bluuebunny/crossref_metadata_2025_split huggingface dataset:
Actual Status of Inhibitor Hemophilia Management in Russia
Authors: Zozulya Nadezhda , Andreeva Tatiana , Vdovin Vladimir , Perina Farida , Parovichnikova Elena N. , Savchenko Valeri 
Published: October 2019

Abstract
Background: The number of inhibitor hemophilia patients in Russia is presently about 200 patients (4% of all hemophilia patients). Current medical community is moving towards increasing the efficacy and safety of treatment, as well as the standardization of services and improvement of standards of care in rare diseases area. Patient registries and databases are key tools for the improvement of patient care, healthcare resources planning as well as social, economical and quality of life outcomes. Due to the lack of data on clinical course of disease, treatment approaches and outcomes, National Hematology Society initiated continuously running prospective, multi-center, open-label, non-interventional, observational Registry of INhibitor Hemophilia patients (RING) in July 2014.
Aims: to document the natural history of hemophilia A or B disease in subjects with any type of inhibitors and to describe long-term treatment related outcomes in routine clinical practice.
Methods: Data of all inhibitor patients are collected by physicians from patients medical records documenting the routine patient's examination and reflecting the medical practice at the particular treatment center. All data are entered into e-database using on-line access secured by the Operator.
Results: In total 183 patients (101 adult and 82 children) from 47 centers were included in the Registry by June 2015. The majority of patients (90%) have severe hemophilia. The age of inhibitors diagnostics was: ≤10 years - 83%, 11-20 years - 3%, 21- 50 years - 5%, ≥51 years - 3% of patients. Chronic viral infections are diagnosed in 55 (30%) patients: 1 - HIV, 5- HBV and 49 - HCV. In 28% of patients annual bleeding rate (ABR) are ≥ 16, in 42% - 6-15, in 21% - ≤ 5. Approximately 25% of patients have zero joint ABR. Life threatening bleedings were registered in 25 patients (14%). Only 15 (8%) patients undergo immune tolerance induction (ITI); treatment with bypassing agents is administered in 151 (82%) patients. Due to the fact that activated prothrombin complex concentrate (aPCC) is not supplied by the State guarantees program 83% of patients receive rFVIIa (on-demand treatment - 50% and prophylaxis - 33%) and only 17% of patients have an access to aPCC treatment. The current data show that 34 (27%) patients with rFVIIa treatment need ≥3 injections to stop a bleeding episode thus they are identified as non-responders.
Conclusion: The clear view of current treatment situation based on available Registry data demonstrates the necessity of administering alternative treatment approaches for non-responding to rFVIIa inhibitor patients, which would optimize resources of the State guarantees program.

Disclosures
No relevant conflicts of interest to declare.
Lysosomal Newborn Screening in a Cohort in Mexican Population-
Authors: Navarrete Juana Ines 
Published: October 2019

Abstract
INTRODUCTION: The goal of newborn screening is an early detection of inborn erros of metabolism diseases. In Mexico we began newborn screening since 1977 with very few inborn errors of metabolism such as phenylketonuria, galactosemia, congenital hypothyroidism, sickle cell anemia and cytic fibrosis (1). Since that date we have been increasing our newborn screening our newborn screening slowly and now a days we screen in most states of the country 15 inborn errors of metabolism(2). In 2012 we started with some patients through out the country a wider neonatal screening that include 5 lysosomal storage diseases.
MATERIAL AND METHODS: Petróleos Mexicanos is a big governmental institution with approximately 10,000 workers and their families. Since 2005 a larger newborn screening has been done to all newborns in this institution through all the country. We test for most aminoacidopathies, including acidurias, hemoglobinopathies, G6PD deficiency, adrenal hyperplasia, cystic fibrosis and biotinidase deficiency; since August 2012 we included 6 lysosomal storage diseases; Gaucher disease, Fabry disease, Hurler disease, Pompe disease, Niemann-Pick type A and B disease and Krabbe disease.
RESULTS: Up to date we have screened 10,853 newborns, we have found 9 patients with lysosomal storage diseases. We found 4 newborns with mutations for Fabry disease, 4 newborns with Pompe disease, three were pseudodeficiencies and one was combined heterozygous for a late onset presentation and pseudodeficiencies and 1 patient with Hurler disease (Table 2). We present here our clinical correlation between genotype-phenotype in these patients. We found a frequency in our population of 1 in 2713 newborns for both Fabry and Pompe disease.
DISCUSSION: Newborn screening is a major public health achievement that has improve the morbidity and mortality of inborn errors of metabolism. The introduction of newborn screening for lysosomal storage diseases presents new challenges. This is the first latinamerican study of early detection of lysosomal storage diseases made by neonatal screening there are about 11 similar international studies. It is important point out that the most common lysosomal storage disease found in our study was Pompe diseases the pseudodeficiency type and Fabry disease type II with a frequency of 1 in 2713 newborns for both diseases. Spada et al; and Hwu et al; have reported frequencies of 1 in 1250 to 3100 male newborns. The mutation most commonly found was c.1088G>A, (p.R363H) for Fabry disease and c.1726G>A(p.G576S) for Pompe disease.
References: 1. Nakamura K, Am J Med Genet Part C, 2011; 157, 63-71. 2. Zhou et al, J. Pediatr 2011 159 1 7-13. 3. Alterescu GM, Clin. Genet 2001:60:46-51. 2001. 4. Desnick R. J.: Enzyme Replacement Therapy and Enhancement therapies for Lysosomal Storage Diseases. J. Inher Metab Dis 2004; 27:385-4013.

