bluuebunny/crossref_metadata_2025_split huggingface dataset:
Case Report on a Female Patient with Congenital Haemophilia a and An Acquired Type-2 Inhibitor to FVIII
Authors: Kurnik Karin , Bidlingmaier Christoph 
Published: April 2020

Abstract
Only rare cases of female haemophilia have been reported. Moreover, no case of congenital haemophilia A associated with an acquired type-2 inhibitor to FVIII in a female child has been described in literature yet. We report the case of a 15-year old girl, diagnosed with severe haemophilia A (FVIII:C &lt;1%) at the age of 13 months. Her father suffered from severe haemophilia A (FVIII:C &lt;1%) due to an intron 22 inversion. In the girl both an intron 22 inversion and a non-random X-inactivation were found. At 3 years the girl received prophylactic treatment, and recombinant FVIII (3 × 40 IU/kg/week) from 6 years of age. Bleeding episodes were rare. Aged 13 she developed two large spontaneous haematomas. In-depth analysis confirmed decreased FVIII-recovery, –half-life and an inhibitor titre of 7.6 BU following type-2 kinetics (haemophilic autoantibodies). All of the few reported children with an aquired FVIII inhibitor presented with other autoimmune diseases or had used penicillin. Our patient showed none of this, but interestingly experienced menarche 4 weeks after inhibitor detection. This lets us speculate that the ethiology of the inhibitor formation in our patient at the time of hormonal change might be similar to that in pregnant or postpartum women.
Taking into account the recommendations of Ma et al. (2006) regarding inhibitor development during pregnancy in healthy women, we initiated a modified ITI with 75 IU/kg rFVIII concentrate (4,000 IU per infusion) every other day without additional immunosuppressive therapy. After 6 weeks of ITI, inhibitor titres declined and became negative after 8 months, resulting in a normal FVIII half-life till now.
Treatment Patterns and Splenectomy Failure Rates in Patients with Chronic Immune Thrombocytopenic Purpura in Canada: A Retrospective Chart Review
Authors: Mikhael Joseph , Tinmouth Alan , Merali Tazmin , Amin Mo , Lam Wendy 
Published: April 2020

Abstract
Background: In clinical practice there is little consensus on treating patients with chronic immune thrombocytopenic purpura (ITP) beyond first line therapy with steroids.
Objectives:
Describe the demographic and disease characteristics of adult ITP patients who receive treatment; Obtain the treatment approach for patients with ITP beyond first line treatment with steroids, with an emphasis on splenectomy; and Evaluate adverse bleeding outcomes associated with ITP.
Methods: A retrospective chart review of ITP patients after initial therapy with steroids was conducted. Ten physicians (oncologists and hematologists) were recruited from across Canada and each physician provided at least 5 ITP cases for review. A total of 51 cases were reviewed and patients (&gt; 18 years old) were required to have had more than 1 course of steroids as treatment to be eligible.
Results: The average age of patients at diagnosis was 42 (range 3 to 82 years); 37 (73%) of the patients were female, and 37 (73%) were Caucasian. The median platelet count upon presentation was 5×109/L. Median lines of therapy after initial therapy was 3 (range 0 to 6). Second line therapies varied, but most commonly patients underwent splenectomy (43%), followed by continued steroid treatment (16%), steroids plus IV Ig (16%), IV Ig alone (14%), immunosuppressant alone (2%), and anti-D plus steroids (2%). Of the patients reviewed, 40 (78%) eventually underwent splenectomy. Of the 40 splenectomized patients, 27 were splenectomized within the first year of diagnosis and 35 underwent splenectomy during the first 5 years. In addition, 62 of the splenectomies were laparoscopic, and the median hospital stay for all procedures was 5 days (range 1 to 60 days). Immediate failure of splenectomy occurred in 18% of patients, and the one- and five- year relapse/failure rates (defined as platelet count &lt; 30×109/L) were 38% and 55%, respectively. Other therapies following splenectomy included IV Ig, azathioprine, cyclosporine, danazol, mycophenolate, rituximab, vincristine, cyclophosphamide, and anti-D. Only 6% of IV Ig use was for chronic maintenance therapy and 74% of use was for urgent therapy. A total of 61 bleeding events occurred, of which 10 were major (upper GI, mouth, rectal, and nose). Major bleeding events required hospitalization of the patient with an average length of stay of 5 days (range 2 to 14 days).
Conclusions: Notwithstanding several limitations, the retrospective chart review suggests that there is wide variation in long-term therapy for patients with chronic ITP in Canada. Splenectomy was the most widely used second line therapy, although 18% of the patients were non-responders and the five-year relapse/failure rate was 55%.
Refractory Severe Thrombocytopenia Associated with Intravenous Treprostinil Use: a Case Report
Authors: Kuo Sherry , Tandon Rajive , Boggio Lisa 
Published: April 2020

