bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pyrimidine 5′ Nucleotidase Deficiency: Clinical and Molecular Characterization of Two New Italian Patients.
Authors: Fermo Elisa , Marcello Anna , Bianchi Paola , Viglio Simona , Chiarelli Laurent R. , Valentini Giovanna , Zanella Alberto 
Published: April 2020

Abstract
Hereditary pyrimidine 5′ nucleotidase deficiency (P5′N) is the most frequent abnormality of the red cell nucleotide metabolism causing hereditary non-spherocytic hemolytic anemia. The disorder is characterized by mild-to-moderate hemolytic anemia associated with reticulocytosis and hyperbilirubinemia and the accumulation of high concentrations of pyrimidine nucleotides within the erythrocyte. P5′N-1 gene is localized on 7p15-p14; eighteen mutations have been so far identified in 27 unrelated families, 6 of them of Italian origin. The aim of this study is to describe the hematological, biochemical and molecular characteristics of two new Italian patients affected by P5′N deficiency. Case1: The propositus was a 37 yrs old woman of Northern Italian origin affected by chronic hemolytic anemia with Hb levels ranging from 8.2 to 10.5 g/dL. At the time of the study Hb was 8.4 g/dL, reticulocytes 300x109/L, unconjugated bilirubin 3.2 mg/dL. Peripheral blood smear examination showed basophilic stippling and purines/pyrimidines ratio (OD260/280) was decreased (1, ref. values 1.4 – 2.98). P5′N activity, measured by capillary electrophoresis, was undetectable. Molecular analysis of P5′N-1 gene showed the presence of a new homozygous deletion of two bp (ag) at the splice junction between intron 7 and exon 8, which probably results in a splicing alteration and in the absence of a functional protein. Case2: The propositus, a 37 yrs old woman of Northern Italian origin carrying the hemoglobin variant HbD Punjab, had an history of chronic hemolytic anemia since childhood; at the age of 14 yrs splenectomy and colecystectomy were performed. The patient needed blood transfusions because of exacerbation of anemia (Hb 4.7g/dL) during parvovirus B19 infection. Iron status parameters were increased requiring desferrioxamine treatment. At the time of the study Hb was 9.3 g/dL, reticulocytes 752x109/L, unconjugated bilirubin 13.3 mg/dL. Serum ferritin was 1980 mg/mL and transferrin saturation 115%. The propositus was found to be homozygous for Gilbert’s syndrome and heterozygous for mutation H63D of HFE gene. Basophilic stippling (6%) was observed in peripheral blood smear. Pur/pyr ratio was 0.8 and residual P5′N activity was 40% of normal. Complete sequencing of P5′N-1 gene showed the presence of the frameshift mutation ins GG710-711, already described in Italian and Turkish patients, and the new in-frame aminoacidic deletion of Gln 143.
Mutations in the RNA Component of Telomerase, TERC, in Children with Aplastic Anemia and Myelodysplastic Syndrome: An NMDP Study.
Authors: Field Joshua J. , Mason Philip J. , Barnes Yvonne J. , King Allison A. , Bessler Monica , Wilson David B. 
