bluuebunny/crossref_metadata_2025_split huggingface dataset:
Plasma Thromboplastin Antecedent (PTA) Deficiency: Clinical, Coagulation, Therapeutic and Hereditary Aspects of a New Hemophilia-like Disease
Authors: ROSENTHAL ROBERT L. , DRESKIN O. HERMAN , ROSENTHAL NATHAN 
Published: October 2019

Abstract
1. An analysis of the original PTA deficient family, including coagulation studies performed upon 13 members comprising 4 generations, has been presented.
2. PTA deficiency is transmitted as an autosomal dominant trait with a probable high degree of penetrance and variable expression of the gene.
3. PTA deficiency can occur its varying degrees ranging from a severe form with prolonged clotting time and markedly abnormal heparin clotting time and prothrombin utilization to a mild form manifesting a normal clotting time and slightly impaired prothrombin utiliztition.
4. Studies on the treatment of PTA deficiency reveal that the defect is corrected by the administration of stored plasma with the effect gradually disappearing over the period of one week.
5. Various properties of PTA are discussed and compared with AHG and PTC.
Splenic Aspiration in Clinical and Experimental Hematology
Authors: WATSON R. JANET , SHAPIRO HENRY D. , ELLISON ROSE RUTH , LICHTMAN HERBERT C. 
Published: October 2019

Abstract
1. The findings obtained by the study of splenic aspirations in 140 patients are presented. Diagnostically they were most helpful in myeloid metaplasia, lymphoma, kala-azar, and Gaucher’s disease.
2. The findings were of particular theoretical interest in multiple myeloma and sickle cell anemia. This method of investigation could be extended to elucidating further the role of the spleen in other diseases.
3. No morbidity was encountered in this series. If certain precautions are observed, splenic puncture is a safe procedure, and should be undertaken in patients with undiagnosed splenomegaly.
Acquired Circulating Anticoagulants in Systemic "Collagen Disease"
Authors: FRICK PAUL G. , WEIMER MARY K. 
Published: October 2019

Abstract
1. A circulating anticoagulant was demonstrated in three patients with "collagen disease." In all cases it was associated with a false positive blood serology and a four plus cephalin cholesterol flocculation.
2. In one instance, the anticoagulant was transferred to a newborn infant where it persisted for seven weeks. The infant also demonstrated the abnormal serologic and turbidimetric tests during the first six months of life.
3. The anticoagulant probably acted as anti-thromboplastin.
4. The occurrence of a clotting inhibitor in association with other manifestations of hypersensitivity and its transplacental transfer strongly suggest that the mechanism of development is immunologic in type.
5. This study included a total of thirty patients with lupus erythematosus disseminatus and associated conditions conventionally grouped under the term "collagen disease." The incidence of circulating anticoagulants was 10 per cent.
Mutations in the Gene for Neutrophil Elastase in Severe Congenital Neutropenia Are Heterozygous and of Male Origin.
Authors: Stein Steven , Paddock Marcia , Bolyard Audrey Anna , Raymond Chris , Olson Maynard , Aprikyan Andrew , Dale David C. 
Published: October 2019

Abstract
Severe congenital neutropenia (SCN) occurs sporadically and as an autosomal-dominant or autosomal-recessive inherited disorder. Most sporadic and autosomal dominant cases are caused by mutations in the gene for neutrophil elastase (NE or ELA2). The gene for this potent serine protease is located on chromosome 19p13.3 and has 5 exons. In collaboration with the Severe Chronic Neutropenia International Registry (SCNIR), we have sequenced the NE gene from blood, bone marrow or other tissues samples for more than 300 patients with the clinical diagnosis of SCN using Big Dye Terminator, version 3.1 and ABI 3700. These studies have included 21 families with 2 or more affected members and 15 families with sporadically occurring SCN, i.e., two healthy parents with normal blood counts having an affected child. In 15 families with sporadic cases of SCN attributable to 13 different mutations in the NE gene, we sought to determine the parent of origin for the NE mutation. The 5 exons of the NE gene were sequenced twice in the parents and the affected child using Big Dye Terminator, version 3.1 and ABI 3700. After confirming the mutation in each patient and the absence of mutations in each parent, PCR was used to scan the adjacent region to the NE gene in the parents and the child to identify polymorphisms that could be used to distinguish between maternally and paternally inherited chromosomes. A fosmid library of 2-4X coverage for each patient’s DNA sample was made which successfully separated the patient’s genome into its constituent’s haplotypes. The fosmids containing the NE gene were identified by PCR and genotyped at the site of the mutant allele and the parent-informative marker. In all cases, the new NE gene mutation in the affected child was on the paternal allele.
Combination of the mTOR Inhibitor Rapamycin and Revlimid™ (CC-5013) Has Synergistic Activity in Multiple Myeloma (MM).
Authors: Raje Noopur , Kumar Shaji , Hideshima Teru , Ishitsuka Kenji , Chauhan Dharminder , Mitsiades Constantine , Podar Klaus , Le Gouill Steven , Richardson Paul , Munshi Nikhil C. , Stirling David I. , Antin Joseph H. , Anderson Kenneth C. 
Published: October 2019

