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Knee, Kelly M., Amey Barakat, Lindsay Tomlinson, Lila Ramaiah, Zane Wenzel, Brendon Kapinos, Youngwook Ahn, et al. "Sickle Cell Disease Model Mice Lacking 2,3-Dpg Show Reduced RBC Sickling and Improvements in Markers of Hemolytic Anemia." Blood 136, Supplement 1 (November 5, 2020): 27–28. http://dx.doi.org/10.1182/blood-2020-142052.
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Sickle cell disease (SCD) is a severe genetic disorder caused by a mutation in hemoglobin (b6Glu-Val), which allows the mutant hemoglobin to assemble into long polymers when deoxygenated. Over time, these polymers build up and deform red blood cells, leading to hemolytic anemia, vaso-occlusion, and end organ damage. A number of recent therapies for SCD have focused on modulating the mutant hemoglobin directly, however, reduction or elimination of 2,3-DPG to reduce Hb S polymerization and RBC sickling has recently been proposed as a therapeutic strategy for SCD. Current clinical studies focus on activation of pyruvate kinase to reduce 2,3-DPG, however, direct targeting of the enzyme which produces 2,3-DPG; Bisphosphoglycerate Mutase (BPGM) may also be possible. In this study we evaluate the impact of elimination of 2,3-DPG on SCD pathology by complete knockout of BPGM in Townes model mice. Animals with complete knockout of BPGM (BPGM -/-) have no detectable 2,3-DPG, while animals that are heterozygous for BPGM (BPGM -/+) have 2,3-DPG levels comparable to Townes mice. Western Blot analysis confirms that BPGM -/- animals completely lack BPGM, while BPGM -/+ animals have BPGM levels that are nearly equivalent to Townes mice. As expected from the lack of 2,3-DPG, BPGM -/- animals have increased oxygen affinity, observed as a 39% decrease in p50 relative to Townes mice. Complete elimination of 2,3-DPG has significant effects on markers of hemolytic anemia in BPGM -/- mice. Mice lacking 2,3-DPG have a 60% increase in hemoglobin (3.7 g/dL), a 53% increase in red blood cell count, and a 29% increase in hematocrit relative to Townes mice. The BPGM -/- mice also have a 57% decrease in reticulocytes, and a 61% decrease in spleen weight relative to Townes animals, consistent with decreased extramedullary hematopoiesis. Consistent with the reduction in hemolysis, BPGM -/- animals had a 59% reduction in red blood cell sickling under robust hypoxic conditions. BPGM -/+ animals had hemoglobin, RBC, and hematocrit levels that were similar to Townes animals, and a similar degree of RBC sickling to Townes mice. Liver phenotype was similar across all variants, with areas of random necrosis observed in BPGM -/-, BPGM -/+ and Townes mice. Higher percentages of microcytic and/or hyperchromic RBCs were observed in BPGM -/- animals relative to BPGM -/+ or Townes animals. These results suggest that modulation of 2,3-DPG has a positive effect on RBC sickling and hemolytic anemia, which may have therapeutic benefits for SCD patients. However, the lack of improvement in organ damage suggests that modulation of 2,3-DPG alone may not be sufficient for complete elimination of SCD phenotypes, and further investigation of this therapeutic avenue may be necessary. Disclosures No relevant conflicts of interest to declare.
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Aljahdali, Anfal S., Faik N. Musayev, John W. Burgner, Mohini S. Ghatge, Vibha Shekar, Yan Zhang, Abdelsattar M. Omar, and Martin K. Safo. "Molecular insight into 2-phosphoglycolate activation of the phosphatase activity of bisphosphoglycerate mutase." Acta Crystallographica Section D Structural Biology 78, no. 4 (March 11, 2022): 472–82. http://dx.doi.org/10.1107/s2059798322001802.
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Bisphosphoglycerate mutase (BPGM) is an erythrocyte-specific multifunctional enzyme that is responsible for the regulation of 2,3-bisphosphoglycerate (2,3-BPG) in red blood cells through its synthase and phosphatase activities; the latter enzymatic function is stimulated by the endogenous activator 2-phosphoglycolate (2-PG). 2,3-BPG is a natural allosteric effector of hemoglobin (Hb) that is responsible for decreasing the affinity of Hb for oxygen to facilitate tissue oxygenation. Here, crystal structures of BPGM with 2-PG in the presence and absence of 3-phosphoglycerate are reported at 2.25 and 2.48 Å resolution, respectively. Structure analysis revealed a new binding site for 2-PG at the dimer interface for the first time, in addition to the expected active-site binding. Also, conformational non-equivalence of the two active sites was observed as one of the sites was found in an open conformation, with the residues at the active-site entrance, including Arg100, Arg116 and Arg117, and the C-terminus disordered. The kinetic result is consistent with the binding of 2-PG to an allosteric or noncatalytic site as well as the active site. This study paves the way for the rational targeting of BPGM for therapeutic purposes, especially for the treatment of sickle cell disease.
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Qiang, Qingfen, Jeanne M. Manalo, Hong Sun, Yujin Zhang, Anren Song, Alexander Q. Wen, Y. Edward Wen, et al. "Erythrocyte adenosine A2B receptor prevents cognitive and auditory dysfunction by promoting hypoxic and metabolic reprogramming." PLOS Biology 19, no. 6 (June 17, 2021): e3001239. http://dx.doi.org/10.1371/journal.pbio.3001239.
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Hypoxia drives aging and promotes age-related cognition and hearing functional decline. Despite the role of erythrocytes in oxygen (O2) transport, their role in the onset of aging and age-related cognitive decline and hearing loss (HL) remains undetermined. Recent studies revealed that signaling through the erythrocyte adenosine A2B receptor (ADORA2B) promotes O2 release to counteract hypoxia at high altitude. However, nothing is known about a role for erythrocyte ADORA2B in age-related functional decline. Here, we report that loss of murine erythrocyte–specific ADORA2B (eAdora2b−/−) accelerates early onset of age-related impairments in spatial learning, memory, and hearing ability. eAdora2b-/- mice display the early aging-like cellular and molecular features including the proliferation and activation of microglia and macrophages, elevation of pro-inflammatory cytokines, and attenuation of hypoxia-induced glycolytic gene expression to counteract hypoxia in the hippocampus (HIP), cortex, or cochlea. Hypoxia sufficiently accelerates early onset of cognitive and cochlear functional decline and inflammatory response in eAdora2b−/− mice. Mechanistically, erythrocyte ADORA2B-mediated activation of AMP-activated protein kinase (AMPK) and bisphosphoglycerate mutase (BPGM) promotes hypoxic and metabolic reprogramming to enhance production of 2,3-bisphosphoglycerate (2,3-BPG), an erythrocyte-specific metabolite triggering O2 delivery. Significantly, this finding led us to further discover that murine erythroblast ADORA2B and BPGM mRNA levels and erythrocyte BPGM activity are reduced during normal aging. Overall, we determined that erythrocyte ADORA2B–BPGM axis is a key component for anti-aging and anti-age–related functional decline.
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Lemarchandel, V., V. Joulin, C. Valentin, R. Rosa, F. Galacteros, J. Rosa, and M. Cohen- Solal. "Compound heterozygosity in a complete erythrocyte bisphosphoglycerate mutase deficiency." Blood 80, no. 10 (November 15, 1992): 2643–49. http://dx.doi.org/10.1182/blood.v80.10.2643.2643.
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Abstract Erythrocyte bisphosphoglycerate mutase (BPGM) deficiency is a rare disease associated with a decrease in 2,3-diphosphoglycerate concentration. A complete BPGM deficiency was described in 1978 by Rosa et al (J Clin Invest 62:907, 1978) and was shown to be associated with 30% to 50% of an inactive enzyme detectable by specific antibodies and resulting from an 89 Arg-->Cys substitution. The propositus' three sisters exhibited the same phenotype, while his two children had an intermediate phenotype. Samples from the family were examined using polymerase chain reaction and allele-specific oligonucleotide hybridization and sequencing techniques. Amplification of erythrocyte total RNA from the propositus' sister around the 89 mutation indicated the presence of two forms of messenger RNAs, a major form with the 89 Arg-->Cys mutation and a minor form with a normal sequence. Sequence studies of the propositus' DNA samples indicated heterozygosity at locus 89 and another heterozygosity with the deletion of nucleotide C 205 or C 206. Therefore, the total BPGM deficiency results from a genetic compound with one allele coding for an inactive enzyme (mutation BPGM Creteil I) and the other bearing a frameshift mutation (mutation BPGM Creteil II). Examination of the propositus' two children indicated that they both inherited the BPGM Creteil I mutation.
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Lemarchandel, V., V. Joulin, C. Valentin, R. Rosa, F. Galacteros, J. Rosa, and M. Cohen- Solal. "Compound heterozygosity in a complete erythrocyte bisphosphoglycerate mutase deficiency." Blood 80, no. 10 (November 15, 1992): 2643–49. http://dx.doi.org/10.1182/blood.v80.10.2643.bloodjournal80102643.
