docling/tests/data/groundtruth/docling_v2/10-1055-s-0043-1775965.nxml.itxt

item-0 at level 0: unspecified: group _root_
  item-1 at level 1: title: Differential Effects of Erythrop ... xpression on Venous Thrombosis in Mice
  item-2 at level 1: paragraph: Sven Stockhausen; Medizinische K ... ximilians-Universität, Munich, Germany
  item-3 at level 1: text: Background Deep vein thrombosis  ...  and potential therapeutic strategies.
  item-4 at level 1: section_header: Introduction
    item-5 at level 2: text: Red blood cells (RBCs) are the p ... geted prevention and treatment of DVT.
    item-6 at level 2: text: The mechanism of RBC-mediated DV ...  DVT has not been conclusively proven.
    item-7 at level 2: text: In this project we used a transg ... so identified in this mouse strain. 15
    item-8 at level 2: text: The spleen is responsible for RB ...  DVT in vivo remains unclear. 20 25 26
  item-9 at level 1: section_header: Mouse Model
    item-10 at level 2: text: C57BL/6 mice were obtained from  ... nich) and were authorized accordingly.
  item-11 at level 1: section_header: Statistics
    item-12 at level 2: text: Statistical analysis was conduct ... re compared using the chi-square test.
  item-13 at level 1: section_header: Chronic EPO Overproduction Leads to Increased DVT in Mice
    item-14 at level 2: text: To investigate the impact of chr ... ailable in the online version]). 28 29
    item-15 at level 2: text: Based on clinical observations i ...  L [available in the online version]).
  item-16 at level 1: section_header: High RBC Count Leads to a Decrea ... elet Accumulation in Venous Thrombosis
    item-17 at level 2: text: Having observed a correlation be ... y thinner fibrin fibers ( Fig. 3A-C ).
    item-18 at level 2: text: To quantify platelet accumulatio ...  from EPO transgenic mice ( Fig. 2C ).
    item-19 at level 2: text: As mentioned previously, inflamm ... brinogen and platelets were decreased.
  item-20 at level 1: section_header: Short-Term Administration of EPO Does Not Foster DVT
    item-21 at level 2: text: Due to the significant impact of ...  G [available in the online version]).
    item-22 at level 2: text: To further investigate the under ...  unchanged after 2 weeks of treatment.
    item-23 at level 2: text: To analyze the impact of 2-week  ... ollowing EPO treatment ( Fig. 3D, E ).
  item-24 at level 1: section_header: Splenectomy Does Not Affect Venous Thrombus Formation
    item-25 at level 2: text: As the data suggested a qualitat ... [available in the online version]). 13
    item-26 at level 2: text: To investigate the role of splen ...  obtained from nonsplenectomized mice.
    item-27 at level 2: text: Finally, we analyzed the impact  ...  of enhanced or normal erythropoiesis.
  item-28 at level 1: section_header: Discussion
    item-29 at level 2: text: Here, we present evidence for a  ... ouse model through chronic hypoxia. 37
    item-30 at level 2: text: In our analyses, we observed tha ... ing to a 70% reduction in lifespan. 15
    item-31 at level 2: text: During the ageing process, RBCs  ... easing the risk of DVT formation. 5 71
    item-32 at level 2: text: Clearance of RBCs primarily occu ... bsence of the spleen after removal. 15
    item-33 at level 2: text: Besides their activating effect  ... subjected to short-term EPO injection.
  item-34 at level 1: section_header: Figures
  item-36 at level 1: picture
    item-36 at level 2: caption: Fig. 1 EPO-overexpressing mice experience an increased incidence of DVT formation. (A) Comparison of RBC count between EPO-overexpressing Tg(EPO) mice (n = 12) and control (WT) mice (n = 13). (B) Comparison of platelet count between EPO-overexpressing Tg(EPO) (n = 7) mice and control (WT) mice (n = 5). (C) Comparison of thrombus weight between Tg(EPO) mice (n = 9) and WT (n = 10); mean age in the Tg(EPO) group: 19.8 weeks; mean age in the WT group: 20.1 weeks. (D) Comparison of thrombus incidence between Tg(EPO) mice (n = 9) and WT mice (n = 10); NS = nonsignificant, *p < 0.05, **p < 0.01, ***p < 0.001. DVT, deep vein thrombosis; EPO, erythropoietin; RBC, red blood cell; WT, wild type.
