bluuebunny/crossref_metadata_2025_split huggingface dataset:
Establishment of a Murine Model of Allogeneic Umbilical Cord Blood Transplantation
Authors: Wang Yi , Depei Wu , Chen Guanghua , Sun Aining 
Published: April 2020

Abstract
This study was undertaken to establish a murine model of unrelated allogeneic umbilical cord blood transplantation (UCBT). To investigate the potential possibility to reconstitute the immunohematopietic system in adult mice recipients across major histocompatibility complex by mixed 2 near term cord blood transplantation: 1×106 umbilical cord blood mononuclear cells from 20 day old C57BL/6 (H2b) pups and 1×106 umbilical cord blood mononuclear cells from 20 day old C3H (H2k) mixed pups delivered by cesarean were injected into lethally irradiated (8.0Gy) BALB/c (H2d) adult mice in the same time. After one month, anti-H2b and anti-H2k antibody were used to investigate donor chimera in recipients. Results showed that all of the control mice that reconstituted with only 1×106 umbilical cord blood mononuclear cells from 20-day-old C57BL/6 (H2b) pups died with in three weeks after radiation. 70% recipients that reconstituted with mixed umbilical cord blood mononuclear cells survived in the observation period of three months. Two donors mixed chimera was demonstrated by using anti-H2b and anti-H2k antibody in the same recipient. In vitro mixed lymphocyte response showed immune tolerance. The results confirmed that two kinds of umbilical cord blood can form mixed chimera in the same adult mice recipients. Mixed umbilical cord blood transplantation can reconstitute the immunohematopietic system in adult mice recipients without causing serious GVHD. This model may be suitable to address many questions raised in clinical mixed umbilical cord blood transplantation.
Blood Cell Versus Vascular Endothelial Cell Contributions to Nitrite Homeostasis and Blood Pressure Regulation
Authors: Wood Katherine C , Liu Virginia B , Noguchi Audrey , Wang Xunde , Raghavachari Nalini , Gladwin Mark T. 
Published: April 2020

Abstract
Mice genetically deficient in constitutive nitric oxide synthase (eNOS) are hypertensive compared to normal C57Bl6 (wild type) mice, indicating the importance of constitutively produced nitric oxide (NO•) to blood pressure regulation and vascular homeostasis. The objective of this study was to use molecular methodologies to determine the contribution of eNOS in circulating blood cells to the intravascular pool of nitrite, a metabolite and storage form of nitric oxide (NO•) with powerful vasorelaxant activity, and to the regulation of blood pressure under physiological conditions. We used bone marrow transplant to create chimeric mice genetically deficient in eNOS in either circulating blood cells (−/+) or vascular endothelium (+/−), confirmed by flow cytometry, western blot, RT-PCR and immunohistochemistry. Nitrite concentrations in whole blood and plasma of chimeras were quantified using ozone-based reductive chemiluminescence. Mean arterial and diastolic blood pressures of chimeras were assessed in the absence/presence of NOS stimulation with oral L-Arginine or NOS inhibition with oral L-Name. A highly significant inverse correlation between plasma nitrite concentrations and blood pressures was noted across all groups of chimeric mice. Importantly, in agreement with higher blood pressures in −/+ chimeras compared to eNOS positive controls (+/+ chimeras globally competent for eNOS), significantly reduced whole blood and plasma nitrite concentrations were also measured. Blood pressure responses to NOS inhibition or stimulation were intact in all chimera groups except eNOS negative controls (−/− chimeras globally deficient for eNOS) and, importantly, blunted in −/+ chimeras compared to eNOS positive controls. These findings indicate a functional blood cell eNOS that is a major contributor to circulating nitrite concentrations and plays a critical role in vascular homeostasis.
Characterization of NK1.1+ CD8 T Cells in Allogeneic Hematopoietic Cell Transplantation Recipients
Authors: Wang Yi , Wang Hui , Wang Shumei , Sykes Megan , Yang Yong-Guang 
Published: April 2020