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Erythrocyte bisphosphoglycerate mutase (BPGM) deficiency is a rare disease associated with a decrease in 2,3-diphosphoglycerate concentration. A complete BPGM deficiency was described in 1978 by Rosa et al (J Clin Invest 62:907, 1978) and was shown to be associated with 30% to 50% of an inactive enzyme detectable by specific antibodies and resulting from an 89 Arg-->Cys substitution. The propositus' three sisters exhibited the same phenotype, while his two children had an intermediate phenotype. Samples from the family were examined using polymerase chain reaction and allele-specific oligonucleotide hybridization and sequencing techniques. Amplification of erythrocyte total RNA from the propositus' sister around the 89 mutation indicated the presence of two forms of messenger RNAs, a major form with the 89 Arg-->Cys mutation and a minor form with a normal sequence. Sequence studies of the propositus' DNA samples indicated heterozygosity at locus 89 and another heterozygosity with the deletion of nucleotide C 205 or C 206. Therefore, the total BPGM deficiency results from a genetic compound with one allele coding for an inactive enzyme (mutation BPGM Creteil I) and the other bearing a frameshift mutation (mutation BPGM Creteil II). Examination of the propositus' two children indicated that they both inherited the BPGM Creteil I mutation.
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Luque, J., M. D. Delgado, P. Rodriguez-Horche, M. T. Company, and M. Pinilla. "Bisphosphoglycerate mutase and pyruvate kinase activities during maturation of reticulocytes and ageing of erythrocytes." Bioscience Reports 7, no. 2 (February 1, 1987): 113–19. http://dx.doi.org/10.1007/bf01121874.
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An increase in bisphosphoglycerate mutase (BPGM) and a decrease in pyruvate kinase (PK), i.e. a decrease in PK/BPGM ratio, was observed in red cell populations from anemic rats containing 95% down to 3% reticulocytes in blood. Such a ratio has been used to study the fractionation of recticulocytes, according to their degree of maturation, after counter-current distribution of those cell populations in dextrahpoly (ethylene glycol) two-phase systems. When applying this procedure to the fractionation according to age of erythrocytes from normal rats, the decrease of PK with cellular age was observed without a significant variation in BPGM activity.
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Yang Liu, Bin Wu, Hongxu Wang, and Pengjiang Ma. "BPGM: A big graph mining tool." Tsinghua Science and Technology 19, no. 1 (February 2014): 33–38. http://dx.doi.org/10.1109/tst.2014.6733206.
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Roberts, Bruce R., Chris Wolverton, and Lauren Janowicz. "The impact of substrate and irrigation interval on the post-transplant root growth of container-grown zinnia and tomato1." Journal of Environmental Horticulture 35, no. 1 (March 1, 2017): 1–5. http://dx.doi.org/10.24266/0738-2898-35.1.1.
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Abstract Substrate type and irrigation interval were studied to determine their impact on the post-transplant root growth of ‘Thumbelina' zinnia (Zinnis elegans Jacq.) and ‘Celebrity Hybrid' tomato (Lycopersicon esculentum Mill.). Seeds of both species were planted in 80 cm3 (2.7 fl oz) plug cells containing either Metromix 360™ (MM360) or Ball Professional Growing Mix™ (BPGM) and, following germination, the seedlings were transplanted into 450 cm3 (27.5 in3) plastic pots containing the same substrate. Evapotranspiration (ETO) was measured gravimetrically each day and the water lost via ETO added back to the substrate at intervals of 24, 48 or 96hr. For zinnia, root growth was consistently better for seedlings grown in BPGM, a substrate with greater water holding capacity and air-filled porosity. For plants grown in BPGM and irrigated every 48hr, root dry weight was significantly greater than it was for any of the remaining treatments. For tomato, root growth was greater for seedlings grown in BPGM and for transplants irrigated at 96 hr intervals; but, unlike zinnia, no significant interactions between substrate type and irrigation interval were observed. The results of this study show that root growth of plug-grown transplants can be improved by selecting a substrate with high porosity that allows for optimum oxygen and water exchange, and by extending the irrigation cycle to 48 hr (zinnia) or 96 hr (tomato). Index words: irrigation scheduling, moisture stress, plant establishment, soilless growing media, transplant production Species used in this study: ‘Thumbelina' zinnia (Zinnia elegans Jacq.); ‘Celebrity Hybrid' (Lycopersicon esculentum Mill.).
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Zhang, Jian, Chi Huang, Zehan Liu, Shuai Ren, Zilong Shen, Kecheng Han, Weiguang Xin, Guanyi He, and Jianyu Liu. "Screening of Potential Biomarkers in the Peripheral Serum for Steroid-Induced Osteonecrosis of the Femoral Head Based on WGCNA and Machine Learning Algorithms." Disease Markers 2022 (February 10, 2022): 1–17. http://dx.doi.org/10.1155/2022/2639470.
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Background. Steroid-induced osteonecrosis of the femoral head (SONFH) has produced a substantial burden of medical and social experience. However, the current diagnosis is still limited. Thus, this study is aimed at identifying potential biomarkers in the peripheral serum of patients with SONFH. Methods. The expression profile data of SONFH (number: GSE123568) was acquired from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) in SONFH were identified and used for weighted gene coexpression network analysis (WGCNA). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to investigate the biological functions. The protein-protein interaction (PPI) network and machine learning algorithms were employed to screen for potential biomarkers. Gene set enrichment analysis (GSEA), transcription factor (TF) enrichment analysis, and competing endogenous RNA (ceRNA) network were used to determine the biological functions and regulatory mechanisms of the potential biomarkers. Results. A total of 562 DEGs, including 318 upregulated and 244 downregulated genes, were identified between SONFH and control samples, and 94 target genes were screened based on WGCNA. Moreover, biological function analysis suggested that target genes were mainly involved in erythrocyte differentiation, homeostasis and development, and myeloid cell homeostasis and development. Furthermore, GYPA, TMCC2, and BPGM were identified as potential biomarkers in the peripheral serum of patients with SONFH based on machine learning algorithms and showed good diagnostic values. GSEA revealed that GYPA, TMCC2, and BPGM were mainly involved in immune-related biological processes (BPs) and signaling pathways. Finally, we found that GYPA might be regulated by hsa-miR-3137 and that BPGM might be regulated by hsa-miR-340-3p. Conclusion. GYPA, TMCC2, and BPGM are potential biomarkers in the peripheral serum of patients with SONFH and might affect SONFH by regulating erythrocytes and immunity.
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Petousi, N., R. R. Copley, T. R. J. Lappin, S. E. Haggan, C. M. Bento, H. Cario, M. J. Percy, et al. "Erythrocytosis associated with a novel missense mutation in the BPGM gene." Haematologica 99, no. 10 (July 11, 2014): e201-e204. http://dx.doi.org/10.3324/haematol.2014.109306.
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Sayama, Seisuke, Anren Song, and Yang Xia. "Maternal Erythrocyte ENT1-Mediated AMPK Activation and Oxygen Delivery: A Missing Component Counteracting Placental Hypoxia, Dysfunction, and Fetal Growth Restriction." Blood 134, Supplement_1 (November 13, 2019): 341. http://dx.doi.org/10.1182/blood-2019-123053.
