bluuebunny/crossref_metadata_2025_split huggingface dataset:
Recurrent Vascular Access Site Thrombosis in Patients on Hemodialysis - a Problem of Thrombophilia?.
Authors: Klamroth Robert , Seibt Frank , Rimpler Hartmut , Landgraf Helmut 
Published: October 2019

Abstract
Introduction: Vascular access site thrombosis in patients receiving hemodialysis is a major cause of hospital admission and recurrent surgery. The underlying pathologic cause is often stenosis of the venous vessel due to fibromuscular hyperplasia. But in the case of early failure occasional studies have investigated that hypercoagulability could play an important role in this context.
Aim of the study: Is there a higher prevalence of hereditary and acquired thrombophilic risk factors in patients with vascular access thrombosis in comparison to patients without?
Patients: In 2002 and 2003 we examined 52 consecutive patients (mean age 66,1 years) receiving hemodialyisis. 27 patients (pts) in group 1 had a history of vascular access site thrombosis and 25 pts in group 2 had not and an open vascular access for longer than at least six months. All pts in group 1 had a history of at least two occlusions of vascular access. 10/27 pts in group 1 with prosthetic grafts had a history of thrombosis of arteriovenous fistula before implantation of PFTE graft.
Methods: In every patient hereditary and acquired thrombophilic risk factors were determined including antithrombin (AT), protein C (PC), protein S (PS), factor V-G1691A-mutation (FVM), prothrombin-G20210A-mutation (FIIM), homocysteine, lipoproteine (a) (Lpa), lupus anticoagulant (LA), cardiolipin antibodies IgG and IgM (ACA), fibrinogen and factor VIII. Platelet hyperreactivity was studied by light transmittance aggregometry in platelet rich plasma (Aggregometer PAP 4, moelab inc.). Aggregation was recorded as the maximum percentage change in light transmittance from baseline using platelet poor plasma as a reference. We defined sticky platelets as platelet aggregation > 30% after induction with different concentrations of ADP (10, 1 and 0,5 μmol) in platelet rich plasma.
Results: We found in 14/27 pts with vascular access site thrombosis antiphospholipd antibodies (LA and/or ACA) in comparison to only 2/25 in pts without thrombosis. Activated platelets like the sticky platelets syndrome was shown in 11/27 pts in group 1 and 4/25 pts in group 2. In both groups hyperhomocysteinaemia (23/27 pts and 21/25 resp.), factor VIII elevation (21/27 pts and 22/25 resp.), fibrinogen elevation (22/27 pts and 21/25 resp.) and high levels of Lpa (7/27 pts and 6/25 resp.) were quite similar. There were no significant differences in the number of hereditary risk factors like AT, PC, PS, FVM and FIIM in both groups.
Conclusions: In patients receiving hemodialysis we found a high prevalence of acquired thrombophlic risk factors like elevation of factor VIII, homocysteine and fibrinogen. There seems to be causal relation between vascular access site thrombosis and espacially antiphospholpid antibodies and activated platelets (sticky platelets syndrome). The evaluation of these thrombophilic risk factors in patients with recurrent vascular access site thrombosis could lead to an improved antithrombotic therapy.
Manifestations of the Antiphospholipid Syndrome in Patients with Malignancies.
Authors: Miesbach Wolfgang A. , Asmelash Geneth , Puetz Birgit , Boehm Martina , Scharrer Inge 
Published: October 2019

Abstract
The presence of antiphospholipid antibodies has been reported in a large variety of malignancies. It is not clear, however, if the antiphospholipid antibodies are related to thrombotic associations of the antiphospholipid syndrome (APS) in these patients. We investigated the frequency of thrombotic manifestations in 58 patients with the presence of antiphospholipid antibodies and a history of neoplasia, including haematologic and lymphoproliferative malignancies.
Methods Antiphospholipid antibodies were detected by clotting assay (lupus anticoagulant, LA) or by enzyme-linked immunosorbent assay (anticardiolipin antibodies). LA were tested by more than 2 different methods according to the proposed criteria of the SSC of the ISTH.
Results 39/58 patients suffered from solid tumours mostly from carcinoma of the breast, prostate, and colon and 19/58 patients from malignant haematologic or lymphoproliferative diseases mostly from Non-Hodgkin lymphoma. One patient was suffering simultaneously from two carcinomas of the prostate and the testicle and a Non-Hodgkin’s lymphoma.
Among the patients with solid tumours 18/39 (46 %) patients had thromboembolic complications of the antiphospholipid syndrome. Among the patients with haematologic and lymphoproliferative malignancies only 6/19 (32 %) suffered from thromboembolic complications. Thrombotic manifestations were more common on the arterial than the venous site. There was no relation between the titres of aCL antibodies and the rate of clinical manifestations. In two patients aPL disappeared after the effective treatment of the tumor. Especially patients with very high titres did not present any thromboembolic manifestation.
Conclusion The presence of antiphospholipid antibodies may identify a subset of cancer patients with high risk of developing thrombotic complications but the frequency of thrombosis is lower in aPL positive patients with lymphoproliferative and haematological malignancies. Especially in these patients very high titres of aCL antibodies do not seem to be associated with clinical manifestations of the APS.
Age May Modify the Relationship between Factor V Leiden Mutation - but Not G20210A Prothrombin Gene Variation - and Idiopathic Venous Thromboembolism: An Hospital-Based Case-Control Study.
Authors: Le Gal Gregoire , Lacut Karine , Couturaud Francis , Oger Emmanuel , Mottier Dominique 
Published: October 2019

