bluuebunny/crossref_metadata_2025_split huggingface dataset:
Puerto Rican Child with Combined Heterozygocity for Hemoglobin S and Porto Alegre.
Authors: Lojo Liliana , Santiago Pedro J. , Rivera Enid , Rivera Doris E. , Renta Jessica , Cadilla Carmen L. 
Published: April 2020

Abstract
Hemoglobin S is the most common abnormal hemoglobin detected in newborn screening in Puerto Rico. The purpose of this report is to present laboratory and clinical data from a newborn proven to have combined heterozygocity for hemoglobin S and a very rare hemoglobin. Abnormal newborn screening results using isoelectric focusing prompted us to further study an infant with an unknown hemoglobin. Testing with cellulose acetate and citrate agar electrophoresis revealed that this patient had, besides hemoglobin S, a rare fast moving band. We consulted the Haemoglobinopathy Screening Service at St. James’s University Hospital in Leeds, UK where our patient’s hemoglobin was identified as hemoglobin Porto Alegre by means of mass spectrometry. DNA sequencing at the molecular genetics laboratory in the Medical Sciences Campus, University of Puerto Rico confirmed our patient’s hemoglobin to be Hb Porto Alegre. The patient’s father was found to be heterozygous for hemoglobin S and her mother a carrier of hemoglobin Porto Alegre. Our patient has been followed-up for the last two years. She has remained asymptomatic, except for usual mild childhood diseases. Laboratory results up to 18 months of age have revealed Hgb 11.4–11.6 g/dl, MCV 87.5 fl, MCH 28.7 pg, reticulocyte count 2%, normal total bilirubin and LDH levels. Hemoglobin Porto Alegre involves a mutation in ß9(A6)Ser→Cys which promotes hemoglobin polymerization via disulfide bonds (Baudin-Creuza 2002). This mutation does not seem to alter hemoglobin function and apparently does not increase tendency for sickling. Tondo (1963) has reported that in vivo hemoglobin Porto Alegre exists in its normal tetrameric form, with no associated hemolysis. Martinez (1977) has suggested that hemoglobin Porto Alegre can exist in its tetrameric functional form by means of high levels of glutathione in red blood cells. Therefore, patients with combined hemoglobin S and Porto Alegre would be expected to be asymptomatic. Hemoglobin Porto Alegre has been documented in a few patients from Brazil, Portugal, Argentina, Cuba, and the Canary Islands. To our knowledge this is the first reported double heterozygote for hemoglobin S and Porto Alegre. We are following our patient closely for the development of any significant hematologic or clinical symptoms.
The Prevalence of Renal Dysfunction in Young Adults with Sickle Cell Disease.
Authors: Osarogiagbon Raymond U. , McHugh Laura M. , Kronish Lori E. , Chismark Elisabeth A. 
Published: April 2020

Abstract
Sickle cell disease is typified by dramatic acute painful episodes, which consume the majority of patients’ and healthcare providers’ attention. However, the cumulative damage to vital organs that occurs in this disease is often the determinant of early death. The kidneys are a major target organ in sickle cell disease and renal failure is a highly morbid event. It is also strongly associated with early death. Glomerular hyperfiltration is often the earliest sign of renal injury. This is followed by return to apparently normal filtration rates, then successively lower rates, culminating in end-stage renal disease. Microalbuminuria follows hyperfiltration and progresses to frank proteinuria. We evaluated the prevalence of renal dysfunction in a cohort of patients with sickle cell disease managed at the University of Tennessee Cancer Institute’s adult sickle cell program. At entry into the program, patients underwent a routine battery of tests, including 24 hour urine collection for measurement of creatinine clearance and urine protein. Our findings are summarized in the table below.