Abstract
Background: Intravenous epoprostenol was the first drug approved by the U.S. Food and Drug Administration for the treatment of pulmonary arterial hypertension. Treprostinil, a tricyclic benzidene prostocycline analog of epoprostenol has been favored over epoprostenol due to its more advantageous mode of administration and similar efficacy. All prostocyclines are known to cause thrombocytopenia, but severe platelet reductions are rare below 100,000/μL.
Aim: We report the first case of severe thrombocytopenia associated with intravenous treprostinil with subsequent recovery of a normal platelet count following transition to epoprostenol.
Case Report: A 55 year-old Caucasian woman diagnosed with idiopathic pulmonary artery hypertension noted significant improvement in symptoms and exercise tolerance following initiation of intravenous treprostinil. However, she developed new onset severe thrombocytopenia with her platelet count falling to 51,000/μL (150,000–399,000/μL) and recurrent anterior epistaxis after receiving intravenous treprostinil for six months. She had no known hematologic disorders or history of recurrent bleeding and was not receiving any other new medications. There was no evidence of disseminated intravascular coagulation, hemolysis, coagulation factor deficiency, or splenic sequestration. Empiric treatment with corticosteroids was started for positive antiplatelet IgG direct antibody, but the platelet count further declined to 25,000/μL. The patient was subsequently transitioned from intravenous treprostinil to epoprostenol under the assumption that her severe thrombocytopenia was an idiosyncratic reaction rather than a class effect of prostocyclines. Her platelet count had returned to normal two weeks after the transition.
Conclusion: In practice, patients developing thrombocytopenia from epoprostenol are frequently switched to treprostinil hoping to attenuate the degree of thrombocytopenia. This case is the first report of severe thrombocytopenia associated with intravenous treprostinil, as well as its successful resolution following transition to another prostocycline analog, epoprostenol. Although the exact mechanism of thrombocytopenia is unknown, it raises a question if the cyclohexane ring on treprostinil compared to the lactone ring on epoprostenol can potentially lead to more severe thrombocytopenia. Although at this point, it is uncertain if severe thrombocytopenia is an idiosyncratic reaction to treprostinil, this report will hopefully draw attention to this potential adverse effect and identify the mechanism of this reaction.
Successful Management of Refractory Severe Thrombocytopenia Associated with Antiphospholipid Syndrome Using rituximab
Authors: Jaouni Soad Al 
Published: April 2020

Abstract
Background: Rituximab, a monoclonal antibody against the pan B-cell antigen CD20, that induces a rapid in vivo depletions of normal B lymphocytes. Although this agent was originally developed for use in patients with B-cell-NHL, its use has been successfully extended to many autoimmune disorders. Thrombocytopenia associated with antiphospholipid syndrome can be mild to severe. Severe cases can be associated with significant morbidity and mortality if refractory to the usual therapy for autoimmune thrombocytopenia.
Aim: To report an effective regimen in treating a case of severe bleeding thrombocytopenia associated with antiphospholipid syndrome.
Patient and Methods: A 49 year old female was diagnosed with thrombocytopenia associated with antiphospholipid syndrome and antibodies to double-stranded DNA, controlled over the last 15 years. Unfortunately, over the last two years the patient has had frequent admissions for uncontrolled nose bleed, soft palate petechiae and generalized bruising.
Many treatments have been tried for this case of difficult refractory bleeding with platelets in the range 2.0–6.0 K/uL. These include systemic treatments such as corticosteroids, high pulse therapy of methylprednisolone, Imuran, i.v. immunoglobulin infusion and anti Rh-D intravenous therapy.
We report an adult female patient with severe refractory bleeding thrombocytopenia associated with antiphospholipid syndrome and successful treatment with Rituximab. Her platelets count have been maintained above 200.0K/uL over a 14 months period. Unfortunately, this patient has had side effects of steroid therapy.
In conclusion: Rituximab may be effective treatment on patients with refractory, severe thrombocytopenia associated with antiphospholipid syndrome and significant bleeding complications. Rituximab is a promising alternative option for the eradication of the autoantibodies and restoration of normal hemostasis while avoiding the use of high-dose steroid in refractory bleeding thrombocytopenia.
Assessment of Bone Marrow Mesenchymal Stem Cells Biological Characteristics in Patients with Polycythemia Vera
Authors: Zhang Xiaoyan , Li Jianyong , Lu Hua , Wu Yujie , Xia Jun , Duan Limin , Zhang Yaping 
Published: April 2020