Abstract
Interleukin-6 (IL-6) and insulin-like growth factor-1 (IGF-1) mediate growth of MM cells via activation of the mitogen-activated protein kinase (MAPK), JAK2/STAT3, and phosphatidylinositol 3′-kinase/Akt kinase (PI3-K/Akt) signaling cascades. We have previously demonstrated the in vitro and in vivo activity of Revlimid™ (CC-5013), an immunomodulatory analog (IMiD) of thalidomide, in MM. In the present study, we have examined the anti-MM activity of rapamycin, a specific mTOR inhibitor, combined with Revlimid™. This combination was highly synergistic at 0.1nmol/L of rapamycin with 0.1mmol/L of Revlimid™, and remained synergistic at higher concentrations. Based on the Chou-Talalay method, combination indices of < 1 were noted for all dose ranges of Revlimid™ and rapamycin, suggesting strong synergism. Importantly, this combination was able to overcome drug resistance when tested against MM cell lines resistant to conventional (doxorubicin, melphalan, dexamethasone) chemotherapy. Moreover, the combination, but not rapamycin alone, was able to overcome the growth advantage conferred on MM cells by IL-6, IGF-1, or adherence to bone marrow stromal cells (BMSCs). Cytotoxicity triggered by a combination of rapamycin with Revlimid™ resulted in apoptosis of MM cells. Furthermore, differential signaling cascades, including the MAPK and PI3-K/Akt pathways, were targeted by these drugs individually and in combination, suggesting the molecular mechanism by which they interfere with MM growth and survival. These studies therefore provide the framework for the clinical evaluation of targeted agents like mTOR inhibitors combined with immunomodulatory agents like Revlimid™ to improve patient outcome in MM.
The Natural History of Idiopathic Erythrocythosis: A Cohort Study of 74 Patients.
Authors: Guido Finazzi , Marco Ruggeri , Monica Marconi , Francesco Rodeghiero , Tiziano Barbui 
Published: October 2019

Abstract
Patients with absolute erythrocytosis not due to a detectable cause and not fulfilling the criteria for diagnosis of polycythemia vera (PV) are descriptively classified as Idiopathic Erythrocytosis (IE). Based on scanty and retrospective data, this disease is considered to be an heterogeneous entity, including “early” PV, unrecognized secondary erythrocytosis and other miscellaneous conditions. However, appropriate prospective studies to evaluate the natural history of patients with IE are not available. We report here the results of a cohort study of 74 patients with IE (66 males, 8 females, median age 56 years, range 14–82) followed in two Italian institutions. By definition, at baseline all IE patients had increased hematocrit (median 54%, range 48–68%) and increased red blood cell mass (> 25% above mean normal predicted value), but normal leukocyte (median values 8.1 x 109/L, range 2.3–12) and platelet counts (median values 197 x 109/L, range 117–467), as well as normal erythropoietin level, arterial O2 saturation, chest X ray and abdominal ultrasound scanning (i.e. no splenomegaly). Granulocyte PRV-1 expression was also normal in 29 patients (39%) analyzed. At diagnosis, 12 patients (16%) reported a previous history of major thrombosis (7 ischemic cardiopathies, 4 cerebral ischemic events and 1 deep vein thrombosis). All IE patients were treated with phlebotomy to maintain a target hematocrit <45% and 24 patients (32%) were given aspirin, 100 mg/die, for previous thrombosis or microvascular symptoms. No cytotoxic drugs were given. The IE cohort was followed in the outpatient clinic with physical examination and full blood count at least every three months for a median period of 3.5 years (range 1–23). Twentythree patients (31%) were followed for more than 8 years. No patient was lost to follow-up. During the observation period, no disease potentially associated with secondary eryhtrocytosis emerged and no hematological transition into overt PV, myelofibrosis or acute leukemia occurred; two patients had a major thrombotic event (1 cerebral ischemia and 1 deep vein thrombosis) with an estimated incidence of thrombotic complications of 0.8% patient-year. The incidence of thrombosis was significantly lower than observed in 205 patients with overt PV followed during the same period in one of the two institutions (Bergamo, 3.49% patient-year, p<0.05). This study indicates that: a. the natural history of patients with IE, at least in the first years, is characterized by a remarkable and unexpected homogeneity without appearance of overt PV or diseases associated with secondary erythrocythosis; b. the diagnosis of IE identifies a group of absolute erythrocythoses at lower risk of thrombotic complications not requiring cytotoxic drug therapy; c. the diagnostic work-up of patients with absolute erythrocythosis should carefully distinguish IE from PV because the natural history and management of the two diseases is different.
The Hox a Locus Is Strongly Targeted by MMLV in an E2a-PBX1 Induced B-Cell Leukemia Model.
Authors: Bijl Janet J. , Sauvageau Martin , Thompson Alex , Sauvageau Guy 
Published: October 2019