  item-38 at level 1: picture
    item-38 at level 2: caption: Fig. 2 Chronic overproduction of EPO in mice leads to a decrease in the accumulation of classical drivers of DVT formation, including platelets, neutrophils, and fibrinogen. (A) The proportion of RBC-covered area in the thrombi of EPO-overexpressing Tg(EPO) mice (n = 4) was compared to control (WT) (n = 3) by immunofluorescence staining of cross-sections of the IVC 48 hours after flow reduction. (B) The proportion of fibrinogen-covered area in the thrombi of EPO- overexpressing Tg(EPO) mice (n = 3) was compared to control (WT) (n = 3) using immunofluorescence staining of cross-sections of the IVC 48 hours after flow reduction. (C) The proportion of platelet-covered area in the thrombi of EPO-overexpressing Tg(EPO) mice (n = 3) was compared to control (WT) (n = 3) by immunofluorescence staining of cross-sections of the IVC 48 hours after flow reduction. (D) Quantification of neutrophils was performed by immunofluorescence staining of cross-sections of the IVC 48 hours after flow reduction in EPO-overexpressing Tg(EPO) mice (n = 3) compared to control (WT) (n = 3). (E) Immunofluorescence staining of cross-sections of the IVC 48 hours after flow reduction from EPO-overexpressing Tg(EPO) mice (top) was compared to control (WT) (bottom) for TER119 in red (RBC), CD42b in green (platelets), and Hoechst in blue (DNA). The merged image is on the left, and the single channel image is on the right. Scale bar: 50 µm. (F) Immunofluorescence staining of cross-sections of the IVC 48 hours after flow reduction from EPO-overexpressing Tg(EPO) mice (top) was compared to control (WT) (bottom) for TER119 in red (RBC), fibrinogen in green, and Hoechst in blue (DNA); the merged image is on the left, and single-channel images are on the right. Scale bar: 50 µm.; NS = nonsignificant, *p < 0.05, **p < 0.01, ***p < 0.001. DVT, deep vein thrombosis; EPO, erythropoietin; IVC, inferior vena cava; RBC, red blood cell; WT, wild type.
  item-40 at level 1: picture
    item-40 at level 2: caption: Fig. 3 The interaction of RBC with fibrinogen leads to the formation of a branched fibrin structure. (A) Immunofluorescence staining of cross-sections of the IVC 48 hours after flow reduction from EPO-overexpressing Tg(EPO) mice (left) was compared to control (WT) for fibrinogen (green). Scare bar: 50 µm. (B) High-resolution confocal images of immunofluorescence staining of cross-sections of the IVC 48 hours after flow reduction from EPO-overexpressing Tg(EPO) mice (left) compared to control (WT) for fibrinogen (green), RBC (red), and DNA (blue). Scale bar: 2 µm. (C) The mean diameter of 2,141 fibrin fibers was measured in cross-sections of thrombi from Tg(EPO) mice (left), and compared to 5,238 fibrin fibers of WT thrombi. (D) The mean diameter of 3,797 fibrin fibers was measured in two cross-sections of thrombi from 2-week EPO-injected mice (left), and compared to 10,920 fibrin fibers of three cross-sections from control (2-week NaCl-injected mice). (E) High-resolution confocal images of immunofluorescence staining of two cross-sections of the IVC 48 hours after flow reduction from 2-week EPO-injected mice (left) were compared to control (2 week NaCl-injected mice) for fibrinogen (green), RBC (red), and DNA (blue). Scale bar: 2 µm. EPO, erythropoietin; IVC, inferior vena cava; RBC, red blood cell; WT, wild type.
  item-42 at level 1: picture
    item-42 at level 2: caption: Fig. 4 Two-week EPO injection leads to thrombocytopenia without an impact on the bone marrow. (A) RBC count in peripheral blood after 6 × 300 IU EPO treatment of C57Bl/6J mice (n = 10) was compared to control (6 × 30 µL NaCl injection) (n = 9). (B) Platelet count in peripheral blood after 6 × 300 IU EPO treatment of C57Bl/6J mice (n = 10) was compared to control (6 × 30 µL NaCl injection) (n = 10). (C) Area of RBC-positive area in the bone marrow of 6 × 300 IU EPO-treated C57Bl/6J mice (n = 4) was compared to control (6 × 30 µL NaCl injection) (n = 4). (D) Immunofluorescence staining of cross-sections of the bone marrow after 2-week EPO injection (top) was compared to NaCl injection (bottom) stained for TER119 (violet) and Hoechst (white). Scale bar: 100 µm. (E) Number of megakaryocyte count in the bone marrow of 6 × 300 IU EPO-treated C57Bl/6J mice (n = 4) was compared to control (6 × 30 µL NaCl injection) (n = 4). (F) Immunofluorescence staining of cross-sections of the bone marrow after 2-week EPO injection (top) compared to NaCl injection (bottom) stained for CD41 (violet) and Hoechst (white). Scale bar: 100 µm. (G) Platelet large cell ratio in peripheral blood of 6 × 300 IU EPO-treated C57Bl/6J mice (n = 10) compared to control (6 × 30 µL NaCl injection) (n = 9). (H) Thrombus weight of 6 × 300 IU EPO-treated C57Bl/6J mice (n = 10) and NaCl-injected control mice (n = 10). (I) Thrombus incidence of 6 × 300 IU EPO-treated C57Bl/6J mice (n = 10) and NaCl-injected control mice (n = 10). NS = nonsignificant, *p < 0.05, **p < 0.01, ***p < 0.001. EPO, erythropoietin; RBC, red blood cell.