Abstract
Objective: To isolate and culture bone marrow mesenchymal stem cell (MSCs) from polycythemia vera (PV) patients and identify their in-vitro biological characteristics.
Methods: Bone marrow was extracted from 9 patients with PV and 6 normal individuals. MSCs were isolated by density gradient centrifugation. The adherent cells were cultured and passaged. The cultured cells harvested from the third passage were assessed. The morphology of the cells was observed by Wright’s staining and the growth curve of the cells was plotted by cell counting. The ultrastructure of MSCs was observed with electron microscopy. The cell cycle and immunophenotype of the expanded MSCs were detected by flow cytometry (FCM). Different agents were used to induce MSCs to differentiate into osteocyte and adipocyte. Von Kossa staining and oil-red staining were used to examine the differentiation. Conventional cytogenetics (CC) was carried out to detect karyotype. Alleles specific polymerase chain reaction (AS-PCR) was used to detect JAK2V617F mutation. RT-PCR were used to analyze the expression of multiple hematopoietic growth factors at mRNA level for MSCs.
Results: MSCs from patients and normal individuals displayed a fibroblast-like morphology adhering to the culture plate and the doubling time of expand MSC was about 43h. The cells were CD73, CD90 positive, and CD34, CD45, CD133, HLA-DR negative. Under suitable conditions, these MSCs could differentiate into osteocytes and adipocytes. MSCs from patients and normal individuals showed normal karyotype. And JAK2V617F mutation was negative in both MSCs. The same expression pattern was seen in patients and normal individuals that they expressed mRNA of SCF, Flt3-ligand, TPO, LIF, IL-6, IL-11 and did not express G-CSF, GM-CSF and IL-3.
Conclusion: MSCs from patients with PV are similar to that from normal individuals.
Thiotepa in the Conditioning Regimen Decreases Rejection after HLA Identical Allogeneic Marrow Transplantation in Children with Beta Thalassemia Major Aged Less Then 4 Years
Authors: Polchi Paola , Gaziev Javid , Lucarelli Guido , Sodani Pietro , Alfieri Cecilia , De Angelis Gioia , Gallucci Cristiano , Isgro Antonella , Marziali Marco , Roveda Andrea , Paciaroni Katia 
Published: April 2020

Abstract
BMT in children with beta thalassemia major is the only form of definitive cure with correction of the genetic disease in a relevant proportion of patients, and long term thalassemia free survival. Lucarelli and coll. showed since 1986 that transplant performed early in the course of disease gives high chance of cure, and a system of class definition was reported in 1990 identifying as in class 1 patients without any evidence of organ damage, based on hepatomegaly, presence of liver fibrosis and history of regular or irregular iron chelation. Majority of children aged less then four years were then in class 1 and occasionally in class 2. Thiotepa has been introduced in the conditioning regimen of children less then 4 y.o. in the attempt to decrease the rejection rate in this category of good prognosis patients. Out of 519 children affected by not advanced Beta Thalassemia Major, either in class 1 or 2 of risk, with an HLA identical sibling donor, 25 patients aged 1 to 4 yo received an intensified conditioning regimen including thiotepa 10 mg/kg given in one day, in addition to standard BUS-CY regimen called PC 6. This was based on the observation that children aged below 4 transplanted after the standard PC 6 had significantly higher relapse rate with return to thalassemic pre transplant condition (OS, TFS, REJ were 87%; 86%; 4% vs 87%; 73%; 14% in children aged above or below 4 yo respectively (n=398 =&gt;4; n=96 &lt; 4). The increased relapse rate was more relevant in children treated with standard PC 6 and GVHD prophylaxis performed using short MTX plus CSA versus those who received cyclosporine alone. The GVHD prophylaxis protocol including sMTX + CSA +Pred, used after 1999 for all patients in our practice significantly improved OS, lowered GVHD rate, but had a trend toward increased rejection rate. This trend was more evident in children aged less than 4 despite an increased dose of Busulfan (16 mg/kg total) was already in use for younger children. Therefore, since 2003 we introduced Thiotepa for children less then 4 yo, maintaining the GVHD prophylaxis with sMTX and CSA. In the last group of 25 children aged below 4 receiving the intensified protocol, 1 patient rejected the graft and had thalassemia recovery, one died of GVHD and infection, and 23 are alive and cured. Actuarial probability of OS, TFS, Rejection were 96%; 91%, 5%. These results favourably compare to the previous results in this category of patients, confirming that Thiotepa exerts additional immunosuppressive and eradicating action, counteracting the effect of GVHD prophylaxis with sMTX+CSA, that exerts an action favouring rejection.
Optimization of a BCR-ABL-Specific DNA Vaccine against Ph+ ALL Using a Nonviral Vector System and Nanoparticle Delivery
Authors: Rott Yvonne , Arndt Stefanie , Green Jordan , Anderson Daniel , Stripecke Renata , Wittig Burghardt , Schmidt Manuel , Köchling Joachim 
Published: April 2020