Abstract
We previously developed a B-ALL model from E2a-PBX1 transgenic mice (Bijl et al. Abstract 469 ASH 2003 and paper in preparation). We now exploit this model to identify oncogenic collaborators to E2a-PBX1. To achieve this, 18 newborn E2a-PBX1 transgenic mice and 23 control littermates were intraperitonally injected with MMLV. The occurrence of B-ALL in E2a-PBX1 transgenic animals was significantly accelerated when compared to control littermates (mean survival 162 ± 31 versus 191 ± 50 days, respectively, P=0.03), suggesting the presence of E2a-PBX1 specific collaborators. Inverse PCR was performed on genomic DNA isolated from seven E2a-PBX1 and six control tumors. Seventy-two different retroviral insertion sites were recovered from the tumors. Six common integration sites (CIS) for MMLV were identified; including two novel CIS, i.e. Pde4d, a phosphodiesterase, and a hypothetical phospholipase A930027K05Rik. Strikingly, the Hox a locus was targeted in six of seven E2a-PBX1 tumors, and only one of six control tumors. MMLV integrants in the Hox a locus were detected in a 19kb region located between Hoxa6/Hoxa7 and Hoxa10. Two tumors were targeted twice in this locus. Q-PCR analysis for expression of all Hox a cluster genes identified a significant increase for the Hoxa3 to Hoxa10 genes in the transgenic tumors when compared to the expression in a control MMLV induced B-cell tumor (see table, below). Hoxa7 and Hoxa6 were the most frequently and strongly activated (6/6 and 5/6 transgenic tumors, respectively), showing values of 5700 and 6600 fold over expression in one particular tumor. Interestingly, a Hox a locus insertion in a control tumor (table: last row) did not have any effect on gene activation. In this study we demonstrate a preference for MMLV to target the Hox a locus in an E2a-PBX1context. The resultant gene activation suggests that several members of the Hox a locus are functional collaborators to E2a-PBX1 in the induction of B-cell leukemia.
Fold difference in expression for Hox a cluster genes in Hox a targeted tumors compared to a control tumor.
Insertions a3 a4 a5 a6 a7 a9 a10 a9/a10 −1 1 4 3 138 2 4 a3 17 55 1067 5700 6608 171 62 a10 3 1 11 98 38 3 −4 a9/a10 1 −1 2 21 9 2 −2 a9/a10 2x 13 6 19 221 191 85 2 a7/a9 + a10 8 4 11 66 44 9 149 a10 −2 −6 2 2 −2 −8 −5
GPIbα Dimer Formation and Cell Surface Expression: A Filamin Binding Domain Is Essential for Dimer Formation but Not for Assembly of a GPIb/IX Complex.
Authors: Kanaji Taisuke , Hamasaki Naotaka , Izuhara Kenji , Ware Jerry 
Published: October 2019