Disclosures
No relevant conflicts of interest to declare.
Acute Promyelocytic Leukemia, Are US Doctors and Hospitals Prepared? Tools and Know How, Yes; But, What about Readiness?
Authors: Morris Daniel 
Published: October 2019

Abstract
Acute Promyelocytic Leukemia (APL) is a hematologic emergency or urgency. It is a potential disaster that is also potentially curable. My research attempts to ascertain the readiness to care for patients who present with this (suspected) diagnosis and require urgent therapy.
I polled a sample of approximately 140 (United States) hematologist-oncologists as to the availability of Tretinoin/all-trans retinoid acid (ATRA) on a stat basis at the hospitals at which they practice and 18 responded. In other words, is the medicine on the shelf and can be given when ordered.
Available on the shelf and ready to give ATRA at all hospitals in which practiced: 2/18
Do not know: 3/18
Not available at all hospitals practiced: 13/18
Amazingly, one of the physicians stated that the family is given a prescription to get ATRA filled, and another responded that he had to "beg" to get the hospital to obtain it.
I posted two questions on a social media site in the Emergency Medicine category; This social media web site states that the total physician membership is 343,000 (verified). The responses are as follows:
Is ATRA on the shelf and available at the hospital they practice (17 responses to this question)
13/17 Do not know
2/17 On the shelf and available
2/17 Not on the shelf and would need to be ordered
Is APL a possible question for board exam in Emergency Medicine? (16 responses to this question)
9/16 No
4/16 Not sure
3/16 Yes
The price for ATRA according to www.uptodate.com is (Tretinoin Oral) 10 mg (100): $2988.16. Issues related to this topic include the cost, the supply (sold in quantities of 100), shelf life/expiration date, the agggregate cost to maintain it at every hospital based ER , and consideration that the patients are not all acutely ill and sometimes can wait a day before starting therapy. Even if this approach of not having ATRA in stock is not optimal, this approach is occurring.
Whether or not Joint Commission or the more voluntary Commission on Cancer should require that ATRA be on the shelf at a hospital, or available within a certain amount of time may merit further discussion.
Finding out how prepared we are to treat APL is not an easy undertaking and my effort seems to indicate that we probably do not have the wherewithal in many cases. Obtaining more data on the scope of the problem and perhaps devising a "Practice Guideline" by experts in the APL area dealing with the timeliness of availability of ATRA would seem to be in order. The lifesaving nature of the intervention would seem to justify the consideration of having hospitals maintaining a supply at all times. If the manufacturer would be willing to sell a supply of #30 (currently available in bottles of 100 per Facts and Comparison 2015 edition), this might be useful in implementing wider availability.

