bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparable Survival and Transplant-Related Mortality Following Allogeneic Hematopoietic Cell Transplantation from HLA Allele-Matched Unrelated and HLA-Identical Sibling Donors.
Authors: Cao Thai M. , Tsai Schickwann , Kelley Linda , Alder Stephen C. , Fuller Thomas C. , Pulsipher Michael A. 
Published: October 2019

Abstract
Comprehensive analyses of unrelated donor (URD) and recipient HLA-matching for allogeneic hematopoietic cell transplantation (AHCT) have demonstrated better outcomes when allele typing is performed using high-resolution nucleotide sequence-based techniques. To evaluate survival following myeloablative AHCT using allele-level HLA-matched URD as compared with HLA-identical sibling donors, we analyzed outcomes for 430 patients treated at our center between March 1991 and April 2005. Sequence-based allele typing was retrospectively performed for HLA-A, B, C, DR and DQ when not done at time of AHCT for URD (n = 124; 29%) and non-sibling related donors (n = 19; 4%). Donors were HLA-identical siblings (n = 276; 64%), HLA allele-matched URD (n = 52; 12%), single HLA-locus mismatched donors (n = 52; 12%), or > 1 locus mismatched donors (n = 50; 12%). The median age at transplant was 23.4 years (range: 0.2 – 61). The most common diagnoses were AML (n = 107; 25%), CML (n = 90; 21%), ALL (n = 86; 20%) and MDS (n = 50; 12%). Total body irradiation-based preparative regimens were used for 283 patients (66%). Bone marrow (BM) was the graft for 388 patients (90%) and GCSF-mobilized peripheral blood stem cells (PBSC) for the remaining 42 (10%). Graft-versus-host disease (GVHD) prophylaxes were cyclosporine and methotrexate (n = 327; 76%), long methotrexate (n = 42; 10%), T-cell depletion (n = 19; 4%), or other regimens (n = 42; 10%). With a median follow-up of 4.8 years (range: 0.2 – 12.1), the 5-year estimate of overall survival (OS) for the entire group was 48.2% (95% CI: 45.7 – 50.7) and transplant-related mortality (TRM) was 31.4% (95% CI: 28.8 – 34). As shown in the Table, OS and TRM were indistinguishable between AHCT performed with HLA-identical siblings compared with HLA allele-matched URD. There was also no difference in grade III – IV acute GVHD (P = .46) between these two groups whereas there was a trend towards more extensive chronic GVHD (HR 1.8; 95% CI: 0.9 – 3.6; P = 0.12) for the URD recipients. Using a multivariate analysis to adjust for advanced disease, age (> vs ≤ 30 years), graft (BM vs PBSC) and female-to-male gender mismatch, there remained no difference in OS between HLA-identical siblings and HLA allele-matched URD (P = 0.67). These results demonstrate that key outcomes (OS, TRM, and severe acute GVHD) are equivalent in recipients of grafts from either allele-level 10/10 HLA-matched URD or HLA-identical siblings.
Overall Survival TRM Number Hazard Ratio 95% CI P value Hazard Ratio95% CI P value HLA-ID Sibling 276 1 - - 1 - - HLA-ID URD 52 1.1 0.7 – 1.7 0.67 0.8 0.4 – 1.6 0.58 1 Locus MM 52 1.3 0.9 – 2.0 0.19 1.4 0.8 – 2.4 0.25 > 1 Locus MM 50 2.0 1.4 – 2.9 < 0.001 2.6 1.7 – 4.1 < 0.001
HLA Mismatched/Haploidentical Haematopoietic Stem Cell Transplantation for the Treatment of Chronic Myelogenous Leukemia.
Authors: Huang Xiao Jun , Xu Lan-ping , Liu Kai-yan , Liu Dai-hong , Chen Huan , Han Wei , Chen Yu-hong , Wang Jing-zhi , Chen Yao , Zhang Xiao-hui , Shi Hong-xia , Lu Dao-pei 
Published: October 2019

Abstract
Backgroud: Allogeneic marrow transplantation remains the only proven curative therapy for CML, but many patients have no suitably HLA matched related donor. Recently, we developed a new method for HLA-mismatched /haploidentical transplantation without in vitro T cell depletion. Conditioning including antithymocyte globulin followed by un-manipulated HLA-mismatched/haploidentical blood and marrow achieved comparable outcomes to HLA identical sibling transplantation. So good outcomes of HSCT from mismatched family donor for CML patients could be expected.
