at a time and ready for immediate implantation into any patient. The HAV is currently being evaluated in Phase II and Phase III clinical trials in the U.S., E.U. and Israel as an arteriovenous vascular access graft for hemodialysis in patients with end-stage renal failure and as an arterial substitute for patients in need of vascular bypass for peripheral arterial disease or vascular trauma. Following clinical implantation, we have observed repopulation and remodeling of the manufactured vessel with the hosts' own cells. We hypothesize that the biological composition of the HAV, compared to synthetic vascular grafts, promotes its physiological integration into host tissue including support of normal host cell infiltration and function. Host cells that identify histologically similar to vascular smooth muscle cells appear to repopulate the middle of the vessel and recipient cells characterized as endothelial cells appear to cover the luminal surface of the implanted vessel. Clinical observations of the Phase II trials have demonstrated excellent vessel durability and a freedom from both early and delayed infection. Based on the success of the Phase II studies, a Global Phase III study is underway for patients in need of dialysis access shunts and vascular programs for vascular (arterial) bypass and trauma are expanding.

Disclosures
Lawson: InnaVasc: Patents & Royalties; Humacyte, Inc.: Employment.
Biology of LAG3 Checkpoint Blockade
Authors: Vignali Dario A.A. 
Published: April 2021

Abstract
Dr. Vignali will discuss the biology of the inhibitory receptor LAG3, and its contribution to T cell exhaustion and immune regulation on CD8+ and CD4+ intratumoral T cells in mouse models of cancer and autoimmunity, and in a variety of human malignancies. He will also discuss the impact of LAG3 on regulatory T cell function. Given the considerable interest in inhibitory receptors in general, and LAG3 recently, as immunotherapeutic targets, he will also discuss how differential LAG3 expression could impact responsiveness to immunotherapy.

Disclosures
Vignali: Potenza Therapeutics: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties: Licensed IP, Research Funding; Tizona Therapeutics: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties, Research Funding; Oncorus: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees; Pieris: Membership on an entity's Board of Directors or advisory committees; FStar: Membership on an entity's Board of Directors or advisory committees; BMS: Equity Ownership; Merck: Equity Ownership; Crescendo: Consultancy; Intellia: Consultancy; MPM: Consultancy; Onkaido/Moderna: Consultancy; Servier: Consultancy.
Clonal Evolution During Stress Hematopoiesis
Authors: Link Daniel C. 
Published: April 2021

Hematopoietic stem and progenitor cells (HSPCs) acquire somatic mutations with age resulting in a heterogeneous cell population, with each HSPC possessing its own unique set of private mutations. HSPCs that acquire somatic mutations that confer a competitive fitness advantage relative to their normal counterparts may clonally expand. Indeed, several groups have documented the presence of clonal hematopoiesis in healthy individuals. Although originally thought to be limited to older individuals, a recent study using an ultra-sensitive sequencing technique showed that expanded hematopoietic clones are detectable in the majority of healthy 50-60-year-old individuals. With some notable exceptions, the same genes that are commonly mutated in clonal hematopoiesis also are somatically mutated in myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). However, only a small fraction of individuals with clonal hematopoiesis subsequently develop a myeloid malignancy and the size of the mutant clone in clonal hematopoiesis can remain stable for years without disease progression. These observations raise several questions that will be addressed in this presentation. What drives expansion of hematopoietic clones? What role do external hematopoietic stressors, such as exposure to chemotherapy, play in the development of clonal hematopoiesis? Why do so few people with clonal hematopoiesis develop a myeloid malignancy, and are there certain mutations that confer a higher risk of transformation?

Disclosures
No relevant conflicts of interest to declare.
The fatty acid elongase ELOVL6 regulates bortezomib resistance in multiple myeloma
Authors: Lipchick Brittany C. , Utley Adam , Han Zhannan , Moparthy Sudha , Yun Dong Hyun , Bianchi-Smiraglia Anna , Wolff David W. , Fink Emily , Liu Liang , Furdui Cristina M. , Lee Jingyun , Lee Kelvin P. , Nikiforov Mikhail A. 
Published: April 2021