Abstract
NKT cells from naïve mice are mainly CD4+ or CD4−CD8−. However, it has been reported that CD8+ NKT cells can be expanded in vitro from splenocytes, bone marrow cells and thymocytes of C57BL/6 (B6) mice by stimulation with anti-CD3 mAb and cytokines, and that the expanded CD8+ NKT cells mediate strong graft-vs.-leukemic (GVL) effects without severe GVHD after adoptive transfer into allogeneic mice. We now describe the presence of CD8+NK1.1+ cells in recipient livers (approximately 2–6%), but not in other tissues (spleen, lung, bone marrow, thymus and PBMC), in various allogeneic hematopoietic cell transplantation (allo-HCT) models. The generation of CD8+NK1.1+ cells is likely a consequence of alloresponses, as these cells were not detected in the liver of syngeneic HCT controls. Flow cytometric analysis confirmed that these cells are CD1d-independent, TCRαβ+ T cells with a memory phenotype (CD44+ and CD62L−), and do not express CD49b, Ly-49C/I, Ly-49G2, or Ly-49D. In a sublethally (6 Gy)-irradiated B6-to-B6D2F1 allo-HCT model, NK1.1+ CD8 T cells became detectable by week 2, increased in number until approximately week 8, and gradually declined thereafter but were still detectable in the liver at day 100 after allo-HCT. By using CD45.1 and CD45.2 congeneic donors, we determined that the majority of NK1.1+ CD8 T cells were derived from the donor splenocytes. Furthermore, depletion of CD8+, but not NK1.1+, cells from the donor splenocytes prior to transplantation prevented the generation of NK1.1+ CD8 T cells, indicating that these cells were derived from donor NK1.1−CD8+ splenic T cells. Our data demonstrate that donor CD8 T cells can acquire NK1.1 expression upon activation in allo-HCT recipients, and that these NK1.1+ CD8 T cells maintain a memory phenotype and persist in the recipients with preferential accumulation in the liver. Studies are currently in progress to determine the role of activated donor NK1.1+ CD8 T cells in GVHD and GVL effects.
Mesenchymal Stem Cell Therapy in a Rat Model for GvHD
Authors: Zinöcker Severin , Wang Mengyu , Nestvold Janne , Vaage John Torgils , Kvalheim Gunnar , Rolstad Bent 
Published: April 2020

Abstract
Graft-vs-host-disease (GvHD) is an important hindrance for the usage of allogeneic bone marrow cell transplantation (alloBMT) as a treatment strategy for leukemias and other hematopoietic malignancies. Mesenchymal stem cells (MSC) can interact with the immune system and inhibit alloreactive cells. Infusions of MSC into GvHD patients have alleviated disease severity and overall survival and in recent clinical trials.
We are studying the immunoregulatory potential of MSC as a putative therapy for GvHD in an experimental animal model. Bone marrow-derived MSC from PVG rats are highly efficient in blocking PVG lymphocytes stimulated by allogeneic BN lymphocytes in mixed lymphocyte reactions. We have found a substantial contribution of indirect (without cell contact) inhibition of allostimulation by MSC in vitro, and have examined some of the possible mediating factors among rat cytokines. In our in vivo model we have transplanted BN host rats with 30×106 PVG donor bone marrow (depleted for T cells), and induced GvHD by injection of 1×106 peripheral T cells from PVG donors two weeks later. We have successfully reversed the lethality of GvHD in a number of diseased animals by pre-emptive and repeated infusions of MSC, improving the overall survival rate by 30%. We are furthermore able to trace injected MSC in vivo, and thereby hope to obtain important information on the migration and behaviour of those cells in the development of GvHD. Additional transplantation experiments to improve the effectiveness of the cellular treatment and test more MSC lines from different MHC-congenic rat strains are underway. Moreover, we will include a rat myelogenic leukemia (BNML) in our GvHD model based on our existing protocols and investigate the long-term outcome of MSC therapy (mortality and disease recurrence) with respect to both graft-versus-leukemia and graft-versus-host immunity.
Significant Expansion of Myeloid Derived Suppressor Cells in Patients with High- Risk Breast Cancer Treated with Dose Dense Adjuvant Chemotherapy
Authors: Talmadge James E , Reed Elizabeth , Cowan Kenneth , Gabrilovich Dmitry , Warkentin Phyllis , Janssen William E. , Wisecarver James , Anderson James , Mailliard Mary , Maas Sarah , Turken Orhan , Coskun Uger 
Published: April 2020