Published: April 2020

Abstract
Mutations in TERC, the RNA component of telomerase, result in autosomal dominant dyskeratosis congenita (DC), a rare bone marrow failure syndrome. DC is clinically heterogeneous and TERC mutations have been detected in a subset of patients previously diagnosed with idiopathic aplastic anemia (AA) and myelodysplastic syndrome (MDS). Unrecognized TERC mutations are clinically relevant as patients with DC respond poorly to immunotherapy and have an increased risk of complications following conventional conditioning for stem cell transplant (SCT). We aimed to determine the frequency of TERC mutations in pediatric patients with AA and MDS who require a SCT. We obtained 315 blood or bone marrow samples from the National Donor Marrow Program Registry from children under age 18 with bone marrow failure who underwent an unrelated stem cell transplant. We screened these samples for mutations in the TERC gene using direct DNA sequencing. To exclude polymorphisms, we also screened 537 racially diverse healthy controls. The study group was composed of patients with MDS (n=151), AA (n=123), and juvenile myelomonocytic leukemia (JMML) (n=41), which may be difficult to distinguish from MDS. The mean age at the time of transplant was 9 years. We found sequence alterations in the promoter region of TERC in 2 patients. A 2 base pair deletion (-240delCT) was identified in a 4 year-old child with MDS and a 1 year-old child with JMML was found to have a point mutation (-99C→G), which was identified previously in an 18 year-old patient with paroxysmal nocturnal hemoglobinuria and is known to affect the Sp1 binding site. The pathogenicity of this mutation is unclear. In summary, our findings suggest that screening for TERC gene mutations is unlikely to diagnose occult DC in children with severe bone marrow failure who require a stem cell transplant but have no clinical features or history to suggest a familial bone marrow failure syndrome.
The “Thomas Plot”: Its Usefulness in the Assessment of Iron Status in Anemia.
Authors: Canals Carmen , Sardà Pilar , Piazuelo Jose María , Remacha Ángel F. 
Published: April 2020

Abstract
A diagnostic plot to assess iron status in anemia of chronic disorders (ACD) has recently been developed (Thomas C, Thomas L. Lab Hematol 2005). The plot is based on the relation between the ratio of the soluble transferrin receptor value to the logarithm of the ferritin value (sTfR/log sFt) and the RET-He parameter -reticulocyte hemoglobin equivalent-. A different cut-off point for sTfR/log sFt is proposed for patients with a CRP ≤5mg/L or with a CRP &gt;5mg/L. The main aim of our study was to validate the usefulness of the “Thomas Plot” in assessing iron status. We also ascertained whether a modification on the sTfR/log sFt cut-off point improved the results. CRP was determined in 306 patients, 198 with iron deficiency (ID) and 108 with ACD. All of them were anemia cases studied in accordance with our standard protocol. RET-He, sTfR and sTfR/log sFt were calculated in all samples. The Thomas Plot was applied to all the cases and the results were compared. A discrepancy was considered relevant when it implied a change in patient treatment. Using logistic regression analysis, we investigated the optimum cut-off point of sTfR/log sFt in our series. Overall, the Thomas Plot correctly classified 223/306 patients (73%). There were clinically relevant discrepancies in 29 patients (9.5%), in 5 patients with CRP ≤5mg/L (5%) and in 24 patients with CRP &gt;5mg/L (12%). It failed to identify ID in 12/198 patients (6%) but classified 17 cases with ACD (16%) as latent ID or IDA. The optimum sTfR/log sFt cut-off point in our series was 2.4 for patients with CRP ≤5mg/L and 2.39 for patients with CRP&gt;5mg/L. As a consequence, we investigated a modified “Thomas Plot” using a sTfR/log sFt cut-off point of 2.4 for all the patients. The modified algorithm correctly classified 77% of the patients. There were clinical relevant discrepancies only in 17 patients (5%). It failed to identify ID in 12/198 patients (6%) and only classified 5 cases with ACD as latent ID or IDA (4.6%). The sensitivity of the Thomas Plot to identify ID was very high but the specificity was lower. A cut-off point of 2.4 for sTfR/log sFt is useful for patients with or without inflammatory traits and yields more accurate results.
A Nationwide Survey of Immunosuppressive Therapy for Idiopathic Acquired Pure Red Cell Aplasia in Japan and a Proposal for First-Line Treatment: Report of the PRCA Collaborative Study Group.