Abstract
Introduction:
Factor V Leiden is the most common inherited risk factor for venous thromboembolism (VTE). A four- to sevenfold increased risk of VTE for the heterozygous state has been reported by numerous epidemiological studies but most of them did not include patients over 70 years. Surprisingly, we found in a previous study no association between Factor V Leiden and VTE in patients over 70 years.
Methods
Therefore we conducted a large hospital-based matched case-control study to test the hypothesis of an interaction between age and the factor V mutation, as well as G20210A prothrombin gene variation.
Results:
We analysed 392 patients experiencing VTE not related to a major acquired risk factor and their matched controls. Factor V Leiden was not associated with VTE in patients aged 80 years and over: odds ratio 0.8 (95%CI 0.2-3.4). There was a significant interaction between age and the mutation for VTE risk (p=0.03). Conversely, the association between the G20210A variant and VTE was consistent across age-groups: odds ratio 2.8 (95%CI 1.4–5.8).
In conclusion, age may modify the relation between factor V Leiden and VTE. The prevalence of the factor V mutation decreased with increasing age among patients with VTE but not among controls.
Rapid Detection of 871G>A Mutation by Sequence Specific PCR in ABO*A301 Blood Donors.
Authors: Novaretti Marcia C.Z. , Domingues Alexandre E. , Pares Madalena M.N.S. , Dorlhiac-Llacer Pedro E. , Chamone Dalton A.F. 
Published: October 2019

Abstract
Background: A3 blood subgroup is heterogeneous at molecular level. To date only two cases of ABO*A301 allele containing a single substitution 871G>A (Asp291Asn) were found in A3B individuals exclusively. The purpose of this study was to use a sequence specific PCR (PCR-SSP) to detect 871G>A mutation in A3 and A3B samples.
Material and methods: Twenty blood samples from unrelated donors were classified serologically as A3 and A3B using tube test technique and gel-test simultaneously. Agglutination of red cells by lectins was performed according to manufacturer’s instructions using anti-A1 lectin from Dolichos biflorus and anti-H from Ulex europaeus. Genomic DNA was prepared from peripheral blood cells by a salting-out method. ABO genotyping was performed using a polymerase chain reaction method with sequence specific primers: 95s (5′-CAC CAG GCC ATG ATG GTC A-3′) for consense G detection, 96s (5′-CAC CAG GCC ATG ATG GTC A-3′) for mutation A detection and 244as (5′-CGC AGT GAA CCT CAG CTT C-3′) for both detections on gene ABO exon VII (Seltsam, Transfusion 2003, 43(4): 428-39). PCR amplification was carried out under the following conditions: 5 initial cycles at 94°C for 10 minutes and 65°C for 1 minute; 20 cycles of 94°C for 10 seconds and 65°C for 1 minute; 20 final cycles of 94°C for 20 seconds, 61.5°C for 50 seconds and 72°C for 30 seconds. PCR amplification of 0.1 μg of genomic DNA was performed in a total volume of 25 μL containing 0.2 mM of each dNTP, 5 units Taq DNA polymerase in the buffer supplied, 2.5 mM MgCl2 and 12.5 pmoles of each primer. Fragment of 177 bp derived from each mutation or consense assay was separated for 90 minutes at 102V using 0.5 μg.mL−1 ethidium bromide-stained 2.5% agarose gels and visualized in a UV light apparatus (Eagle Eye II).
Results: We found 871G>A mutation in heterozygosis in three blood donors (15%), two of them typed as A3B and one as A3.
Conclusion: The 871G>A mutation is relatively commom in A3B and A3 individuals and can be easily detect by PCR-SSP.
Autoantibody Formation and Alloimunization in Sickle Cell Disease Patients.
Authors: Novaretti Marcia C.Z. , Jens Eduardo , Pagliarini Thiago , Rodrigues Andreia L. , Dorlhiac-Llacer Pedro E. , Chamone Dalton A.F. 
Published: October 2019