Abstract
The hereditary bleeding disorder Bernard-Soulier syndrome (BSS) is caused by an absent or dysfunctional GPIb/IX complex. Lopez et al. have reported that three subunits of the complex (GPIbα, GPIbβ and GPIX) are all necessary for efficient translocation to the cell surface. In contrast, Meyer et al. have demonstrated that GPIbα alone can incorporate into the cell surface. We have established a cell culture system with inducible levels of GPIbβ and GPIX. Using these cell lines, we characterize an intrinsic ability of GPIbα to form dimers and the relevance of specific GPIbα sequences for dimer formation and the assembly of a GPIb/IX complex.
A stable CHO cell line was established that expresses GPIbβ and GPIX under the control of Tet-responsive elements. This cell line was then transfected with WT GPIbα and truncated GPIbα mutants. Expression was monitored by Western blot analysis and flow cytometry.
We observed that expression of GPIbα, by itself, targets GPIbα to the cell surface as a dimer albeit at reduced levels. The GPIbα binding domain for filamin (residues 545–605) is essential for forming a dimer but not for assembly of a GPIb/IX complex. We also observed GPIbα dimer formation in a transgenic mouse that expresses human GPIbα and a mouse chimeric GPIb-IX complex suggesting dimer formation can occur in the platelet. To investigate the role of filamin in the dimerization process, we established a stable cell line deficient in filamin by targeting filamin expression with RNAi. We observed GPIbα dimers in the absence of filamin expression. These results illustrate the importance of expressing all three subunits of the GPIb/IX complex, but also document as intrinsic potential within GPIbα for surface expression via intermolecular homodimerization.
ATP Release from Red Blood Cells Is Regulated by a Negative Feedback Pathway where ADP Acts on P2Y13 Receptors.
Authors: Olsson Martin L. , Wang Lingwei , Olivecrona Goran , Gotberg Matthias , Amisten Stefan , Winzell Maria Sorhede , Erlinge David 
Published: October 2019

Abstract
Background: Red blood cells regulate tissue circulation and O2 delivery by releasing the vasodilator adenosine triphosphate (ATP) in response to hypoxia. When released extracellularly, ATP is rapidly degraded to adenosine diphosphate (ADP) in the circulation by ectonucleotidases. ATP and ADP activate subtypes of the large P2 receptor family (15 subtypes). Here we show that ADP acting on P2Y13 receptors on red blood cells serves as a negative feedback pathway for the inhibition of ATP release.
Methods: mRNA was quantified with real-time PCR. Western blot was used to detect P2 receptors with available antibodies. cAMP levels were determined with an enzyme immunoassay. ATP release was measured in incubated red blood cells using microdialysis and a luciferase assay. In a pig model, catheters were inserted through the carotid artery to place a catheter in the left coronary artery, and through the jugular vein to place a microdialysis probe in the coronary vein. 2-MeSADP was injected in the artery and ATP levels were measured in the coronary vein.
Results: mRNA of the ADP receptor P2Y13 was highly expressed in human red blood cells and reticulocytes, whilst other ADP receptors were not (Fig.1).
Figure Figure
The stable ADP analogue 2-MeSADP decreased ATP release from red blood cells by inhibition of cAMP. The P2Y12 and P2Y13 receptor antagonist AR-C67085 (30 mM), but not the P2Y1 blocker MRS2179, inhibited the effects of 2-MeSADP. At doses where AR-C67085 only blocks P2Y12 (100 nM), it had no effect. AR-C67085 and the nucleotidase apyrase increased cAMP per se, indicating a constant cAMP inhibitory effect of endogenous extracellular ADP. 2-MeSADP reduced plasma ATP concentrations in an in vivo pig model. Furthermore, a missense polymorphism in the coding region of P2Y13 has been found that is in total disequilibrium with 5 polymorphisms in P2Y12 (the important ADP receptor in platelets) forming a haplotype that could contribute to vascular disease.
Conclusion: Our results show that P2Y13 is selectively expressed in human red blood cells. The ATP degradation product ADP inhibited ATP release by acting on this receptor. This negative feedback system could be important in the control of plasma ATP levels and tissue circulation. Because blood consists of approximately 40% red blood cells, containing a 1000-fold higher ATP concentration than plasma (mM vs. uM), even a minor release of ATP from the high intracellular concentrations could have major circulatory effects. A negative system may therefore be of great physiological importance to mitigate ATP release. In addition, this finding could be of interest for efforts to preserve intracellular ATP in red blood cells during storage.
Hypoxic Vasodilation by Red Blood Cells and Impairment in Vascular Disorders.
Authors: Pawloski John R. , McMahon Timothy J. , Ahearne Greg , Piantadosi Claude A. , Singel David J. , Stamler Jonathan S. 
Published: October 2019