Authors: Hirokawa Makoto , Sawada Ken-ichi , Motegi Mutsuhito , Mamiya Shigeo , Teramura Masanao , Nakao Shinji , Ozawa Keiya , Urabe Akio , Omine Mitsuhiro 
Published: April 2020

Abstract
We have conducted a nationwide survey to define the first-line immunosuppressive treatment for idiopathic acquired pure red cell aplasia (PRCA) in Japanese adult patients diagnosed from January 1990 to December 2004. Questionnaires were sent to forty-seven medical centers and eighty-seven patients with acquired chronic PRCA from forty-seven institutions were registered in this retrospective study. Thirty-four patients (39%) were classified into idiopathic PRCA. Cyclosporine A (CsA), prednisolone (PSL) and cyclophosphamide (CY) were initially given to 20, 10 and 3 patients with idiopathic PRCA, respectively. One patient was treated with anabolic steroid as first-line therapy. The response rates with CsA and PSL were 80 and 60 %, respectively. All patients who had been resistant to PSL and two out of three CY-resistant patients were responsive to CsA. An anabolic steroid resistant patient also responded to CsA. Thus, CsA was used in twenty nine patients as a form of primary or salvage treatment with an 86 % clinical response rate. Nine out of 25 (36%) CsA-responders eventually relapsed and 12.5% of the patients relapsed during therapy. The median remission duration in the CsA responders was seven years. All PSL-responders were predicted to eventually relapse within 2 years, and four out of seven patients on prednisolone relapsed during maintenance therapy. All patients responded to CsA during their second and/or third remission induction with or without PSL. Overall survival was not different between the CsA- and PSL-responders. Although the difference in remission duration between both responders was not statistically significant (p&lt;0.114), which was probably due to the limited number of patients included in the study, CsA may be a best first-line treatment option for Japanese idiopathic acquired PRCA.
Clinical Relevance of Cytogenetics Abnormalities in Adult Patients with Acquired Aplastic Anaemia.
Authors: Gupta Vikas , Brooker Carol , Tooze Jennifer A. , Yi Qi-long , Sage Deborah , Turner David , Kangasabapathy Pamela , Marsh Judith C.W. 
Published: April 2020

Abstract
The clinical relevance of cytogenetics abnormalities in aplastic anaemia (AA) patients at time of diagnosis is unclear. We evaluated the clinical course of 81 AA patients with successful cytogenetics at diagnosis and treated with immunosuppressive therapy (IST) from January 1993 to March 2004. A cytogenetic study was considered to be successful if there were a minimum of 15 evaluable metaphases in the absence of a clonal abnormality. Response to IST, survival and later clonal complications in patients with an abnormal karyotype (n=10) was compared to those with a normal karyotype (n=71).
The cytogenetic abnormalities at diagnosis consisted of trisomy 6 (n=2), trisomy 8 (n=2), trisomy 15 (n=2), monosomy 7 (n=1), add(10) (n=1), t(3;11) (n=1) and t(4;6) (n=1). Four out of five evaluable patients with a trisomy responded to a first or subsequent course of IST. One patient with monosomy 7 achieved a complete response and later developed haemolytic PNH but with no recurrence of the monosomy 7. None of the patients with a non-numerical karyotypic abnormality responded to IST. No significant differences in 4-year event-free survival (EFS) (54% vs. 30%, p=0.15), overall survival (OS) (84% vs. 80%, p=0.33) or later clonal disorders (PNH, MDS and AML) were observed between the patients with a normal karyotype and those with an abnormal karyotype. Advanced age (≥60 years) was the only independent poor prognostic factor for survival in a multivariate analysis.
Among the patients with a normal karyotype (n=71), 6 patients later developed a clonal cytogenetic abnormality with a cumulative risk of 10% at 4 years. These abnormalities were trisomy 15 (n=2), trisomy 6(n=1), monosomy 7 (n=2) and t(13;15) (n=1). None of the three patients who acquired trisomies developed any clinically significant problem, while acquisition of monosomy 7 was associated with a transformation to MDS/AML.
Our data show that AA patients with a trisomy cytogenetic clone at diagnosis show a similar response to IST, evolution to later clonal abnormalities and survival, compared to those AA patients with a normal karyotype.
Results of Immunosuppressive Therapy in Children with Acquired Severe Aplastic Anaemia (SAA). Report Polish Paediatric Leukaemia and Lymphoma Study Group.