Abstract
Myeloid derived suppressor cells (MDSCs) have been reported to be expanded in cancer patients, following growth factor administration and after chemotherapy. These cells have been associated with a loss of T-cell number and function and provide one mechanism of immune evasion. We examined the effect of dose dense chemotherapy on immune phenotypes and function in patients with breast cancers 4 cms or larger and/or four or more involved nodes. The adjuvant therapy was dose-dense doxorubicin, cyclophosphamide (AC) followed by paclitaxel (P), then 33 doses of radiation (R). Blood samples were obtained and studied prior to therapy, 1 week post AC and 1, 15 and 21 weeks post P and then 3, 6 and 12 months later. Flow cytometric analyses of cellular phenotypes were done on these blood specimens and compared to the levels prior to therapeutic intervention and to normal age and sex matched donors. Twenty-three pts have been followed a median of 29 months (range 5.5–50.5 months) from study entry. Two patients relapsed 8 and 23 months after diagnosis. T-cell CD-4 numbers declined following AC from an average of 4.9±0.5 ×106/ml to 1.7±0.3×106/ml, but increased to an average of 2.7± 0.3 × 106/ml, 21 weeks after P or 12 weeks after R. In this study the MDSCs were defined as Lin- (CD3, CD19, CD14 and CD13), HLA-DR- and CD33+. The numbers of MDSCs, which in normal donors were 0.62±0.16×106/ml and in the cancer patients at diagnosis were 11.8±9.6×106/ml increased to 58.4±25.9×106/ml 15 weeks after R. They remained significantly elevated through one year after diagnosis when they were 27.3±12.3×106/ml. The majority of the MDSCs had a high side scatter and forward scatter by flow analysis suggesting a granulocytic commitment rather than a monocytic commitment. The increase in MDSC numbers was apparently associated with R as the numbers of MDSCs were not significantly increased by AC (15.7±13.5×106/ml) or P (10.9±6×106/ml) one week following completion of each cycle of dose dense therapy. In association with the increase in MDSCs there was a significant decrease in PHA proliferation by the peripherial blood mononuclear cells (MNCs) and suppressive activity by irradiated MNC for allergenic lymphocytes.
Hemophagocytic Lymphohistiocytosis (HLH) in Adults Should Be Treated through Intensive Immune Suppression: 2 Case Reports
Authors: Xie Shuangfeng , Yin Songmei , Ma Liping , Nie Danian , Li Yiqing , Wang Xiuju , Liang Dongmei 
Published: April 2020

Abstract
Background: HLH includes the great majority of patients with macrophage-related disorders. Although HLH is considered as a reactive disease, it is an invariably fatal disease. HLH have 2 types: familial hemophagocytic lymphohistiocytosis and secondary hemophagocytic syndrome. The HLH-2004 protocol made by Histiocytosis Association of America (HAA) was for patients aged &lt;18 years. There was no standard protocol for adults patients with HLH. But the HLH cases was not rare in patients aged &gt;18 years, especially in China. We had 2 HLH cases recently, the 2 patients had different prognosis through different treatment.
Patients and treatment: Case 1, a 20 years old man, he had mild infection 3 weeks ago, and was cured by antibiotics. Then 2 weeks ago, he got persistent fever, lymphadenectasis, hepatosplenomegaly and pancytopenia. Bone marrow smear discovered 3.5% hematophagocyte, no malignant cells. His direct bilirubin (DBIL) increased to 229.9μmol/L, fibrinogen decreased to 0.14g/L, albumin decreased to 24.6g/L in 5 days. Then he was treated with the HLH-2004 protocol (VP-16+dexamathsone+ciclosporin A). After 10 days treatment, his DBIL became 149.7μmol/L, fibrinogen increased to 1.73g/L, albumin increased to 31.2g/L, The pancytopenia was improved too. Now he was still in recover state. Case 2, a 33 years old woman, she was diagnosed enteric T-cell Non- Hodgkin’s Lymphoma 3 weeks ago. She had normal blood cell analysis and liver function at that time. She had colon perforation because of the lymphoma before the diagnosis and the abdominal inflammatory was severe. Also 1 week after her diagnosis, she had HLH. We try to improve her state by supportive treatment. No chemotherapy was used. Soon, severe pancytopenia appears, her fibrinogen decreased to 0.84g/L, albumin decreased to 23.1g/L, alanine transarninase (AST) increased to 201U/L. Her state aggravated very fast and died of hemorrhea and shock. Almost all adult patients with HLH had poor prognosis without the HLH-2004 protocol treatment. HLH-2004 protocol might be a hope for adult patients with HLH. We need to investigate the effects of HLH-2004 protocol through treating more adult patients with HLH. Also, after the diagnosis of HLH, the patient should be treated as soon as possible, because the multi-organ failure could be developed very fast. The chance of treatment might be lost because of multi-organ failure.
Twenty Units of Blood and Counting
Authors: Brownrigg Emma , Carr Debbie , Copeman Michael C. , Creighton Kim , Demasi Catherine , Domanski Julie , Harrison Susan , Hodges Cherry , Kanakis Joanne , Kelleher Sharyn , McCosker-Rees Joanna , Prall Rachel , Richardson Dawn , Ryan Patricia , Swan Craig , Terry Susan , Tovey Julie , Van Kuilenberg Rosita , Wills Melissa , Slader Cassandra 
Published: April 2020