  item-44 at level 1: picture
    item-44 at level 2: caption: Fig. 5 Splenectomy does not affect the blood count as well as DVT formation. (A) WBC count in C57Bl/6J mice without treatment (n = 5), 48 hours after induction of DVT (n = 7), 5 weeks after splenectomy (n = 3), and 5 weeks after splenectomy with an additional 48-hour induction of DVT (n = 9). (B) Platelet count in C57Bl/6J mice without treatment (n = 6), 48 hours after induction of DVT (n = 6), 5 weeks after splenectomy (n = 3), and 5 weeks after splenectomy with an additional 48-hour induction of DVT (n = 9). (C) RBC count in C57Bl/6J mice without treatment (n = 6), 48 hours after induction of DVT (n = 6), 5 weeks after splenectomy (n = 3), and 5 weeks after splenectomy with an additional 48-hour induction of DVT (n = 9). (D) Thrombus weight in C57Bl6 wild-type mice without splenectomy (n = 6) and with splenectomy (n = 6) (E) Thrombus incidence in C57Bl/6J wild-type mice without splenectomy (n = 6) and with splenectomy (n = 6). (F) Thrombus weight in EPO-overexpressing Tg(EPO) mice without splenectomy (n = 9) and with splenectomy (n = 6) compared to control WT mice without splenectomy (n = 10) and with splenectomy (n = 11). (G) Thrombus incidence in EPO-overexpressing Tg(EPO) mice without splenectomy (n = 9) and with splenectomy (n = 6) compared to control WT mice without splenectomy (n = 10) and with splenectomy (n = 11). NS = nonsignificant, *p < 0.05, **p < 0.01, ***p < 0.001. DVT, deep vein thrombosis; RBC, red blood cell; WT, wild type.
  item-45 at level 1: section_header: References
  item-46 at level 1: list: group list
    item-47 at level 2: list_item: G Ramsey; P F Lindholm. Thrombos ... ansfusions. Semin Thromb Hemost (2019)
    item-48 at level 2: list_item: M A Kumar; T A Boland; M Baiou.  ... noid hemorrhage. Neurocrit Care (2014)
    item-49 at level 2: list_item: R Goel; E U Patel; M M Cushing.  ... th American Registry. JAMA Surg (2018)
    item-50 at level 2: list_item: C Wang; I Le Ray; B Lee; A Wikma ... venous thromboembolism. Sci Rep (2019)
    item-51 at level 2: list_item: B S Donahue. Red cell transfusio ... ic risk in children. Pediatrics (2020)
    item-52 at level 2: list_item: M Dicato. Venous thromboembolic  ... g agents: an update. Oncologist (2008)
    item-53 at level 2: list_item: C L Bennett; S M Silver; B Djulb ...  cancer-associated anemia. JAMA (2008)
    item-54 at level 2: list_item: E Chievitz; T Thiede. Complicati ... ycythaemia vera. Acta Med Scand (1962)
    item-55 at level 2: list_item: V R Gordeuk; J T Prchal. Vascula ... lycythemia. Semin Thromb Hemost (2006)
    item-56 at level 2: list_item: S Ballestri; E Romagnoli; D Ario ... m: a narrative review. Adv Ther (2023)
    item-57 at level 2: list_item: M L von Brühl; K Stark; A Steinh ... osis in mice in vivo. J Exp Med (2012)
    item-58 at level 2: list_item: G D Lowe; A J Lee; A Rumley; J F ... rgh Artery Study. Br J Haematol (1997)
    item-59 at level 2: list_item: J Vogel; I Kiessling; K Heinicke ... gulating blood viscosity. Blood (2003)
    item-60 at level 2: list_item: F T Ruschitzka; R H Wenger; T St ... ietin. Proc Natl Acad Sci U S A (2000)
    item-61 at level 2: list_item: A Bogdanova; D Mihov; H Lutz; B  ... xpressing erythropoietin. Blood (2007)
    item-62 at level 2: list_item: R E Mebius; G Kraal. Structure a ...  of the spleen. Nat Rev Immunol (2005)