Abstract
Background: Alloantibody and autoantibody formation to red blood cell (RBC) antigens is one of the observed complications in sickle cell disease patients (SCD). The incidence of alloimmunization and autoantibodies in this selected group of patients is particularly high, although the clinical implication of autoantibodies in sickle cell disease patients is not clear. The purpose of this study is to evaluate the rate of alloantibody and autoantibody formation in SCD patients.
Study design and methods: A retrospective analysis of transfused sickle cell disease patients followed at Fundacao Pro-Sangue Hemocentro de Sao Paulo between 1988 and 2004 were retrieved. Data on transfusion history, were correlated with development of alloantibodies and autoantibodies.
Results: The study group was composed by 43 sickle cell disease patients followed for a mean of 89 months (22–116). The number of RBC units transfused (mean) was 64 (4–208). The development of the first alloantibody was detected after a mean of 40 months (1–107) after the first transfusion in our institution. Out of these patients, 31 (72.1%) were identified with RBC alloantibodies; 9 of these patients (21%) had both allo and autoantibodies to RBC antigens, whereas 5 (55.6%) developed autoantibodies after alloimmunization. The one remainder had only autoantibodies.
Conclusion: The alloimmunization rate was extremely high (72.1%) and can be partially explained because of the extended time of follow-up (mean of 89 months). Different from the literature the development of autoantibodies preceeded alloantibodies in 44.4%. The impact of this observation in clinical practice warrants further investigation.
High Frequency of HPA-2B Allele in Brazilian Apheresis Blood Donors Detected by Sequence Specific PCR.
Authors: Novaretti Marcia C.Z. , Bonifacio Silvia L. , Manhani Reinaldo , Dorlhiac-Llacer Pedro E. , Mendrone Alfredo , Chamone Dalton A.F. 
Published: October 2019

Abstract
Background: Human platelet antigens (HPA), mainly HPA-1 and HPA-2, play a significant role in alloimmune thrombocytopenic syndromes. Accurate HPA typing is important for the diagnosis and therapy of these patients. HPA gene frequencies vary in different populations. Thus, it is relevant to determine HPA genotype in distinct populations. The purpose of this study was to perform HPA-1,-2 genotyping in apheresis Brazilian blood donors.
Material and Methods: Genomic DNA was prepared from whole blood of 57 unrelated repeated apheresis Brazilian blood donors using a commercial DNA isolation kit (Qiagen,. GmbH, Germany). Sequency specific PCR (SSP-PCR) was performed amplifying HPA-1A/HPA-1B and HPA-2A/2B genes using a pair of primers for each allele as described elsewhere (HPA-1A ACT TAC AGG CCC TGC CTC T; HPA-1B ACT TAC AGG CCC TGC CTC C; HPA-1AS AGC CGG AGT GCA ATC CTC TG; HPA-2A CCC CCA GGG CTC CTG AC; HPA-2B CCC CCA GGG CTC CTG AT and HPA-2AS GCC AGC GAC GAA AAT AGA GG). A fragment of the human growth hormone gene (HGH) served as internal positive control. PCR was carried out in a final volume of 20 μL, containing 0.2 mg of genomic DNA, 0.2 mM of each dNTP, 5% of glycerol, 1.5 units of platinum Taq polymerase (Invitrogen, Brazil) in the buffer supllied, 5.0 mM MgCl2 and 10.0 pmoles of each primer. Fragments of 196 bp derived from HPA-1A/1B mutation and of 241 bp for HPA-2A/2B were separated for 90 minutes at 102V using 0.5 μg.mL−1 ethidium bromide-stained 2.0% agarose gels and visualized in a UV light apparatus (Eagle Eye II, EUA).
Results: The genotype frequencies presented 77.2% of HPA-1A/1A, 19.3% of HPA-1A/1B and 3.5% of HPA1B/1B; for HPA-2 the frequencies were 56.1% of HPA-2A/2A, 35.1% of HPA-2A/2B and 8.8% HPA-2B/2B. The gene frequencies in apheresis blood donors are 0.87 for HPA-1A, 0.13 for HPA-1B, 0.74 for HPA-2A and 0.26 for HPA-2B.
Conclusion: In this studied population we found high gene frequency homozygous HPA-2B when compared with other data already published. (Castro et al, European Journal of Immunogenetics,1999;26:355–60).
Mobilisation and Harvest of Peripheral Blood Stem Cells from 187 Healthy Donors. A Comparison of Two Different Apheresis Machines and Three Apheresis Programs.
Authors: Kumlien Gunilla , Bergstrom Gunilla , Ayoub Amal , Kanogo Liisa , Hagglund Hans 
Published: October 2019