Abstract
Physiological O2 gradients are principal regulators of blood flow in the microcirculation: position-to-position changes in hemoglobin (Hb) O2 saturation are coupled to regulated vasodilation (“hypoxic vasodilation”). The mechanism by which graded changes in O2 content of blood evoke this response has been a great challenge to understand. A new role for red blood cells (RBCs) in hypoxic dilation of blood vessels and inhibition of platelet activation involving release of nitric oxide (NO) bioactivity is described. We show that NO groups can be transferred within hemoglobin (Hb) from hemes to highly-conserved cysteine thiols (β-Cys93) to form bioactive S-nitrosohemoglobin (SNO-Hb), and that efficient production of SNO-Hb requires selective processing of NO within the β-subunits. Bioactive SNO-Hb is localized primarily to the RBC membrane through interaction with Band 3, the transmembrane anion-exchanger 1 protein (AE1). Upon deoxygenation, transfer of the NO group from β-Cys93 of Hb to a cysteine thiol within AE1 serves the RBC vasodilator activity. In this way, O2 binding in Hb modulates the release of NO bioactivity. We further show that RBC NO bioactivity is inversely proportional to pO2 and impaired in disease. In an aortic ring bioassay sparged with variable concentrations of O2, addition of normal human RBCs elicited graded responses from relaxation at tissue pO2 (~3–7 mm Hg, hypoxic vasodilation), to loss of relaxation and progressively greater contractions at pO2’s of 10–63 mm Hg (hyperoxic vasoconstriction). Notably, RBC SNO-Hb levels and hypoxic vasodilation are impaired in several diseases characterized by vascular dysfunction. For example, in RBCs from patients with pulmonary arterial hypertension (PAH), we found decreased (13% of control) SNO-Hb content (assessed by photolysis-chemiluminescence) and impaired O2-dependent vasodilation (bioassay). RBCs from patients with other ischemic disorders have also been examined: RBCs demonstrate a pathogenesis-based impairment in their ability to mediate hypoxic vasodilation by NO. These results confirm the (patho)physiologic importance of RBC NO, and suggest that RBC dysfunction may contribute to impaired blood flow in diseases of the heart, lung and blood.
Regulation of SCL Expression by the Homeodomain Protein Otx-1 and the Erythroid Transcription Factor GATA-1.
Authors: Martin Richard , Sanguin-Gendreau Virginie , Tremblay Mathieu , Levantini Elena , Magli Christina , Hoang Trang 
Published: October 2019

Abstract
Members of the bicoid homeodomain-containing proteins are important in establishing left-right asymmetry and the antero-posterior axis, suggesting that they could also be involved in asymmetric determination within the hematopoietic system. We have previously shown that Otx1, a member of the bicoid homeodomain-containing proteins, is co-expressed with the SCL transcription factor in hematopoietic pluripotent and erythroid progenitor cells and Otx1-deficiency impairs the erythroid compartment in mice, associated with decreased SCL levels. In the present study, we provide molecular and functional evidence that SCL is a direct transcriptional target of Otx1. First, we show by chromatin immunoprecipitation that Otx1 and GATA-1 are specifically bound to the SCL proximal promoter in erythroid cells. Second, Otx-1 synergizes with GATA-1 to activate transcription from the SCL proximal promoter and this activity depends on the integrity of the proximal GATA site of the SCL promoter 1a. At the molecular level, we show that this synergy occurs via a physical interaction between Otx-1 and GATA-1 in erythroid cells, which maps to the homeodomain of Otx-1. Furthermore, a gain of function of Otx1 in primary hematopoietic cells gives rise to a 6-fold increase in endogenous SCL levels, an increase in TER119-positive erythroid cells and a decrease in the number of CD11b-positive myeloid cells. Finally, a gain of function of SCL rescues the erythroid deficiency in Otx1−/− mice, consistent with the view that SCL operates downstream of Otx1. Taken together, our observations indicate that Otx1, GATA-1 and SCL operate within the same genetic pathway to specify the erythroid fate during hematopoiesis.