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Background: Insufficient oxygen supply is associated with the pathophysiology of fetal growth restriction (FGR). Although the erythrocyte (RBC) is the most abundant and only cell type to deliver oxygen, its function and regulatory mechanism in FGR remains unknown. Recently, adenosine uptake by equilibrative nucleoside transporter 1 (ENT1), a key adenosine transporter expressed in RBCs, was reported to be crucial for RBCs to deliver oxygen. We aimed to investigate the involvement of RBCs' oxygen delivering capacity in maintaining fetal growth by focusing on RBC ENT1. Methods and Results: The mating strategy was to delete ENT1 only on the maternal RBCs but not in the placentas or fetuses to assess the effect of maternal RBC ENT1 on fetal growth. Specifically, EpoR-Cre+ (EPO) female mouse was used as a control and Ent1f/f-EpoR-Cre+ (E1FE) female mouse as an experimental mouse and mated with WT male mouse. As a result, E1FE dams showed FGR phenotype with reduction of 12.9% in fetal weight compared to EPO group. The maternal RBCs showed decrease in p50 and 2,3-BPG in E1FE, indicating decreased oxygen delivery in E1FE RBCs. To determine the molecular basis underlying the FGR phenotype seen in EIFE dams, we conducted a metabolomics screening of the RBCs isolated from controls and EIFE dams. It showed that adenosine metabolism inside the RBCs is the most impacted pathway. Specifically, it showed decrease in adenosine, AMP, and hypoxanthine, but adenine, ADP, and ATP did not show any reduction, implicating that ENT1-mediated uptake of adenosine is largely converted to AMP. We then incubated either WT or ENT1 KO RBCs with isotopically 13C15N labeled adenosine and traced the metabolism of intracellular adenosine derived from labeled adenosine. Indeed, adenosine was rapidly phosphorylated to AMP upon uptake, and 13C15N labeled AMP levels were lower in the ENT1 KO RBCs compared to controls. These findings provide evidence that 1) the most affected metabolic pathway in the RBCs of EIFE dam is adenosine metabolism; 2) ENT1-mediated uptake of extracellular adenosine is largely converted to AMP but not ATP. We hypothesized that decreased AMP/ATP ratio underlies the reduced 2,3-BPG production by lowering AMPK activity and subsequently decreasing BPG mutase (BPGM) activity. We measured AMPK phosphorylation and BPGM activity in the RBCs from E1FE and EPO dams. Both AMPK and BPGM activity were decreased in RBCs of E1FE dams compared to controls. Thus, we conclude that i) adenosine derived from uptake via ENT1 is largely converted to AMP; ii) lack of maternal RBC ENT1 lowers AMP/ATP ratio and activity of AMPK and BPGM in maternal RBC. We conducted immunofluorescence staining to assess hypoxia in the placentas, and confirmed the increased expression of HIF-1α in the placentas from E1FE dams. To determine functional changes of these placentas, we conducted metabolomics profiling in both the placenta and maternal plasma. Of all the metabolites altered, amino acids (AA) were the most reduced metabolites in E1FE placentas. In contrast, AA were the most accumulated in maternal plasma. We then injected isotopically labelled 13C15N AA mix in both controls and EIFE dams 24 hours prior to sacrifice. 13C15N AA level was decreased in the placentas in EIFE compared to the controls, while it was accumulated in plasma of EIFE, indicating reduced AA transporter function in the placentas of EIFE. Finally, we performed real time PCR to quantify the mRNA of the known main transporters of AA in the mouse placenta. It showed reduction of LAT1 mRNA in E1FE placenta, where there was no difference in LAT2, SNAT1, or SNAT2. Western blot of the placenta lysates confirmed the expression of LAT1 was indeed reduced. To validate our mouse finding and determine if HIF-1α elevation directly induces LAT1 mRNA in humans, we treated cultured human trophoblast cell line (HTR-8/SVneo cells) with or without DMOG, a cell permeable prolyl-4-hydroxylase inhibitor. After confirming DMOG upregulated HIF-1α, we found stabilized HIF-1α induced LAT1 mRNA levels. Thus, we conclude that elevated HIF-1α underlies the reduction of LAT1 mRNA in cultured human trophoblasts. Conclusion: Our findings suggest that maternal RBCs' oxygen delivering capacity mediated by ENT1 is essential for maintaining adequate placental oxygenation to support fetal growth via AA transporter function. Strategies to improve RBCs' function to deliver oxygen may provide new therapeutic possibilities for FGR. Figure Disclosures No relevant conflicts of interest to declare.
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Walter, MNM, SY Chan, M. Vatish, and MD Kilby. "PF.07 Expression of 2, 3-Bisphosphoglycerate Mutase (BPGM) in Human Placenta." Archives of Disease in Childhood - Fetal and Neonatal Edition 98, Suppl 1 (April 2013): A6.2—A6. http://dx.doi.org/10.1136/archdischild-2013-303966.019.
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Wang, Gang, Yan Huang, Ningning Zhang, Wenhu Liu, Changnan Wang, Xiaoyan Zhu, and Xin Ni. "Hydrogen Sulfide Is a Regulator of Hemoglobin Oxygen-Carrying Capacity via Controlling 2,3-BPG Production in Erythrocytes." Oxidative Medicine and Cellular Longevity 2021 (February 13, 2021): 1–16. http://dx.doi.org/10.1155/2021/8877691.
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Hydrogen sulfide (H2S) is naturally synthesized in a wide range of mammalian tissues. Whether H2S is involved in the regulation of erythrocyte functions remains unknown. Using mice with a genetic deficiency in a H2S natural synthesis enzyme cystathionine-γ-lyase (CSE) and high-throughput metabolomic profiling, we found that levels of erythrocyte 2,3-bisphosphoglycerate (2,3-BPG), an erythroid-specific metabolite negatively regulating hemoglobin- (Hb-) oxygen (O2) binding affinity, were increased in CSE knockout (Cse-/-) mice under normoxia. Consistently, the 50% oxygen saturation (P50) value was increased in erythrocytes of Cse-/- mice. These effects were reversed by treatment with H2S donor GYY4137. In the models of cultured mouse and human erythrocytes, we found that H2S directly acts on erythrocytes to decrease 2,3-BPG production, thereby enhancing Hb-O2 binding affinity. Mouse genetic studies showed that H2S produced by peripheral tissues has a tonic inhibitory effect on 2,3-BPG production and consequently maintains Hb-O2 binding affinity in erythrocytes. We further revealed that H2S promotes Hb release from the membrane to the cytosol and consequently enhances bisphosphoglycerate mutase (BPGM) anchoring to the membrane. These processes might be associated with S-sulfhydration of Hb. Moreover, hypoxia decreased the circulatory H2S level and increased the erythrocyte 2,3-BPG content in mice, which could be reversed by GYY4137 treatment. Altogether, our study revealed a novel signaling pathway that regulates oxygen-carrying capacity in erythrocytes and highlights a previously unrecognized role of H2S in erythrocyte 2,3-BPG production.
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Lazana, Ioanna, Azim Mohamedali, Frances Smith, Hugues Lavallade, Donal McLornan, and Kavita Raj. "Uniparental disomy (UPD) of a novel bisphosphoglycerate mutase ( BPGM ) mutation leading to erythrocytosis." British Journal of Haematology 192, no. 1 (November 20, 2020): 220–23. http://dx.doi.org/10.1111/bjh.17223.
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Dracopoli, Nicholas C., David M. Feltquate, Brigitta Sam, and Manfred Schartl. "Taql and Mspl RFLPs are detected by the human 2,3-biphosphoglycerate mutase (BPGM) cDNA." Nucleic Acids Research 18, no. 7 (1990): 1928. http://dx.doi.org/10.1093/nar/18.7.1928.
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Barichard, Fernande, Virginie Joulin, Isabelle Henry, Marie-Claude Garel, Colette Valentin, Raymonde Rosa, Michel Cohen-Solal, and Claudine Junien. "Chromosomal assignment of the human 2,3-bisphosphoglycerate mutase gene (BPGM) to region 7q34→7q22." Human Genetics 77, no. 3 (November 1987): 283–85. http://dx.doi.org/10.1007/bf00284487.
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Lim, M., H. M. Brown, K. L. Kind, J. Breen, M. R. Anastasi, L. J. Ritter, E. K. Tregoweth, D. T. Dinh, J. G. Thompson, and K. R. Dunning. "Haemoglobin expression in in vivo murine preimplantation embryos suggests a role in oxygen-regulated gene expression." Reproduction, Fertility and Development 31, no. 4 (2019): 724. http://dx.doi.org/10.1071/rd17321.
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Haemoglobin expression is not restricted to erythroid cells. We investigated the gene expression of the haemoglobin subunits haemoglobin, alpha adult chain 1 (Hba-a1) and haemoglobin, beta (Hbb), 2,3-bisphosphoglycerate mutase (Bpgm) and the oxygen-regulated genes BCL2/adenovirus E1B interacting protein 3 (Bnip3), solute carrier family 2 (facilitated glucose transporter), member 1 (Slc2a1) and N-myc downstream regulated gene 1 (Ndrg1) in the murine preimplantation embryo, comparing invivo to invitro gene expression. Relatively high levels of Hba-a1 and Hbb were expressed invivo from the 2-cell to blastocyst stage; in contrast, little or no expression occurred invitro. We hypothesised that the presence of haemoglobin invivo creates a low oxygen environment to induce oxygen-regulated gene expression, supported by high expression of Slc2a1 and Ndrg1 in invivo relative to invitro embryos. In addition, analysis of an invitro-derived human embryo gene expression public dataset revealed low expression of haemoglobin subunit alpha (HBA) and HBB, and high expression of BPGM. To explore whether there was a developmental stage-specific effect of haemoglobin, we added exogenous haemoglobin either up to the 4-cell stage or throughout development to the blastocyst stage, but observed no difference in blastocyst rate or the inner cell mass to trophectoderm cell ratio. We conclude that haemoglobin in the invivo preimplantation embryo raises an interesting premise of potential mechanisms for oxygen regulation, which may influence oxygen-regulated gene expression.
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Iskandar, Burhanuddin, Bambang Madiyono, Sudigdo Sastroasmoro, Sukman T. Putra, Mulyadi M. Djer, and Anis Karuniawati. "Comparison of minimal inhibitory and bactericidal capacity of oral penicillin V with benzathine penicillin G to Streptococcus beta--hemolyticus group A in children with rheumatic heart disease." Paediatrica Indonesiana 48, no. 3 (September 26, 2016): 152. http://dx.doi.org/10.14238/pi48.3.2008.152-5.
