bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mu Opioid Receptor (OPRM-1) Polymorphisms among Patients with Sickle Cell Disease.
Authors: Mehta Sharmila , Campbell Nicholas , Bailey Lakiea , Kutlar Ferdane , Elam Dedrey , Clair Betsy , Kutlar Abdullah 
Published: April 2020

Abstract
Opioid analgesics form an important component of the management of acute and chronic pain in patients with sickle cell disease (SCD). Chronic opioid use, sometimes associated with dependence and addiction in a subset of patients, may pose difficult management problems. This adds to the sometime antagonistic relationship that can develop between providers and patients with SCD. Pain management can become a sociological and psychological issue in the management of SCD. The Mu opioid receptor (OPRM-1) is the primary site of action of endogenous opioid peptides (enkephalins and endorphins) and of opioid analgesics, such as morphine, methadone, fentanyl, heroin, and related compounds. Rapid activation of the mu receptor results in a euphoric effect, conferring the reinforcing or rewarding effects of opioids and thus contributing to the development of addiction. It has been known that there is variation between individuals in sensitivity to opioids suggesting potential variation in the receptor protein and the gene. Some recent data have shown that polymorphisms in the OPRM-1 gene affect pain threshold and tolerance as well as opioid requirements for optimal pain control. In an effort to unravel the complex issues surrounding pain frequency, pain tolerance, opioid usage, and opioid addiction in this patient population, we conducted a study of our adult sickle cell patients for the frequency of the two common cSNPs and another SNP in the IVSII (G691C) of the OPRM-1 gene. DNA samples from randomly selected patients were used in this study. The OPRM-1 gene was PCR amplified and subjected to cycle sequencing on an ABI Prism automated sequencer. The results showed that of the 97 adult SCD patients screened for the C17T polymorphism (GenBank accession #AY292291 and 292290), 67 (69.7%) had the wild type (CC), 28 (29.1%) were heterozygous (CT), and 11 (11.4%) were homozygous (TT) for the SNP. This represents an allele frequency for the T of 0.26. Of the 39 sequenced samples for the A1189G, all (100%) showed the wild type (AA). These results closely resemble those reported by Bond et al (PNAS, 95;9608,1998) for the frequency of the two cSNPs in the African-American population (allele frequency of 0.21 for the C17T and 0.016 for the A118G). Of the 16 samples screened for IVS II G691C polymorphism (GenBank accession #AY299483), 6 (37.5%) had the wild type (GG), 9 (56.3%) were heterozygous (GC), and 1 (6.2%) was homozygous (CC). Our results among SCD patients show a high frequency of C17T polymorphism in the OPRM-1 gene. Detailed studies in higher numbers of SCD patients and clinical correlations with pain frequency and threshold, opioid usage, opioid abuse and drug seeking behavior will be undertaken. It is expected that these studies will clarify the role of OPRM-1 polymorphisms as a genetic modifier associated with pain frequency, with tolerance as well as opioid use and abuse.
Haptoglobin Genotypes among Pediatric and Adult Sickle Cell Patients: An Emerging Genetic Modifier?.
Authors: Hughes Heather Y. , Kutlar Ferdane , Clair Betsy , Elam Dedrey , McKie Kathleen M. , Kutlar Abdullah 
Published: April 2020

Abstract
The role of hemolysis and its end product, cell free hemoglobin, as a Nitric Oxide (NO) scavenger in the pathogenesis of some complications of sickle cell disease (SCD) such as pulmonary hypertension has recently been recognized. Hemoglobin (Hb) released from red blood cells as a result of hemolysis complexes with haptoglobin (Hp). Hb-Hp complexes are removed from the circulation by tissue macrophages in the liver and spleen through CD163, the newly identified hemoglobin scavenger receptor expressed on the monocytes and macrophages. Hp is a polymorphic protein with two well-known alleles, Hp-1 and Hp-2 with worldwide distribution. Although the worldwide frequencies of these two allelles are approximately equal, there is significant ethnic and geographic variation in the distribution of Hp-1 and Hp-2 among different populations. Hp locus is on chromosome 16q22. each Hp monomer consists of one α-chain and one β-chain. An intragenic duplication event during evolution resulted in an elongated α-chain thus generating the Hp-2 allele. Thus individuals homozygous for the long α-2 chain