Disclosures
No relevant conflicts of interest to declare.
Exposure to New Therapies and Treatment Delay in Multiple Myeloma: Experience from a Safety Net Hospital
Authors: O'Brien Timothy , Ruda Barbara , Cluckey Andrew 
Published: October 2019

Abstract
Introduction:
Treatments for myeloma have evolved over the past several years with resultant improvements in survival. Two new medications, bortezomib (B) and lenalidomide (L), have played a major role in this development. Each has received FDA approval in the past 10 years. Both medications are relatively expensive and, due to its teratogenic effects, lenalidomide requires a pt training and education program before pharmacies may fill the prescription. In addition, specialty pharmacies need to be identified and prior approval is required from the insurance carrier. This study addresses whether or not a relatively underinsured, indigent population was able to receive these medications and in a timely fashion.
Methods:
Electronic medical records of all pts diagnosed with myeloma from 1/2005-12/2014 at MetroHealth Medical Center (a large safety net institution in Cleveland, OH) were reviewed.
88 pts were identified. Med age was 59 (35 -87); 50% were African American (AA), 43% caucasian (C), 1% Hispanic and 6% unknown. Health insurance status: 26% private; 35% medicare; 17% medicaid; and 22% were uninsured.
Results:
77% AA received either B or L or both vs 82% C pts (NS).
Overall, 17/88 (19%) never received either L or B. This included: 5 due to pt preference or co-morbidities which precluded treatment,; 2 who died before they could receive any therapy; 3 lost to follow-up before treatment could be started; 6 who received thalidomide and then either transferred care elsewhere, died or did not require another treatment; and 1 who transferred to another facility for stem cell transplant evaluation before treatment was initiated. Therefore, all 71 pts who were candidates and w/treatment data available received B or L. Of the 17 who did not get B or L: 6 were uninsured, 5 private insurance, 4 medicare, and 2 medicaid.
Overall, 66 pts received B; 31 received L.
Ave time from prescription to pt start of medication: 19.1 days for L (med 14.5d, range 0-65d) vs 1.56d for B (med 0d, range 0-20d), p<0.00001. The pts treated with L were not overrepresented from any particular insurance status (29% were uninsured; 19% medicaid; 29% medicare; 23% private). This payor mix was similar for those who received B (21% uninsured; 13% medicaid, 36% medicare and 30% private).
Conclusions:
In this underinsured, indigent population, all appropriate pts with myeloma received at least one of the 2 novel drugs. There was a statistically significant delay in treatment for those pts who were prescribed lenalidomide as compared to bortezomib (19.1 days L vs 1.56 days for B). It is not clear that this is a clinically significant difference but suggests that bortezomib containing regimens may be a better option in cases where urgent therapy is needed, particularly in this population.

Disclosures
No relevant conflicts of interest to declare.
COL4A1 is a Novel Causative Gene Responsible for Congenital Hemolytic Anemia, Representing Characteristic Clinical Course in Infants
Authors: Ogura Hiromi , Ohga Shouichi , Aoki Takako , Utsugisawa Taiju , Takahashi Hidehiro , Iwai Asayuki , Watanabe Kenichiro , Okuno Yusuke , Yoshida Kenichi , Miyano Satoru , Ogawa Seishi , Kojima Seiji , Kanno Hitoshi 
Published: October 2019

Abstract
We have been working on the differential diagnosis of congenital hemolytic anemia, but even with extensive analysis of hemoglobin, red cell membrane and enzymes, approximately 40% of patients remained to be diagnosed. In this study, we analyzed 17 undiagnosed hemolytic anemia subjects under the age of 1 by whole-exome sequencing, and identified COL4A1 gene mutations in 5 cases (29.4%). All patients were de novo cases without family histories and exhibited moderate to severe neonatal hemolytic anemia: Hgb, 5.2-9.3 g/dl; MCV, 90.0-126.9; MCHC, 29.9-32.7; and reticulocyte count, 9.2-33.0%. Either schizocytes or poikilocytes were observed in peripheral blood smears of 3 cases, suggesting that the microangiopathy was attributable to hemolysis. Previous reports showed that mutation of COL4A1 accounts for brain small-vessel disease characterized by stroke and eye abnormalities and the most characteristic complications of the present cases were congenital anomaly in the central nervous system, such as porencephaly, schizencephaly, congenital hydrocephalus, cataracts or paraventricular calcification, as reported previously. Hemolytic anemia became less severe within 2 months after birth, and all cases no longer required red cell transfusion after Day 50. COL4A1 encodes subtype 1 of type IV collagen, which is most abundantly expressed in basement membranes, including the vasculature. The COL4A1 gene mutations identified in the cases were all novel missense mutations except one, located in exons 26, 27, 37, 38 and 51. Although the pathophysiological significance of the mutations remains unclear, COL4A1 is the first identified causative gene responsible for congenital hemolytic anemia without intrinsic defects of red blood cells, and mutation of COL4A1 is the most prevalent cause of neonatal hemolytic anemia.

