bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Effect of Combined Chelation Therapy in the Treatment of Severe Myocardial Iron Loading in Beta Thalassaemia Major.
Authors: Tanner Mark A. , Galanello Renzo , Dessi Carlo , Westwood Mark A. , Pennell Dudley J. 
Published: April 2020

Abstract
Introduction: Thalassaemia major ™ is an inherited haemoglobinopathy resulting from mutations of the β-globin gene. Survival is dependent upon lifelong blood transfusions and results in progressive total body iron overload. Cardiac failure secondary to myocardial iron loading is by far the commonest cause of death with only 50% of patients in the UK in recent cohorts surviving beyond the age of 35 years. Conventional treatment with the parenteral iron chelator deferoxamine improves morbidity and mortality but fails to clear the heart of iron in approximately two-thirds of patients. More recently, the oral iron chelator deferiprone has been demonstrated to remove myocardial iron and it has been proposed that in combination with deferoxamine it may have an additive effect. Through the T2* multi-echo cardiac magnetic resonance (CMR) technique, myocardial iron can now be non-invasively and reproducibly quantified, thereby providing a means to assess myocardial iron loading in response to chelation treatment.
Purpose: To report the changes in myocardial T2* and cardiac function in patients with severe myocardial iron loading (myocardial T2* &lt;8ms) following treatment with the un-blinded addition of deferiprone to continued deferoxamine chelation therapy.
Methods: A mobile CMR scanner (1.5T Siemens Sonata) was transported to Cagliari, Italy. The myocardial T2* was assessed in 167 patients with TM aged 18v42 (30 +/− 5.3years). Of these, 22 patients were identified as having severe myocardial iron loading (T2* &lt;8ms), 15 of whom (at their clinician’s discretion) were subsequently given deferiprone in addition to continued deferoxamine therapy. Patients underwent further CMR assessment at baseline, 6 and 12 months.
Results: Over a period of 12 months myocardial T2* improved significantly (6.1+/− 1.0ms to 8.2ms +/−2.5ms, p=0.007) in association with an improvement in cardiac function (LV ejection fraction 52 +/−12% to 65% +/−6.6%, p&lt;0.0001). See Figures 1 and 2.
Conclusion: This prospective study shows that patients with severe myocardial iron loading show significant improvement in iron loading and myocardial function with the addition of deferiprone to continued parenteral deferoxamine.
Figure 1 Figure 1. Figure 2 Figure 2.
Genotype Phenotype Correlation in Thalassemia Syndromes and Their Correlation with Xmn-1 Polymorphism.
Authors: Sachdeva Anupam , Raina Aditya , Khanna Varinder Kumar , Arya Subhash Chander , Yadav Satya Prakash , Verma Ishwar 
Published: April 2020

Abstract
Among the inherited disorders of blood, thalassemia constitutes a major bulk of genetic diseases in India. It causes a high degree of morbidity. In a study conducted in India it has been estimated that there is a frequency of 1:2700 at the time of birth. Thus on an average 9000 new thalassemics are born every year in India.. The present study has been aimed at investigating the clinical and hematological spectrum in the above syndrome. It also assessed the prevalence of Xmn -1 polymorphism its relationship with various mutations and its role in modifying the clinical manifestations. The study was conducted on fifty patients representing 49 families and consisting of 33 males and 17 females who were homozygous for beta thalassemia and ranged in age from 3 months to 32 years. The patients were screened for common Indian mutations and their Xmn polymorphism status and this was correlated with their clinical parameters. Apositive correlation in presence of Xmn-1 polymorphism and IVS1-1 mutation was noted. There was also a correlation between age at diagnosis and also the age at first transfusion. There was a correlation between Xmn polymorphism and IVS1-1 mutation. This was found most commonly in the Punjabi Khatri community. None of the patients with blood group O had positivity of Xmn polymorphism. There was no correlation between Xmn and hemoglobin at diagnosis, HbF at diagnosis, MCV at diagnosis, amount of blood in mL/Kg/year and sex and religion.
Atypical Hemoglobin H Disease Due to Compound Heterozygosity for -α3.7 and --SEA Deletions and Heterozygosity for the β-Chain Variant Hemoglobin Raleigh (β1 VAL→ALA).
Authors: Kutlar Ferdane , Knovich Mary Ann , Elam Dedrey , Jobe Daniel B. , Buss David H. , Harbin Jeanette , Davis Deborah H. , Sabio Hernon , Kutlar Abdullah 
Published: April 2020