By the age of 25 years, almost 80% of patients with sickle cell disease showed significant abnormality of glomerular filtration and 40% had significant proteinuria. This worsens with age. By a median of 37 years, 2 in 3 patients have developed significant proteinuria and hypofiltration has become the predominant pattern of glomerular filtration abnormality. Since renal injury is reversible in the early stages, more emphasis needs to be placed on aggressive early screening, surveillance and intervention. Intervention at the stage of hyperfiltration and microalbuminuria is easier and much more likely to be successful. Patients and their healthcare providers need to be educated on the high prevalence of renal damage. Efforts at education, screening, surveillance and, where necessary, treatment should begin well before adolescence.
Expression of High Levels of Human γ-Globin in Adult Mice Carrying a Transgene of the Brazilian Type of Hereditary Persistence of Fetal Hemoglobin.
Authors: Cunha Anderson F. , Brugnerotto Ana F. , Melo Monica B. , Corat Marcus A.F. , Gimenes Ana P. , Passos Luiz A.C. , Devlin Emily E. , Bodine David M. , Saad Sara T.O. , Costa Fernando F. 
Published: April 2020

Abstract
The term, hereditary persistence of fetal hemoglobin (HPFH), describes a hereditary benign disease, characterized by an increase in fetal hemoglobin (HbF) during adult life. Non-deletional forms of HPFH are characterized by single-base mutations in the promoter region (most of them between −114 and −202 from the cap site) of either the Gγor Aγ-globin gene, resulting in an increase of HbF ranging from 3 to 20% in heterozygotes. Many point mutations in this region have been described, including the Aγ-195 C →G mutation that causes the Brazilian type of HPFH (HPFH-B). A previous study showed that the -195 mutations alone was not able to increase gene expression in vitro and a modest increase was observed when a LCR element fragment (HS2) was introduced in the construction. This study showed too, that the the mechanism of HbF elevation by the -195 mutation was neither mediated by the Sp-1 transcription factor nor by the creation of a CACCC box, as described for the -198 mutation. Thus, other proteins may be involved in the over-expression of the γ-globin chain and/or may depend on DNA structure and these mechanisms remain to be clarified. To better understand this mechanism we have developed HPFH-B transgenic mice. The promoter with mutation was amplified using the genomic DNA of a HPFH-B patient and cloned in a μLCRAγψβδβ cosmid. This construct, containing the micro-LCR and other essential elements of human beta-globin gene cluster was microinjected into single cell mouse embryos. To detect the differences in developmental regulation of the human gamma-globin gene expression in the transgenic mice, we analyzed the yolk sac derived embryonic blood at embryonic day 10.5 (E10.5) and the fetal liver of mouse embryos at E13.5 of both mutated and non-mutated transgenic mice by RNAse Protection Assay (RPA) and Real time PCR (RT-PCR). Levels of expression of murine alpha-globin mRNA were used as internal controls in the RPA experiment and levels of murine-GAPDH were used in RT-PCR. mRNA levels of human gamma-globin in fetal liver of transgenic mice containing mutation were clearly higher, as compared with control transgenic mice bearing cosmid construct with wild a type sequence gamma promoter. Additionally a nearly 10-fold increase in human gamma-globin expression was detected in fetal livers of HPFH transgenic mice at E13.5 compared to the yolk sac. Thus, our data indicate that the - 195 mutation is the unique cause of elevation of HbF in Brazilian HPFH, but the exact mechanism needs to be clarified.
Automated Sorting of Monocytes from Whole Blood for Reproducible Gene Expression Profiling.
Authors: Poggel Carsten , Adams Timo , Martin Sabine , Pickel Carola , Prahl Nicole , Schmitz Jurgen , Bosio Andreas 
Published: April 2020