Disclosures
No relevant conflicts of interest to declare.
Deep Proteomic Analysis of Human Erythropoiesis
Authors: Ducamp Sarah , Gautier Emilie-Fleur , Leduc Marjorie , Salnot Virginie , Dussiot Michael , Guillonneau François , Giarratana Marie-Catherine , Douay Luc , Lacombe Catherine , Verdier Frederique , Zermati Yael , Mayeux Patrick 
Published: October 2019

Abstract
*the first two authors are co-first authors
Introduction. Erythropoiesis is a complex process starting from pluripotent medullary progenitors and leading to the production of highly specialized and enucleated erythrocytes. Two successive phases are generally distinguished: an amplification phase with intense proliferation of morphologically similar progenitors and a terminal differentiation phase with few cell divisions and strong cellular modifications. Although erythropoiesis is a continuous process, these modifications allow the identification of specific maturation stages and the passage from one stage to the following one seems to correlate with a cell division. Several transcriptomic analyses of erythroid differentiation have been published but only few and very limited proteomic studies have been reported. Since post transcriptomic modifications are responsible for a large part of the proteome variations, a direct proteomic analysis of the erythroid differentiation is required to accurately assess the modifications that occur during this process.
Results. For this study, we used CD34+ cord blood progenitors and an optimized three step cell culture method allowing the production of highly synchronized cell populations of erythroid cells at various differentiation stages. Several cellular populations from erythroid progenitors up to reticulocytes were analyzed by a label-free analysis and mass spectrometry that led to the absolute quantification of more than 6000 proteins with a false discovery rate of less than 1% (n=3). Moreover, the relative expression of well-known stage-specific erythroid markers such as TFRC, BAND3 or GLUT1, transcription factors, heme biosynthesis enzymes followed the expected pattern.
To complete this study, we performed a quantitative analysis of the repartition of proteins between the generated reticulocytes and the expelled nucleus (pyrenocyte). To do that, pyrenocytes and reticulocytes were sorted by FACS according to size, Hoechst 33342 and glycophorin A labelling. Equal numbers of reticulocytes and pyrenocytes were used to prepare peptides that were analyzed by mass spectrometry after iTRAQ labelling. These experiments allowed the quantitative repartition of 1153 proteins including most erythrocyte-specific membrane proteins.
Conclusion. All these results significantly increase our knowledge of the protein expression pattern during erythropoiesis and should constitute a valuable data base for subsequent studies regarding both physiological and disordered erythropoiesis.

Disclosures
No relevant conflicts of interest to declare.
Induction of Fetal Hemoglobin and Reduction of Disease Pathology in Sickle Cell Mice By a Synthetic Zinc Finger Gamma-Globin Activator
Authors: Peterson Kenneth R , Makala Levi C , Takezaki Mayuko , Barbas Carlos F , Pace Betty 
Published: October 2019

Abstract
A plethora of research has established that the most effective treatment for sickle cell disease (SCD) is increased fetal hemoglobin (HbF). Fetal hemoglobin normally accounts for less than 0.5% of total hemoglobin in adults; increasing levels to approximately 10% alleviates much of the pathophysiology associated with SCD. Hydroxyurea is the only FDA-approved treatment for SCD that results in enhanced HbF production, but this drug is highly pleiotropic in its action and does not exclusively modulate γ-globin gene expression. Thus, research has focused on identifying agents that specifically reactivate γ-globin gene expression during adult definitive erythropoiesis, with minimal off-target effects. Artificial transcription factors (ATFs) are synthetic proteins designed to bind at a specific DNA sequence and modulate gene expression. The artificial zinc finger gg1-VP64 was designed to target the -117 region of the A γ-globin gene proximal promoter and activate expression of this gene. Previous studies demonstrated that HbF levels were increased in K562 cells, murine chemical inducer of dimerization (CID)-dependent bone marrow cells carrying a human β-globin locus yeast artificial chromosome (β-YAC) transgene, in CD34+ erythroid progenitor cells from normal donors and β-thalassemia patients, and in vivo, in gg1-VP64 β-YAC double transgenic (bigenic) mice. Transgenic mice with enforced expression of the gg1-VP64 fusion protein only in the erythroid-megakaryocytic compartment were crossed into the Townes sickle cell knock-in mouse (Jackson Laboratory) background. Compared with control sickle cell (HbSS) mice, gg1-VP64 ATF sickle cell (gg1-HbSS) mice had hematological values at levels found in wild-type homozygous or heterozygous adult hemoglobin (HbAA or HbAS, respectively) mice. For example, average RBC (106/mm3) was 11.7 for wild-type mice and 12.9 for gg1 HbSS, compared to 8.2 for HbSS mice. Average HGB (g/dl) was 15.1 for wild-type mice and gg1 HbSS mice, versus 10.0 for HbSS mice. Average HCT was 52.5% for wild-type mice, 53.7% for gg1 HbSS mice, but only 41.5% for HbSS mice. Finally, average WBC (103/mm3) was 9.4 for wild-type mice, 9.0 for gg1 HbSS mice and 91.0 for HbSS mice. HPLC and Western blot analysis to determine the effect of gg1-VP64 on HbF synthesis are underway. In addition, we are examining mice for numbers of HbF-positive cells, mature cells, and reticulocytes, as well as looking at organ damage. Our results demonstrate that the ATF class of reagent may be an effective gene therapy for treatment of SCD.

