bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flippase Activity Decreases throughout Erythrocyte Life-Span in Normal and Sickle Mice.
Authors: DeJong Kitty , Bhagat Amrita , Emerson Renee K. , Kuypers Frans A. 
Published: October 2019

Abstract
Phosphatidylserine (PS) exposure contributes to recognition and removal of aged erythrocytes. PS exposure can only arise in cells in which the aminophospholipid translocase, or flippase, is defunct, or otherwise PS would be transported back into the inner cell monolayer. To provide evidence that flippase activity is reduced in old erythrocytes, we studied mouse erythrocytes that were biotinylated in vivo. This way we were able to distinguish the aging, biotinylated, RBC population from the newly formed, unbiotinylated RBC. Detection of the biotinylation with fluorescent streptavidin was combined with a flow cytometric assay for flippase activity using NBD-PS. We found that flippase activity decreased over time during the life-span of the RBC, as shown by a decrease in flippase activity of the biotinylated RBC compared to the newly formed RBC. A particularly high flippase activity was found in CD71-positive reticulocytes. Murine sickle cells show a much higher erythrocyte turnover than normal erythrocytes, with a 25–50% newly formed population each day and complete elimination of all biotinylated cells in 10 days. As in control mice, flippase activity decreased during the (short) life-span of sickle RBC and at all times, a subpopulation of sickle cells had a markedly reduced flippase activity. High flippase activity was only found in newly released erythrocytes. Our data indicate that the flippase activity decreases throughout the life-span of erythrocytes and can be markedly different in subpopulations of RBC in vivo. This decrease reduces the cell’s potential to maintain its phospholipid asymmetry, which results in PS exposure as a removal signal for the damaged cell. We conclude that the inability of sickle RBC to maintain flippase activity is related to the decreased survival time of those cells.
Long-Term Vitamin D Supplementation Improves Bone Mineral Density (BMD) in Patients with Sickle Cell Disease.
Authors: Adewoye Adeboye H. , Holick Michael F. , McMahon Lillian C. , Ma Qanli , Steinberg Martin H. 
Published: October 2019

Abstract
Among the many complications of sickle cell disease (SCD) are osteopenia and osteoporosis. We and others have previously documented a high prevalence of low serum 25-OH vitamin D level with accompanying osteopenia and osteoporosis in patients with SCD; however the long-term effects of treatment on BMD have not been reported. We hypothesized that supplementation with oral vitamin D and calcium will result in an improvement in BMD and accordingly, treated 6 adult subjects with SCD for 12 months with oral supplementation with vitamin D and calcium and studied the effects of treatment on BMD. Subjects with serum 25-OH D levels <20 ng/ml were placed on 50,000 IU of oral vitamin D weekly for 8 weeks, followed by 50,000 IU every other week for 44 weeks; subjects also received 1000 mg of elemental calcium daily. We evaluated BMD at weeks 1 and 52. Subjects had a mean pre-treatment plasma 25-OH vitamin D of 9.1 ng/ml. BMD was abnormal in all subjects; 4 subjects had osteoporosis with a Z-score <-2.5, and 2 subjects had osteopenia with a Z-score −1.9 to −2.5 (in the lumbo-sacral vertebrae). All subjects had mildly elevated intact PTH levels which were within the reference range but declined further post-treatment. Following one year of treatment with vitamin D and calcium, we observed a mean post-treatment increase in BMD of 5.4% (p value <0.02) and a 176% increase in plasma 25-OH D level (from 9.1ng/ml to 28ng/ml; p value <0.009). All patients with SCD we have studied have hypovitaminosis D that in adults is associated with low BMD. In patients followed for one year who received supplementation with calcium and vitamin D, 25-OH D level increased in all and BMD improved in four of six subjects. These results suggest that this treatment regimen can be effective is restoring BMD. The reasons for apparent unresponsiveness in some patients must be further evaluated. Clinical trials to document the effectiveness of treatment for this ubiquitous problem should be instituted.
Mean change in 25-OH vitamin D level Mean change in 25-OH vitamin D level Mean Change in BMD: Pre and Post Treatment Mean Change in BMD: Pre and Post Treatment
Genomic Instability in Sickle Cell Disease Patients Using Hydroxyurea Assessed by Micronucleus Assay.
Authors: Friedrisch Joao R. , Maluf Sharbel W. , Bittar Christina M. , Silva Maria A. , Silla Lucia M. 
Published: October 2019