Abstract
Background:
Between January 1995 and July 2004, mobilisation and harvest of peripheral blood stem cells for allogeneic transplantation was performed in 136 related and 51 unrelated healthy donors. Four related and one unrelated donor were harvested on two separate occasions after renewed G-CSF mobilisation. For logistic reasons, automated harvest programs were used until recently. Automated apheresis programs for peripheral blood stem cell harvest are less labour intensive but unfortunately also less efficient. One of the potential health hazards for donors is the depletion of platelets, thus it is essential to minimize platelet depletion.
Method:
Between 1995 and 1999 CS3000 (Baxter) cell separator with an automated stem cell harvest program was used. Since 1995 Cobe Spectra (Gambro) cell separator is used, initially with the automated program AutoPBSC and since March 2004 with the “manual” program MNC. The 187 medical reports were analysed retrospectively.
Results:
Median age was 40 years (range 14–71), median weight was 75 kg (range 42–130) and the male/female ratio was 1.3 (107/80). In a majority of donors (97%) antecubital veins were used as access to the circulation. In 4% of donors antecubital veins were unsuitable, and a femoral catheter was therefore employed.
Donors were mobilised with G-CSF 10 ug/kg and harvested on days five and six. Transplantation dose was median 6,3 x 10(6) CD34+ cells/kg recipient (range 1,2–98,6). Harvest yield was median 5,8 x 10(6) CD34+ cells/kg donor (range 1,4–16,3). Donor platelet count after second apheresis was median 129 x 10(9)/L (range 57–268) (normal range 150–400).
There were no significant differences regarding yield of CD34+ cells between male and female donors or between the two automated apheresis programs CS3000 and CobeSpectra AutoPBSC but the shift to CobeSpectra MNC-program led to a significantly increased yield only after six (three male and three female) donors (Mann-Whitney Test p<0,005) and median yield doubled to 10 x 10(6) CD34+ cells/kg donor.
There was no significant difference between CS3000 and Cobe Spectra MNC regarding platelet depletion but Cobe Spectra AutoPBSC was significantly less platelet depleting compared both to CS3000 (Mann-Whitney Test p<0,001) and Cobe Spectra MNC (Mann-Witney Test p<0,001).
Conclusion:
No serious side effects were reported during G-CSF mobilisation or apheresis. Cobe Spectra MNC program is significantly more efficient regarding yield of CD34+ cells than the two automated programs. Cobe Spectra AutoPBSC is significantly less platelet depleting than both CS3000 and Cobe Spectra MNC.
Gene Expression Profiles, Differentiation and Lineage Commitment of Human Myeloid Progenitors.
Authors: Edvardsson Louise , Dykes Josefina , Olofsson Tor 
Published: October 2019