Abstract
Microarray-based gene expression profiling has been used to develop clinically relevant molecular classifiers for many different diseases. Furthermore, it has been shown for various chronic diseases that specific gene expression patterns are reflected at the level of blood cells. However, blood is a complex tissue comprising numerous cell types. Therefore, the contribution of rare cell types to a whole blood expression profile might not be detected and a substantial proportion of what is usually reported as “up-regulation” or “down-regulation” might actually be the result of a shift in cell populations and not of a true regulatory process. In order to circumvent these problems, several techniques have been established to analyze purified subpopulations rather than whole blood samples. Previously, it has been shown, for example, that reproducible gene expression profiles can be generated by positive selection of blood cell subsets from PBMCs1. As the preparation of PBMCs by, for example, Ficoll is time-consuming, inconvenient, and not amenable to automation, we have set up a combined direct whole blood cell separation and gene expression profiling protocol. By using Whole Blood CD14 MicroBeads in combination with the autoMACS Pro™ Separator, the separation protocol generally allowed enrichment of monocytes from whole blood within 30 min with purities higher than 90%. In combination with the depletion of neutrophils, the major source of contaminating RNA, purities increased to over 95% for all tested blood donors. Monocytes included the CD14bright/CD16− as well as the CD14dim/CD16+ populations. To assess the reproducibility of gene expression profiles and the influence of several experimental parameters, monocytes were sorted from 5 ml whole blood. RNA was extracted and hybridized to microarrays and the Pearson correlation coefficients of pairwise comparisons were calculated. Technical repeats of monocyte analysis from blood donated at different days showed a higher correlation coefficient than whole blood RNA. Blood storage at room temperature resulted in a strong deregulation of many genes, whereas blood stored at 4°C showed minimal changes, which is in agreement with previous studies. Skipping the centrifugation step, which is used to remove unbound MicroBeads did not alter the gene expression profiles. Incubation of sorted cells in PrepProtect™ Stabilization Buffer showed no alteration of gene expression thus enabling the shipping of cells without liquid nitrogen. Monocytes play a crucial role in diseases like atherosclerosis. Our rapid and simple protocol for combined direct cell sorting from whole blood and gene expression profiling of monocytes might help to ease the discovery of new biomarkers and to screen and monitor patients. 1 Lyons et al., BMC Genomics (2007), 8:64.
Mycophenolic Acid Causes Dysfunction of Human Polymorphonuclear Neutrophils and Dendritic Cells during the Interaction with Aspergillus fumigatus.
Authors: Loeffler Juergen , Mezger Markus , Einsele Hermann 
Published: April 2020

Abstract
Immunosuppressive drugs are a variety of substances to prevent transplant rejection. There is increasing data that immunosuppressants not only modulate lymphocyte function but also interfere with other immune effector cells. In this study, we analyzed the influence of Mycophenolic acid (MPA) on the activity of human polymorphonuclear neutrophils (PMNs) and dendritic cells (DCs) during the interaction with the mold Aspergillus fumigatus. PMNs were obtained from blood using Bicoll separation. Release of reactive oxygen species (ROS) was detected photometrically by using dichlorofluorescein. Monocytes were isolated by magnetic-associated cell sorting followed by differentiation into DCs using GM-CSF and IL-4. DCs were either co-cultivated with MPA (10 μg/ml), with A. fumigatus germ tubes (MOI=1) or both stimuli. Whole-genome microarray analyses (Affymetrix U133A) or quantitative real-time PCR assays were performed to quantify differentially regulated genes. MPA could be identified as an enhancer of the A. fumigatus induced oxidative burst of PMNs while fungal killing efficiency was not affected. Incubation of fully differentiated Mo-derived DCs in the presence of MPA did not alter gene expression patterns. However, if DCs were treated during early differentiation processes, DC development was influenced by MPA resulting in increased apoptosis. In addition, we revealed impaired expression of a variety of pro-inflammatory immune response genes (TNF-α, CXCL10, IL-12). In conclusion, MPA influences PMN and DC function during immune defense against fungi. MPA triggers an acute inflammatory syndrome by enhancing the A. fumigatus induced oxidative burst. Furthermore, innate and adaptive immune responses might be impaired because of a reduced number of DCs and lower levels of pro-inflammatory cytokines and chemokines.
The Role of Nramp1 in Erythrophagocytosis.
Authors: Soe-Lin Shan , Sheftel Alex D. , Ponka Prem 
Published: April 2020