Abstract
The co-existence of Hemoglobin H (Hb H) disease and heterozygosity for β-chain structural variants is a rare occurrence. Hb H disease has been reported in conjunction with Hb E, Hb C, Hb S, and Hb Hamilton. The combination of Hb H disease with Hb C and Hb S reportedly results in a mild hemolytic anemia without detectable Hb H. We present a new case of atypical Hb H disease that was also heterozygous for the rare β-chain variant Hb Raleigh. The patient is a 27-year-old Cambodian female referred for the evaluation of microcytic anemia unresponsive to iron. She had a lifelong history of generalized fatigue, exertional dyspnea, and weakness in her legs. Physical exam was unremarkable except for pallor of mucuous membranes. There was no hepatosplenomegaly. She had a Hb of 9.7, HCT 30.8, MCV 56, MCH 17.6, MCHC 31.5, ferritin 92. Hb analysis on IEF revealed Hb A, Hb A2, and an abnormal band slightly more anodic to Hb A. No Hb H was observed. On cation exchange HPLC, she had 49.7% Hb A, 48.1% Hb X, and 2.2% Hb A2. Reverse phase HPLC revealed a βx chain eluting immediately before βA. Oxygen affinity was slightly reduced. PCR amplification and sequencing of the β-globin gene revealed heterozygosity for Hb Raleigh (Exon 1, codon 1, GTG→GCG, VAL→ALA). The patient was also found to be a compound heterozygote for -α3.7 and --SEA deletions. This case represents a novel interaction of a structural β-chain variant with Hb H disease. Hb Raleigh has previously been reported in Caucasians and in two Swedish families. It has decreased oxygen affinity. This is the first report of this variant in a Cambodian population. The absence of any detectable Hb H likely results from the inability of variant β-chains to form a viable tetramer with a resultant decrease in βA. The low oxygen affinity did not negatively impact on the degree of anemia. This case, like some others reported previously, shows that the accurate diagnosis of Hb H disease in association with structural β-chain variants can be established by molecular methods, and the detection of Hb H on electrophoretic and chromatographic analyses may not always be reliable.
How To Optimize the Detection of the Left Shift with the Instrument COULTER LH750®.
Authors: Capel-Casbas Maria J. , Duran Jose J. , Naranjo Dolores , Pastor Isabel , Berlanga Rosalia , Masset Maria J. , Piqueras Josep , Pujol-Moix Nuria 
Published: April 2020

Abstract
The Coulter LH750® is a last generation automated hematology analyzer. In 1999 we evaluated the optimization of immature granulocytes/bands (IG/B) detection in the Coulter GEN*S (Blood Vol.94 n° 10 sup.1, 1999 page 45 b), in which we observed that the Mean Volume of the Neutrophil population (M NE V) and the standard deviation of the neutrophil volume (SD NE V) were the most useful parameters to improve the detection of the presence of IG/B. Recently the utility of SD NE V has been proposed in the diagnosis of septic patients.(ISLH 05). The purpose of this study is to confirm that the NE morphometric parameters in the LH750 have the same values and cut-off as the GEN*S to discriminate the cases with lefts shift and if it is possible to create rules/flags that use these parameters to allow the automated detection of IG/B with cut-off levels specific to the origin (emergency room, outpatients, intensive care etc.) of the patient sample.
In addition we investigated if the parameters are useful when taking into account not only the % of IG/B, but also their absolute number as recommended by the NCCLS. The overall intent being to optimize the efficiency of our laboratory in the detection of IG/B by decreasing the number of smears, but without increasing the number of false negatives. We have selected from the routine samples of a 2000 beds hospital, 340 consecutive cases on the LH750® that triggered the bands /Immature granulocytes flags, Immature NE 1 (NE1) or Immature NE 2 (NE2). We considered the samples with more than 0,9 IG/B (103/mL) as positive. Blood smears were performed on all the positive samples (340) and a WBC differential was performed according the NCCLS protocol (H 20A). The percentage and absolute number of IG/B were collected. The statistical analysis was performed using Medcalc® version 8.1.0.0. A ROC Curve and the cut-off for the detection of positive samples with immature IG/B using the SD NE V and M NE V was calculated.
RESULTS: For the flag NE1 the best parameter to select the postive cases with left shift was the SD NE V with a cut-off of &gt;24,2. (although this flag is actually defined for a moderate left shift). For the flag NE2 the best parameter to select the postive cases with left shift was the M NE V with a cut-off of &gt; 161.
CONCLUSION: Before the study performed in 1999 we validated the Imm NE1 and Imm NE2 flags according to the NE% and NE# or the WBC values, that statistically demonstrated limited utility. Parameters which permitted to optimize the detection of IG/B, were the Neutrophile morphological parameters; M NE V and SD NE V. The values of the LH 750 are slightly different to those of the GEN*S. We will subsequently study their utility and advantages compared to the IG/B counting in the detection of bacterial infections and sepsis due to better reproducibility.
Rituximab in the Treatment of Secondary Autoimmune Neutropenia.
Authors: Pivkova Aleksandra A. , Cevreska Lidija L. , Stojanoski Zlate Z. , Panovska Irina I. , Siljanoski Nikola N. , Stavrik Sonja S. Genadieva , Balkanov Svetlana S. Krstevska , Trajkova Sanja S. , Georgievski Borce B. 
Published: April 2020