    item-63 at level 2: list_item: M A Boxer; J Braun; L Ellman. Th ... ctomy thrombocytosis. Arch Surg (1978)
    item-64 at level 2: list_item: P N Khan; R J Nair; J Olivares;  ... ytosis. Proc Bayl Univ Med Cent (2009)
    item-65 at level 2: list_item: R W Thomsen; W M Schoonen; D K F ...  cohort study. J Thromb Haemost (2010)
    item-66 at level 2: list_item: G J Kato. Vascular complications ... or hematologic disorders. Blood (2009)
    item-67 at level 2: list_item: E M Sewify; D Sayed; R F Abdel A ... bocytopenic purpura. Thromb Res (2013)
    item-68 at level 2: list_item: M K Frey; S Alias; M P Winter. S ... of thrombosis. J Am Heart Assoc (2014)
    item-69 at level 2: list_item: D Bratosin; J Mazurier; J P Tiss ... acrophages. A review. Biochimie (1998)
    item-70 at level 2: list_item: A T Taher; K M Musallam; M Karim ... ia intermedia. J Thromb Haemost (2010)
    item-71 at level 2: list_item: M Seki; N Arashiki; Y Takakuwa;  ... nt erythrocytes. J Cell Mol Med (2020)
    item-72 at level 2: list_item: M F Whelihan; K G Mann. The role ... thrombin generation. Thromb Res (2013)
    item-73 at level 2: list_item: T Frietsch; M H Maurer; J Vogel; ... tosis. J Cereb Blood Flow Metab (2007)
    item-74 at level 2: list_item: O Mitchell; D M Feldman; M Diako ... hronic liver disease. Hepat Med (2016)
    item-75 at level 2: list_item: Y Lv; W Y Lau; Y Li. Hyperspleni ... nd current status. Exp Ther Med (2016)
    item-76 at level 2: list_item: K F Wagner; D M Katschinski; J H ... xpressing erythropoietin. Blood (2001)
    item-77 at level 2: list_item: C Klatt; I Krüger; S Zey. Platel ... t for thrombosis. J Clin Invest (2018)
    item-78 at level 2: list_item: J Shibata; J Hasegawa; H J Sieme ... uences of erythrocytosis. Blood (2003)
    item-79 at level 2: list_item: E Babu; D Basu. Platelet large c ... unts. Indian J Pathol Microbiol (2004)
    item-80 at level 2: list_item: F Formenti; P A Beer; Q P Croft. ... n-of-function mutation. FASEB J (2011)
    item-81 at level 2: list_item: H M Ashraf; A Javed; S Ashraf. P ... mia. J Coll Physicians Surg Pak (2006)
    item-82 at level 2: list_item: D P Smallman; C M McBratney; C H ... and Military Academies. Mil Med (2011)
    item-83 at level 2: list_item: M Li; X Tang; Z Liao. Hypoxia an ... ability at high altitude. Blood (2022)
    item-84 at level 2: list_item: T P McDonald; R E Clift; M B Cot ... thrombocytopenia in mice. Blood (1992)
    item-85 at level 2: list_item: X Jaïs; V Ioos; C Jardim. Splene ...  pulmonary hypertension. Thorax (2005)
    item-86 at level 2: list_item: J M Watters; C N Sambasivan; K Z ... e state after trauma. Am J Surg (2010)
    item-87 at level 2: list_item: S Visudhiphan; K Ketsa-Ard; A Pi ... boembolism. Biomed Pharmacother (1985)
    item-88 at level 2: list_item: T P McDonald; M B Cottrell; R E  ... production in mice. Exp Hematol (1987)
    item-89 at level 2: list_item: Y Shikama; T Ishibashi; H Kimura ... is in vivo in mice. Exp Hematol (1992)
    item-90 at level 2: list_item: C W Jackson; C C Edwards. Biphas ... ypobaric hypoxia. Br J Haematol (1977)
    item-91 at level 2: list_item: T P McDonald. Platelet productio ... ansfused mice. Scand J Haematol (1978)
    item-92 at level 2: list_item: Z Rolović; N Basara; L Biljanovi ... normobaric hypoxia. Exp Hematol (1990)
    item-93 at level 2: list_item: R F Wolf; J Peng; P Friese; L S  ... atelets in dogs. Thromb Haemost (1997)
