bluuebunny/crossref_metadata_2025_split huggingface dataset:
A134T, G156A and G1333A SNPs in the CD177 Gene Are Associated with Atypical Expression of Human Neutrophil Antigen 2a (HNA-2a)
Authors: Moritz Elyse , Chiba Akemi Kuroda , Albuquerque Dulcinéia M , Costa Fernando F , Yamamoto Mihoko , Bordin Jose Orlando 
Published: April 2020

Abstract
Introduction: The HNA-2a has gained clinical importance for being involved in immune neutropenias, febrile transfusion reactions and transfusion-related acute lung injury (TRALI). The HNA-2a shows the unique feature of being expressed only on a neutrophil subpopulation. The gene encoding the HNA-2a, CD177, is located on chromosome 19 at 19q13.2 and the cDNA consists of 2364 bp (RefSeq: NM 020406) coding for 437 amino acids including a signal peptide of 21 amino acids. In this study we investigated the presence of SNPs in the CD177 and its association with atypical expressions of HNA-2a.
Methods: After evaluating the expression of HNA-2a in 135 randomly selected Brazilians by flow cytometry (FACSCalibur, BD) using two pure monoclonal antibodies (TAG4 and 7D8), we observed that some subjects revealed an atypical expression of the antigen (a non reactive cell population plus two distinct reactive cell populations). Subjects were grouped according to cytometric results as having high (≥80%), low (≤50%) or atypical expression of HNA-2a. We isolated granulocyte RNA from the selected individuals and reverse transcription was performed according to kit protocol (SuperScript III First-Strand Synthesis, Invitrogen). Full-length cDNA was amplified and then sequenced according the manufacturer’s protocol (Mega BACE, GE). Data were analyzed using the Bioedit program.
Results: The flow cytometry analysis revealed that 131 (97%) subjects were HNA-2a positive, 4 (3%) HNA-2a negative, while 8 subjects (5.9%) showed an atypical expression of HNA-2a with levels ranging from 49 to 80%. Sequencing analysis of subjects with high (n=14), low (n=22) or atypical (n=8) HNA-2a expression identified six polimorfic sites (C49G, A134T, G156A, A793C, G1084A and G1333A), five of them predicting amino acid exchanges (except the SNP G156A). Comparing the cDNA sequences between individuals with typical (n=36) or atypical expression (n=8), we observed three polimorphisms (A134T, G156A and G1333A) that showed a significantly higher frequency in the group with atypical expression comparing to the group of individuals with typical expression (P= 0.004; 0.006; and &lt;0.0001, respectively; Fisher exact test).
Conclusions: The analysis of individuals with two distinct positive cell populations revealed three SNPs strongly related to this atypical HNA-2a expression, but do not seem to be related to the size of the HNA-2a-expressing neutrophil subpopulations. Matsuo et al (Transfusion2000;40:654) and Caruccio et al (Transfusion2003;43:357) have described atypical reactions with HNA-2a-specific MoAbs, however this is the first study in the literature that gives some clues to the molecular basis of the atypical expression of HNA- 2a in otherwise healthy individuals.
Primary Prophylaxis Using Peg-Filgrastim Is Useful Preventing Severe Infections and Neutropenia in Patients Diagnosed with Non Hodgkin Lymphoma (NHL). Report of Latin America International Multicenter Study
Authors: Villela Luis , Villalobos Alberto , Reyes-Brena Gustavo , Duque Jorge , Solís-Poblano Juan Carlos , Reynoso Eduardo , Ovilla Roberto , Camacho Alejandro , Vazquez Norberto , Garces Oscar , Pedrasa Enrique , Silva Salvador 
Published: April 2020