Abstract
Autoimmune cyclical neutropenia is a rare syndrome with periodically severe but temporary neutropenia with occurrence of 3–4 weeks, but not rarely in 2 or 5 week intervals. Antibodies with specificity for the neutrophils has been demonstrated in a number of different neutropenic disorders and is often associated with systemic autoimmune disease. Since the role of monoclonal antibodies in the treatment of autoimmune disorders is not clearly optimized, we evaluated the effectiveness of Rituximab in the treatment of immune neutropenia.
We present two cases (male 66 years and 42 years female) with immune neutropenia, associated with systemic autoimmune disease, Chron’s disease in male patient and rheumatoid arthritis in female patient. Bone marrow biopsy revealed reduction of granulopoesis and absence of neutrophils in the peripheral smear-agranulocytosis. Immunological investigations revealed increased polyclonal IgG, positive indirect granulocyte immunofluorescence test. Both patients had a clinical course with continuously intermittent neutropenia every 2 weeks, presented with recurrent febrile episodes and serious lower respiratory tract infections. Treatment with G-CSF was immediately started in both patients to reach neutrophil recovery, but after one year of periodically persistent neutropenia it was decided to start treatment with Rituximab, an infusion with standard single agent 375 mg/m2 weekly for 4 weeks. After 4 infusions neutrophils reached normal levels and the treatment continued with every 2 weeks infusion of monoclonal antibody in duration of 2 months and now once a month as maintenance treatment. The patients are one year and six months after beginning treatment with monoclonal antibodies, in stable clinical and laboratory condition. We can conclude that B cells play a central role in the autoimmune process and Rituximab treatment has been used in patients with autoimmune neutropenia. Further randomized studies will define optimal role of Rituximab in the treatment of autoimmune diseases.
Human Cytomegalovirus Inhibits Granulopoiesis Via Direct Infection and Inducing Apoptosis.
Authors: Wang Qing Wen , He Zheng Xian , Li Xiao Feng , Xiong Wei , Chen Mei Lian , Li Karen Kwai Har , Yang Mo 
Published: April 2020