    item-94 at level 2: list_item: J K Fraser; A S Tan; F K Lin; M  ... use megakaryocytes. Exp Hematol (1989)
    item-95 at level 2: list_item: H Sasaki; Y Hirabayashi; T Ishib ... ion of receptor mRNAs. Leuk Res (1995)
    item-96 at level 2: list_item: R D McBane; C Gonzalez; D O Hodg ...  thrombi. J Thromb Thrombolysis (2014)
    item-97 at level 2: list_item: M Buttarello; G Mezzapelle; F Fr ...  limitations. Int J Lab Hematol (2020)
    item-98 at level 2: list_item: S Guthikonda; C L Alviar; M Vadu ... tery disease. J Am Coll Cardiol (2008)
    item-99 at level 2: list_item: M S Goel; S L Diamond. Adhesion  ...  polymerized from plasma. Blood (2002)
    item-100 at level 2: list_item: P Hermand; P Gane; M Huet. Red c ... IIbbeta 3 integrin. J Biol Chem (2003)
    item-101 at level 2: list_item: A Orbach; O Zelig; S Yedgar; G B ... ragility. Transfus Med Hemother (2017)
    item-102 at level 2: list_item: C C Helms; M Marvel; W Zhao. Mec ... et activation. J Thromb Haemost (2013)
    item-103 at level 2: list_item: J Villagra; S Shiva; L A Hunter; ...  by cell-free hemoglobin. Blood (2007)
    item-104 at level 2: list_item: S Gambaryan; H Subramanian; L Ke ... O synthesis. Cell Commun Signal (2016)
    item-105 at level 2: list_item: S Krauss. Haptoglobin metabolism in polycythemia vera. Blood (1969)
    item-106 at level 2: list_item: A Vignoli; S Gamba; P EJ van der ... a vera patients. Blood Transfus (2022)
    item-107 at level 2: list_item: J H Lawrence. The control of pol ... of 172 patients. J Am Med Assoc (1949)
    item-108 at level 2: list_item: T C Pearson; G Wetherley-Mein. V ... iferative polycythaemia. Lancet (1978)
    item-109 at level 2: list_item: J F Fazekas; D Nelson. Cerebral  ... hemia vera. AMA Arch Intern Med (1956)
    item-110 at level 2: list_item: D J Thomas; J Marshall; R W Russ ... ebral blood-flow in man. Lancet (1977)
    item-111 at level 2: list_item: A D'Emilio; R Battista; E Dini.  ... on and busulphan. Br J Haematol (1987)
    item-112 at level 2: list_item: J E Taylor; I S Henderson; W K S ...  haemodialysis patients. Lancet (1991)
    item-113 at level 2: list_item: J J Zwaginga; M J IJsseldijk; P  ... y uremic plasma. Thromb Haemost (1991)
    item-114 at level 2: list_item: F Fabris; I Cordiano; M L Randi. ...  haemodialysis. Pediatr Nephrol (1991)
    item-115 at level 2: list_item: T Akizawa; E Kinugasa; T Kitaoka ... emodialysis patients. Nephron J (1991)
    item-116 at level 2: list_item: G Viganò; A Benigni; D Mendogni; ... remic bleeding. Am J Kidney Dis (1991)
    item-117 at level 2: list_item: J A Rios; J Hambleton; M Viele.  ... tivation treatment. Transfusion (2006)
    item-118 at level 2: list_item: S Khandelwal; N van Rooijen; R K ... om the circulation. Transfusion (2007)
    item-119 at level 2: list_item: G J Bosman; J M Werre; F L Wille ... s for transfusion. Transfus Med (2008)
    item-120 at level 2: list_item: W H Crosby. Normal functions of  ... ed blood cells: a review. Blood (1959)
    item-121 at level 2: list_item: R Varin; S Mirshahi; P Mirshahi. ... cacy of rivaroxaban. Thromb Res (2013)
    item-122 at level 2: list_item: F A Carvalho; S Connell; G Milte ... on human erythrocytes. ACS Nano (2010)
    item-123 at level 2: list_item: N Wohner; P Sótonyi; R Machovich ... . Arterioscler Thromb Vasc Biol (2011)