bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mitochondrial DNA Microsatellites as New Markers for the Quantitative Determination of Hematopoietic Chimerism after Allogeneic Stem Cell Transplantation.
Authors: Shin Myung-Geun , Kim Hyeoung-Joon , Kim Hye-Ran , Lee Il-Kwon , Lee Je-Jung , Kook Hoon , Hwang Tai-Ju , Cho Duck , Kee Seung-Jung , Shin Jong-Hee , Suh Soon-Pal , Ryang Dong-Wook 
Published: April 2020

Abstract
Variable number tandem repeat (VNTR) and short tandem repeat (STR) markers of nuclear DNA are increasingly used for monitoring the engraftment of donor cells after stem cell transplantation (SCT). Mitochondria are the only organelles of animal cells other than the nucleus that contain DNA as well as their own machinery for RNA and protein synthesis. Mitochondrial DNA (mtDNA) is present in multiple copies (usually 103 to 104 copies per cell) (Blood2004;103:4466–77). We reported new markers using the analysis of mtDNA control region and microsatellites (mtMS) for monitoring donor cell engraftment in marrow transplant recipients. MtDNA control regions (nucleotide position (np) 16024 to 16569 and 1 to 576) and six mtMS (np 303–315 poly C, np 16184–16193 poly C, CA repeat starting at np 514, np 3566–3572 poly C, np 12385–12391 poly C and np 12418–12426 poly A) from total DNA samples from 25 sets of cases (donor, recipient and after SCT) were amplified using designated specific primers and PCR. We directly sequenced the control region which includes hypervariable regions, then carried out a qualitative and quantitative profiling mtDNA length heteroplasmies of six mtMS using size-based PCR product separation by capillary electrophoresis (ABI 3100 Genetic Analyzer and ABI Prism Genotyper version 3.1). The results were compared to those from six VNTR markers (D12S391, D1S80, F13A1, HUM RENA-4, HUM FABP2 and D18S51–5). MtDNA control region polymorphisms lead to the identification of an informative marker in 88% (22 cases) of all cases. Among six mtMS markers, the result of informativeness from np 303–315 and np 16184–16193 poly C mtMSs was 63% and 67% respectively. The combination of mtDNA control region direct sequencing, np 303–315 and np 16184–16193 poly C length heteroplasmies completely distinguished donor cells from recipients. In data from a typical mixing experiment for the determination of sensitivity, the detection limit (DL) of gene scan analysis in a mtDNA mixture was visible at 1% heteroplasmy in np 303–315 poly C mtMS marker; however DL from D12S391 in the same mixing experiment was 5–10% heteroplasmy. MtDNA control region sequencing and the length heteroplasmies of np 303–315 and np 16184–16193 poly C tracts can provide sensitive, accurate and quantitative determination of mixed chimerism after SCT.
Allogeneic Hematopoietic Stem Cell Transplantation after Combination Therapy with Imatinib and Intensive Chemotherapy for Newly Diagnosed Philadelphia Chromosome Positive (Ph+) Acute Lymphoblastic Leukemia.
Authors: Usui Noriko , Yano Shingo , Takahara Shinobu , Asai Osamu , Dobashi Nobuaki , Osawa Hiroshi , Takei Yutaka , Ogasawara Yoji , Otubo Hiroko , Yamaguchi Yuko , Saito Takeshi , Minami Jiro , Kaito Ken , Kobayashi Masayuki 
Published: April 2020

Abstract
[Background] In JALSG Ph + ALL protocol(IDEAMOP) study, combination therapy with conventional intensive chemotherapy with imatinib (600mg/day) provided high quality of hematologic and cytogenetic (94%) and molecular (77%) complete remission (CR) for the patients with newly diagnosed Ph + ALL(Blood2004;104:748a #2736). However, a few of patients attained molecular CR relapsed subsequently, and allogeneic hematopoietic stem cell transplantation (allo-HSCT) should be considered for younger patients as possible as they can. In order to investigate a role of allo-HSCT post IDEAMOP therapy, three patients treated in our institute were analyzed efficacy and toxicity of allo-HSCT.