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Background Injection ofbenzatine penicillin G (BPG) every 28days is still the drug of choice for secondary prevention of rheu-matic heart disease (RHD). BPG sometimes poses problems dueto pain at the injection site, possible anaphylaxis, and is not alwaysavailable. Some centers choose oral penicillin over BPG.Objectives To compare minimal inhibitory capacity (MIC) andminimal bactericidal capacity (MBC) of oral penicillin V serumwith those of BPG among SGA infected RHD.Methods This was a clinical trial with crossover design study tocompare MIC of penicillin V and BPG. Outcome measures wereMIC and MBC. Statistical analysis was performed using pairedt-test and wilcoxon test.Result There were 32 subjects consisted of 17 males and 15females. The mean value of MIC and MBC serum of penicillinV were 0.031 and 0.125. The mean value of MIC and MBCserum of BPG3 were 0.094 and 0.031. Respectively the MICof penicillin V was similar to that of BPGy The mean value ofMIC and MBC of BPG4 were 0.125 and 0.250. Respectively theMIC of penicillin V was significantly higher than that of BPG 4.The MBC of penicillin V was significantly higher than that ofBPG 4. The MIC ofBPG 3 was similar to that ofBPG 4• The MBCof BPG 3 was similar to that of BPG 4.Conclusions The MIC of penicillin V was similar to that ofBPG 3,the MBC of oral penicillin V was higher than that ofBPG 3• TheMIC and MBC of penicillin V was higher than those of BPG 4.
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E, Guoji, Binda Sun, Bao Liu, Gang Xu, Shu He, Yu Wang, Lan Feng, et al. "Enhanced BPGM/2,3-DPG pathway activity suppresses glycolysis in hypoxic astrocytes via FIH-1 and TET2." Brain Research Bulletin 192 (January 2023): 36–46. http://dx.doi.org/10.1016/j.brainresbull.2022.11.002.
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Wu, X., S. K. Davis, S. L. F. Davis, D. S. Gallagher Jr, Y. Song, R. A. Brenneman, C.-C. Yeh, and J. F. Taylor. "Assignment of laminin B1 polypeptide (LAMB1 ) and 2,3 bisphosphoglycerate mutase (BPGM ) to the physical and genetic maps of BTA4." Animal Genetics 31, no. 4 (August 2000): 282–83. http://dx.doi.org/10.1046/j.1365-2052.2000.00629.x.
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Nguyen, Jean-Michel, Philippe Juin, Daniel Antonioli, Alexandre Moreau-Gaudry, Mario Campone, and Pascal Jézéquel. "Identification of paclitaxel resistance through a new statistical approach based on a random forest of perfect trees classifcation." Journal of Clinical Oncology 38, no. 15_suppl (May 20, 2020): e13513-e13513. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.e13513.
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e13513 Background: Predictors of paclitaxel sensitivity in breast cancer published ten years ago, are still pending. The authors showed that paclitaxel pathological complete response (pCR) was in one hand, encountered in aggressive breast tumor with immune response and in another hand, paclitaxel resistance in less aggressive tumor. We have developed a new analysis paradigm, mixing neurons into nodes of trees classification and news class of statistical information based on free-error trees classification. We proposed to reanalyze the Bauer et al’s dataset using this novel approach. Methods: GES22513 dataset including 14 duplicated observations and 54675 anonymized probes was analyzed. A random forest of one million trees whose nodes were composed of neurons including 15 probes, was developped. We selected probes for which a free-error classification was obtained and ranked them according to the inverse of the probability of being a confounding factor and to the inverse of the probability of interacting with another probe. We compared the sets of probes which were necessary to obtain an error-free classification between those associated with a decrease and those associated with an increase of the probability of pCR. Results: Our 15 best ranked predictors were free-error classification for all observations. This includes gene expression of TLCD2, BRCC3, CHI3L2 and PROX1. Their over-expressions were associated with an increase in the probability of pCR, and gene expression of APH1B, ARFGEF1, ARID2, BPGM, CAMK2N1, CCNY, PARM1, PHKA1, PSMD9, SUDS3 (two probes) whose over-expressions are associated with a decrease in the probability of pCR. Ten out of these probes were concordant with Bauer et al’s conclusion. Four probes ( BPGM, PHKA1, CCNY and ARFGEF1) are in contradiction with it. The limited biological information were available for TLCD2. The statistical analysis also showed that TLCD2, BBRCC3, CHI3L2 and PROX1 were altogether positively modulated by eight genes/probes ( CKS1B, ADIG, NCR3, RIN3, NIPAL1, 234422_at, DCLRE1C, SLC17A4). At the opposite, the modulation of genes associated with a decrease in the probability of pCR, was rather heterogeneous and involves many more genes. Conclusions: This preliminary work shows that our statistical approach allows a perfect classification of tumors with and without pCR. Also, it proves that the selected probes/genes are respectively associated with aggressiveness/basal and less aggressiveness/luminal phenotypes. These results need to be validated on an independent cohort.
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Weisenhorn, Erin M., Thomas Raife, Thomas J. van't Erve, Nicholas M. Riley, John Hess, and Joshua J. Coon. "Multi-Omics Evidence for Inheritance of Energy Pathways in Red Blood Cells." Blood 128, no. 22 (December 2, 2016): 3838. http://dx.doi.org/10.1182/blood.v128.22.3838.3838.
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Abstract Introduction Each year over 90 million units of blood are transfused worldwide. Our dependence on this blood supply requires optimized blood collection and storage. During storage, red blood cells (RBCs) undergo degenerative processes resulting in altered metabolic characteristics. In the past decade numerous studies have implicated longer storage of RBCs in adverse patient outcomes. The post-storage ATP level in blood is the single best predictor of transfused RBC in vivo recovery. Although the rate of ATP decline is highly variable between individuals, post-storage ATP levels are primarily determined by inheritance. Understanding the effect of storage on energy metabolism pathways is thus of vital importance to maintaining a safe and effective blood supply. Methods We performed comprehensive metabolomics and proteomics studies of mono- and di-zygotic twin pairs to measure heritability of molecules and identify correlations with ATP and other markers in energy metabolism. Metabolite levels were measured at six time points from 0-56 days to elucidate changes that occur during storage. An obstacle for RBC proteomics is the massive quantity of hemoglobin, constituting 97% of protein material. This was avoided by preparing RBC membrane fractions, which mitigated the need for hemoglobin depletion. All proteomics data was collected on an Orbitrap Elite hybrid ion trap-orbitrap mass spectrometer (Thermo Fisher Scientific). Metabolomics data was collected by Metabolon Inc. and was collated with proteomics results to give a complete view of RBC metabolism. Preliminary Data Our optimized method for collecting proteomics data in RBCs has yielded the greatest depth of coverage observed without the use of commercial hemoglobin depletion. Purified RBCs were lysed and centrifuged to collect membrane fractions allowing us to identify 1280 proteins and 330 metabolites from mono and di-zygotic twins. Of these, 146 proteins and 148 metabolites were found to be over 30% heritable. We observe a high degree of heritability in metabolites involved in energy metabolism, especially glycolysis. This is supported by the heritability in key regulatory enzymes including phosphofructokinase (PFK) (57%) and bisphosphoglycerate mutase (BPGM) (50%). Additionally we observe high correlations between both glycolytic proteins and metabolites suggesting that this crucial energy metabolism pathway is inherited en blocat various levels. A number of the correlations we observed can be combined to produce a model to predict post-storage ATP levels. Five key parameters in this model include PFK, carbonic anhydrase 1 (CA1), band 3, BPGM, and pH. Strikingly, concentrations of all protein components of this model were at least 45% heritable. Band 3, BPGM, and CA1 correlate negatively with post storage ATP levels and together shuttle flux away from glycolysis and ATP production. We also observe a positive correlation between pH and post-storage ATP. A negative correlation observed between CA1, which is 84% heritable, and post-storage ATP, is especially significant in that it provides a hypothetical model for the heritability of ATP decline during storage. Our model proposes that RBC units, which are stored in gas permeable bags allowing CO2 to diffuse into the bag, are subject to genetically determined, CA1-mediated production of carbonic acid, resulting in inhibition of PFK. This model is further supported by negative correlations between CA1 and pH during storage. We propose that heritable concentrations of CA1 negatively influence pH, which allosterically inhibits PFK and impedes energy metabolism and subsequently ATP production. We conclude that individuals inherit a phenotype composed of higher or lower concentrations of key energy metabolism proteins that regulate flux through glycolysis during RBC storage. Heritability of energy metabolism can result in markedly different RBC storage profiles and knowledge of heritable RBC energy metabolism can be used to improve and individualize RBC storage methods. Disclosures Hess: ASH: Patents & Royalties: 4 US patents related to RBC storage solution AS-7.
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Oliveira, Jennifer L., Lori A. Frederick, Lea M. Coon, Molly Susan Hein, Mrinal M. Patnaik, Ayalew Tefferi, Animesh Pardanani, Stefan K. Grebe, David S. Viswanatha, and James D. Hoyer. "Spectrum of Mutations Associated with Hereditary Erythrocytosis." Blood 126, no. 23 (December 3, 2015): 2140. http://dx.doi.org/10.1182/blood.v126.23.2140.2140.