AbstractResistance to the proteasome inhibitor bortezomib (BTZ) represents a major obstacle in the treatment of multiple myeloma (MM). The contribution of lipid metabolism in the resistance of MM cells to BTZ is mostly unknown. Here we report that levels of fatty acid elongase 6 (ELOVL6) were lower in MM cells from BTZ-nonresponsive vs BTZ-responsive patients and in cultured MM cells selected for BTZ resistance compared with parental counterparts. Accordingly, depletion of ELOVL6 in parental MM cells suppressed BTZ-induced endoplasmic reticulum (ER) stress and cytotoxicity, whereas restoration of ELOVL6 levels in BTZ-resistant MM cells sensitized them to BTZ in tissue culture settings and, as xenografts, in a plasmacytoma mouse model. Furthermore, for the first time, we identified changes in the BTZ-induced lipidome between parental and BTZ-resistant MM cell lines underlying a functional difference in their response to BTZ. We demonstrated that restoration of ELOVL6 levels in BTZ-resistant MM cells resensitized them to BTZ largely via upregulation of ELOVL6-dependent ceramide species, which was a prerequisite for BTZ-induced ER stress and cell death in these cells. Our data characterize ELOVL6 as a major clinically relevant regulator of MM cell resistance to BTZ, which can emerge from the impaired ability of these cells to alter ceramide composition in response to BTZ.
The effect of platelet G proteins on platelet extravasation and tumor growth in the murine model of ovarian cancer
Authors: Cho Min Soon , Li Jiasong , Gonzalez-Delgado Ricardo , Lee Hani , Vasquez Matthew , He Tiancheng , He Yunjie , Liu Kai , Sasano Tomoyuki , Nürnberg Bernd , Sood Anil K. , Wong Stephen T. C. , Afshar-Kharghan Vahid 
Published: April 2021

Abstract
We and other investigators have shown that platelets promote metastasis and the growth of tumors. Our rationale for conducting this study is that platelets’ prometastatic and progrowth effects depend on a close encounter between platelets and cancer cells. This interaction occurs inside blood vessels with circulating tumor cells and outside blood vessels with cancer cells residing in the tumor parenchyma. Our hypothesis was that platelet extravasation is required for the effect of platelets on tumor growth. Platelets respond to environmental stimuli by activation of G protein–coupled receptors on their surface. We investigated the impact of various platelet G proteins on the growth of ovarian cancer tumors and platelet extravasation. We used mice with platelet-specific deficiency of Gαi2 (Gi), Gα13 (G13), or Gαq (Gq) in a syngeneic ovarian cancer model. We measured the total weight of tumor nodules resected from tumor-bearing mice. We developed methods for automated whole-slide image acquisition and unbiased computerized image analysis to quantify extravasated platelets. We compared the number of platelets inside tumor nodules of platelet G protein–deficient tumor-bearing mice. We found that deficiency of Gi and G13, but not Gq, in platelets resulted in smaller tumors compared with those in corresponding littermates. Deficiency of Gi and G13 in platelets reduced the number of extravasated platelets by &gt;90%, but deficiency of Gq did not reduce the number of extravasated platelets significantly. The lack of Gi or G13 in platelets reduced platelet extravasation into the tumor and tumor growth.
New Guinean orogenic dynamics and biota evolution revealed using a custom geospatial analysis pipeline
Authors: Toussaint Emmanuel F. A. , White Lloyd T. , Shaverdo Helena , Lam Athena , Surbakti Suriani , Panjaitan Rawati , Sumoked Bob , von Rintelen Thomas , Sagata Katayo , Balke Michael 
Published: April 2021

Abstract
Background
The New Guinean archipelago has been shaped by millions of years of plate tectonic activity combined with long-term fluctuations in climate and sea level. These processes combined with New Guinea’s location at the tectonic junction between the Australian and Pacific plates are inherently linked to the evolution of its rich endemic biota. With the advent of molecular phylogenetics and an increasing amount of geological data, the field of New Guinean biogeography begins to be reinvigorated.