express large multimeric molecules (Hp 2-2). Over the past 5 years, Hp-2 allele has emerged as a major susceptibility gene for development of vascular complications such as coronary artery restenosis and risk of cardiovascular disease, particularly in diabetic patients. It has been reported that the Hb-Hp-2 complexes have a 10-fold greater affinity for the CD163 receptor compared to Hb-Hp-1, and the interaction of Hb-Hp-2 complexes with CD163 on macrophages generates a more powerful inflammatory response with the release of pro-inflammatory cytokines such as IL-1β, IL-6, and GM-CSF. These observations led us to examine the role of Hp polymorphisms in sickle cell disease. We performed genotyping of the Hp-alleles with PCR in 21 pediatric patients (age 4 mo-10 years) and 41 adults patients (age 16–60). The results of this genotyping are shown in the table below. The Hp-1 allele frequency was 0.24 among pediatric patients and 0.52 among adults; while that of Hp-2 was 0.76 in the pediatric age group and 0.48 in adults. This difference was significant (p=0.0023). The significantly higher allele frequency of Hp-2 in pediatric patients compared to adults may indicate a selection against this genotype and suggests that Hp-1 allele may be associated with a survival advantage. This is in keeping with the deleterious effects of Hp-2 allele reported in the literature and suggests yet another mechanism whereby hemolysis may lead to adverse outcomes associated with Hp polymorphisms, in addition to the NO scavenging effects of cell free hemoglobin. Detailed studies are currently being undertaken to elucidate the mechanism whereby Hp polymorphisms may impact on the pathophysiology of SCD.
PERCENTAGE OF HAPTOGLOBIN GENOTYPES BETWEEN ADULT AND PEDIATRIC SICKLE CELL PATIENTS ADULT PEDS sHp 1–1 41.4% 9.5% Hp 1–2 22.5% 28.6% Hp 2–2 36.6% 62.0%
K-Cl Cotransport and MCHC in HbE Transgenic Mice and Human HbE.
Authors: Romero Jose R. , Rivera Alicia , Etzion Zipora , Suzuka Sandra M. , Vichinsky Elliott , Nagel Ronald L. , Fabry Mary E. 
Published: April 2020

Abstract
In HbE, HbC, and HbOArab a glutamic acid is mutated to a lysine resulting in a cumulative increase in positive charge of +4 units per tetramer when compared to HbA. These mutations occur at β−26, β−6, and β−121, respectively. We have reported an increase in both K-Cl cotransport and in red cell density (that is directly proportional to MCHC) for HbC and HbOArab. In the case of HbC, we demonstrated that transgenic mice expressing HbC also have these properties. We also previously reported that human homozyous HbE red blood cells (hRBC) have normal red cell density; that is, they have the same MCHC as HbA hRBC. We now report that RBC from mice (mRBC) expressing HbE have elevated K-Cl cotransport and normal MCHC and that these properties have been found in HbE hRBC as well. In mice expressing exclusively human α and βE, K-Cl cotransport in mRBC with HbE versus C57Bl was 10±0.9 versus 2.0±0.9 mmol/L cells x hr (FU), respectively. Mice expressing exclusively HbE have mRBC that are microcytic vs C57 (MCV=32.9±1.5 vs 47.5±2.0 fl, N=5, p&lt;0.0003), have low MCH vs C57 (MCH=9.8±0.1 vs 14.7±0.8 pg, N=5, p&lt;0.0004), but have normal MCHC vs C57 (CHCM=29.7±1.1 vs 30.9±1.9 g/dl, p=NS). In a human patient with HbE β0 thalassemia, we found elevated K-Cl cotransport (10±0.9 versus 2.0±0.9 FU) and red cell indices vs HbA that were analogous to those seen in HbE transgenic mice: (MCV=63.8 vs 88.1±2.3 fl), (MCH=20.9 vs 30.9±1.0 pg), but a normal MCHC vs HbA (CHCM=33.0 vs 34.9±1.2 g/dl). Because this patient suffers from severe thalassemia and hence has an elevated reticulocyte count (17%), the elevation in K-Cl cotransport may be due, in part or totally, to the presence of reticulocytes. However, reticulocytes have a lower MCHC than mature RBC, and blood with a high percent of reticulocytes generally has a low average MCHC; however, we observed a normal MCHC for this patient and others in whom K-Cl was not measured. We attribute the normal MCHC observed in both human patients with HbE and in our transgenic mouse model to the effect of K-Cl cotransport on a red cell with a thalassemic background. In conclusion, the low MCHC anticipated from the well-known thalassemic nature of the HbE mutation is not realized due to enhanced K-Cl cotransport. This demonstrates that the mouse model reproduces the hematologic features of the human disease and will provide a suitable model for gene therapy and other therapeutic approaches for HbE hemoglobinopathies.