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Abstract Background: Mechanisms of hereditary erythrocytosis have been elucidated recently. These include high oxygen affinity (HOA) hemoglobin (Hb) variants, bisphosphoglycerate mutase (BPGM) deficiency, abnormalities in the erythropoietin receptor (EPOR) and oxygen-sensing pathway (OSP) proteins prolyl hydroxylase domain-2 (PHD2), hypoxia-inducible factor-2 alpha subunit (HIF2A), and von Hippel-Lindau (VHL). We present our experience with these disorders. Methods: Evaluation of erythrocytosis patients' blood samples for HOA Hb variants by protein or DNA sequencing methods has been performed in our laboratory for over 30 years. Testing has included alkaline/acid electrophoresis, isoelectric focusing, capillary electrophoresis, high performance liquid chromatography, mass spectrometry and Sanger sequencing. BPGM deficiency was determined by enzyme activity assay and/or genetic testing. Since 2012 we have evaluated the EPOR, EGLN1 (PHD2), EPAS1(HIF2A), and VHL genes. Reflexive evaluations, which include testing for p50 and pertinent Sanger sequencing of HBB, HBA1, HBA2, EPOR (exon 8), EGLN1 (exons 1-5), EPAS1 (exon 12) and VHL, have been performed on a subset of cases. Genetic results were correlated with clinical and phenotypic data. Results: Retrospective database review identified 62 confirmed HOA Hb variants (48β, 14α) with clinical erythrocytosis (Table 1). Multiple Hb variants were silent on at least one protein method but showed decreased p50 values. One homozygous BPGM mutation case was identified. Of the 394 cases evaluated for EPOR and OSP abnormalities, 39 cases (10%) contained genetic alterations. Of these, 11 were known pathogenic mutations and 28 were novel alterations including 4 pathogenic, 12 likely pathogenic and 12 variants of unknown significance (VUS). Eighteen EGLN1, 10 EPAS1, and 7 EPOR alterations were detected, all heterozygous. Four VHL mutation cases were identified: 3 homozygous/compound heterozygous known mutations and one novel heterozygous VUS (p.R200Q). All EPOR mutations resulted in a premature stop codon. Most of the EPAS1 alterations were located near proline 531, although two predicted splice site disturbances were found. The EGLN1 alterations were novel, variable, and affect the enzyme's catalytic domain. All 4 VHL cases involved the R200 amino acid position in at least one allele. Serum Epo levels varied: decreased in EPOR (5/5); normal in PHD2 (11/11) and most HIF2A (6/7) and VHL (3/4) cases; increased in 1 HIF2A and 1 VHL case. p50 was normal in EPOR and OSP cases (31/31 tested). Conclusion: A spectrum of abnormalities was identified. Salient points include: 1) Diagnostically, a high index of suspicion for HOA Hb variants is required as they can give false negative results by routine protein detection methods. p50 and Epo levels were useful for triage. 2) Phenotypically, serum Epo levels were low in EPOR and normal in PHD2 and HIF2A cases. Unexpectedly, the two classic Chuvash polycythemia VHL cases were associated with normal Epo levels. 3) Mechanistically, the alterations were truncating in EPOR, variable and scattered in EGLN1 and clustered near proline 531 in EPAS1. VHL mutations were rare and consistently affected the R200 amino acid position in at least one allele. Novel alterations were frequent, especially in EGLN1. 4) Clinically, many patients were asymptomatic but a subset demonstrated recurrent headaches and one had chest pain. Clotting complications included cerebrovascular accident in one EPAS1 and EGLN1 case each and portal vein thrombosis in one EGLN1 case. Table 1. Clinically significant high oxygen affinity Hb variants at Mayo Clinic n Beta Alpha >30 Malmo Tarrant 10 - 20 Bethesda, San Diego, Andrew-Minneapolis, M-Saskatoon, Olympia, Syracuse, Abruzzo Dallas 5 - 10 Ty Gard, Ypsilanti, Alberta, Coimbra, North Chicago J-Cape Town 2 - 4 Brigham, Kempsey, Little Rock, Puttelange, Wood, Brisbane, Cowtown, Creteil, Linkoping, Osler, Potomac, Providence, Ranier, Sparta, Vanderbilt, Heathrow, Helsinki, Hiroshima, Homozygous HPFH, McKees-Rocks, Pierre Benite, Regina Chesapeake, Ethiopia, Chiapas, Columbia Missouri, Burlington, Legnano 1 Alcorn County*, Bologna-St. Orsola, Bunbury, Cambridge-MA, Cardarelli, Chandigarh, Johnstown, Nantes, Nebraska, Olomouc, Palmerston North, Pitie-Salpetriere, Saint-Jacques, South Milwaukee Linwood, Longview*, Milledgeville, Sarasota Springs, Voorhees * novel Disclosures Pardanani: Stemline: Research Funding.
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Fedorova, M. S., I. Y. Karpova, A. V. Lipatova, E. A. Pudova, Z. G. Guvatova, D. V. Kochetkov, A. V. Chaika, et al. "Knockdown of hexokinase 2 results in a decreased expression level of the glycolytic enzymes PFKP, BPGM, and GPI in RKO cell line." Vavilov Journal of Genetics and Breeding 21, no. 8 (January 1, 2017): 932–36. http://dx.doi.org/10.18699/vj17.315.
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Men, Lihui, Wenting Hui, Xin Guan, Tongtong Song, Xuan Wang, Siwei Zhang, and Xia Chen. "Cardiac Transcriptome Analysis Reveals Nr4a1 Mediated Glucose Metabolism Dysregulation in Response to High-Fat Diet." Genes 11, no. 7 (June 29, 2020): 720. http://dx.doi.org/10.3390/genes11070720.
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Obesity is associated with an increased risk of developing cardiovascular disease (CVD), with limited alterations in cardiac genomic characteristics known. Cardiac transcriptome analysis was conducted to profile gene signatures in high-fat diet (HFD)-induced obese mice. A total of 184 differentially expressed genes (DEGs) were identified between groups. Based on the gene ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs, the critical role of closely interlocked glucose metabolism was determined in HFD-induced cardiac remodeling DEGs, including Nr4a1, Fgf21, Slc2a3, Pck1, Gck, Hmgcs2, and Bpgm. Subsequently, the expression levels of these DEGs were evaluated in both the myocardium and palmitic acid (PA)-stimulated H9c2 cardiomyocytes using qPCR. Nr4a1 was highlighted according to its overexpression resulting from the HFD. Additionally, inhibition of Nr4a1 by siRNA reversed the PA-induced altered expression of glucose metabolism-related DEGs and hexokinase 2 (HK2), the rate-limiting enzyme in glycolysis, thus indicating that Nr4a1 could modulate glucose metabolism homeostasis by regulating the expression of key enzymes in glycolysis, which may subsequently influence cardiac function in obesity. Overall, we provide a comprehensive understanding of the myocardium transcript molecular framework influenced by HFD and propose Nr4a1 as a key glucose metabolism target in obesity-induced CVD.
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Girodon, Francois, Fabrice Airaud, Garrec Céline, Pacault Mathilde, Dumont Solenne, Ricordeau Ingrid, Bézieau Stéphane, and Betty Gardie. "Next Generation Sequencing Is a Useful Tool for the Diagnosis of Congenital/Idiopathic Erythrocytoses." Blood 128, no. 22 (December 2, 2016): 2434. http://dx.doi.org/10.1182/blood.v128.22.2434.2434.
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Abstract Introduction: Erythrocytoses are characterized by an elevated red cell mass. The most widely studied disease is Polycythemia Vera (PV), a myeloproliferative neoplasm due to the acquired JAK2-V617F mutation. However, other types of erythrocytoses exist and are of major importance. They can be either inherited (Congenital Erythrocytosis-CE) or diagnosed in adult patients with no family history (Idiopathic Erythrocytosis-IE). CE/IE are not associated with myeloproliferation but they can be associated with severe thrombo-embolic or haemorrhagic events, pulmonary arterial hypertension and, rarely, tumours. The 8 genes identified so far as causing CE lie at the crossroads of major biological pathways (metabolism, inflammation, oncogenesis) and are implicated in multiple diseases. These genes are involved (i) in the regulation of the hypoxia pathway, PHD2 (also called EGLN1), HIF-2A (EPAS1), VHL, (ii) in proliferation and differentiation of erythroid progenitors (EPOR), or (iii) in mature cell function, haemoglobins (HBB, HBA1, HBA2) or bisphosphoglyceratemutase (BPGM). However, in 80% of cases the cause remains unknown meaning that no proper diagnosis can be made, no prognosis or advice can be provided to CE/IE patients and their families, and no curative treatment exists. Method: We created and developed a national network in France to (i) identify, (ii) collect and (iii) analyze the genomic abnormalities in patients suspected of CE/IE. The selection of patients was performed using a clinical and biological data sheet including mandatory further tests in order to exclude patients with PV or obvious secondary erythrocytosis related to lung, cardiac or renal disorder. Next generation sequencing (NGS) has been used to analyse the presence of mutations in 17 genes (VHL, PHD1, PHD2 and PHD3, HIF1A, HIF2A, HIF3A, FH, BPGM, and 8 other candidate genes). SureDesign software (Agilent, Santa Clara, CA) was used to design the custom HaloPlex capture assay. For sequence capture, HaloPlex Target Enrichment System Kit (AgilentR), for Illumina sequencing was used, according to the manufacturer's instructions. Results: To date, samples from 103 patients have been recorded, among whom 46 have been tested using NGS approach. Variants in 10 (21%) patients [9 males and 1 female ; median age 50 y. (12-71)] with unknown significance have been detected, including 4 in PHD genes, 5 in HIF genes, and 1 in JAK2 gene. In patients with variants, a familial history of erythrocytosis was noted in 2. No independent thrombotic complication was reported in the 10 patients. The proportion of variants detected (21%) was close to the classical rate of genomic abnormalities usually observed in CE/IE. In 2 patients (one with a PHD2 and one with a JAK2 variants), the erythropoietin was low, whereas for the others, the erythropoietin was normal. Of note, the median age of the patients was surprisingly high, suggesting that the diagnostic was not previously performed due to the absence of available tests. Indeed, the diagnostic approaches using NGS techniques led to a considerable time gain and facilitated the identification of certain molecular abnormalities associated with CE/IE Conclusion: NGS is a useful tool to explore mutations in CE/IE, but identifies genetic variants in only 20% of patients with such disorder. In vitro, in cellulo and in vivo (including zebrafish models) functional studies are currently performed to validate the clinical relevance of these variants. Further exams including whole exome sequencing are planned to achieve a right diagnosis in the 80% remaining CE/IE patients without identified genomic abnormalities. Disclosures No relevant conflicts of interest to declare.