Abstract
Hydroxyurea (HU) is an antineoplastic drug, which also plays an important role in treatment of sickle cell disease patients (SCD). Short-term HU toxicities primarily include transient myelosuppression, but long-term HU risks have not been defined. The mutagenic and carcinogenic potential of HU is not established, although HU has been associated with an increased risk of leukemia in some patients with myeloproliferative disorders.
In the present study we analyzed the presence of DNA damage in patients with SCD treated with HU, in peripheral blood lymphocytes, using micronucleus assay. We analyzed 36 patients with SCD (16 males and 20 females), aged 2–59 years (mean 25,75 ± 14,5), received oral HU median dose of 26,5 mg/Kg/day, for a period of 0.6– 11,8 years (mean 5,01). The control group was composed of 34 healthy individuals (16 males and 18 females), aged 4–52 years (mean 26,61 ± 7,20). The results revealed that frequency of micronucleus was significantly higher in SCD patients using HU than in controls(p=0,0182). This study indicates a possible genotoxicity of the HU, although further works are necessary to evaluate its mutagenicity.
Exercise Capacity and Pulmonary Function in Sickle Cell Patients with Pulmonary Arterial Hypertension.
Authors: van Beers Eduard J. , van der Plas Mart N. , Bogaard Harm J. , Biemond Bart J. 
Published: October 2019

Abstract
Pulmonary arterial hypertension (PAH) is increasingly recognised as a complication of sickle cell disease (SCD), with a prevalence of approximately 30%. In SCD, a tricuspid regurgitant jet velocity (TRV; determined by cardiac ultrasound) of 2.5 m/s or higher is used as a non-invasive marker of PAH and is associated with a poor prognosis. Since PAH in SCD is usually mild, it remains unclear whether PAH is directly responsible for the early death of these patients. Exercise capacity is closely related to survival in heart failure and other chronic cardiopulmonary diseases. We hypothesized that if PAH in SCD determines survival, it should be related to exercise capacity. The aim of this study was to determine whether exercise capacity is reduced in SCD patients with an increased TRV. In addition, it was determined whether exercise capacity is related to pulmonary function in these patients. Cardiac ultrasound, pulmonary function testing and a symptom-limited maximal cardiopulmonary exercise tests(CPET) using cycle ergometry were performed in 42 consecutive patients with SCD. PAH was present in 9/42 (21%) of the patients.
Exercise capacity was decreased in all patients, with a trend to a lower peak oxygen uptake (VO2peak) and oxygen uptake at the anaerobic threshold (VO2peak/AT) in patients with PAH compared to patients without PAH (74±18% and 57±18% versus 63±14% and 46±10%, respectively, p=0.16 and p=0.095). After matching for blood hemoglobin content, differences in exercise capacity disappeared. Pulmonary function was also impaired in both groups, with much larger differences between PAH and non-PAH patients: vital capacity (%predicted) 72±13% vs 85±15% and total lung capacity 70±11% vs. 81±11% (p=0.025 and p=0.020). No differences in FEV1 or membrane diffusion capacity were demonstrated. Multiple regression analysis, with the use of VO2peak as dependent variable and age, SCD phenotype, hemoglobin, spirometric parametres and TRV identified only vital capacity and hemoglobin as a significant independent correlates of VO2peak and not TRV. In conclusion, PAH in SCD is not related to exercise capacity, which could mean that it has no direct effect on mortality. SCD is associated with a restrictive pulmonary function impairment, especially in the group of patients with PAH. Interestingly, low VO2peak in SCD appears to be determined by the severity of the anemia instead of the prevalence of PAH.
Monocyte Chemoattractant Protein-1 and Interleukin-8 Levels in Acute Inflammation Induced by Prolonged Brisk Exercise.
Authors: Papassotiriou Ioannis , Tsironi Maria , Skenderi Katerina , Chrousos George 
Published: October 2019