Methods: 81 consecutive patients received mismatche family donor HSCT for CML were analyzed. There are 40 cases in CP1, 10 cases in CP2 after BC or AP, 14 cases in AP and 17 cases in BC pre-HSCT. Patients received the modified BuCy2 regimen consisting of cytarabine, busulfan, cyclophosphamide, Me-CCNU and ATG. Both of donor G-BM (G-CSF immobilize ) and G-PB were harvested and infused to recipients. All patients were given CsA/MTX and MMF for prophylaxis of GVHD.
Results All patients achieved full donor chimerism. The cumulative incidence of aGVHD is 62.71% (CI 57.3– %−68.2%), grade III–IV is 24.69% (CI 28.7 %−30.7 %). 2 yrs cumulative incidence of cGVHD was 56.48 % (CI 49.6 %–−53.4%). 2 years OS is 71.33%(CI 66%–−76.7%), 2-year LFS were 61.23% (CI 54.66%–−67.8 %). The cumulative incidence of relapse is 17.74% (CI 11.3%–−14.2 %). The incidence of relapse in CML-CP1, CP2, AP and BC group pre-HSCT were 10.8% (CI 2.67%–18.95%), 0%, 9.2% (CI 0.5%–− 17.9%) and 42.71% (28.47%–56.95%%), with P=0.0150. The OS in CML-CP1, CP2, AP and BC group pre-HSCT were 67.66% (59.93%–75.39%), 83.33% (68.12%±098.54%), 77.38% (65.88%±88.90%), 69.68 % (58.32%±81.04 ) with p=0.7876.
Conclusion For patients with CML without an HLA identical sibling donor, haploidentical family members can be an alternative donor for HSCT. In our HSCT protocol, survival of HSCT for CML in advanced stage was no worse than that in stable stage. The optimal time of HSCT for CML patients from MM-family donors need to be further studied.
Assessment of the Immune Reconstitution by Cylex Immuknow™ Assay in Patients after Allogeneic Stem Cell Transplantation.
Authors: Gesundheit Benjamin , Azar Yehudit , Bringer Rachel , Steinberg-Prop Schivti , Israeli Moshe , Shapira Michael Y. , Resnick Igor , Samuel Simcha , Slavin Shimon , Or Reuven 
Published: October 2019

Abstract
After engraftment of allogeneic stem cell transplantation (allo-SCT) patients are severely immuno-suppressed and are susceptible for recurrent opportunistic infections on one hand; on the other hand, particularly without immunosuppressive treatment over-activity of the graft can lead to uncontrolled acute or chronic graft-versus-host disease (GvHD); optimal immunological activity of the graft is necessary to control the underlying malignant disease and to prevent relapse. Therefore, accurate immune monitoring is crucial for the assessment and appropriate management of the immune-compromised patients after allo-SCT. Since October 2005 up to 6 consecutive blood samples were collected from 51 patients undergoing allo-SCT in Hadassah Medical Center following engraftment. Immune function was analyzed by Cylex Immuknow™ assay, an FDA approved immune cell function test for the assessment of cell-mediated immunity. ATP activity from magnetically separated and lysed CD4 lymphocytes is measured by light intensity, thereby reflecting the current immune function of the patient. From other clinical applications of this test, the following stratification of the immune response was established according to the following levels of ATP activity: Low (0–225 ng/mL), moderate (225–525 ng/mL) and high activity (>525 ng/mL) of immune response. Our preliminary results from allo-SCT patients are in keeping with their clinical course and the known stratification of the ATP level:The gradual increase to moderate ATP levels over several months after allo-SCT are in keeping with normal immune reconstitution with an uneventful post transplant clinical course;high ATP levels were observed in patients prior to the clinical presentation of acute GVHD;low ATP levels were associated with recurrent infections and relapse.
Remarkably, there was no correlation with the recovery of the white cell count after allo-SCT. Our results with consecutive tests are the first study of Cylex Immuknow™ assay in patients after allo-SCT and are in keeping with other currently known clinical applications of this assay for other indications. Our preliminary observations indicate a promising contribution of Cylex Immuknow™ assay as a simple and fast monitoring technique for patients undergoing allo-SCT, which might also predict clinical complications. Our future follow-up studies aim to confirm these preliminary results in order to gain confidence in the assay’s clinical contribution as a possible standard test for patients after allo-SCT.