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MULQUINEY, Peter J., William A. BUBB, and Philip W. KUCHEL. "Model of 2,3-bisphosphoglycerate metabolism in the human erythrocyte based on detailed enzyme kinetic equations1: in vivo kinetic characterization of 2,3-bisphosphoglycerate synthase/phosphatase using 13C and 31P NMR." Biochemical Journal 342, no. 3 (September 5, 1999): 567–80. http://dx.doi.org/10.1042/bj3420567.
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This is the first in a series of three papers [see also Mulquiney and Kuchel (1999) Biochem. J. 342, 579-594; Mulquiney and Kuchel (1999) Biochem. J. 342, 595-602] that present a detailed mathematical model of erythrocyte metabolism which explains the regulation and control of 2,3-bisphosphoglycerate (2,3-BPG) metabolism. 2,3-BPG is a modulator of haemoglobin oxygen affinity and hence plays an important role in blood oxygen transport and delivery. This paper presents an in vivo kinetic characterization of 2,3-BPG synthase/phosphatase (BPGS/P), the enzyme that catalyses both the synthesis and degradation of 2,3-BPG. Much previous work had indicated that the behaviour of this enzyme in vitro is markedly different from that in vivo. 13C and 31P NMR were used to monitor the time courses of selected metabolites when erythrocytes were incubated with or without [U-13C]glucose. Simulations of the experimental time courses were then made. By iteratively changing the parameters of the BPGS/P part of the model until a good match between the NMR-derived data and simulations were achieved, it was possible to characterize BPGS/P kineticallyin vivo. This work revealed that: (1) the pH-dependence of the synthase activity results largely from a strong co-operative inhibition of the synthase activity by protons; (2) 3-phosphoglycerate and 2-phosphoglycerate are much weaker inhibitors of 2,3-BPG phosphatase in vivo than in vitro; (3) the Km of BPGS/P for 2,3-BPG is significantly higher than that measured in vitro; (4) the maximal activity of the phosphatase in vivo is approximately twice that in vitro, when Pi is the sole activator (second substrate); and (5) 2-phosphoglycollate appears to play no role in the activation of the phosphatase in vivo. Using the newly determined kinetic parameters, the percentage of glycolytic carbon flux that passes through the 2,3-BPG shunt in the normal in vivo steady state was estimated to be 19%.
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Santana, André Felipe Lemos, and Carina Frota Alves. "BPMG – Um Modelo Conceitual para Governança em BPM – Aplicação numa Organização Pública." iSys - Brazilian Journal of Information Systems 9, no. 1 (May 16, 2016): 139–67. http://dx.doi.org/10.5753/isys.2016.304.
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Governança em BPM é um aspecto apontado como fundamental para o sucesso efetivo do gerenciamento de processos de negócio. Isso parece ser especialmente verdade no contexto do setor público, onde a efetividade, prestação de contas e transparência (três princípios básicos da governança) precisam ser coordenadas. No entanto, as abordagens dessa governança têm se caracterizado por conceitos complexos e pouco precisos, e modelos em alto nível que oferecem pouca ajuda sobre como implementá-la. Além disso, há um número reduzido de trabalhos empíricos com relatos práticos neste tema. Este trabalho relata a construção de um Modelo Conceitual para Governança em BPM e sua aplicação numa organização pública brasileira, utilizando uma abordagem de design-science. A avaliação sobre a aplicação do modelo apontou que ele foi percebido como significativamente útil para criar um entendimento comum bem como para apoiar a estruturação da governança em BPM na organização pesquisada.
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Kim, Jung-whan, Karen I. Zeller, Yunyue Wang, Anil G. Jegga, Bruce J. Aronow, Kathryn A. O'Donnell, and Chi V. Dang. "Evaluation of Myc E-Box Phylogenetic Footprints in Glycolytic Genes by Chromatin Immunoprecipitation Assays." Molecular and Cellular Biology 24, no. 13 (July 1, 2004): 5923–36. http://dx.doi.org/10.1128/mcb.24.13.5923-5936.2004.
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ABSTRACT Prediction of gene regulatory sequences using phylogenetic footprinting has advanced considerably but lacks experimental validation. Here, we report whether transcription factor binding sites predicted by dot plotting or web-based Trafac analysis could be validated by chromatin immunoprecipitation assays. MYC overexpression enhances glycolysis without hypoxia and hence may contribute to altered tumor metabolism. Because the full spectrum of glycolytic genes directly regulated by Myc is not known, we chose Myc as a model transcription factor to determine whether it binds target glycolytic genes that have conserved canonical Myc binding sites or E boxes (5′-CACGTG-3′). Conserved canonical E boxes in ENO1, HK2, and LDHA occur in 31- to 111-bp islands with high interspecies sequence identity (>65%). Trafac analysis revealed another region in ENO1 that corresponds to a murine region with a noncanonical E box. Myc bound all these conserved regions well in the human P493-6 B lymphocytes. We also determined whether Myc could bind nonconserved canonical E boxes found in the remaining human glycolytic genes. Myc bound PFKM, but it did not significantly bind GPI, PGK1, and PKM2. Binding to BPGM, PGAM2, and PKLR was not detected. Both GAPD and TPI1 do not have conserved E boxes but are induced and bound by Myc through regions with noncanonical E boxes. Our results indicate that Myc binds well to conserved canonical E boxes, but not nonconserved E boxes. However, the binding of Myc to unpredicted genomic regions with noncanonical E boxes reveals a limitation of phylogenetic footprinting. In aggregate, these observations indicate that Myc is an important regulator of glycolytic genes, suggesting that MYC plays a key role in a switch to glycolytic metabolism during cell proliferation or tumorigenesis.
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Kriete, Alicia S., Katsiaryna Prudnikova, and Michele S. Marcolongo. "Modulating physical properties of porcine urethra with injection of novel biomimetic proteoglycans ex vivo." Interface Focus 9, no. 4 (June 14, 2019): 20190013. http://dx.doi.org/10.1098/rsfs.2019.0013.
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Urinary incontinence is a significant challenge for women who are affected by it. We propose augmenting the tissue structure to restore normal biomechanics by molecularly engineering the tissue using a novel family of biomimetic proteoglycans (BPGs). This work examines the ability of BPGs to modulate the mechanical and physical properties of porcine urethras ex vivo to determine the feasibility of BPGs to be implemented as molecular treatment for stress urinary incontinence (SUI). We investigated compliance by performing a unique radial expansion testing method using urethras from six- to nine-month-old pigs. The urethras were injected with 0.5 ml BPG solution at three sites every approximately 120° (conc.: 25 mg ml −1 , 50 mg ml −1 and 75 mg ml −1 in 1× phosphate-buffered saline (PBS); n = 4 per group) and compared them with PBS-injected controls. Young's modulus was calculated by treating the urethra as a thin-walled pressure vessel. A water uptake study was performed by soaking 10 mm urethra biopsy samples that were injected with 0.1 ml BPG solution (conc.: 50 mg ml −1 , 100 mg ml −1 and 200 mg ml −1 in 1× PBS; n = 6 per group) in 5 ml PBS for 24 h. Although there was no significant difference in Young's modulus data, there were differences between groups as can be seen in the raw radial expansion testing data. Results showed that BPGs have the potential to increase hydration in samples, and that there was a significant difference in water uptake between BPG-injected samples and the controls (100 mg ml −1 samples versus PBS samples, p < 0.05). This work shows that BPGs have the potential to be implemented as a molecular treatment for SUI, by restoring the diminished proteoglycan content and subsequently increasing hydration and improving the compliance of urethral tissue.