Abstract
Background: Severe infectious events after chemotherapy (CHT) are potentially life-threatening. This is directly related to neutropenia. Granulocyte colony-stimulating factor is used as supporting therapy to prevent neutropenia. Peg-filgrastim has been available in Mexico since 2005 and there is not any report about the use of this drug in Latin America.
Aim: To evaluate grade 4 neutropenia and infections related to neutropenia while using peg-filgrastim.
Methods: Patients between the ages of 18–65, with NHL (de novo or relapsed) with ≤ 30% bone marrow involvement were eligible. CHT was selected by each institution. Complete blood counts were obtained on days 0 (day before chemotherapy), 7, and 14.
Outcomes: Fifty three patients with NHL were included (one withdrew from study after the 2nd cycle of CHT). Three hundred and fourteen cycles were analyzed. Clinical characteristics: Age (mean ±SD): 51.8(±13.5), male: 47%, ECOG ≥ 2:32%), Advance stage (III/IV Ann Arbor): 53%, adjusted IPI (Intermediate/High):60.4%, First-line CHT 72%, second-line CHT 28%. Median neutrophil count (ANM) on day 0 was 8.69 ×109/L (IC95%: 6.44 to 10.94, previous each CHT), ANM on day +7 was 7.9×109/L (IC95%: 5.34 a 10.45) and day +14 ANM was 8.61 ×109/L (IC95%: 6.74 a 10.48) see figure 1. Neutropenia grade 4 was observed in 12.8% of the cycles, being more frequent on day 7 (90%) it means chemotherapy nadir, see figure 2. Severe infectious events were registered in 0.63% of the global cycles (two pneumonias, one of them when ANC was over 2 ×109/L) and non-severe in 1.91% being more frequent superior tract infections. No delays in CHT administration or deaths were reported.
Conclusion: Peg-filgarstim could decrease the frequency of grade 4 neutropenia the day before the next administration of CHT as well as the incidence of severe and non-severe infections in young patients.
Anemia Prevalence in a Population of Over One Thousand School Children in Northeastern Mexico
Authors: Escoboza Jose R Borbolla , Ugalde Patricia Argentina , Elizondo Luz Leticia , Patiño Beatriz Eugenia , Villela Luis M , Scott Sean P , Diaz Fernando Cristobal , Perfecto Yocanxochitl , Garcia Maria Isabel , Rocha Carlos , Kim Jina 
Published: April 2020

Abstract
Introduction: Anemia is a common finding in most developing countries. Of all populations, children are the ones who suffer most from anemia due to the long-term effects it can have on their growth and development. Large scale public programs to prevent anemia should always be based on epidemiological studies from significant sample sizes.
Patients and Methods: We included elementary school children from the state of Nuevo León, in northern Mexico (pop 4.2 million). We selected the children from eight public schools, six from Monterrey’s metropolitan area (pop 3.5 million) and two from the suburban area (pop. 0.5 million), mimicking the population distribution in our state. Three of these elementary schools give out school lunches consisting of milk and peanuts. Hemoglobin levels were obtained by using a Beckman-Coulter HMX® automated system (Miami, FL). We defined anemia as less than 12 g/dL of hemoglobin.
Results: After obtaining informed consent from the parents, we studied 1,137 children (550 males, 587 females), all of them between 6 and 12 years of age. Mean hemoglobin was 13.18 g/dL for females and 13.27 g/dL for males. We found 53 cases of anemia: 21 males and 32 females. Among the 53 anemic patients, 50 had hemoglobin between 11–12 g/dL, 1 between 10–11g/dL and 2 between 9–10 g/dL. We found significant differences in mean leukocyte numbers (7,200 vs 6,500/uL P=0.018); Mean Corpuscular Volume (82.9 vs 82.10 fL P=0.39); Mean Corpuscular Hemoglobin (29 vs 28.6 ug P=0.001), between the non-anemic and the anemic patients. Comparing the number of anemic children between schools with (28.7%) and without (71.3%) school lunches, the proportion of anemic children was significantly increased (26 vs 27 P=0.001) in schools that do not provide this school lunch.
Conclusions: Our results show a very accurate picture of the status of anemia in the urban and suburban areas of the state of Nuevo Leon in northeastern Mexico. Although one anemic child is one to many, our results show marked improvement when compared with our national means. Even though there are other factors that influence the absence of anemia in the different schools, the administration of the school lunch appears to have beneficial effects on anemia in our population.
Physicians and Patients Perception of a Problem as Ethically Relevant in Oncohematology: Only a Question of Perspective?
Authors: Giordano Giulio , Sacchini Dario , Farina Giuliana , Piano Silvia , de Paula Ignacio Carrasco , Storti Sergio 
Published: April 2020