Results
We inferred a comprehensive dated molecular phylogeny of endemic diving beetles to test historical hypotheses pertaining to the evolution of the New Guinean biota. We used geospatial analysis techniques to compare our phylogenetic results with a newly developed geological terrane map of New Guinea as well as the altitudinal and geographic range of species (https://arcg.is/189zmz). Our divergence time estimations indicate a crown age (early diversification) for New Guinea Exocelina beetles in the mid-Miocene ca. 17 Ma, when the New Guinean orogeny was at an early stage. Geographic and geological ancestral state reconstructions suggest an origin of Exocelina ancestors on the eastern part of the New Guinean central range on basement rocks (with a shared affinity with the Australian Plate). Our results do not support the hypothesis of ancestors migrating to the northern margin of the Australian Plate from Pacific terranes that incrementally accreted to New Guinea over time. However, our analyses support to some extent a scenario in which Exocelina ancestors would have been able to colonize back and forth between the amalgamated Australian and Pacific terranes from the Miocene onwards. Our reconstructions also do not support an origin on ultramafic or ophiolite rocks that have been colonized much later in the evolution of the radiation. Macroevolutionary analyses do not support the hypothesis of heterogeneous diversification rates throughout the evolution of this radiation, suggesting instead a continuous slowdown in speciation.

Conclusions
Overall, our geospatial analysis approach to investigate the links between the location and evolution of New Guinea’s biota with the underlying geology sheds a new light on the patterns and processes of lineage diversification in this exceedingly diverse region of the planet.
Safety application of muscle relaxants and the traditional low-frequency ventilation during the flexible or rigid bronchoscopy in patients with central airway obstruction: a retrospective observational study
Authors: Li Jing-Jin , Li Nan , Ma Wei-Jia , Bao Ming-Xue , Chen Zi-Yang , Ding Zheng-Nian 
Published: April 2021

AbstractBackgroundBronchoscopy treatments of central airway obstruction (CAO) under general anesthesia are high-risky procedures, and posing a giant challenge to the anesthesiologists. We summarized and analyzed our clinical experience in patients with CAO undergoing flexible or rigid bronchoscopy, to estimate the safety of skeletal muscle relaxants application and the traditional Low-frequency ventilation.MethodsClinical data of 375 patients with CAO who underwent urgent endoscopic treatments in general anesthesia from January 2016 to October 2019 were retrospectively reviewed. The use ratio of skeletal muscle relaxants, dose of skeletal muscle relaxants used, the incidence of perioperative adverse events, adequacy of ventilation and gas exchange, post-operative recovery between rigid bronchoscopy and flexible bronchoscopy therapy, and risk factors for postoperative ICU admission were evaluated.ResultsOf the 375 patients with CAO, 204 patients were treated with flexible bronchoscopy and 171 patients were treated with rigid bronchoscopy. Muscle relaxants were used in 362 of 375 patients (including 313 cisatracurium, 45 rocuronium, 4 atracurium, and 13 unrecorded). The usage rate of muscle relaxants (96.5% in total) was very high in patients with CAO who underwent either flexible bronchoscopy (96.6%) or rigid bronchoscopy (96.5%) therapy. The dosage of skeletal muscle relaxants (Cisatracium) used was higher in rigid bronchoscopy compared with flexible bronchoscopy therapy (10.8 ± 3.8 VS 11.6 ± 3.6 mg, respectively,p < 0.05). No patient suffered the failure of ventilation, bronchospasm and intraoperative cough either in flexible or rigid bronchoscopy therapy. Hypoxemia was occurred in 13 patients (8 in flexible, 5 in rigid bronchoscopy) during the procedure, and reintubation after extubation happened in 2 patients with flexible bronchoscopy. Sufficient ventilation was successfully established using the traditional Low-frequency ventilation with no significant carbon dioxide accumulation and hypoxemia occurred both in flexible and rigid bronchoscopy group (p > 0.05). Three patients (1 in flexible and 2 in rigid) died, during the post-operative recovery, and the higher grade of American Society of Anesthesiologists (ASA) and obvious dyspnea or orthopnea were the independent risk factors for postoperative ICU admission.ConclusionThe muscle relaxants and low-frequency traditional ventilation can be safely used both in flexible and rigid bronchoscopy treatments in patients with CAO. These results may provide strong clinical evidence for optimizing the anesthesia management of bronchoscopy for these patients.
Seasonality in extra-pulmonary tuberculosis notifications in Germany 2004-2014- a time series analysis
Authors: Charles Tanja , Eckardt Matthias , Karo Basel , Haas Walter , Kröger Stefan 
Published: April 2021

Abstract
Background
Seasonality in tuberculosis (TB) has been found in different parts of the world, showing a peak in spring/summer and a trough in autumn/winter. The evidence is less clear which factors drive seasonality. It was our aim to identify and evaluate seasonality in the notifications of TB in Germany, additionally investigating the possible variance of seasonality by disease site, sex and age group.