A Comparative Analysis of the Leukaemic Potential of Mature T Cells Versus T Cell Precursors.
Authors: Newrzela Sebastian , Baum Christopher , Li Zhixiong , Hansmann Martin-Leo , Hartmann Sylvia , Hartmann Marianne , Cornils Kerstin , Fehse Boris , von Laer Dorothee M. 
Published: October 2019

Abstract
After the report of two cases of leukaemia caused by insertional mutagenesis of a retroviral vector in children with SCID, it became clear that safety issues of therapeutic gene transfer must be addressed more thoroughly. We analysed whether gene transfer into mature T cells and haematopoietic stem cells bear the same risk of generating T cell leukaemia through activation of specific T cell oncogenes, such as LMO2, TCL1 and ΔTrkA. To address this issue, we used the Rag-1 mouse model, which allows long term analysis of transplanted T cells and haematopoietic stem cells. We were able to transduce mature T cells and haematopoietic stem cells of C57BL/6 (Ly5.1) donor mice with oncoretroviral vectors expressing LMO2, TCL1 and ΔTrkA. Transduction efficacies of up to 70% were achieved for mature T cells and approximately 90% for haematopoietic stem cells. After transplantation into Rag-1-deficient recipients, stem cell transplanted animals developed T cell lymphomas/leukemia for all investigated oncogenes after characteristic incubation times, mostly of a CD8+CD4+ double positive phenotype. T cell lymphomas were characterised by gross thymic mass, splenomegaly and heavily enlarged lymph nodes, although none of the control- vector- transduced mice developed lymphoma/leukaemia. LM PCR analysis revealed mono- or oligoclonality of the tumours. T cell transplanted animals showed no signs of leukaemia development so far. However, after several attempts, one immortalized T cell progenitor clone could be generated after transduction with LMO2. Our results so far indicate that mature T cells are less susceptible to transformation by known T cell proto-oncogenes, but the studies are still ongoing.
Intravenous Immunoglobulin (IVIG) Diminishes Immune-Mediated Clearance of Platelets Expressing an Integrin αIIbβ3 Transgene Product That Restores Hemostasis in a Canine Model for Glanzmann Thrombasthenia.
Authors: Fang Juan , Jensen Eric S. , Du Lily M. , Boudreaux Mary K. , Wilcox David A. 
Published: October 2019

Abstract
While individual inherited platelet disorders are rare, in aggregate, these diseases can account for a bleeding diathesis in ~1:20,000 individuals. One classic disease is Glanzmann thrombasthenia (GT), a rare autosomal-recessive bleeding disorder that is characterized by genetic defects of the platelet-specific integrin, αIIbβ3. Molecular abnormalities in either the αIIb- or β3-subunit can disrupt receptor synthesis, assembly, and/or function thereby preventing platelets from binding the receptors major adhesive ligands (von Willebrand factor and fibrinogen) to form platelet aggregates as a primary response to vascular injury. As a model to develop methods for gene therapy of platelet defects, GT is especially challenging because platelet integrin level as well as genetic transfer efficiency play a crucial role in restoring normal hemostasis. Developing effective treatment for animal models for this disease should ensure that a similar strategy would be effective for the treatment of other inherited platelet disorders. In experiments to be presented, SCF/G-CSF mobilized CD34+ peripheral blood cells (PBC) from integrin αIIb-deficient dogs were transduced with a lentivirus vector encoding the human αIIb integrin subunit driven by the tissue-specific αIIb promoter. GT dogs received an autologous transplant of lentivirus-transduced PBC using low dose (100–300 cGy) irradiation as transplant preconditioning and three scheduled doses of post-transplant in vivo drug treatment to enrich for transduced hematopoietic stem cells. Fluorescent immunocytometric analysis detected a moderate level (10%) of circulating blood platelets expressing αIIbβ3 following the initial transplant, which increased to 20–30% of normal levels following drug selection with O6BG and BCNU. Remarkably, we observed a significant improvement in a clinical sign of the GT phenotype with reduced bruising on the animals’ legs when αIIbβ3 was restored to the surface of only a fraction of circulating platelets. Further analysis showed restored ability of lentivirus-transduced platelets to aggregate and retract a fibrin clot in vitro using peripheral blood collected from these animals. Treatment of two animals with intravenous immunoglobulin effectively diminished platelet clearance due to an antibody response to newly expressed αIIbβ3 receptor. Reduced bleeding times were measured indicating the feasibility for targeting platelets with genetic therapies for better management of patients with inherited bleeding disorders. These studies should help us to define molecular and cellular events necessary for targeting therapeutics to platelets, which will contribute to eventual, safe, effective, and technologically viable strategies for correcting inherited bleeding disorders in humans.