Abstract
Ethical issues are connatural with the medical activity. Moral questions regarding some medical specialities as obstetrics or pre or perinatal medicine are well known. Nevertheless few is known about ethical problems perceived as remarkable from physicians and patients in onco-hematology. This is a retrospective monocentric descriptive study. Its purpose is to recognize, in oncohematology, what are the problems perceived as ethically relevant both from physicians and patients. Ethical issues regarding onco-hematologic patients were recognized consulting clinical diary of 100 patients treated in our institution. Patients have casually been selected in the temporal period from January 2004 to June of 2008. Ethical questions were listed in two groups: those perceived as remarkable for the physicians and those for the patients. Median age of the patients was 54.5 years (R 33–86). M/F was 55/45. Sixty-one patients were affected by chronic or indolent hemopathies. Sixty-six patients received 1st line therapy, instead 34 2nd line or superior. Seventy-five patients had primary school instruction, 22 secondary, 3 had got a degree. Eighty patients were assisted by a relative. Twenty-nine patients perceived a problem as ethically relevant (5 compliance to treatments; 5 refusal of treatment; 7 intolerance to hospitalization; 9 scarce trust in caregivers; 3 difficulty to understand the therapeutic plan). In 38 cases physicians perceived a problem as ethically relevant (8 over treatment; 7 type of treatment to adopt; 8 correct timing of therapy; 11 correct use of resources; 4 adequacy of treatment). Physicians perceived these problems mainly about patients requiring 2nd line treatment or further (28 cases out of 38). In 12 cases only patients perceived a problem, in 21 cases only physicians perceived a problem, while in 17 cases a problem was perceived as ethically relevant by physicians and patients simultaneously. In our study, in onco-hematology the main issue perceived by patients as ethically relevant is the scarce trust in caregivers, while for physicians it is the correct use of resources. Physicians perceive an ethical problem mainly considering patient candidate to 2nd line treatment or further. In 24% of cases, a problem perceived as ethically relevant by patients is underestimated by physicians. In this contest the possibility of an ethical consultation within a clinical counselling might create a convergence between physician and patient perspective.
Improving the Sensitivity and the Specificity of HLA-B27 Typing by Replacing Serological Tests with a TaqMan-PCR Assay
Authors: Weng Xiaoduan , Robin Louise , Audet Marie-lise , Hébert Linda , Lapointe Ginette , Major Micheline , Lemieux Bernard , Biron Guy , Bélanger Karl , Soulières Denis 
Published: April 2020

Abstract
HLA-B27 is a strong diagnostic biomarker as it is found in 90– 95% of ankylosing spondylitis. Routinely, the false-positive results generated by cross-reactivity of HLA-B27 monoclonal antibodies (mAbs) with some other type of HLA-B antigens are problematic. The present study aims at improving the sensitivity and specificity of typing for HLA-B27 by using a more accurate DNA assay. A real time sequence specific primer polymerase chain reaction (PCR-SSP) is performed by using MGB TaqMan oligoprobe and an ABI PRISM™ 7500 sequence detection system. The TaqMan PCR-SSP assay is compared with Immunophenotyping by flow cytometric antigen assay (BD HLA-B27-Kit, clone GS145.2). The results are finally confirmed by sequencing (ABI PRISM ® 3100). As summarised in the table, three methods are identical for clearly positive and negative results. There is 90% concordance for borderline negative results obtained by immunophenotyping with TaqMan PCR-SSC and/or sequencing. However, only 15.4% of borderline positives obtained by immunophenotyping are confirmed as true HLA-B27 positives by sequencing as well as TaqMan PCR-SSC. TaqMan PCR-SSP DNA assay completely correlates with sequencing (gold standard) with 100% PPV and NPV, compared to a PPV of 15% and a NPV of 98% for borderline results by immunophenotyping.
Conclusion: when compared with flow cytometric antigen assay, typing HLA-B27 by TaqMan PCR-SSP clearly demonstrates an advantage to better establish the genotype of the patient. Specifically, there is a marked difference for ambitious positive results from immunophenotyping. Moreover, compared to immunophenotyping, there is no need of fresh samples, can test a larger number of samples concomitantly, and reduces technical time as well as cost.
N = 52 Immunophenotyping TaqMan PCR-SSP Sequencing Immuno vs Sequencing TaqMan PCR-SSP vs Sequencing Positive 11 11 11 100% 100% Negative 17 17 17 100% 100% Positive (borderline by Immunophenotyping) 13 2 2 15% 100% Negative (borderline ) by Immunophenotyping) 10 9 9 98% 100%
A Mitochondrially Targeted Nitroxide JP4-039 Protects and Mitigates against Total Body Irradiation Induced Hematopoietic Syndrome
Authors: Epperly Michael W , Pierce Joshua G , Dixon Tracy , Franicola Darcy , Nie Suhua , Zhang Xichen , Vlasova Irina I , Wipf Peter , Kagan Valerian , Greenberger Joel 
Published: April 2020