Abstract
Erythrocytes contain the majority of the body’s iron in the form of heme, a component of the red blood cell pigment hemoglobin. Erythrocytes transport oxygen bound to hemoglobin from the lungs to the tissues, and become progressively more damaged as they age. Macrophages are responsible for the detection and degradation of these senescent red blood cells, and play a central role in the recycling of hemoglobin-derived iron via transferrin to the bone marrow in order for erythropoiesis to occur. This recycling process, and the influence of inflammatory mediators on it, are poorly understood. Nramp1 is a protein expressed within macrophages, and is an iron transporter discovered to be mutated in mice susceptible to tuberculosis and leprosy infections. As Nramp1 localizes to phagosomal membranes following phagocytosis, we speculate that Nramp1 may play a role in the recycling of hemoglobin-derived iron following the internalization of effete erythrocytes. To test this possibility, we have utilized a model of erythrophagocytosis using both Nramp1 replete and deficient RAW264.7 murine macrophages, and macrophages elicited from wild type and Nramp1-knockout 129SV mice. We have found that Nramp1 protein is increased in primary macrophages following internalization of opsonized murine erythrocytes. This increase peaks at 6 hours of erythrocyte exposure, and decreases to baseline levels overnight. Our model of erythrophagocytosis did not induce macrophage activation as no nitric oxide production was observed. This therefore discounts the possibility that inflammatory factors are the cause of the increase in Nramp1 protein levels. Using the fluorescent iron-binding probe, calcein, we monitored changes in the macrophage labile iron pool (LIP) following erythrocyte loading. This transit pool of loosely bound, chelator-accessible iron is thought to reflect the overall iron status of the cell. We have found that, following erythrophagocytosis of opsonized murine erythrocytes, Nramp1-replete RAW264.7 macrophages experience a greater-fold increase in labile iron as compared to Nramp1-deficient macrophages. Furthermore, we have also found that primary peritoneal macrophages elicited from wild type 129SV mice phagocytose more erythrocytes when compared to macrophages elicited from their Nramp1-knockout counterparts. Interestingly, we have also observed that treatment with erythropoietin, the hormone responsible for the promotion of erythrocyte precursor survival and ultimately the increase in red blood cell production, resulted in a modest increase in Nramp1 protein levels. Our observations show distinct differences in iron metabolism between Nramp1 replete and deficient macrophages undergoing erythrophagocytosis. We speculate that Nramp1 may serve to promote a more efficient intracellular transport of iron following red blood cell engulfment.
Multi-Scale Analysis of Granulopoiesis: A Bottoms-Up Approach.
Authors: Lee Daniel W. , Thompson Christopher , Obeyesekere Mandri N. , Corey Seth J. 
Published: April 2020

Abstract
Granulopoiesis constitutes a dynamical system, able to respond acutely to internal or external stimuli, with production of neutrophils. Because neutrophils are short-lived (&lt;24 hr) in the peripheral circulation, a major component of bone marrow activity is devoted to their generation. The hematopoietic growth factor, granulocyte colony-stimulating factor (G-CSF), is the most important growth factor for the production of granulocytes. When it or its cognate receptor, the G-CSF Receptor, is genetically ablated in mice, severe neutropenia results. The two most commonly expressed isoforms are the normal full-length G-CSF Receptor (class I) and a truncated, differentiation-defective receptor (class IV). We hypothesized that expression of these two receptor isoforms critically control granulopoiesis. We have developed a quantitative PCR assay and have found that class IV expression is variably upregulated in primary AML and leukemic cell lines. Receptor endocytosis rates differ between class I and class IV, which contributes to proximal phosphoprotein signaling profile (e.g. Lyn, Erk1/2, Akt). Aberrant expression of class IV leads to profound changes in intracellular signaling of both positive and negative effectors, resulting in proliferation without differentiation. Using a multi-scale approach, we have developed a mathematical model containing ordinary differential equations (ODE) to explain the kinetic properties of this dynamical system. This model involves three major steps: probability of receptor dimerization, single cell fate decision making, and population distribution of differentiated and non-differentiated cells. Computer simulation of receptor isoform expression and endocytosis revealed a decreased ratio of class I to class IV over time until steady state (∼120 min). Further validation is underway. This model can be applied to better understand disease states such as congenital neutropenia and acute myeloid leukemia, where aberrant G-CSF Receptor signaling can occur.
Human Meprin Alfa Metalloprotease Is Differentially Expressed in Peripheral Blood Cells.
Authors: Dunham Kimberly , Yura Renee , Bao Lei , Bond Judith , Sun Qi 
Published: April 2020