[Patients and Methods] Those three newly diagnosed Ph + ALL patients (M/F:2/1 Age:30,31,38 y.o.) were treated with IDEAMOP between Jan 28,2003 and Feb 13,2004. All patients attained CR after single course of induction therapy and median time of CR was 21 (19–27) days. During consolidation therapies, two of them found HLA-matched unrelated donors and one patient chose to receive cord blood transplantation. As conditioning regimen for allo-HSCT, all of them received total body irradiation, high-dose cyclophosphamide and high-dose etoposide before allo-SCT.
[Results] Allo-HSCT was undergoing during their first CR(CR1). Regimen related toxicity and complications of the transplantation were well tolerated. The reconstitutions of bone marrow function were similar to other patients with Ph-negative ALL treated by allo-HSCT. Although two patients presented with either limited or extensive chronic GvHD, they were clinically controlled with or without steroid. Minimal residual disease (MRD), measured by RQ-PCR assay, was detected in two of three patients before allo-HSCT. However, MRD was not detected in any of the patients after allo-HSCT. All of three patients remain in CR at a median follow-up time of 25 (18–31) months.
Allo-HSCT post JALSG-Ph+ALL (IDEAMOP)protocol Patient Status at HSCT MRD pre HSCT Cell Source MRD postHSCT cGvHD Outcome OS(M) 30Y M CR1 yes Cord Blood no limited alive 31+ 31Y M CR1 no MUD no extensive alive 25+ 38Y F CR1 yes MUD no extensive alive 18+
[Conclusion] Although the number of patients are small, imatinib (600mg/day) with intensive chemotherapy as remission induction for newly diagnosed Ph + ALL may have benefit for rapid control of disease, and may provide better outcome of allo-HSCT without additional severe toxicities.
Population Pharmacokinetic of Oral Busulfan in Japanese Undergoing Hematopoietic Stem Cell Transplantation.
Authors: Sasaki Noriaki , Ogata Kentaro , Yamauchi Keita , Takamatsu Yasushi , Suzumiya Junji , Tamura Kazuo , Yukawa Eiji , Hara Shuuji , Ono Nobufumi 
Published: April 2020

Abstract
We previous evaluated busulfan (BU) pharmacokinetics (PK) in seven Japanese adult patients who underwent allogenic hematopoietic stem cell transplantation (HSCT) at Fukuoka University Hospital. We showed that the average plasma BU concentrations at steady state (Css) ranged from 745 to 2422 ng/mL. BU population PK (PPK) parameters for Caucasian have already been reported. However, few data are available for Asian people.
We therefore investigated the PPK parameters of BU in 82 Japanese adult patients (51 males and 31 females) who underwent HSCT following a BU-containing conditioning regimen. Their average age was 43.4 years, ranging from 16 to 68 years. Blood samples were collected in heparinized tubes at 0.5,1,2,4 and 6 hours after the oral administration of BU, and the plasma BU concentrations were measured by high performance liquid chromatography (HPLC). A one compartment open model with first-order absorption was used. We estimated individual BU doses by using Bayesian, and PK parameters by using nonlinear mixed effects modeling (NONMEM) computer program.
Since Oral clearance (L/h/kg) (CL/F) and distribution volumes (L/kg) (Vd/F) were proportional to total body weight, so we set the following formulas. The final PPK parameters were CL/F = 0.139•TBW•1.14GEN •1.09VPA •1.52TBIL≥1.2, Vd/F = 0.76•TBW•0.902GEN, absorption rate constant (h−1) (ka) = 2.58. TBW is total body weight (kg), VPA = 1 for concomitant administration of valproic acid and 0 for another anticonvulsant, GEN = 1 for patients gender female and 0 for male, TBIL≥1.2 = 1 for total bilirubin above 1.2 mg/dL and 0 for below.