Abstract
Nitroxides are efficient scavengers of free radicals and have been proposed for use as radioprotective agents. To circumvent the need for high concentrations for in vivo effects, and to target drugs to the mitochondria to prevent ionizing irradiation-induced apoptosis, we tested a family of compounds based on combining a hemi-gramicidin linker to a nitroxide. One small molecule (JP4-039) produced a 32-fold increased mitochondrial nitroxide localization in 32D cl 3 cells in vitro after one hour incubation at 37°C compared to the unlinked nitroxide tempo at 10uM. The cells were lyzed by rapidly freezing then thawing, followed by centrifugation to yield the nuclear, cytoplasmic, and mitochondrial fractions. EPR was used to quantitate nitroxide in each JP4-039 or tempo treated cell fraction. There was a 32-fold increase in the nitroxide signal for JP4-039 accumulation in the mitochondria compared to the nitroxide tempo. Irradiation survival curves were performed by incubating 32D cl 3 murine hematopoietic progenitor cells in JP4-039 or tempo (1 μM) for one hour before irradiation or by addition immediately following irradiation. The cells were irradiated to doses ranging from 0 to 8 Gy, suspended in methylcellulose-containing media, incubated for seven days at 37°C and colonies of greater than 50 cells were scored. The data was analyzed using linear quadratic and single-hit, multi-target models. Incubation of 32D cl 3 cells in JP4-039 for one hour prior to irradiation resulted in a significant increase in Do to 2.25 ± 0.11 Gy compared to 1.19 ± 0.13 Gy for 32D cl 3 cells alone or 1.32 ± 0.09 Gy for 32D cl 3 cells incubated in tempo (p=0.0042 or 0.0073, respectively). Similar results were achieved giving JP4-039 after irradiation with a Do of 1.97 ± 0.01 Gy (p &lt; 0.0001). To test effectiveness in vivo, C57BL/6NHsd female mice were injected intraperitoneally with JP4-039 or tempo (10 mg/kg) 10 minutes before (or four hours after) total body irradiation to the LD 75/30 dose of 9.75 Gy. Mice were monitored and those moribund from the hematopoietic syndrome were sacrificed. Mice injected with 10 mg/kg JP4-039 before irradiation had a significant increase in survival of 60% compared to 28% for control irradiated mice (p = 0.0005) or 44% for tempo treated mice (p = 0.2252). JP4-039 given after irradiation had a survival of 53% at 30 days after irradiation (p = 0.0474). Thus, JP4-039, a mitochondrial targeted nitroxide, is an effective radioprotector, and mitigator.
Extracellular HMGB1 Enhances Proliferation, Differentiation and Migration of Human CD34+ Hematopoietic Stem Cells in Vitro
Authors: Wang Xingbing , Chen Xin , Ren Weihua , Xu Xiucai , Song Kaidi , Liu Huilan , Sun Zimin , Huang Shiang 
Published: April 2020

Abstract
High-mobility group box 1 protein (HMGB1) is a chromatin protein and acts as a cytokine involved in inflammation, cell proliferation, differentiation, migration and stem cell recruitment. So far, little is known about its effect on hematopoietic stem cells (HSCs). In this study, we investigated whether receptors for HMGB1 are expressed on human CD34+ HSCs, and whether HMGB1 could affect HSCs proliferation, differentiation and migration in vitro. As examined by FACS analysis and RT-PCR, cord blood CD34+ HSCs express the HMGB1 receptors RAGE (receptor for advanced glycation end products), TLR2 (Toll-like receptor 2) and TLR4. To study the effects of HMGB1 on CD34+ HSCs proliferation and differentiation, freshly isolated cord blood CD34+ HSCs were cultured for 7 days in medium alone or in the presence of HMGB1. Flow cytometric analysis showed that HMGB1 (50ng/ml) can induce the differentiation of CD34+ HSCs along the granulo-monocytic (CD14+, CD13+) lineage and erythropoiesis (CD71+). In contrast, HMGB1 did not induce the expression of CD41, a marker for megakaryocyte lineage. The numbers of cells cultured in the presence of HMGB1 were always increased in comparison with controls. Furthermore, higher numbers of granulomonocytic progenitors (CFU-GM), erythroid progenitors (BFU-E), and CFU-MIX were confirmed by CFC assays in a HMGB1 dose dependent manner after 14-day culture. The results suggest that HMGB1 enhances CD34+ HSCs proliferation and differentiation. We next assessed the effects of HMGB1 on HSCs migration by chemotaxis assay using Boyden chambers. HMGB1 dose-dependently increased the chemotactic migration of CD34+ HSCs. A neutralizing anti-RAGE antibody significantly blocked the HMGB1-induced migration of HSCs, whereas neutralizing TLR2 and TLR4 antibodies did not significantly influence HMGB1-stimulated HSCs migration, suggesting that the migratory effect of HMGB1 on human HSCs is predominantly mediated by RAGE. In summary, our results provide the first report of HMGB1 receptors expression profile of human cord blood CD34+ HSCs and demonstrate that HMGB1 can increase the proliferation and migration of HSCs and directly induce of HSCs along myeloid differentiation and erythropoiesis in vitro. Further studies will be needed to clarify the mechanism of HMGB1 activation and the physiological function of HMGB1 in HSCs in vivo.
Expression of Functional Toll-Like Receptor 2 on Mouse Embryonic Stem Cells
Authors: Taylor Tammi , Derbigny Wilbert , Kim Young-June , Broxmeyer Hal E. 
Published: April 2020