Ten Year-Old Male with Hemoglobin Lufkin/Beta-Zero Thalassemia Compound Heterozygote Who Exhibited Beta-Thalassemia Major Characteristics.
Authors: Blutreich Ahna M. , Shu Peihong , Bragdon Jeremy S. , Kurtin Paul J. , Hoyer James D. , Karayalcin Gungor 
Published: April 2020

Abstract
Hemoglobin Lufkin is a rare and mildly unstable hemoglobin variant with increased oxygen affinity. Since 1977, two cases of hemoglobin Lufkin trait and one hemoglobin Lufkin/hemoglobin S have been described. This report is the first case of hemoglobin Lufkin/beta-zero thalassemia in a 10 year-old Caucasian male of Irish/Italian/German background. The patient presented with jaundice, splenomegaly and thalassemia major facies. On blood smear examination, RBC morphology showed hypochromia, microcytosis, many target cells, some spherocytes and basophilic stippling of RBC’s. On both alkaline hemoglobin (Hb) electrophoresis and isoelectric focusing (IEF) there was an absence of Hb A with a predominant band slightly anodal to the Hb A position. Hb A2 (4.8%)and Hb F (5.3%) were elevated as measured by high performance liquid chromatography. The heat unstable Hb test was abnormal. DNA sequencing of the beta globin gene confirmed a GCC to GAC mutation at codon 29 (gly to asp) consistent with Hb Lufkin. DNA sequence analysis also revealed a beta-zero thalassemia mutation, IVS-1-1, (G to A). The mother’s sample also showed the same beta-thalassemia mutation. Neither hemoglobin Lufkin nor beta-thalassemia were identified in the father; further studies are being done.
Thalassemia Major, Intermedia & Sickle-Thal Patients in Lebanon.
Authors: Taher Ali , Shamseddeen Wael , Arnaout Samir , Gharzuddin Walid , Zeineh Nabil , Alam Samir , Isma’eel Hussein 
Published: April 2020

Abstract
Introduction: Cardiac dysfunction in thalassemia remains of primary concern. The prevalence of these abnormalities varies in different phenotypic/genotypic groups of thalassemia.
Objective: To describe the difference in cardiac function among Thalassemia Major ™, Thalassemia Intermedia (TI), and Sickle-Thalassemia (ST) patients.
Methods: 66 TM, 41 TI and 17 ST patients were randomly selected from a population of 450 TM,100 TI and 26 ST patients respectively. The patients were interviewed and their charts reviewed and all had an echocardiography. Pulmonary Hypertension (PHT) was defined as Pulmonary Artery Systolic Pressure&gt; 30mmHg.
Results: 58.8%, 50% and 41.5% of ST,TM and TI patients respectively were males. Mean age was 24.4 ± 11.6, 19.4 ± 7.6, 19.5 ± 10.9 year old for TI, TM and ST. Tricuspid Regurgitant jet was detected in 47%, 30% and 82% of TI, TM & ST patients respectively. PHT was found in 83%, 18% and 10% of TI, ST, TM patients. Deceleration Time (DT) was significantly longer in TI than in TM and ST, with no significant difference between TM and ST. End diastolic dimension (EDD) was significantly larger in TI than TM and ST (for more details about cardiac parameters refer to tables 2a and 2b). Thrombotic events occurred in 6% of TI patients and in 0 TM and ST patients.
Discussion: PHT is primarily prevalent in TI patients which further supports the notion that a different pathophysiological process leads to this than in TM and ST patients. Moreover these results stress the need for better method of assessment of PHT, because of the limited number of patients in whom a TR jet could be detected. A prolonged DT is one of the earliest sign of diastolic dysfunction in thalassemics. TI patients have revealed themselves to have more impaired diastology by both prolonged DT and Isovolumic relaxation time (IVRT). Splenectomy was performed more in Lebanese TI than that reported in the literature (87% vs 65%), because most of our TI patients were managed as TM. This was before the currently recognized association of splenectomy and PHT and thrombosis. In addition chamber sizes (EDD) was significantly more in TI than both TM and ST patients. This can be attributed to the more profound chronic dilutional volume overload secondary to uncorrected anemia. Finally as is evident TI patients are developing more complications than both TM and ST. Transfusion/ chelation therapy to avoid PHT, diastolic dysfunction and thrombosis in TI patients has been proposed as a solution. If the pathophysiology of these complications is iron-mediated then prophylactic transfusion/ chelation therapy would be more advisable than waiting for them to develop. Especially in view of the morbidity associated with thrombosis, and limitation of exercise tolerance secondary to PHT.