Abstract
With the objective to further elucidate the mechanism behind commitment to erythroid and neutrophil lineages, we isolated by cell sorting common myeloid progenitors (CMPs), granulocyte/monocyte progenitors (GMPs) and megakaryocyte/erythrocyte progenitors (MEPs), based on the surface expression of CD123 (IL3R-α) and CD45RA on human CD34+ bone marrow cells (first described by Manz et al. PNAS, 2002;99:11872). Methylcellulose cultures supporting the growth of myeloid and erythroid progenitors, and real-time RT-PCR mapping the gene expression of Flt3, c-kit, TpoR, GATA-2, GATA-1, SCL, NF-E2, EpoR, ABO, β-globin, GPA, PU.1, C/EBPα, C/EBPε, G-CSFR, proteinase 3 (PR3) and lactoferrin, were used to validate and characterize the progenitors and their progeny. Cell sorted progenitors were labeled with CFDA, SE to track cell division and cultured in suspension to induce neutrophil or erythroid differentiation (SCF+G-CSF for neutrophil and Epo+IL–3+GM–CSF for erythroid culture). After 3–5 days, cells that had gone through 1–8 divisions were sorted and changes in clonogenicity and gene expression were studied. The CMP-population retained some clonogenicity after as many divisions as were tested (at the most six divisions in erythroid and eight in neutrophil culture) and the CMPs differentiated along the lineage defined by the culture system, as evidenced both by the methylcellulose cultures and an increasing expression of GATA-1 and EpoR in erythroid and PU.1, G-CSFR and PR3 in neutrophil cultures, respectively. On the other hand, the GMP-population displayed granulocyte/monocyte (G/M)-differentiation irrespective of the culture system used, although it divided fewer times and lost its clonogenic capacity faster in erythroid culture. Little or no clonogenicity remained after 4–5 divisions in erythroid culture, while some colony-forming capacity remained even after seven divisions in neutrophil culture (maximum number tested). The increased expression of the granulopoiesis-associated genes was also less pronounced in the erythroid culture. The MEP-population dominated by erythroid differentiation capacity retained colony-forming capacity for at least 6–7 divisions in both erythroid and neutrophil cultures, although with a higher overall clonogenicity in erythroid culture. Unexpectedly, however, MEPs were restricted to G/M-differentiation when cultured in neutrophil culture. In cells from erythroid culture the expression of GATA-1, EpoR and β-globin increased, while a corresponding pattern was seen for PU.1, G-CSFR and PR3 in neutrophil culture. Overall, our data support the progenitor classification, based on the surface expression of CD123 and CD45RA, with regard to CMP and GMP populations but question it with regard to the MEP-population. The change in differentiation course for the MEPs in neutrophil culture could be a result of an initially present G/M-potential or a less strict commitment susceptible to cytokine-induced redifferentiation.
In Vitro Hematopoietic Lineage Interconversion from Human Bone Marrow Stem and Progenitor Cells.
Authors: Chen Ling , Jean-Noel Stephanie , Hall Kevin , Shi Ying , Rodgers Griffin P. 
Published: October 2019

Abstract
The cell surface antigen, CD133, marks a fraction of hematopoietic stem and progenitor cells and has been successfully used to study their differential biology. To evaluate the differentiating capacity of stem/progenitor cells, we cultivated purified normal human bone marrow CD133 selected cells for 2 weeks with erythropoietin (EPO) or granulocyte colony-stimulating factor (G-CSF) to induce erythroid or myeloid differentiation, respectively. After the second week of cultivation, we reversed the seeding environment of the two populations by placing EPO treated cells into a G-CSF environment and G-CSF treated cells into an EPO environment for an additional 2-week culture. The cells produced in the culture were phenotypically defined by morphology and flow cytometry, and genotypically by RNA and proteomic analyses. Three-color flow cytometry was used for identifying CD133+ progenitors, CD36+ erythroid and CD13+ myeloid cells, as shown in Table 1. The morphology of the cultured cells, assessed by Wright-Giemsa staining, is consistent with the conversion of cellular specific markers. Rapid analysis of gene expression demonstrated co-expression of 76% of 266 genes analyzed among the erythroid and myeloid lineages. Furthermore, proteomic analysis exhibited the sharing of 33% of 9518 expressed protein spots assayed in the two populations after the first 2-week culture, and 32% after 2 weeks of the switch culture. Our data clearly demonstrate that the committed erythroid and myeloid precursors are able to change their fate and can switch into the opposite cell type by a conversion pathway under a specifically defined condition. We termed this switch as interconversion, considering conversion of hematopoietic cells to non-hematopoietic cells. Furthermore, the observations presented in this study show that cytokines used can improve the conversion. We are developing a mathematical model describing the kinetics of hematopoietic stem/progenitor cell transitions into specific lineages, along with the conversion of committed cells based on multiple potential energy wells corresponding to different cell states and cytokines.
Table 1. Expression of cell surface markers after 4-week culture
D0 1 week 2 weeks 4 weeks CD expression (%) E G E G E2w →G2w → G2w E2w Data are presented as a mean of at least 2 experiments. E: EPO; G: G-CSF; E2w or G2w: EPO or G-CSF treatment for two weeks. CD133+ 96.19 15.74 13.6 0.24 0.36 0.01 0.63 CD36+ 0 60.37 27.39 96.37 25.87 45.41 68.54 CD13+ 0.43 35.41 57.29 24.41 92.1 85.87 37.76 CD133+ / CD36+ 0.44 22.24 15.97 0.12 0.18 1.55 7.65 CD133+ / CD13+ 1.24 19.43 13.36 0.36 1.09 13.31 14.92 CD36+ / CD13+ 0.09 41.25 17.80 23.69 54.1 46.60 79.41