Abstract
Embryonic stem (ES) cells have the capacity to produce all cell types of the body. Understanding murine ES (mES) cell proliferation, survival, differentiation, and self renewal will enhance knowledge of developmental biology and essential use of ES cells. Recently, Toll Like Receptor (TLR) activation has been shown by others to play a role in influencing the differentiation of hematopoietic stem cells. Previous studies have also shown that TLR activation prevents mesenchymal stem cell differentiation into adipocytes, chondrocytes, and osteocytes and plays a role in bone repair. We hypothesized that certain TLR’s would be expressed on mES cells and that the ligands for these expressed TLR’s would induce functional activity in the mESC’s. Therefore, we wanted to determine if TLRs are expressed on mES cells and if so, are they functional. Three different mES cell lines (R1, CGR8, and E14) were used to determine if TLRs are expressed at the mRNA level using primers for murine TLR1-9 mRNA. We found that TLR’s 1, 2, 3, 6, and 9 were expressed at the mRNA level, but TLR’s 4, 5, 7, and 8 were not. Based on the availability of antibodies to TLR’s, and using flow cytometry, we found expression of TLR2 but not TLR 4 on the surface of all three mES cell lines. TLR ligands were used to treat mES cells in the presence of leukemia inhibitory factor (LIF) for an hour. Activation of TLR2 by its ligand Pam3Cys, a synthetic tri-acyl lipoprotein, on mES cells induced NF-κβ nuclear translocation when compared to ES cells not stimulated with TLR ligands. LPS, the ligand for TLR4 did not induce NF-κβ nuclear translocation on ES cells, consistent with lack of cell surface expression of TLR4 on mES cells. TLR expression and TLR ligand interaction were not associated with changes in the morphology of the mES cells or expression of Oct-4, SSEA-1, KLF-4, or Sox-2, markers for maintenance of the undifferentiated state of mES cells. This suggests that the cells remain in an undifferentiated state even after TLR activation by Pam3Cys in the presence of LIF. Thus our study has identified functionally active TLR2 on the surface of mES cells, information that may be of use to further defining a role for TLR’s on ES cells, and for manipulation of other ES cell functions.
Study on the Mechanism of ‘ei’formation in the Bone Marrow Smears of the Patients with BMMNC-Coombs Test(+) Hematocytopenia
Authors: Fu Rong , Wang Yi hao , Shao Zonghong , Chen Jin , Li Lijuan , Liu Hui , Wang Huaquan , Xing Limin , Liu Hong 
Published: April 2020

Abstract
Objective To explore the mechanism of ‘erythroblastic island(EI)’ formation in the bone marrow of the patients with BMMNC-COOMBS( {)hematocytopenia.
Methods The category of BMMNC (hematopoietic stem cells, nucleated erythrocytes, granulocytes) -Ab in 48 patients with BMMNC-COOMBS( {) hematocytopenia were detected with FACS. The autologous antibody in the“EI” of these cases were explored with immuonhistofluorescence(IF). Clinical and other laboratory characteristics of these cases were also analyzed retrospectively.
Results Mouse anti-hIgG could be detected in the ‘EI’on the BM smear of 14 cases(29.17%), autologous antibody mainly deposited at the edge/membranes between macrophage and erythroblasts rather than in the cell plasma. Positive reaction were seen in all the cases with BMMNC-GlycoA IgG(+). The initial manifestation of the 14 patients was anemia. The percentage of reticulocyte (1.98%) and the serum level of IBIL of these cases (9.39 mmol/l)(5–24.90 mmol/l) were elevated in the 14 patients; Hypercellularity with higher percentage of erythroid (44.14%) in BM was seen in most patients. These cases had good reponse to high dosage of IVIgG (HDIVIG) or/and Glucocorticoids. There were significant difference of these indices between the IF(+), the IF(−)group and control group.
Conclusion Macrophage was connected with erythroblasts through autologous IgG in the‘EI’s of some patients with BMMNC-Coombs test(+) Hemocytopenia. These ‘EI’s were the places where macrophages devoured and destroyed erythroblasts rather than the places where erythroid development and differentiation. The pathogenic mechanism of this kind of disease might be associated with macrophages phagocyting and destroying more BM hematopoietic cells.