CL/F in Japanese and Caucasian were 0.139 and 0.149, respectively, Vd/F were 0.76 and 0.64, respectively, and ka were 2.58 and 1.68, respectively.
These data showed that CL/F and Vd/F were almost same between Japanese and Caucasian, but ka was larger. We also found that high serum levels of total bilirubin and administration of VPA induced the increase of CL/F at 52 % and 9 %, respectively. The interindividual variabilities in CL/F, Vd/F and ka were 21.9%, 20.9% and 82.3%, respectively, and the residual variability was 18.4% as coefficient of variation.
This study represented that oral BU PPK parameters in Japanese were different from that in Caucasian. BU PPK parameters were influenced by gender, serum levels of total bilirubin and administration of VPA. Our study showed that BU PPK differed between Japanese and Caucasian.
Differential Regulation of bFGF- and VEGF-Induced Angiogenesis in Mice Lacking Junctional Adhesion Molecule-A.
Authors: Naik Meghna U. , Naik Ulhas P. 
Published: April 2020

Abstract
Angiogenesis, the formation of new blood vessels from preexisting vessels, is a complex process involving vascular endothelial cell activation, proliferation, migration, and tube formation. Several growth factors have been shown to induce this process. Among them vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are most widely studied. These two growth factors are believed to induce angiogenesis through distinct signaling pathways. Recently, we have shown that signaling through junctional adhesion molecule-A (JAM-A) is required for bFGF-induced angiogenesis. Here, we present evidence that JAM-A differentially regulates both bFGF-induced and VEGF-induced angiogenic responses. Using human umbilical cord vein endothelial cells (HUVEC) we found that bFGF-induced but not VEGF-induced migration was inhibited by function blocking anti-JAM-A antibody. We also found that mutant JAM-A blocked bFGF-induced but not VEGF-induced MAP kinase activation. To conclusively determine the role of JAM-A in angiogenic signaling, we generated JAM-A null mice. Endothelial cells isolated from JAM-A null mice failed to migrate in response to bFGF stimulation, but showed enhanced cell migration in response to VEGF. Further, using aortic ring and Matrigel® plug assays, we found that bFGF failed to induce blood vessel formation in JAM-A null mice. Interestingly, an increase in the number of blood vessels was observed in response to VEGF in these mice suggesting that in the absence of JAM-A bFGF-induced angiogenic pathway is impaired, but VEGF-induced pathway is upregulated to compensate. We next determined the effect of increased angiogenesis on tumor growth. We found that B16F0 melanoma growth was enhanced at least by two-fold in JAM-A null mice compared to wild-type mice. To understand the mechanism of hypersensitivity of JAM-A null mice to VEGF, we determined the expression of VEGF receptors on endothelial cells. We found that surface expression of VEGFR-2 is upregulated in endothelial cells isolated from JAM-A null mice. These results suggest that signaling through JAM-A is essential for the bFGF-induced pathway and when this pathway is impaired, the VEGF-induced pathway is upregulated which leads to enhanced angiogenesis and tumor growth.
Pulse Cyclophosphamide in the Treatment of Liver GvHD Unresponsive to Steroids.
Authors: Mayer Jiri , Krejci Marta , Doubek Michael , Brychtova Yvona 
Published: April 2020

Abstract
Introduction: Steroids provide an active but inadequate therapy for acute GvHD. Corticosteroid-resistant acute GvHD is extremely difficult to manage and is associated with very high morbidity and mortality. Cyclophosphamide (Cy) has been used for a long time as an immunosuppressive drug for the treatment of many autoimmune disorders, as a cytotoxic drug, and Cy is also a part of both classical conditioning regimens, Cy+TBI and Cy+busulfan. Pulse Cy at a dose of 1 g/m2 is now an established mainstay in the treatment of lupus nephritis. Therefore, we hypothesized that this pulse treatment can be used also for steroid-refractory GvHD. In a previous work, we showed that intestinal GvHD responded poorly to pulse Cy, whilst liver, skin, and oral GvHD responded very well. Since steroid-refractory acute skin GvHD is not frequent, in this report, we analyzed our new data concerning liver GvHD.