Abstract
Although 40–50% of adults and 70–80 % of children with acute lymphoblastic leukemia (ALL) can be cured by poly chemo therapy, the prognosis of patients with Philadelphia chromosome positive (Ph+) ALL remains poor. Therefore, new relapse prevention strategies are needed for patients with Ph+ ALL during remission. We have shown previously, that vaccination of mice with leukemia cell lines modified to express costimulatory molecules and cytokines induce a systemic immunity against the syngeneic BCR-ABLp185 expressing leukemia cell line BM185. However, the difficulties to culture and transfect human leukemia cells limit the clinical application of leukemia cell based vaccines. Thus, we evaluated the immunization of mice with DNA-based vaccines subsequently challenged by the cell line BM185. Combinations of minimalistic immunogenically defined gene expression (MIDGE) vectors encoding a BCR-ABLp185 fusion specific peptide, GM-CSF, IL12, IL27 or CD40L were used for in vivo transfection of murine skin. In addition, we used natural DNA-based double stem-loop immunomodulators (dSLIM), containing three CpG-motifs as non-specific immune adjuvant. In order to increase transfection efficacy, MIDGE-vectors were microencapsulated into poly(β-aminoester) nanoparticles with diameters of 200 nm.
Mice immunized with the BCR-ABL/GM-CSF/dSLIM vaccine showed a significant longer mean tumor-free (p=0.019) and overall survival (p=0.008) compared to nonvaccinated mice. BCR-ABL specific sequences were required to prevent Ph+ acute lymphoblastic leukemia. Furthermore, CTL assays showed that specific lysis was significantly higher after vaccination with BCR-ABL/GM-CSF/dSLIM compared to GMCSF/dSLIM (p&lt;0.05) and to naïve mice (p&lt;0.005). The vaccine efficacy was clearly dosedependent. Microencapsulation of MIDGE vectors increased the efficacy of the vaccine compared to the naked DNA-vaccine. Mice immunized with the microencapsulated vaccine BCR-ABL/GM-CSF/dSLIM showed a significant longer mean tumor-free (p&lt;0.0001) and overall survival (p&lt;0.0001) compared to non-vaccinated mice and 70% survived and never developed leukemia. Cotransfection with IL27 or IL12 lead to significant longer tumor free (IL27: p=0.02; IL12: p&lt;000.1) and overall survival (IL-27: p=0.03; IL12: p&lt;000.1) compared to the vaccine BCR-ABL/GM-CSF/dSLIM. The best protection with a survival rate of 91% was observed in mice which received the vaccine BCR-ABL/GMCSF/IL12/dSLIM. We have shown previously in T-cell depletion studies that CD8+ T cells were the effector cells in the BM185 cell-based vaccine model and currently we evaluate whether CD8+ T cells also play a major role in the BM185 DNA-based vaccine model. In conclusion, we provide survival and functional data that show immunization and protection of mice with optimized leukemia specific DNA-vaccines.
Efficient Gene Transfer into Human CD34+ Peripheral Blood Progenitor Cells Using Pseudotyped Recombinant Adeno-Associated Viral Vectors
Authors: Sellner Leopold , Veldwijk Marlon , Stiefelhagen Marius , Kleinschmidt Jurgen A. , Laufs Stephanie , Topaly Julian , Zeller W. Jens , Wenz Frederik , Fruehauf Stefan 
Published: April 2020