Authors: Pawelec K. , Matysiak M. , Niewiadomska E. , Rokicka-Milewska R. , Jackowska T. , Kowalczyk J. , Stefaniak J. , Balwierz W. , Zalecka-Czerpko E. , Chybicka A. , Szmyd K. , Sonta-Jakimczyk D. , Bubala H. , Krauze A. , Wysocki M. , Kurylak A. , Wachowiak J. , Kaczmarek-Karnold M. , Stolarska M. , Karolczyk I. , Krawczuk-Rybak M. , Leszczynska E. , Urasinski T. , Peregud-Pogorzelski J. , Wlazlowski M. , Balcerska A. 
Published: April 2020

Abstract
Introduction: Bone marrow transplantation (BMT) from HLA identical family donors is the treatment chosen for children with (SAA). When no donor is available, combined immunosuppressive therapy (IS) is given.
Material and method: SAA was recognised in 85 children (31 girls, 54 boys) aged 2–17.5 years in ten haematological clinic in Poland between 1993–2003 years. All patients received IS according SAA Working Party of the EBMT protocol: ALG or ATG, CsA, prenisolon and in 79 patients G-CSF or GM-CSF was additionally administered. Haematological response was evaluated on day 180 of therapy.
Results and conclusion: Complete remission (CR) occurred in 43 patients (40.5%), partial remission (PR) on 22 (25.8%), no response (NR) was obtained in 20 children (23.7%). Period of observation was from 12 months to 10.5 years. During this time relapse occurred in 10 patients (8.5%). Patients with NR and relapse were treated second course of IS or BMT from matched bone marrow donors. Unrelated BMT was performed in 11 patients with NR after IS. We observed 17 deaths (10 early during the first 3 months of IS) in all group. At present 54 children (63.5%) is in first remission (CR or PR) with lasts from 12 months to 10 years. During the 10.5 year of observation we notice one case with late clonal complication (PNH). Transformation to neoplasma was not observed in none of our patients after IS.
Phase II Study of Lenalidomide (CC-5013, Revlimid®) for Patients (pts) with Myelofibrosis (MF).
Authors: Cortes Jorge , Thomas Deborah , Verstovsek Srdan , Giles Francis , Beran Miloslav , Koller Charles , Kantarjian Hagop 
Published: April 2020

Abstract
Introduction: Thalidomide has been shown to induce responses in pts with MF but prolonged administration in these pts may be associated with neurotoxicity and other adverse events. Lenalidomide is the lead clinical compound in a new group of drugs called IMiDs®, which have immunomodulatory properties. It affects cytokine expression 200–5000 times more potently than thalidomide but lacks the teratogenecity, sedation and neurotoxicity.
Methods: We thus designed a phase II study for pts with myelofibrosis (primary or associated with myeloproliferative disorders) with a platelet (plt) count of at least 30 x109/L. Pts may have received prior therapies, but were excluded if they had known hypersensitivity to thalidomide. Pts received lenalidomide 10 mg/day orally (5 mg daily for patients with platelet count less than 100,000 at study entry).