Abstract
Human cytomegalovirus (HCMV) infection can cause delayed leukocyte recovery after bone marrow transplantation and often associated with suppression of granulocyte/macrophage progenitor. Leukocyte lineage may be one of the major sites of HCMV infection. However, whether HCMV can interfere with CFU-GM formation and induce apoptosis in granulocyte/macrophage progenitor have not been well investigated. Human bone marrow mononuclear cells (Ficoll), promyelocyte cell line HL-60 and HCMV AD169 strain were co-cultured. Each bone marrow specimen was HCMV DNA and HCMV IgM negative by PCR and ELISA test. Our results showed that HCMV significantly inhibited the formation of CFU-GM as shown in two different concentrations of viral infection groups: 139.26 ± 5.42 (2×105pfu/ml), and 124.19 ± 8.82 (2×106pfu/ml) (colonies/2×105cells/ml, n=26). These were significant differences compared with the blank control group (P&lt;0.01, P&lt;0.05) respectively. HCMV also significantly inhibited the growth of HL-60 cells in three different concentrations of viral infection groups (101pfu/ml, 102pfu/ml, 103pfu/ml). After incubation for 7 days, viability cells in control group and each infection groups (n=20) were 96%, 83%, 73%, and 64%, showing that HCMV infection decreased the viability of HL-60 cells in a dose-dependent manner. The apoptotic effect was also investigated by Annexin V assay using flow cytometry. The percentage of Annexin V -positive cells were 19.60 ± 1.63 (in group of 101pfu/ml) (P&lt;0.05), 28.70 ± 2.61 (102pfu/ml) (P&lt;0.05), and 36.3 ± 2.57 (103pfu/ml) (P&lt;0.01), compared with control group 3.96 ± 1.75 (n=8). The apoptotic cells were further confirmed by morphologic observation and DNA ladder formation. Furthermore, HCMV DNA was positive in HL-60 cells and cells of CFU-GM measured by PCR and IS-PCR analysis respectively. These were negative in blank and mock control groups. Our data demonstrated that: (1) HCMV AD169 strain inhibited the growth of HL-60 and CFU-GM formation in a dose-dependent manner; (2) HCMV transcription occurred in HL-60 and CFU-GM cells; and (3) This virus induced apoptosis in HL-60 cells. HCMV may have an inhibiting effect on granulopoiesis via direct infection and inducing apoptosis on myeloid progenitors.
An Indispensable Role of GM-CSF in the Differentiation of Human Blood Monocytes toward Langerhans Cells.
Authors: Shibasaki Tetsunori , Katayama Naoyuki , Ohishi Kohshi , Monma Fumihiko , Yu Zhaocai , Nishii Kazuhiro , Masuya Masahiro , Shiku Hiroshi 
Published: April 2020

Abstract
We recently demonstrated that GM-CSF, TGFβ1, and Notch ligand Delta-1, all of which are synthesized in the milieu of the skin, instruct human blood monocytes to differentiate into Langerhans cells (LCs) that are characterized by the expression of CD1a, Langerin, CLA, CCR6, and E-cadherin and the presence of Birbeck granules. Previous studies have shown similar biologic activities of GM-CSF and IL-3 on human blood monocytes. In the present study, we investigated whether GM-CSF and IL-3 have a similar action on blood monocytes with respect to their differentiation into LCs. In contrast to CD14+ monocytes cultured for 7 days in the presence of GM-CSF, TGF-β1, and Delta-1, the cells obtained from culture with IL-3, TGF-β1, and Delta-1 were negative for CLA, CCR6, Langerin, and E-cadherin. These cells expressed HLA-ABC, HLA-DR, CD80, CD86, CD40, CD54, and CD11c but not DC-SIGN. The expression level of CD1a declined, while CD14 was upregulated. The difference between cells cultured with GM-CSF, TGF-β1, and Delta-1 and those with IL-3, TGF-β1, and Delta-1 was corroborated by microarray analysis of the gene expression profiles. Thus, GM-CSF and IL-3 exert distinct effects on human blood monocytes in the presence of TGF-β1 and Delta-1. When GM-CSF was added to cultures containing IL-3, TGF-β1, and Delta-1, the phenotype of cultured cells closely resembled that observed with GM-CSF, TGF-β1, and Delta-1. As GM-CSF and IL-3 share common receptor β subunits, the signals mediated through GM-CSF receptor α subunits appear to be required for the development of LCs from monocytes. We also compared the action of GM-CSF with that of IL-3 in terms of the differentiation of monocytes into macrophages and dendritic cells (DCs). When CD14+ monocytes were cultured in the presence of GM-CSF or IL-3 for 7 days, the resulting macrophages obtained from both cultures were phenotypically indistinguishable. Next, we examined whether IL-3 as well as GM-CSF induces CD14+ monocytes to differentiate into DCs in the presence of IL-4. The phenotypic profiles in cells cultured with IL-3 plus IL-4 were parallel to those seen with DCs cultured in the presence of GM-CSF plus IL-4. These data suggest that although IL-3 can substitute for GM-CSF in the differentiation pathway of CD14+ monocytes toward macrophages or DCs, GM-CSF is indispensable for their LC development.
Immunopotent Polysaccharides of Different Sources Selectively Activate Human Dendritic Cells.
Authors: Chan Godfrey ChiFung , Chan W.K. , Law H.K. , Lin Z.B. , Lau Y.L. 
Published: April 2020