Abstract
Iron overload is a condition seen in patients who have received multiple packed red cell transfusions. Generally, patients who have received &gt;20 transfused units will be at risk. Currently in Australia there is no universal method of tracking the number of transfusions a patient has received, and a cumulative figure requires manual calculation. Therefore, identification of at-risk patients is not straightforward.
Aim: Firstly, to review the primary diagnosis of transfused patients in haematology day units. Secondly, to quantify number of transfusions received and serum ferritin levels of transfused patients. Thirdly, to review the use of iron chelation in these patients and document potential reasons for non-chelation including co-morbidities and concomitant medications.
Method: Medical records for outpatients transfused during the 12-week index period (12 consecutive weeks from Human Research and Ethics Commitee/institution approval) were reviewed in this retrospective multicentre audit. Data were captured using a piloted data collection form. Non-parametric statistical test have been used.
Results: To date, 237 patients, aged 0–95 have been reviewed from 10/20 centres. Common underlying conditions necessitating transfusion were: MDS(35%), chemotherapy-induced anaemia(21%) and thalassaemia(14%). The medical record of 26% patients indicated that transfusions had also been given elsewhere. In total, 119 had received &gt;20units, 58(49%) had been prescribed chelation therapy. Fourteen patients who had received &gt;20units had no documented serum ferritin. Patients receiving iron chelation were younger (median 42 cf.73 years, p=0.0004), had received more transfusions(median 60 cf.18, p&lt;0.0001) and more units(median 187 cf.37, p&lt;0.0001). There was no difference in number of concurrent medical conditions(p=0.73) or concomitant medications(p=0.14). Life expectancy was documented for 6 patients only, 3 were chelated.
Conclusion: There is variability in the monitoring and treatment of iron overload in Australian hospitals, with not all at-risk patients receiving chelation. The development and implementation of universal tracking tools for transfusion nurses and patients may be one means of improving identification of at-risk patients.
Characterization and Development of Genomic DNA Quality Controls for Thrombophilia and Warfarin Sensitivity Testing
Authors: Anekella Bharathi , Wu Jainlei , Huang Catherine , Dryga Sergy , Smith Tamara , Howell Renee , Manak Mark 
Published: April 2020