Table 1: Clinical characteristics of patients
Table 2a: Significant differences between TI and ST patients Table 2b: Significant differences between TM and ST patients
Rapid Identification of the β-Thalassemia Mutations by Applying CEL I Nuclease Mutation Detection System and Capillary Electrophoresis.
Authors: Hung Chia-Cheng , Lin Win-Li , Jou Shiann-Tarng , Su Yi-Ning 
Published: April 2020

Abstract
Background: Beta-thalassemia is a common autosomal recessive hereditary disease in the Meditertanean, Asia and Africa area. It is a single gene inheritable disease resulting from one or more of a total of more than 200 different mutations in the beta-globin gene (HBB). For the clinical practice, the detection of beta-globin mutations were mainly depends on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequence technique. A more sensitive, efficient and reliable mutation-screening method is warrant and essential in order to establish appropriate prevention programs for at-risk populations based upon a molecular diagnosis.
Methods: To further enhance the detection efficiency and accuracy, the purpose of this study is to develop a novel genotyping method for common mutations in beta-globin gene. We have developed a rapid and highly-specific mutation screening test for the diagnosis of beta-thalassemia by CEL I nuclease mutation analysis based on the denaturing high performance liquid chromatography (DHPLC) system and ABI PRISM 310 Genetic Analyzer. To illustrate the efficacy of this approach, the exons of the beta-globin gene were amplified by PCR using primers 5′-labeled with fluorescent dyes of two colors. This study will demonstrate the usefulness of CEL I Nuclease as a method to genotype mutations in beta-globin gene for applying in daily clinical practice.
Results: Assays conditions were established based on the analysis of 50 DNA samples from heterozygotes or compound heterozygotes for the mutations in beta-globin gene. Homozygous wild-type DNA samples produced electropherograms containing only single peak, whereas samples heterozygous for a specific mutation displayed two peaks for that mutation site. In the double-blind validation analysis, all 50 DNA samples were showed 100% assay specificity.
Conclusions: Compared to classic approaches of mutation screening, when coupling with the previous established heteroduplex and primer-extension analysis, this method allows a rapid, highly sensitive, cost effective and semi-automated mutational screening of a large number of samples. Therefore, mutational analysis by CEL I Nuclease Mutation Detection System will be helpful as the first-line clinical screening and diagnosis tool of beta-thalassemia.
Antineutrophil Antibodies Lead to Mistaken Identity in Severe Congenital Neutropenia.
Authors: Boxer Laurence A. , Bolyard Audrey Anna , Schwinzer Beate , Glaser Darryl , Dougan Kenneth , Bonilla Mary Ann , Alter Blanche P. , Cham Bonnie , Kinsey Sally E. , Zeidler Connie , Welte Karl , Dale David C. 
Published: April 2020

Abstract
Autoimmune Neutropenia of Infancy (ANI) is generally regarded as a benign condition diagnosed by finding neutropenia and a positive test for antineutrophil antibodies. To determine whether the presence of antineutrophil antibodies in infancy always predicts a benign disease, data were reviewed for young children with positive antineutrophil tests who are enrolled in the Severe Chronic Neutropenia International Registry (SCNIR). A set of identical twins presented with severe neutropenia at age 5 months. Twin A1 had a preceding history of aphthous ulcers, cellulitis, recurrent otitis media, pneumonitis, an episode of bacteremia and a positive test for antineutrophil antibodies&lt;INS cite=mailto:%20 dateTime=2005-08-01T10:53&gt;.&lt;/INS&gt; Twin A2 had a preceding history of cellulitis, otitis media, and pneumonitis. The median absolute neutrophil count (ANC) of twin A1 and twin A2 was 0.280 x 109/L and 0.116 x 109/L, respectfully and bone marrow examination for twin A1 showed maturation arrest at the myelocyte-metamyelocyte level. With the diagnosis of ANI in one twin, both twins were placed on granulocyte-colony stimulating factor (G-CSF), because of frequent fever and recurrent infections. Fifty-one months after the diagnosis of immune neutropenia, twin A1 developed acute myelogenous leukemia. Further studies then revealed that both twins had a mutation in the neutrophil elastase gene (ELA2) at exon 5 resulting in a nucleotide substitution from TGC to TGA, which indicated a diagnosis of severe congenital neutropenia. Another set of fraternal twins were identified to have ANI at six months because of recurrent abscesses, mouth ulcers, otitis media and positive tests for antineutrophil antibodies. The ANC was 0.067 x 109/L in twin B1 and 0.166 x 109/L in twin B2. At age 31 months twin B1 developed a severe Clostridial infection, which led to the loss of his right leg and part of the anterior abdominal wall. Subsequently both twins were placed on G-CSF. Prior to G-CSF treatment, bone marrow studies revealed an arrest at the promyelocyte stage and an ELA2 mutation in exon 4 leading to a TCG to TTG substitution in both twins thereby indicating a diagnosis of severe congenital neutropenia. Upon further review of the SCNIR, eight other congenital patients were identified to have antineutrophil antibodies and bone marrow patterns consistent with severe congenital neutropenia. In conclusion, it is important to be aware that positive antineutrophil antibodies can occur associated with severe congenital neutropenia. Infants with severe neutropenia and repeated or severe bacterial infections should have a bone marrow examination and ELA2 evaluation to establish correctly the underlying pathogenesis of the neutropenia.