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Zhang, Jing, Xu Bai, Jing Yuan, Xiaojing Zhang, Wei Xu, Huiyi Ye, and Haiyi Wang. "Bladder paraganglioma: CT and MR imaging characteristics in 16 patients." Radiology and Oncology 56, no. 1 (December 30, 2021): 46–53. http://dx.doi.org/10.2478/raon-2021-0055.
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Abstract Background Bladder paraganglioma (BPG) is a rare extra-adrenal pheochromocytoma with variable symptoms and easy to be misdiagnosed and mishandled. The aim of the study was to document the imaging features of BPG using computed tomography (CT) and magnetic resonance imaging (MRI). Patients and methods We retrospectively enrolled consecutive patients with pathology-proven BPG, who underwent CT or MRI examinations before surgery between October 2009 and October 2017. The clinical characteristics, CT, and MRI features of the patients were described and analysed. Results A total of 16 patients with 16 bladder tumours (median age 51 years, 9 females) were included. Among them, 13 patients underwent CT examinations and eight patients underwent MRI examinations preoperatively. Tumour diameters ranged from 1.6−5.4 cm. Most of the tumours grew into the bladder cavity (n = 11) with oval shapes (n = 10) and well-defined margins (n = 14). Intratumour cystic degeneration or necrosis (n = 2) was observed. Two lesions showed peripheral tissue invasion, suggesting malignant BPGs. All 13 lesions imaged with CT exhibited slight hypoattenuation and moderate to marked enhancement. Compared to the gluteus maximus, all lesions showed slight h yperintensity in T2-weighted images, hyperintensity on diffusion-weighted images (DWI), hypointensity on apparent diffusion coefficient maps, hyperintensity on T1-weighted images and a “fast in and slow out” enhanced pattern on contrast-enhanced MRI images. Conclusions BPGs are mostly oval-shaped, broadly-based and hypervascular bladder tumours with hypoattenuation on non-contrast CT, T2 hyperintensity, slight T1 hyperintensity compared to the muscle, marked restricted diffusion on DWI. Peripheral tissue invasion can suggest malignancy of the BPGs. All of these features contribute to preoperative decision-making.
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Sung, Tae-Kyung. "Performance Analysis of TH-BPPM and TH-BPAM UWB System and the Applications in Data and Image Transmission." Journal of Navigation and Port Research 31, no. 2 (March 31, 2007): 159–63. http://dx.doi.org/10.5394/kinpr.2007.31.2.159.
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Liu, Lei, Zehan Cui, Yong Li, Yungang Bao, Mingyu Chen, and Chengyong Wu. "BPM/BPM+." ACM Transactions on Architecture and Code Optimization 11, no. 1 (February 2014): 1–28. http://dx.doi.org/10.1145/2579672.
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Yoshinaga, Marcos Y., Lars Wörmer, Marcus Elvert, and Kai-Uwe Hinrichs. "Novel Cardiolipins from Uncultured Methane-Metabolizing Archaea." Archaea 2012 (2012): 1–9. http://dx.doi.org/10.1155/2012/832097.
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Novel cardiolipins from Archaea were detected by screening the intact polar lipid (IPL) composition of microbial communities associated with methane seepage in deep-sea sediments from the Pakistan margin by high-performance liquid chromatography electrospray ionization mass spectrometry. A series of tentatively identified cardiolipin analogues (dimeric phospholipids or bisphosphatidylglycerol, BPG) represented 0.5% to 5% of total archaeal IPLs. These molecules are similar to the recently described cardiolipin analogues with four phytanyl chains from extreme halophilic archaea. It is worth noting that cardiolipin analogues from the seep archaeal communities are composed of four isoprenoidal chains, which may contain differences in chain length (20 and 25 carbon atoms) and degrees of unsaturation and the presence of a hydroxyl group. Two novel diether lipids, structurally related to the BPGs, are described and interpreted as degradation products of archaeal cardiolipin analogues. Since archaeal communities in seep sediments are dominated by anaerobic methanotrophs, our observations have implications for characterizing structural components of archaeal membranes, in which BPGs are presumed to contribute to modulation of cell permeability properties. Whether BPGs facilitate interspecies interaction in syntrophic methanotrophic consortia remains to be tested.
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García, Tamara. "La brecha de productividad entre hombres y mujeres en la ciencia." UMH Sapiens Divulgación Científica 10, no. 25 (October 2019): 23–25. http://dx.doi.org/10.21134/22553568.2019.25.bphm.
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Vitharanage, Imesha, and Amila Thibbotuwawa. "Enterprise Robotic Process Automation." Bolgoda Plains 01, no. 01 (October 2021): 10–12. http://dx.doi.org/10.31705/bprm.2021.2.
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Robotic Process Automation (RPA) is an emerging technology widely used across multiple sectors such as human resources, healthcare, finance, accounting, manufacturing, higher education and supply chain management, etc. RPA, also known as ‘software bots’, replaces manual, rule-based, repetitive tasks humans perform. These software bots are currently in a journey, evolving to be more sophisticated, mimicking human activities and enabling humans to achieve higher-valued tasks. Hence, RPA impacts the overall operational efficiency in organisations through multiple facets by its integration with employees, existing technologies and infrastructure, and business processes. It reduces the burden on IT as it does not disturb the underlying legacy systems. It increases reliability as bots can work 24x7 effectively. It is used as a time and cost reduction technology as it reduces the size of the manual workload. The tasks performed through RPA is accurate as it is less prone to errors. It increases compliances as it follows the rules and keeps audit trails. The productivity rate of organisations increase as the execution time through RPA is faster than tasks being performed by human employees. Furthermore, RPA is introduced as a low code technology that uses drag and drop functionalities with little to no programming knowledge [1].
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Jayaweera, Mahesh, Hasini Perera, Gimhani Dhanushika, and Buddhika Gunawardane. "Consumption of chilled water stored in a PET bottle multiple times: are we quenching thirst or gulping phthalates?" Bolgoda Plains 01, no. 01 (October 2021): 28–31. http://dx.doi.org/10.31705/bprm.2021.8.
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The statistics forecast that the production of polyethylene terephthalate (PET) bottles worldwide in 2016 was about 485 billion, and the same in 2021, has been approximately 583 billion. Although such productions in many countries have the ear of prominent political and social leaders, high production rates still reign the global market. In parallel, revered scientists globally conflate plausible and incontrovertible medical canons against the use of PET bottles for the protection of public health. Nevertheless, unwashed masses worldwide dislodge or disparage such public health doctrine but face a myriad of health hazards. For many years, mainly beneath the public’s ignorance, the solid collective rhetoric expressed by PET-bottle manufacturing companies has not let such medical dogma take hold in the society, instead purposefully manipulated the market with conflating pure baloneys or fallacies.
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Ariyadasa, Thilini U. "Microalgae: A promising bioresource for a sustainable future." Bolgoda Plains 01, no. 01 (October 2021): 06–09. http://dx.doi.org/10.31705/bprm.2021.5.
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Microalgae are highly diverse unicellular photosynthetic organisms found in aquatic environments. Microalgae produce oxygen during their proliferation, contributing to nearly 50% of the total oxygen production in the world. Concurrently, microalgae consume carbon dioxide in the atmosphere, thereby serving as carbon sinks to alleviate the effects of global warming. In comparison to terrestrial plants, microalgae exhibit rapid growth rates, higher photosynthetic efficiency, shorter harvesting time and higher biomass productivities. Moreover, they do not require arable land or potable water to facilitate their growth, hence becoming a more sustainable feedstock as compared to conventional crops. Altogether, microalgae have been identified as a bioresource with great industrial potential due to their ability to accumulate commercially valuable metabolites that can be extracted and subsequently processed into diverse bioproducts such as biofuels, pharmaceuticals/nutraceuticals, biofertilizer and animal feed.
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Karunaratne, Virajini, Gayathri Ranathunga, and Sulari De Silva. "Curating Kandyan traditional clothing in the UNESCO world cultural heritage in digital paradigm." Bolgoda Plains 01, no. 01 (October 2021): 36–39. http://dx.doi.org/10.31705/bprm.2021.10.
Full textAbstract:
Historical notions of cultural values in the heritage sector have been identified by holders of curatorial expertise based at institutions with large collections of artifacts. However, the rise of new digital technologies has facilitated not only active two-way engagement with heritage, but also a broadening of what we mean by heritage and how it can be accessed, through the co-production of exhibitions, oral histories, and other forms of display and archive based on personal remembrance, recollection and interactivity.
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Karunananda, Asoka S. "Enhancing the power of understanding." Bolgoda Plains 01, no. 01 (October 2021): 13–15. http://dx.doi.org/10.31705/bprm.2021.3.
Full textAbstract:
Power of understanding is a rewarding cognitive capacity required for all of us from early childhood to the highest level of intellectual settings. Among other things, the concept of understanding plays a vital role in education. When I was a second-year undergraduate, I was so curious to know why some of my colleagues could understand subjects much faster than the others, and this curiosity compelled me to research on how understanding manifests in our minds. My literature review revealed that the ultimate happiness/truth stated in Buddhism is a matter of understanding the world differently from the way we do it generally. Literature also showed that many people in Buddha’s time understood the ultimate truth while listening to the discourse of the Buddha. Those who could not understand a matter then and there had to develop certain cognitive skills through various cognitive tasks such as further listening, discussing, thinking, and meditating. This is equally applicable to our educational settings as well because some students understand the subject matter during the lecture itself, while the others need involve in additional reading activities, discussions, tutorial work, and so on.