Results: We have treated 41 pts. Their median age was 65 (range, 42 to 83). Thirty-six pts (88%) had received prior therapy for MF, including thalidomide 13 (32%), hydroxyurea 12 (29%), interferon 11 (27%) (pegylated 7), anagrelide 10 (24%), and erythropoietin 6 (15%). Baseline hemoglobin was &lt;10g/dl in 19 (46%) (13, 32% transfusion dependent); platelets &lt;100 x109/L in 12 (29%); neutrophils &lt;1.5 x109/L in 2 (5%), and 20/34 (59%) had splenomegaly (median 10cm BCM, range 1 to 30) (7 prior splenectomy). Twenty pts are evaluable for response and toxicity (i.e., received at least 30 days of therapy, unless discontinued because of progression or toxicity). One pt discontinued therapy because of toxicity (grade 3 rash). Responses have been observed in 10 (50%) pts. These include CR in 2 pts (normalization of Hgb and WBC, respectively), PR in 2 pts (improvement in plts and hgb, ± spleen), hematologic improvement in 6 pts (improvement in plts 3, spleen 2, WBC 1). Three of 13 transfusion-dependent pts have become transfusion independent. The median time to response was 9 weeks (range, 2 to 22). Responses have been sustained for a median of 14 weeks (range, 2 to 28 weeks). Therapy has been well tolerated. The more common toxicities were rash in 12 (29%), and pruritus in 8 (20%). Grade ≥ 3 toxicities were thrombocytopenia (n=2), rash (n=1), fatigue (n=1), and neutropenia (n=1). Four (10%) pts have required dose reduction and 1 discontinued therapy because of toxicity (rash).
Conclusion: We conclude that lenalidomide has clinical activity in a subset of patients with myelofibrosis with an acceptable toxicity profile. Studies using lenalidomide in combination with other agents are justified.
Clinical and Laboratory Features of Homozygous Sickle Cell Patients in Tanzania; Malaria, Infections and Cerebral Blood Flow Velocity.
Authors: Makani Julie , Meda Elineema , Rwezaula Stella , Williams Thomas , Thein Swee Lay , Marsh Kevin 
Published: April 2020

Abstract
Background: Significant advances have been made in the management of patients with sickle cell disease (SCD), with resultant increase in survival, most notably in the USA where 1,000 children are born with SCD every year. In contrast, in Africa, where it is estimated that 450,000 children are born with SCD every year, there has been little change in the management and minimal reduction in mortality. There is no reason to doubt that similar interventions and consequent improvements in survival can be achieved in Africa. However, the lack of resources mean that it is vital that proposed interventions are based on a clear definition of the problem.
Aims: The study attempts to define causes of morbidity and mortality in SCD in Tanzania focusing on three major areas where it is important to establish clear answers. The aims of the study are to determine the role of Plasmodium falciparum infection in precipitating crises in SCD patients, to describe the major bacterial pathogens associated with admission in SCD patients and to establish the association of increased cerebral blood flow velocities (CBFv) and cerebrovascular accidents (CVA) in SCD patients in East Africa.
Study design: This is an ongoing hospital based, prospective, descriptive cohort study in Muhimbili national hospital, Dar-es-salaam, Tanzania. Patients are seen in outpatient clinic every three months, if there is any acute event requiring hospital attendance and when admitted to the hospital.
Results: From march 2004, there have been 3,650 visits during which clinical and laboratory data has been collected on 885 (male 51.4%) patients, all homozygous for HbS on cellulose acetate electrophoresis. The mean age of the patients is 10 ± 7.2 years, (range 9 months-54 yrs), with 22.7% being under the age of 5 years and 8.28% above 20 years. The frequency of major clinical events, including fits (5.2%), stroke (1.9%) is reported. The prevalence of malaria parasitaemia in outpatients and during admission was 2.9% and 7.7% respectively. 227 patients have been admitted during this period, presenting with pain (44%), anemia (17%) and fever (15%). Transcranial Doppler ultrasonography was done in 400 patients; with 25% having a time averaged maximal mean velocity of 150 cms/second or more, suggesting stenosis.
Conclusion: This study is the initial step in describing the causes of morbidity in patients with SCD disease in Tanzania, and will form the basis of longitudinal studies that will attempt to guide interventional strategies, target research and provide insight into natural history of SCD in East Africa.
Hemoglobin Ville-Marie: A New β-Globin Variant (del480C) Causing Oxydative Hemolysis.