Abstract
Meprin metaloproteases are highly expressed in the apical brush-border of the intestinal and renal proximal tubule epithelia. However, the expression and function of these metaloproteinease in other tissues have not been well established. Using a specific antisera raised against rat but cross-reactive with human Meprin Alfa, we examined the expression of this protein in human peripheral blood cells. Flow cytometry analysis showed that the post-, but not the pre-, immune sera stained strongly for the CD14+ monocytes and moderately for CD56+ NK cells, but not for CD3+ T and CD19+ B cells. To confirm Meprin Alfa expression at the RNA level, peripheral blood cells were fractionated based on the expression of the above surface markers, followed by detection of Meprin Alfa message RNA by semi-quantitative RT-PCR. Consistent with the flow cytometry data, Meprin Alfa messages were detected only in cell fractions containing monocytes and NK cells but not T and B cells. Subsequent sequencing of the PCR amplified fragments confirmed the specificity of the PCR amplification. Interestingly, flow cytometry analysis further revealed that while the expression of Meprin Alfa was maintained in the macrophages differentiated in vitro from isolated monocytes, its expression was reduced in monocytes-derived immature dendritic cells (DC) and diminished in mature DC. The differential expression of Meprin Alfa in the peripheral blood cells suggests important physiological function of this protein in the immune system.
Plasma Cell Dyscrasia in Human Immunodefficiency Disease.
Authors: Jajeh Ahmad , Nukala Suresh 
Published: April 2020

Abstract
Plasmacytosis is commonly seen in the bone marrow biopsies of patients with acquired immunodefficiency disease AIDS and human HIV virus infection. Polyclonal increase of immunoglobulin is very common. However few cases of plasma cell dyscrasia are reported. The purpose of this abstract is to show our institution experience with plasma cell dyscrasia and HIV infection. For the last ten years ten patients were identified based on bone marrow biopsy, immunoelectophoresis and immunofixation, twenty four hours urine collection and urine electrophoresis and immunofixation. Immuno staining for kappa and lambda light chain to check for clonality were done on four cases. Mean age of patients is 49 years (range 33 to 59 years). Seven were black, two hispanic and one white. Nine were males and one female. All patients had plasmacytosis on bone marrow more than 30% and protiurea of more than one gram. five patients had nephrotic range protien urea. one patient had IgA lambda, one patient had Ig G kappa All patients had low CD4 counts except two patients with CD4 above 200 and have long standing HIV infection. One patient presented with hepatosplenomegally and increased IgM level and waldnestrome looking plasmacytoid lymphocytes. One patient had plasmablast and plasma cells syncitia who had rapidly fatal disease. one patient expired of respiratory failure before treatment. One patient, a female presented with plasma cell leukemia and did respond to Doxil, vincristine and dexamethasone therapy. She is alife. Two other patients with long standing HIV infection and CD4 count above 200 are still alife both patients were treated initially with Thalidomide and dexamethasone with excellent response. one of them failed Thalidomide after more than one year response and currently on his fourth cycle of velcaid (proteosome inhibitor). All patients were not on HIV therapy at time of diagnosis. In conclusion plasma cell dyscrasia with HIV infection do respond to treatment with durable response. Repetetive antigenic stimulation with HIV-1 p24 antigen are the proposed mechnism of oncogenesis.
Emerging Epidemics of Babesiosis in Hematology Consultation at a University Hospital.
Authors: Hosur Srilatha , Liu Delong , Seiter Karen , Nelson John , Aguero-Rosenfeld Maria , Brodsky Sergey , Ahmed Tauseef , Wormser Gary 
Published: April 2020

Abstract
Babesiosis was first reported in NY in 1975. 560 cases were reported from 1986–2001 in NY, mostly from Long island. Human babesiosis is a tick- borne disease caused by B.microti in the US. Most of the cases occur between May and August in the north eastern states. The risk factors for severe disease include older age, asplenia, immunocompromised state. As the same tick is the vector for lyme disease and ehrlichiosis, co-infection can occur. In this abstract we report ten cases of babesiosis encountered in hematology consultation since 2005, with 5 cases occurring this year. The patients’ ages ranged between 36–84 yrs. Three patients had neither travel history nor tick bite /outdoor activity. Three patients were immunocompromised (AML, CLL, HIV). Most common presentations were malaise, nausea, and high fever. Two patients had splenomegaly. Six patients had hyperbilirubinemia, anemia and thrombocytopenia. Three patients were also positive for lyme disease. One patient developed splenic rupture. All the patients recovered after treatment with atovoquone and azithromycin. This series of case reports emphasizes the varied atypical presentations of babesioisis including uncommon season of occurrence, absence of a rash or tickbite history, co-existing infection with Lyme disease and complications including splenic rupture.