Left Ventricular Cardiac Function during the Course of a One Year Multicenter Trial of the Safety and Efficacy with ICL670 5–40mg/kg/day and Deferoxamine 20–60 mg/kg/day in Beta-Thalassemia Patients with Transfusional Hemosideosis.
Authors: Kleinert Dorothy A. , Jones Erica , Sison Cristina P. , Marks Peter , Giardina Patricia J. 
Published: April 2020

Abstract
Historically regularly transfused patients (pts) with Thalassemia Major (TM) have received prolonged infusions of subcutaneous deferroxamine (sc DFO) to prevent the lethal cardiac complications of transfusional hemosiderosis. Cardiac function is monitored with annual electrocardiograms (EKG) and 2 Dimensonial Doppler Echocardiogram (Echo). ICL670 (Exjade) is a newly identified oral iron chelator that has completed a one-year worldwide multicenter phase III clinical study of its safety and efficacy in comparison to sc DFO in TM. 17 TM pts in our institution participated in the Novartis ICL670-107 and 108 trials. They ranged in age from 2–36 years (yrs), the youngest being naive to chelation, five pts were previously noncompliant with the use of sc DFO, one of which developed acute heart failure more than 15 yrs prior to the study which resolved with vigorous DFO therapy and in whom all cardiac medications were discontinued 10 yrs prior to study entry. 10 TM pts ranging in age from 2–36 yrs received ICL670 at 10–30mg/kg/day and 7 TM pts ranging in age from 7–33 yrs received sc DFO at 40–60 mg/kg over 8 hour infusions 5 days per week. Each pt underwent a 2 dimensional Doppler Echo as part of their routine care prior to entry and after one year of study as well as regular EKGs before and during the study. Pre and post Liver Iron Concentration (LIC) was performed by biopsy in 15 adolescent/adult patients and 2 children had SQUID LIC. Laboratory analysis of blood counts, serum chemistries and ferritin levels were performed by a central laboratory. No clinically significant EKG changes were observed during the course of the study. Normal Ejection Fractions (EF) = 0.55 ± 0.75 ml/m2 and Shortening Fractions (SF) = &gt;26%. One of the ten pts on ICL670 developed an abnormal EF = 47 and a SF = 24% compared to 0 of 7 pts on sc DFO. None of the pts experienced clinical cardiac decomensation. In conclusion regularly transfused TM pts receiving oral ICL670 for one year had no statistically significant mean changes or relative mean changes in EF or SF on 2 dimensional Dopler Echo in comparison to those pts receiving sc DFO.
Echo Results Mean Pre EF Mean Post EF Mean Change Mean Relative Change ICL670 60.3 ± 5.5 58.6 ± 5.4 −1.7 ± 6.9 −2% ± 12% DFO 63.7 ± 4.6 67.1 ± 4.2 3.4 ± 6.3 6% ± 11% Mean Pre SF Mean Post SF Mean Change Mean Relative Change ICL670 32.4 ± 3.4 31.2 ± 3.3 −1.2 ± 4.3 −3% ± 13% DFO 35.0 ± 3.1 37.4 ± 3.3 2.4 ± 4.6 8% ± 14%