Abstract
Monocyte chemoattractant protein-1 (MCP-1), also known as CCL2, a chemokine that regulates migration and infiltration by monocytes/macrophages, belongs to the CC chemokine subfamily. Interleukin-8 (IL-8), also known as CXCL8, a proinflammatory chemokine with angiogenesis-promoting properties belongs to the CXC chemokine superfamily. Both MCP-1 and IL-8 have important roles in the pathogenesis of many chronic inflammatory disorders, including atherosclerosis and obesity. Their proinflammatory effects are mediated mainly by the CC chemokine receptor 2 (CCR2) and CXCR1/R2, respectively. MCP-1 and IL-8 cause chronic vascular inflammation and induce thrombosis, proliferation and migration of vascular smooth muscle cells, angiogenesis, and oxidative stress. Previous studies indicate that:
MCP-1 production from endothelial cells, smooth muscle cells, and regional leukocytes increases in the presence of endothelial dysfunction and atherosclerosis risk factors; MCP-1 and IL-8 expression is increased in atherosclerotic lesions and injured arteries; and eliminating MCP-1 function decreases neointimal hyperplasia after injury and atheroma formation in mice.
We studied the association between MCP-1 and IL-8 levels and the degree of inflammation in 15 athletes that participated in the ultra-distance foot race of the 246 Km “Sparthathlon”. This race consists of continuous, prolonged, brisk exercise. We reported earlier that Interleukin-6 (IL-6), C-reactive protein (CRP), Serum amyloid A protein (SAA) and free plasma DNA levels markedly increased (by 8000–, 152– 108– and 10-fold, respectively) over the baseline at the end of the race1. However, IL-6 levels returned to normal by 48h, while CRP, SAA and free plasma DNA remained elevated. Circulated levels of MCP-1 and IL-8 were measured by means of a multi-analyte Biochip Array Technology, using the Evidence analyzer (Randox Laboratories, UK). The measurements were performed before (phase I), at the end (phase II) and 48h post-race (phase III). MCP-1 levels at phase I (216.9±48.5 ng/L) (mean±SE), increased significantly at phase II (592.9±115.7 ng/L) and subsequently decreased at phase III (278.1±62.7 ng/L). At the same time period, IL-8 followed a similar pattern (phase I: 9.4±4.5 ng/L, phase II: 28.5±8.8 and phase III: 8.9±4.3 ng/L). A significant positive correlation between MCP-1 and IL-8 was found at phase III (r=0.845, p<0.01), while this correlation was absent in the other two phases, indicating an independent response of each chemokine to inflammatory stimuli in each athlete.
In conclusion, prolonged exercise induces an inflammatory response that is expressed by an increase in circulating MCP-1 and IL-8 levels. Whether these changes have long-term negative effects on the vasculature remains unknown.
HNA−1a, −1b, −2a, −3a and −4a Frequencies in Brazilian Persons.
Authors: Norcia Angela M. , Kimura Elisa Y. , Chiba Akemi K. , Moritz Elyse , Yamamoto Mihoko , Bordin Jose Orlando 
Published: October 2019

Abstract
Granulocyte reactive antibodies have been found to cause clinical disorders such as transfusion related acute lung injury (TRALI), febrile transfusion reactions, alloimmune neonatal neutropenia, immune neutropenia after bone marrow transplantation, refractoriness to granulocyte transfusion, drug-induced neutropenia and autoimmune neutropenia. Using the granulocyte immunofluorescence test (GIFT) by microscopic and flow cytometric analysis, the frequencies of the neutrophil antigens HNA−1a, −1b, −2a, −3a and −4a were determined among 100 random Brazilian blood donors from the Blood Center of Universidade Federal de Sao Paulo, SP, Brazil. Granulocytes were separated from mononuclear cells and red cells by sedimentation with 5% dextran, followed by centrifugation on Ficoll-paque (d = 1.077), and then incubated with anti-sera (anti-HNA−1a, −1b, −2a, −3a, and −4a obtained from American Red Cross, North Central Blood Services, St. Paul, MN) conjugated with fluorescein isothiocyanate (FITC) labeled F(ab’)2 fragments of anti-human IgG. Only cell suspensions containing ≥95% neutrophils with viability ≥90% according to the trypan-blue staining were analyzed. The frequencies of HNA−1a, −1b and −2a were 65%, 83% and 94%, respectively, and for such alloantigens exact same results were observed using either the GIFT performed by microscopy or by flow cytometry. The frequency of HNA-3a was 86% by the microscopic GIFT, and 95% by the flow cytometry analysis; while the frequency of HNA−4a was 93% by the microscopic GIFT, and 94% by the flow cytometric technique. These results indicate that:
GIFT by flow cytometry is more sensitive than the GIFT by microscopy to detect HNA−3a; the phenotypic frequencies found for neutrophil antigens HNA−1a and −1b among Brazilian blood donors are quite similar to those reported among African Americans, but different from those reported for Japanese and Chinese individuals; the phenotype frequencies of the neutrophil antigens HNA−2a, −3a, and −4a in Brazilians are quite similar to those found among Caucasians (Table). (These studies were funded by FAPESP, SP, Brazil - 05/55237–9).
Asians Brazilians Antigen African Americans Chineses Hindus Japaneses Caucasians M FC M, microscopy; FC, flow cytometry; nd, not done HNA-1a 46 – 68 90 44 88 52 – 54 65 65 HNA-1b 78 – 84 52 83 51 – 64 87 – 89 83 83 HNA-2a nd 99 nd 89 87 – 97 94 94 HNA-3a nd nd nd nd 99 86 95 HNA-4a nd nd nd nd 96 93 94
Upregulation of Myeloperoxidase in End Stage Renal Disease and Its Modulation by Hemodialysis. A Paradoxical Paradigm.
Authors: Cunanan Josephine , Hoppensteadt Debra , Bansal Vinod , Fareed Jawed 
Published: October 2019