In Utero Transplantation of Transduced Hematopoietic Stem Cells Results in Deletion of Transgene-Specific T Cells.
Authors: Laje Pablo , Peranteau William H. , Endo Masayuki , Zoltick Philip W. , Flake Alan W. 
Published: October 2019

Abstract
The developing fetal immune system provides a unique opportunity to manipulate normal immunologic development for therapeutic prenatal and anticipated postnatal interventions. In previous studies we have shown that allogeneic in utero hematopoietic cell transplantation (IUHCT) results in donor specific tolerance that can subsequently facilitate non-myeloablative postnatal cellular or organ transplants. It follows that in utero injection of transduced hematopoietic stem cells (HSC) could potentially induce tolerance to a transgene encoded protein. We hypothesized that expression of a transduced antigenic protein by HSC and their progeny would alter thymic T cell development resulting in deletion of antigen specific T-cells. To test this hypothesis, we used the mammary tumor virus (MTV) superantigen system to evaluate the effect of IUHCT of transduced HSC on T cell development. In this system, expression of different MTV oncogenes by different I-E+ strains of mice results in deletion of T cells expressing the relevant Vβ T cell receptor. Specifically, mice which are Mtv7+ delete T cells expressing the Vβ6 T-cell receptor. In this study, CD150+CD48− enriched Balb/c (I-E+ Mtv7−) HSC were transduced with an HIV-based lentivirus expressing MTV7 under an MND promoter. 1.5E+05 transduced cells were injected intravascularly via the vitelline vein into E14 Balb/c fetuses. Non-injected age matched naive Balb/c mice served as the control group. The peripheral blood (PB) and thymuses of injected fetuses and control mice were harvested at day of life (DOL) 10, 20 and 60 and analyzed by flow cytometry for T lymphocyte Vβ6 expression. Additionally, the T cell composition of the thymus was assessed at DOL10 for CD4 and CD8 single positive (SP) and CD4/CD8 double positive (DP) cells. Thymic flow cytometric analysis at DOL10 revealed that greater than 98% of the T cells were CD4CD8 DP, a stage that has not yet undergone negative selection. No significant difference was noted in the percentage of thymic Vβ6+ DP T-cells at this time point or at DOL20 and DOL60. In contrast, there was a significant decrease in the percentage of Vβ6+ peripheral blood SP cells in those mice injected with MTV7 transduced HSC relative to control mice at DOL10, DOL20 and DOL60 (p<0.05) (Fig 1). The current study supports the ability of enriched transduced HSC to induce deletion of transgene specific T cells after IUHCT. In the future, this strategy may be useful to promote tolerance for pre or postnatal cellular or gene therapy.
Figure Figure
Disparity in Diagnosis of Pulmonary Embolism in Hospitalized Patients with Pneumonia.
Authors: Kapoor Avina , Dang Qianyu , Ragni Margaret V. 