Abstract
Background: Genetic tests for diagnosis of clotting disorders and monitoring their treatment are based on identification of single nucleotide polymorphisms (SNPs) that are related respectively to clotting and to drug metabolism crucial to the effectiveness of therapy. Thrombophilia, an inherited condition which predisposes to thromboembolism, is due in part to genetic factors, such as the presence of SNPs in the genes encoding clotting factors. Hyperhomocysteinemia, which poses an extremely elevated risk of thromboembolism to the patient, can be caused by certain mutations to the MTHFR (677CT, 1298AC) gene; genotyping all genes involved in thrombophilia concurrently can offer a more complete picture of the genetic component of thrombophilic risk. Warfarin is the most frequently used oral anticoagulant for treatment and prevention of thromboembolism and requires careful adjustment of therapeutic dose and close monitoring because of its narrow therapeutic index and wide individual variability in response and therapeutic dose. Genetic variations in two genes, CYP2C9 and vitamin K epoxide reductase (VKORC1), have been shown to have a profound effect on warfarin sensitivity. Using genetic tests for identification of these variants clinicians to better estimate the appropriate starting dose for therapy, which may improve drug safety. Currently a number of test systems detect these SNPs. Use of a well-characterized independent quality control sample is important to monitor the accuracy and reproducibility of these tests. We have developed multi-analyte quality controls that contain major SNP variants for both thrombophilia MTHFR and warfarin sensitivity.
Methods: PBMCs from multiple donors were screened for mutations for MTHFR for Thrombophilia and for CYP2C9 and VKORC1 for warfarin sensitivity. Mutations were identified using the Third Wave Technologies INVADER® and Autogenomics INFINITI™ platforms; and ABI TaqMan® SNP assay. All mutations were confirmed by sequencing on an ABI 3130 instrument.
Results: PBMC samples were screened for CYP2C9, VKORC1, and MTHFR mutations. The mutation frequency for CYP2C9*2, CYP2C9*3 and VKORC1 1639GA variants was 28%, 7% and 63% respectively. 100% genotyping concordance was observed in all testing. In addition, heterozygotes for additional VKORC1 SNPs 5808TG, 6484CT, 6853GC, 7566CT, and 9041GA were present in one sample bearing VKORC -1639GA. The mutation frequency for MTHFR 677CT and 1298AC was 17% wt and 29% het for both SNPs. The identification of heterozygotes in several SNPs allowed development of multi-analyte controls for both genes. Data showing reproducibility and accuracy in SNP call rate on various test platforms will be presented.
Conclusion: We have developed multi-analyte controls for thrombophilia and warfarin sensitivity that can monitor extraction, amplification and detection, and evaluated their performance with several test methods.
ENACCTing WELLNESS- a Community Centered Intervention to Enhance Treatment Option Decision- Making among Newly Diagnosed and Recurrent Hematologic Cancer Patients
Authors: Michaels Margo , Golant Mitch , Belkora Jeff , Miller Melissa F , Blakeney Natasha 
Published: April 2020

Abstract
Background: Less than three percent of adult cancer patients participate in cancer clinical trials. This rate is even lower among people of color and the medically underserved who tend to have higher cancer mortality rates than the population as a whole. With timely diagnosis and enhanced access to clinical trials, more patients with blood cancers will have expanded options for treatment and the opportunity to receive quality cancer care.
Methodology: ENACCTing WELLNESS is a pilot project with the Education Network to Advance Cancer Clinical Trials (ENACCT) and The Wellness Community (TWC), which seeks to demonstrate how community-centered education, outreach and support services can both address informational needs and enhance access to care among those living with blood cancers in four communities nationwide. Using an innovative ecological framework directing educational efforts at the intrapersonal, interpersonal, institutional and community perspectives, ENACCTing WELLNESS:
Provides one-on-one treatment decision support with a trained facilitator, using an evidence-based decision-making model, SCOPED (Situation-Choices-Objectives-People-Evaluation-Decisions), to prompt newly diagnosed/newly recurred patients to list questions about their diagnosis and treatment, including the option of receiving their treatment though a clinical trial; Increases awareness of support services for patients, their family members, friends, and caregivers; Educates primary care providers, especially those caring for the medically underserved, about options to refer patients to local clinical trials for first line treatment; and Enhances cultural competency skills of local clinical trial investigators and their teams.
Results: This session will discuss the use of the SCOPED model in the conceptual framework of ENACCTing WELLNESS, and the collaborative effort between two community-centered organizations to enhance patient treatment decision-making options through provider training, referral, and one-on-one patient counseling. We will report lessons learned from phase I of the pilot study and preliminary evaluation data from health care provider training.
Implications: The framework implemented in this pilot may result in increased and timely primary care referral of patients with a suspected hematologic cancer and may assure decision quality as measured by high levels of decision self-efficacy and low levels of decision regret among patients contemplating treatment choices.