Abstract
Myeloperoxidase (MPO), a heme protein released from activated leucocytes mediates the generation of reactive intermediates promoting lipid peroxidation. MPO is secreted by activated neutrophils, monocytes and macrophages. MPO has been reported to be a marker of hemodialysis procedure biocompatibility and oxidative stress with discordant results. Different dialyzer membranes and the type of anticoagulation used contribute to the regulation of MPO in end stage renal disease (ESRD) patients. To validate the hypothesis that MPO is upregulated in ESRD and hemodialysis differentially modulates the MPO in patients with this syndrome, blood plasma samples from 70 patients on maintenance hemodialysis prior to and after hemodialysis session. The pre-dialysis levels of MPO antigen as measured by an ELISA method (Assay Design, ) ranged from 0.5–125 ng/ml (31.1±28.6 ng/ml), in contrast to the aged match normal controls where the range was 0.21–7.5 ng/ml, (2.8±1.3 ng/ml). After dialysis, 10/70 patients did not show any changes in the MPO level, whereas, 4 patients showed a decrease, the remaining 56 patients exhibited marked increase in the circulating MPO level ranging from 9.5–201 ng/ml (74.4±48.2 ng/ml). Progressive atherosclerotic changes are associated with the severity of ESRD where oxidative stress is known to play a major role. While MPO may mediated oxidative stress the beneficial consequences include its depletion from the vascular site. Hemodialysis in ESRD generally results in a downregulation of inflammatory mediators. The increased MPO level in ESRD represents a paradoxical and complex process which may contribute to the pathogenesis of ESRD. The role of different anticoagulants and dialyzer membrane and other factors in regulating circulating MPO level may be important in elucidating the pathogenesis and outcome of this syndrome.
Characterization of Dendritic Cell-Specific Genes p275 and p306.
Authors: Hilgendorf Ingo , Kurz Daniel , Bringmann Anita , Kanz Lothar , Grünebach Frank , Brossart Peter 
Published: October 2019

Abstract
Dendritic cells play an inimitable role in the functioning immune system as they are the most potent antigen presenting cells being able to prime naive T-cells. Their characteristic properties that enable them to take up antigens and present them to leukocytes are due to an expression of specific genes and thus specific proteins that are unique to this subset of antigen-presenting cells. Using a substractive cDNA library based on suppression hybridization between DC cDNA and the reference monocyte cDNA, we identified in DC two differentially expressed genes p275 and p306. p275 codes for a membrane protein and represents a splice isoform of the transport protein NAT-1. The predicted structure of protein p306 is globular, suggesting that the protein is either intracellular or secreted. The expression of both genes was confirmed by RT-PCR using cDNA isolated from peripheral blood monocytes and DC, generated in vitro from monocytes or CD34+ progenitor cells. To further analyze the protein expression polyclonal antibodies were generated by immunization with synthetic peptides deduced from the identified sequences. Interestingly, inhibition of DC differentiation using IL-10 or STI571 (Imatinib) resulted in an impaired expression of both proteins. Utilizing specific primers for two recently described splice variants of p306 we identified a new splice form expressed in DC. While the gene of p306 contains eight exons, splice variant 1 consists of the exons 1,2,4,5,6, and 7 and splice variant 2 contains the exons 1,2,3,4,5,6, a shortened exon 7, and exon 8. The new identified splice form includes the exons 1–7. However, as the open reading frame starts in exon 4, the expressed protein is identical with the one corresponding to splice variant 1. Analyzing different DC populations in peripheral blood we show that p306 is expressed in plasmacytoid, but not myeloid DC. Interestingly, the activation of DC with Toll-like receptor ligands (TLRL) Pam3Cys (TLR2L), Poly I:C (TLR3L), LPS (TLR4L) and R848 (TLR7L) has no influence on the expression of p306. Although the functions of p275 and p306 in DC have yet to be determined, both genes play a role in DC differentiation and are found in different hematopoietic cell populations. Especially p306 might be an interesting marker of plasmacytoid DC as the predicted protein does not resemble any known protein structure.
Enhanced Bacteriocidal Function by WKYMVm in Patients with Acute Leukemia.
Authors: Kim Hawk , Shin Soojin , Park Jae-Hoo , Noh Eui-Kyu , Min Young Joo 
Published: October 2019