Disclosures
Makala: Georgia Regents University: Employment.
Sustained Elevation of Adenosine-ADORA2B Signaling Promotes Chronic Pain through Neuro-Immune Interaction in Sickle Cell Disease
Authors: Adebiyi Morayo G. , Hu Xia , Luo Jialie , Sun Kaiqi , Le Thanh-Thuy T. , Zhang Yujin , Wu Hongyu , Liu Hong , Huang Aji , Kellems Rodney E. , Blackburn Michael R. , Walters Edgar T. , Hu Hongzhen , Xia Yang 
Published: October 2019

Abstract
Sickle cell disease (SCD) is a common inherited hemolytic disorder that can cause severe health complications. One of the main complications of SCD is chronic pain. The molecular mechanism behind SCD chronic pain is poorly understood and effective mechanism-based treatments for SCD chronic pain are extremely limited. Recent studies show that an accumulated adenosine level induces sickling and disease progression by activation of adenosine A2B receptor (ADORA2B) (Zhang et al. 2011, Nature Medicine). However, the functional role of persistently elevated adenosine signaling in SCD chronic pain is undetermined. Here we report that SCD mice have persistent mechanical and thermal hypersensitivity due to sustained elevated circulating adenosine by activation of ADORA2B. Extending from SCD mice, we further demonstrated that elevated circulating adenosine contributes to chronic pain behavior in two additional animal models independent of sickling, mice lacking adenosine deaminase (ADA), an enzyme necessary for the breakdown of adenosine, and mice with Complete Freund's Adjuvant pawn injection, a well-accepted inflammatory chronic pain model, by activation of ADORA2B. Mechanistically, we revealed that adenosine does not directly activates neuronal cells. Instead, elevated adenosine activating ADORA2B on myeloid-derived immune cells causes nociceptor hyperexcitability and promotes chronic pain via IL-6-soluble IL-6 receptor trans-signaling cascade. Overall, our findings have identified that prolonged elevated adenosine promotes the cross talk between immune and neural cells and contributes to chronic pain in three independent animal models. These findings add significant new insight to molecular basis for chronic pain and immediately suggest novel therapeutic possibilities for anti-chronic pain.

Disclosures
No relevant conflicts of interest to declare.
Heterogeneity in the Making of Blood
Authors: Scadden David T. 
Published: October 2019

It is increasingly clear that the bone marrow is comprised of a heterogeneous complex of niches for hematopoietic cells, some for stem cells in the perivascular space and some for progenitors. We have used two approaches to define the role of specific cells in the marrow. First, single cell selection and characterization based on in vivo proximity to HSPC. This method has defined a subset of endosteal lining cells that can be immunophenotypically defined and isolated and reveals IL-18 as a regulator of hematopoietic progenitor quiescence. Second, candidate cell depletion that revealed mature osteolineage cells expressing osteocalcin as regulating the production of thymic emigrants through the expression of Dll4. Deletion of these cells reduces the number and function of T-biased lymphoid progenitors in the marrow space as well as thymic populations and mature T cells in the blood. These data suggest that specific niche subsets can be defined and through them, novel molecular regulators of HSPC function. The bone marrow niche is a heterogeneous composite of distinctive niches.

Disclosures
No relevant conflicts of interest to declare.