Methods
We conducted an integer-valued time series analysis using national surveillance data. We analysed the reported monthly numbers of started treatments between 2004 and 2014 for all notified TB cases and stratified by disease site, sex and age group.

Results
We detected seasonality in the extra-pulmonary TB cases (N = 11,219), with peaks in late spring/summer and troughs in fall/winter. For all TB notifications together (N = 51,090) and for pulmonary TB only (N = 39,714) we did not find a distinct seasonality. Additional stratified analyses did not reveal any clear differences between age groups, the sexes, or between active and passive case finding.

Conclusion
We found seasonality in extra-pulmonary TB only, indicating that seasonality of disease onset might be specific to the disease site. This could point towards differences in disease progression between the different clinical disease manifestations. Sex appears not to be an important driver of seasonality, whereas the role of age remains unclear as this could not be sufficiently investigated.
Second sacral sacralalar‐iliac (S2AI) screw placement in adult degenerative scoliosis (ADS) patients: an imaging study
Authors: Wu Bing , Song Kai , Cheng Junyao , Chi Pengfei , Wang Zhaohan , Wang Zheng 
Published: April 2021

Abstract
Background
The imaging characteristics of sacral sacralalar-iliac (S2AI) screw trajectory in adult degenerative scoliosis (ADS) patients will be determined.

Methods
S2AI screw trajectories were mapped on three-dimensional computed tomography (3DCT) reconstructions of 40 ADS patients. The starting point, placement plane, screw template, and a circle centered at the lowest point of the ilium inner cortex were set on these images. A tangent line from the starting point to the outer diameter of the circle was selected as the axis of the screw trajectory. The related parameters in different populations were analyzed and compared.

Results
The trajectory length of S2AI screws in ADS patients was 12.00 ± 0.99 cm, the lateral angle was 41.24 ± 3.92°, the caudal angle was 27.73 ± 6.45°, the distance from the axis of the screw trajectory to the iliosciatic notch was 1.05 ± 0.81 cm, the distance from the axis of the screw trajectory to the upper edge of the acetabulum was 1.85 ± 0.33 cm, and the iliac width was 2.12 ± 1.65 cm. Compared with females, the lateral angle of male ADS patients was decreased, but the trajectory length was increased (P < 0.05). Compared to patients without ADS in previous studies, the lateral angle of male patients was larger, the lateral angle of female patients was increased, and the caudal angle was decreased (P < 0.05).

Conclusions
There is an ideal trajectory of S2AI screws in ADS patients. A different direction should be noticed in the placement of S2AI screws, especially in female patients.
Waking experience modulates sleep need in mice
Authors: Milinski Linus , Fisher Simon P. , Cui Nanyi , McKillop Laura E. , Blanco-Duque Cristina , Ang Gauri , Yamagata Tomoko , Bannerman David M. , Vyazovskiy Vladyslav V. 
Published: April 2021

Abstract
Background
Homeostatic regulation of sleep is reflected in the maintenance of a daily balance between sleep and wakefulness. Although numerous internal and external factors can influence sleep, it is unclear whether and to what extent the process that keeps track of time spent awake is determined by the content of the waking experience. We hypothesised that alterations in environmental conditions may elicit different types of wakefulness, which will in turn influence both the capacity to sustain continuous wakefulness as well as the rates of accumulating sleep pressure. To address this, we compared the effects of repetitive behaviours such as voluntary wheel running or performing a simple touchscreen task, with wakefulness dominated by novel object exploration, on sleep timing and EEG slow-wave activity (SWA) during subsequent NREM sleep.

Results
We find that voluntary wheel running is associated with higher wake EEG theta-frequency activity and results in longer wake episodes, as compared with exploratory behaviour; yet, it does not lead to higher levels of EEG SWA during subsequent NREM sleep in either the frontal or occipital derivation. Furthermore, engagement in a touchscreen task, motivated by food reward, results in lower SWA during subsequent NREM sleep in both derivations, as compared to exploratory wakefulness, even though the total duration of wakefulness is similar.

Conclusion
Overall, our study suggests that sleep-wake behaviour is highly flexible within an individual and that the homeostatic processes that keep track of time spent awake are sensitive to the nature of the waking experience. We therefore conclude that sleep dynamics are determined, to a large degree, by the interaction between the organism and the environment.