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De Silva, M. Mavin, H. Niles Perera, and Amal S. Kumarage. "Impacts of the first COVID-19 Lockdown on Mobility and Consumption of Households in Sri Lanka: Results of a Survey." Bolgoda Plains 01, no. 01 (October 2021): 06–09. http://dx.doi.org/10.31705/bprm.2021.1.
Full textAbstract:
Sri Lanka was introduced to an unnamed virus on 27th January 2020 when a Chinese tourist was diagnosed with it [1]. Within 8 weeks, the virus named COVID-19 had begun infecting the local population, and the government was taking measures to prevent its spread in Sri Lanka. The government imposed an all-island curfew on 20th March 2020 [2]. Starting from 20th March, the government encouraged Work-from-Home (WFH) and allowed agricultural activities while imposing restrictions on all physical movements [3]. With curfew, the government allowed the distribution of essential items and goods transport through issuing curfew passes. In spite of these measures, people faced many hardships during this time. The department of Transport and Logistics Management, Faculty of Engineering, University of Moratuwa, conducted a survey on 12th April and completed on 20th April 2020, during a period that the country was having curfew almost all the time. The purpose of this survey was to determine the immediate impact that the government’s steps had on the people’s lives and their impressions on the future. Over 1100 respondents from all districts of the country took part in the online and email survey. As this would return a biased sample, we normalised the collected data across the districts and by educational profile to make the representation as accurate as possible [4]. The analysis led to the following 3 noteworthy findings that would be useful for any future emergency or return to curfew if the need so arises.
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Herath, Oshadhi K., Praveen Perera, T. Sivakumar, Buddhi Ayesha, Amal S. Kumarage, and Amal Shehan Perera. "Use of Videography for Traffic Surveys in Sri Lanka." Bolgoda Plains 01, no. 01 (October 2021): 07–08. http://dx.doi.org/10.31705/bprm.2021.6.
Full textAbstract:
Traffic Surveys are crucial for different transport studies like Origin-Destination studies, Traffic Volume estimations, Vehicle Flow characteristics Determinations, Speed and Delay Studies, Turning Movement analysis, Parking Analysis etc. Currently, Sri Lankan researchers are using mostly human-based manual surveys and semi-automated methods for Traffic Surveys. However, there is an issue in cost, effort, the value of time, and the accuracy of data gathering in the above methods. Therefore, under Accelerating Higher Education Expansion and Development (AHEAD) grant, we are researching the possibility of doing surveys using videography and Artificial Intelligence technologies for continuous and accurate data collection to reduce the burden that is currently facing. Using CCTV video cameras, we have done surveys in different locations in Western Province to check
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Sirithunga, Chapa, and Buddhika Jayasekara. "Cognitively Intelligent Models for Human-Robot Interaction with MIRob." Bolgoda Plains 01, no. 01 (October 2021): 04–06. http://dx.doi.org/10.31705/bprm.2021.4.
Full textAbstract:
This research explores how a robot should gather knowledge upon a scenario between a robot and its user and then generate appropriate intelligent responses towards its user. Therefore, cognitive models were developed to act as a robot’s intelligence or the brain to make situation-specific decisions. Such insightful decisions will help the robot act in a social environment without disturbing its user or other humans around.
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Gunawardena, Harinda, and Udaya Wickramasinghe. "Materialising Gender-fluidity through Fashion." Bolgoda Plains 01, no. 01 (October 2021): 08–09. http://dx.doi.org/10.31705/bprm.2021.7.
Full textAbstract:
As the final year comprehensive design project for the Honours Degree of Bachelor of Design, Department of Integrated Design, Faculty of Architecture, University of Moratuwa, I have selected a project which is based upon my own clothing brand. It is an emerging ready-to-wear clothing brand based in Sri Lanka, which was launched in August 2020 through the Colombo Fashion Week named “HARID”. Currently, HARID retails at the Design Collective store in Colombo for a consumer group based upon it. The brand philosophy of HARID is to challenge gender-related stereotypical concepts. As the brand identity, HARID uses heritage craft practices.
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Rodrigo, Thimith. "Increasing the Political Involvement and Political Literacy of Sri Lankan Youth: A Communication Design Aspect." Bolgoda Plains 01, no. 01 (October 2021): 40–41. http://dx.doi.org/10.31705/bprm.2021.11.
Full textAbstract:
Majority of the younger generation of Sri Lanka in particular have a very low regard for the whole subject of politics. The reason for this has been the political dysfunction that they witness in their day-to-day lives. The most common dialogue they hear concerning politics is one where the older generations acknowledge that the political landscape is an utter mess.
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Jagoda, Sathindu, Hirushie Karunathilake, and Janaka R. Gamage. "Life Cycle Thinking for Packaging." Bolgoda Plains 01, no. 01 (October 2021): 50–52. http://dx.doi.org/10.31705/bprm.2021.14.
Full textAbstract:
Unsustainable packaging practices are one of the leading problems in today’s world, leading to unnecessary resource consumption, increased waste generation, environmental pollution, and an overall negative impact on ecosystems. Global statistics show that 8 million metric tons of plastic ends up in the oceans every year. It has been estimated that approximately 79% of plastic produced since 1950 has been sent to landfills or otherwise released to the environment. With the current trends, the United Nations predicts that the plastic content will overweigh the fish in the ocean by 2050 [1].
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Ranaweera, Achini, and Amali Wijekoon. "Is cues of contagious diseases in advertising a friend or foe?" Bolgoda Plains 01, no. 01 (October 2021): 60. http://dx.doi.org/10.31705/bprm.2021.18.
Full textAbstract:
Dr (Mrs.) Achini Ranaweera, a Senior lecturer from the Department of Textile and Apparel Engineering, Faculty of Engineering and Dr (Mrs.) Amali Wijekoon, a Senior lecturer from the Department of Management Technology, Faculty of Business in collaboration with two international researchers from Australia and the UK are all geared up to examine if cues of contagious disease in advertisements can influence consumption behaviour by eliciting negative emotions such as anxiety, disgust, and fear.
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Sewvandi, Galhenage A., and J. T. S. T. Jayawardane. "First-principles calculation on electronic properties of Bismuth-halide inorganic perovskites for solar cell." Bolgoda Plains 01, no. 01 (October 2021): 56–57. http://dx.doi.org/10.31705/bprm.2021.16.
Full textAbstract:
Solar energy is a commonly used alternate source of energy and it can be utilized based on the principle of the photovoltaic effect. The photovoltaic effect converts sun energy into electrical energy using photovoltaic devices (solar cells). A solar cell device should have high efficiency and a long lifetime to be commercially beneficial. Presently, silicon and thin-film solar cells are widely employed. The crystalline solar cells are more efficient but they are also expensive. Thin-film solar cells are formed by placing one or more thin layers of photovoltaic materials on different substrates. Although these cells have a lower cost, they are also less efficient compared to Si-based solar cells. Organic-inorganic hybrid lead halide perovskite solar cells are one of the most promising low-cost power conversion efficiency technologies that could exceed the 26% threshold. However, the lack of environmental stability and of high lead toxicity are the main bottlenecks that impede the future industrialization and commercialization hybrid lead halide perovskite. Hence It is important to achieve high power conversion efficiency while also maintaining stability and non-toxicity in the development of new lead-free perovskite materials.
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Ovitipana, Asanka O. R. B. E. W. D., and Sarath W. S. B. Dasanayaka. "Urban Food Security of the Colombo Metropolitan Region (CMR) in Covid times." Bolgoda Plains 01, no. 01 (October 2021): 46–48. http://dx.doi.org/10.31705/bprm.2021.13.
Full textAbstract:
Colombo Municipality Region (CMR) consists of a highly complex food system that relies on the supply from distant outstations which literally collapsed during the pandemic situation. A requirement exists for empirical research to derive guidelines and recommendations to increase the sustainability and security in the food supply in CMR during a disaster situation.
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Malkanthi, S. N. "An Innovative Approach to Produce Soil-Based Building Products." Bolgoda Plains 01, no. 01 (October 2021): 58–59. http://dx.doi.org/10.31705/bprm.2021.17.
Full textAbstract:
Soil has been used as a building material in different forms, such as mud, adobe, rammed earth, and bricks. Compressed Stabilized Earth Block (CSEB), a form of soil blocks with different additives including cement, fly ash, and lime, is a sustainable building material with many advantages compared to other conventional building materials. The usual practice of past researchers in producing CSEB was to add different materials like sand to the soil to control its clay and silt (finer) content. A high level of finer content is not desirable when it comes to the strength and durability of CSEB. This study proposes to reduce/ extract the finer content in the soil by washing it using a conventional concrete mixing machine.