Abstract
Acute leukemia (AL) is a disease of challenge in that many patients die not only from eventual relapse but also from infection during treatment. In this regard efforts for decreasing infectious mortality will help increasing survival. Our previous study revealed that leukocyte bacteriocidal functions in chemotherapy-treated cancer patients were decreased and were stimulated by a novel hexapeptide, WKYMVm. We evaluated the leukocyte bacteriocidal function in patients with AL and searched whether WKYMVm can enhance the bacteriocidal function in patients with AL. On induction chemotherapy, blood sampling was performed at diagnosis and repeated weekly from start day of chemotherapy until patient died or complete remission was achieved. Tests were done weekly until white blood cell (WBC) count reached up to 1000/mm3 and platelet counts were stable without transfusion after consolidation chemotherapy. Nineteen AL patients and 10 healthy controls were enrolled. Diseases were acute myeloid leukemia (17 patients) and acute lymphoblastic leukemia (2 patients). Median WBC, absolute blast count (ABC) and absolute neutrophil count (ANC) at diagnosis were 5280/mm3 (range 840–315100), 162.9/mm3 (range 0–286741) and 796.4/mm3 (range 126–7336), respectively. Mean values of bacreriocidal activity at diagnosis were increased by concentrations of WKYMVm (13.4 at 0 nM; 24.9 at 1 nM; 33.3 at 10 nM; 38.5 at 100 nM; p<0.001), which were also increased in normal samples (20.6 at 0 nM; 41.6 at 1 nM; 51.6 at 10 nM; 66.4 at 100 nM; p<0.001). At each concentrations of WKYMVm, the bacteriocidal activities were inferior to those of normal control (p=0.029, at 0 nM; p=0.015, at 1 nM; p=0.015, at 10 nM; p=0.015, at 100 nM). The bacteriocidal activities were increased (p=0.008, at 0 nM; p=0.015, at 1 nM; p=0.011, at 10 nM; p=0.021, at 100 nM) compared with the corresponding values when patients achieved complete remission (CR). However, bactericidal activities by stimulation of WKYMVm were inferior to normal control even in CR (p=0.036, at 1 nM; p=0.036, at 10 nM; p=0.036, at 100 nM) although base line value were similar (p=1.0, at 0 nM). After hematological recovery of consolidation chemotherapy, the bacteriocidal activities of patients were similar to those of normal control (p=0.533, at 0 nM; p=0.133, at 1 nM; p=0.133, at 10 nM; p=0.133, at 100 nM). In conclusion, the bacteriocidal activities in AL patients were severely decreased at diagnosis and could be enhanced by WKYMVm. Near normal bacteriocidal activities can be achieved when 1 nM or more concentration of WKYMVm is applied in patients with AL. At the end of consolidation, bacteriocidal activities and stimulation by WKYMVm in patients were almost recovered.
Adult Hemophagocytic Lymphohistiocytosis: Clinical Presentation and Follow-Up in a Consecutive Series of Ten Patients.
Authors: Shirai Keisuke , Montero Alberto , Powell Jesse , Christiansen Lydia , Lazarchick John 
Published: October 2019

Abstract
Hemophagocytic Lymphohistiocytosis (HLH) is very rare in adults but can be fatal without treatment. Reports in adults are limited to case reports and very small case series. Clinically it is characterized by fever, hepatosplenomegaly, lymphadenopathy, severe cytopenias, hepatic dysfunction, coagulopathy, as well as neurological involvement. This syndrome is associated with diverse processes including: infection, rheumatologic, and hematologic malignancies. Presently, the underlying cause of HLH is unknown. We present a cosecutive series of 10 adults with HLH diagnosed at our institution between 2004–2006. All diagnoses were confirmed by pathology. The median age was 59 years (range: 18–73 years), and a male: female ratio of 4:1. All patients uniformly presented with fever. Half of the patients presented with evidence of hepatomegaly or splenomegaly. The most predominant laboratory abnormalities included: leukopenia or thrombocytopenia (100%), and elevation of liver enzymes (50%). EBV IgG was positive in 8 of 10 patients. The underline illnesses associated with HLH were diverse. The underlying causes were as follows; acute leukemia (n=2), infection (n=2), rheumatologic (n=2), post transplant (n=2) sickle cell disease (n=2), unknown (n=2). Mortality rate was 60% with a median survival time since diagnosis of 58 days. One patient is still on maintenance cyclosporin after etoposide treatment. One patient is on steroid and cytoxan. One patient recovered just with supportive care. In conclusion, due to the high morality rate associated with HLH, early treatment with immunosuppressant is warranted and attempts to identify underlying cause.