Abstract
Background: Purified polysaccharides extracted from plants and fungi have been shown to induce immune responses in-vivo and vitro over the past decade. Currently, most of these polysaccharides are found to be glucan but with different branch structure and sizes. Their relative potency and effect on human immune cells remains unknown. This study aims to compare their relative effect on human dendritic cell, the most potent antigen presenting cell.
Materials & Methods: We selected 2 prototypes of purified polysaccharides extracted from: 1) Ganoderma lucidum (GL, Lingzhi, Reishi) mycelium, a widely used herb with long and branching β (1® 3), (1® 6) glucan structure (provided by Prof. Lin ZB, Beijing) and 2) Barley with shorter and different branching β (1® 3), (1® 4) structure (provided by Prof. Cheung VNK, NY). Their characteristics and chemical properties had been reported previously. Human peripheral blood mononuclear cells (PBMCs) proliferation was studied by XTT assay. Human dendritic cells (DCs) were derived from monocytes and maturation of DCs were determined by: a) immunophenotypic shift using flow cytometer; 2) dextran endocytosis assay and 3) mixed lymphocytes reaction. Cytokine secretions were determined by ELISA test. Comparisons between means were by nonparametric Student’s t test (2-tailed).
Results: We found that purified polysaccharides from GL but not barley could induce PBMCs proliferation and maturation of DCs. GL polysaccharides could enhance phenotypic and functional maturation of DCs with significant IL-12 and IL-10 production. DCs were relatively inert to Barley glucans stimulation. However, both polysaccharides did not polarize T cells into the direction of T helper 1, T helper 2 or regulatory T cells.
Conclusions: Our study shown that purified polysaccharides extracted from plants and fungi have different effect on human DCs and their potency and effects are probably affected by their respective sources and structures.
Mast Cell-Specific Gene Targeting.
Authors: Melicoff Ernestina , Thakurdas Shakeel M. , Siddiqi Tanya , Murthy Jayasimha , Sansores-Garcia Leticia , Adachi Roberto 
Published: April 2020

Abstract
For years mast cell (MC) studies have relied on MC lines or bone marrow-derived MC. Regular knockout mice have some disadvantages such as early lethality and developmental defects. Also, the phenotype in MCs from these animals could not be a primary defect but secondary to other cell lines affected by the mutation. The Cre/loxP system allows us to target genetic deletions to specific tissues or cells. Cre, a recombinase from bacteriophage P1, recognizes a DNA sequence motif of 34 bases (loxP). If a DNA segment is flanked by two loxP sites in the same orientation, Cre excises that segment activating or silencing a gene. Many genes have been “floxed” (flanked by two loxP sites), but no MC-specific Cre mouse has been created. We are using promoter regions of MC-specific proteins to drive expression of Cre-recombinase. We produced transgenic mice using the promoter for human alpha-subunit and murine beta-subunit of the high-affinity receptor for IgE (hFcεRIα and mFcεRIβ), both expressed in MCs and basophils. We also used the promoter of the murine mast cell protease mMCP-5, a member of a family of cargo proteins expressed only in MCs. For even more controlled expression, we knocked-in Cre cDNA into the mMCP-5, mMCP-6 and Ras guanine releasing protein 4 (RasGRP4) loci, all of them proteins expressed exclusively in MCs, while knocking-out the respective gene. Heterozygotes will express Cre only in MCs, and homozygotes for mMCP-5, mMCP-6 or RasGRP4 deletions will be regular knockouts that will allow us to study the role of these proteins in mast cells. These three transgenic and three knock-out/knock-in lines have been created and are being crossed with reporter mice. We decided to use a double reporter mouse, the Bgeo/GFP (Jackson Laboratory). This mouse has “floxed” lacZ that is constitutively expressed under the control of the CMV enhancer/chicken actin promoter, and when crossed with our Cre recombinase-expressing mouse, lacZ is excised and enhanced green fluorescent protein (EGFP) is expressed in cells expressing Cre. EGFP can be easily detected in MCs from peritoneal lavage by flow-cytometry, using antibodies against c-kit (CD117) and FcεRI to label the MCs. Results of this screening will be presented at the meeting. After documenting the presence of functional Cre-recombinase in our reporter mice, we will cross them with our “floxed” mice to obtain conditional mast cell-specific knockouts.