Authors: Devaux Celine , Chagnon Pierre , Belisle Claude , Soulieres Denis , Busque Lambert 
Published: April 2020

Abstract
Introduction: A 26 year old male patient was investigated at Maisonneuve-Rosemont Hospital (Montreal, Canada) for chronic hemolysis. The patient was known for many years to have chronic hemolysis of unknow etiology and splenomegaly. He had previously undergone cholecystectomy. There was no family history of hemolytic anemia. The investigation revealed a normal automated CBC but the blood smear showed polychromatophilia, basophilic stippling and a few degmacytes (bite cells). The serum chemistry analysis revealed an increased LDH and bilirubin and a decreased haptoglobin level. The search for Heinz body was positive after exposure to acetyl phenylhydrazine. Unstable hemoglobin was demonstrated using the isopropanol test, but the heat instability test was negative. The dosages of erythrocyte enzymes (G-6PD, pyruvate kinase, gluthation reductase and acethylcholinesterase) were normal. Hb HPLC analysis was done using a VARIANT II, Bio-Rad. Results demonstrated the presence of normal HbA and an abnormal Hb migrating as Hb Hope at a level of 24% of total hemoglobin.
Methods and Results: Blood DNA sample was obtained to perform automated sequencing of the β-globin gene (HBB). The HBB gene was sequenced using primers designed to amplify coding sequences of the three exons, splice junctions, introns and the promoter region. Five samples (the patient and four controls) were analysed. Sequence were compared to the cDNA sequence of the HBB gene (NM_000518) and from the genomic sequence of the region (AC104389). In the case of the propositus, a 1 nucleotide deletion was detected at codon 142 (nucleotide 480 of GenBank entry NM_000518) in exon 3, causing a reading frameshift. This deletion (TGCCCTGGCC [C/del] ACAAGTATCA) was never detected before and eliminates the regular stop codon (TAA) at position 147. A new stop codon is therefore produced after the addition of 14 residues, identical to those observed in Hbs Saveme, Trento and TAK, all of which result from a similar reading frameshift secondary to a deletion. However, a threonine residu is present at position 142 in the case of the propositus.
Conclusion: Hemoglobin Ville-Marie is an elongated C-terminal hemoglobin variant causing mild compensated hemolytic anemia with some degree of hemoglobin instability. Others analysis are being performed to better characterise this new hemoglobin variant, the results of which will be presented at the meeting.
DNA Sequence of Hemoglobin Chico.
Authors: Fond Aaron M. , Neville Stephen , Gotchall John , Liu Enli , Ou Ching-Nan , Rognerud Cheryl L. , Mims Martha P. 
Published: April 2020

Abstract
We were asked to investigate a 54 year old male who was undergoing evaluation for sleep apnea. The patient had a low resting O2 saturation at 91%, but no signs of cardiac or pulmonary disease as indicated by a normal echocardiogram, normal spirometry, and normal high resolution CT of the chest. Arterial blood gases revealed a pO2 of 79 mm Hg on room air, but pCO2 was normal. The patient’s hemoglobin concentration was 13.7 g/dL. The possibility of a mutant hemoglobin was raised and HPLC revealed HbA2 = 2.6%, HbA = 51.4% and an unidentified mutant hemoglobin of 46%. Oxygen dissociation testing demonstrated a P50 of 39 mm Hg (normal 24–32) consistent with a low affinity hemoglobin. The isopropanol test showed that the hemoglobin variant was unstable. Trypsin digestion of the abnormal hemoglobin resulted in a smaller than normal HPLC peak for the β9 segment of β-globin, which is composed of amino acids 67 through 82, and the appearance of a new peak at 193 min. This peak did not match any of the known mutant hemoglobins contained in our reference bank. We PCR amplified and sequenced the β-globin gene. The patient was heterozygous for an A to C mutation in the second position of codon 66. This mutation changed the lysine to threonine and revealed that the hemoglobin variant was Hb Chico. This result was consistent with the loss of ~50% of the β9 peak in the trypsin digest due to loss of a cleavage site after amino acid 66, and the appearance of a new peak which was a hybrid segment of β8 and β9. This represents the first DNA sequence of hemoglobin Chico a mildly unstable hemoglobin variant with decreased oxygen affinity.