Published: October 2019

Abstract
Pulmonary embolism (PE) is a major cause of preventable hospital deaths. Little is known about PE in the setting of pneumonia, as symptoms and signs of PE and pneumonia overlap, confounding diagnosis. With infrequent thromboprophylaxis, PE continues to be a major public health problem. In order to determine the incidence and characteristics of PE in patients with pneumonia, we analyzed statewide discharge data, 1997–2001, collected by the Pennsylvania Health Care Cost Containment Council (PHC4). The analysis included all PE discharges with ICD-9 codes 415.11 and 415.19, excluding pregnant patients and all second or later PE admissions. Infectious disease diagnoses were enumerated by ICD-9 codes, including 030–041, 042, 790.7, 996–998; pneumonia by ICD-9 codes 480–483. 485–487, and 996–998; and diagnostic procedures by revenue code 352 for computerized tomographic (CT), including spiral CT scans. Severity of illness was determined by MediQual Altas Admission Severity Categories. Of 33,036 total PE discharges (all-PE) from Pennsylvania hospital discharges 1997–2001, 5,757 (16.8%) had infection, more than half of which, 3,543 (52.3%), were due to pneumonia (pneumonia-PE). There was a marginal increase in the incidence of pneumonia-PE during this period, 615 (10.3%) in 1997 to 839 (10.5%), p>0.05. Those with pneumonia-PE were typically Caucasian, 77.3%, p<0.01; female, 51.5%, p<0.01; and elderly, 53.1% ≥ 70 years of age, p<0.01, mean age, 66.4±17.3 yr. The proportion of patients undergoing CT scans increased by 15.3%, from 18.9% in 1997 to 21.8% in 2001, p<0.01, an increase 20% lower than than the increase in the all-PE group, 88.9%, from 10.8% in 1997 to 20.4% in 2001, p<0.01. There was a significantly smaller improvement in the severity of illness scores in the pneumonia-PE group, increasing (46.0% to 53.0%) for the three lowest Altas categories and decreasing (46.8% to 42.0%) for the two highest categories, 1997–2001, respectively, compared with the all-PE group, (56.9% to 68.7%) in the three lowest and (35.3% to 26.5%) for the two highest categories, both p<0.01. During this period, mortality rates decreased significantly less in the pneumonia-PE group, by 18.5%, from (21.1% to 17.2%) as compared to a 20% decrease in all-PE, from (11.5% to 9.2%), 1997–2001, p<0.01. The apparent disparity in CT scan frequency and associated lesser improvements in severity of illness and mortality in the pneumonia-PE group, as compared with the all-PE group, suggest early PE diagnosis may have been missed. Modeling the rate of increase in PE diagnosis per increase in CT scan in the all-PE group, we estimate 454 missed cases of PE, which is 12.8% more PE cases among those with pneumonia. We conclude that the lack of increase in CT scans among hospitalized patients with pneumonia is associated with lower rates of PE diagnoses and poorer outcomes. Thromboprophylaxis should be considered in hospitalized pneumonia patients, particularly those at risk for PE.
Figure Figure
Purification of a Single Peptide with Vitamin K Epoxide to Vitamin K and Vitamin K to Vitamin KH2 Activity.
Authors: Chu Pei-hsuan , Huang Teng-yi , Williams Jason , Stafford Darrel W. 
Published: October 2019

Abstract
More than 21 million prescriptions for warfarin are written yearly in the US. Yet, in spite of its importance, vitamin K epoxide reductase (VKOR), the target of warfarin, has resisted purification since its identification in 1972. We report the first successful purification and reconstitution of activity of a recombinant human vitamin K epoxide reductase. A series of detergents were screened to determine that best for solubilization of VKOR from microsomes. Detergents tested that were effective in solubilization of VKOR also led to loss of measurable activity. This loss of activity supports our previous prediction that VKOR is embedded in and requires a membrane environment for enzymatic activity. The short-chain phospholipid, DHPC (1,2-Dihexanoyl-sn-Glycero-3-Phosphocholine) was the detergent of choice to efficiently extract VKOR from the microsomes, even though this reagent completely inhibited enzyme activity. Partial reconstitution was achieved on-column by washing with 0.4 % dioleoylphosphatidylcholine/0.4% deoxycholate. Complete recovery of activity was achieved by removing the deoxycholate through dialysis in the presence of the reducing reagent, THP (Tris(hydroxypropyl)phosphine). During dialysis, the solution became cloudy indicating the formation of membrane-like structure. Purified recombinant VKOR is ~21 kDa (~18.5 kDa + tag); fully active; and over 93% pure. The concentration of warfarin for 50% inhibition is the same for purified protein and microsomes. It has been reported and assumed that VKOR is a multi-subunit enzyme. Our results, however, suggest that a single peptide can accomplish the reaction. The trace amounts of contaminating proteins were identified by mass spectrometry; however, none are apparently relevant to the VKOR reaction. Moreover, the turn-over number of purified VKOR (0.25 sec-1 is approximately two-fold higher than microsomes and about 10 fold higher than the turnover number of gamma-glutamyl carboxylase for CO2 addition. In addition to the vitamin K epoxide to vitamin K reaction, our results also indicate that VKOR can efficiently convert vitamin K to vitamin K epoxide. Our results suggest that ancillary proteins (other than a thioredoxin-like enzyme) are not necessary for full VKOR activity. This purification will allow further characterization of VKOR in relation to other components of the vitamin K cycle and should facilitate its structural determination.