Abstract
Due to their pluripotency, human CD34+ hematopoietic stem cells are targets of interest for the treatment of many acquired and inherited disorders of the hematopoietic system using gene therapeutic approaches. Human CD34+ peripheral blood progenitor cells (PBPC) can be readily mobilized into the blood and harvested by leukapheresis, providing an easily accessible source of hematopoietic progenitor cells. Unfortunately, for gene transfer into CD34+ PBPC, most current vector systems either lack sufficient transduction efficiency or an acceptable safety profile. Standard adeno-associated virus-based vectors have an advantageous safety profile, yet lack the required efficiency. Therefore a panel of pseudotyped recombinant adeno-associated viral (rAAV2/1 - rAAV2/6) vectors expressing the eGFP gene was screened on human G-CSF-mobilized CD34+ PBPC to determine their efficacy. In addition, the difference in transgene expression between conventional single-stranded rAAV and self complementary rAAV (scAAV) vectors was determined. For each vector n≥6 was performed and data are shown as mean ± SD. Of all screened conventional rAAV vectors, rAAV2/6 proved to be the most efficient (13.5% ± 9.8% GFP+ and CD34+ PBPC; p&lt;0.001 vs other vectors) on human CD34+ PBPC, followed by rAAV2/2 (2.6% ± 2.0% GFP+ cells) and rAAV2/1 (1.4% ± 1.2% GFP+ cells). For rAAV2/3, rAAV2/4 and rAAV2/5 no relevant gene transfer efficiency (&lt;1% GFP+ cells) was observed. Furthermore, the relevance of the single-to-double-strand conversion block in transduction of human PBPC could be shown using scAAV vectors. scAAV2/6 and scAAV2/2 (both p&lt;0.001) showed significantly higher gene expression (38.4% ± 12.2% and 11.8% ± 5.7% GFP+ cells, respectively) compared to their conventional counterparts in this cell entity. Similar results were observed for scAAV2/1 vectors (2.8% ± 1.9% GFP+ cells), though the difference was not significant. Of note, as previously observed using AAV peptide library-derived rAAV vectors (Sellner et al., 2008, Exp Hematol. 36), also here inter-patient variances in CD34+ PBPC susceptibility were found. For the first time we were able to obtain clinically relevant gene transfer and expression levels (&gt;10%) with expression rates up to 60% in human CD34+ PBPC using an AAV-based vector system, thereby providing an efficient alternative vector system for gene transfer into this clinically important target cell.
Effects of Proteoglycans on the Maturation of Dendritic Cells Derived from Human Peripheral Blood Monocytes
Authors: Yoshino Hironori , Takahashi Kenji , Kashiwakura Ikuo 
Published: April 2020

Abstract
Dendritic cells (DCs) are a type of antigen-presenting cell which play an essential role in the immune system. The transition from immature DC (iDCs) to mature DCs (mDCs) requires maturation stimuli, such as pro-inflammatory cytokines or pathogen-derived components. Proteoglycans (PGs) are one of main components of the extracellular matrix and are composed of core proteins and glycosaminoglycans that bind to the core proteins. PGs are also constituent elements of bacteria and the role of PGs in the stimulation of DCs has not been elucidated. This study investigated the effects of PGs extracted from the nasal cartilage of a salmon head (S-PG) and the nasal septum cartilage of a whale (W-PG) on the maturation of DCs derived from human peripheral blood monocytes. This study was approved by the Committee of Medical Ethics of Hirosaki University School of Medicine. The human peripheral blood mononuclear cells (PBMCs) were separated from the buffy coat. Furthermore, the monocytes were separated from the PBMCs by allowing them to adhere to a plastic dish. To prepare iDCs, the monocytes were cultured in the presence of 50 ng/ml rhGM-CSF and rhIL-4 for 5 days. The iDCs were stimulated by S-PG or W-PG for 4 days to investigate whether the PGs alone were able to induce the maturation of DCs. In addition, other iDCs were stimulated by a cytokine mixture (rhTNF-α, rhIL-1β, rhIL- 6 and PGE2: MIX) or a combination of MIX+S-PG or W-PG for 48 hours. The surface phenotype of the DCs was analyzed by flow cytometry and the matrix metalloproteinase-9 (MMP-9) activity in the culture supernatants was assayed by zymography. Furthermore, the functions of DCs stimulated by a combination of MIX+S-PG or MIX+W-PG were examined. When the iDCs were stimulated by either S-PG or W-PG alone, the PGs-stimulated DCs did not express the DC-maturation marker CD83, thus indicating that S-PG and W-PG alone could not induce the maturation of DCs. However, the CCR5 expression on DCs stimulated by W-PG was down-regulated. When DCs were stimulated by MIX + 100 μg/ml W-PG, an up-regulation of CCR7 expression was observed. In association with the up-regulation of CCR7 expression, the stimulation by MIX+W-PG actually enhanced the chemotactic responsiveness of DCs to CCR7 ligand MIP-3β. These effects were not observed in the combination of MIX+S-PG. The MMP-9 activity was next examined by zymography, because the degradation of extracellular matrix by MMPs is required for DCs migration. However, neither S-PG nor W-PG promoted MMP-9 secretion. The present study therefore demonstrates that W-PG not but S-PG can selectively regulate the chemotactic activity of DCs in vitro. Further understanding of the mechanism and studies using human PGs is therefore expected to provide valuable insight into the migration of immune cells, including DCs both in vitro and in vivo.