Patients (pts) and methods: This is a retrospective study of 17 pts: 11 pts had acute GvHD, 3 pts had chronic GvHD (albeit progressing from acute GvHD), and 4 pts developed liver GvHD upon DLI. Seven pts had hepatitic variant of liver GvHD (serum aminotransferase ALT or AST elevation &gt;10 times the upper normal limit). Seven pts had steroids plus another previous therapy before receiving Cy pulse.
Results and Conclusions: There were 21 Cy administrations at the dose of about 1 g/m2 (460 mg/m2 – 1500 mg/m2, median dose: 1000 mg/m2). The dose was empirically decreased or increased because of fear of myelosuppression, or previous incomplete response, respectively. There were 47% CR (8/17), 18% (3/17) PR, and 35% (6/17) NR. However, in 4 pts with NR their clinical status stabilized and they responded to another treatment. Four pts died: 2 from intractable liver and intestinal GvHD, 1 from intestinal GvHD with liver GvHD in PR, and 1 from relapsing leukemia. All GvHD related deaths occurred in pts with acute GvHD, whereas all but one pts (relapsing AML) with post-DLI or chronic GvHD survived. Concomitant severe gut GvHD seems to be the risk factor for not reaching CR in liver GvHD, and death. On the other hand, however, 6/7 pts (86%) with hepatitic GvHD reached CR. Pulse Cy exerts a good toxicity profile, except for a manageable, short-term myelosuppression in some patients, and does not appear to increase the risk of opportunistic infections, mixed chimerism, relapses, or posttransplant lymphoproliferative disease. The cost of the drug is negligible.
Risk Factor Analysis for Predicting Chronic Graft-Versus-Host Disease of Progressive or Quiescent Type in a Cohort of Patients with a History of Acute Graft-Versus-Host Disease after Allogeneic Stem Cell Transplantation.
Authors: Sohn Sang Kyun , Kim Dong Hwan , Baek Jin Ho , Kim Jong Gwang , Lee Kyu Bo , Lee Kyoo-Hyung , Lee Je-Hwan , Choi Seong-Jun , Lee Jung-Hee 
Published: April 2020

Abstract
Background: As a series of our previous investigation (Haematologica, 2005. 90: 939–48) identifying poor prognostic factors (lymphocytopenia and visceral involvement) at the onset of acute GVHD (aGVHD) in patients with a history of aGVHD after allogeneic stem cell transplantation (SCT), we tried to identify variables that could predict the development of chronic GVHD of progressive or quiescent type (pq cGVHD) and patients’ outcome after the diagnosis of cGVHD in cohort of 99 patients who experienced aGVHD after allogeneic SCT.
Patients and Methods: We evaluated the risk factors for cGVHD of pq cGVHD with various clinical parameters in patient group with a history of aGVHD and also the prognostic significance of various clinical parameters at diagnosis of cGVHD to determine the prognostic factor for GVHD-specific survival (GSS) in patients with pq cGVHD.
Results: From 118 patient experienced aGVHD of any degree, 99 patients were evaluated for cGVHD. The cumulative incidence of overall and extensive pq cGVHD at 2 years was estimated as 84.4% and 63.1%, respectively. In univariate analyses for risk factors of pq cGVHD, severe grade 3,4 aGVHD, primary treatment failure (PTF), lymphocytopenia (≤100/μl), elevated ALP (&gt;160IU/l), visceral involvement, hepatic or gut involvement were identified. Especially, severe aGVHD (p=0.022 and &lt;0.001), PTF (p=0.009 and 0.010) for overall and extensive pq cGVHD, lymphocytopenia (p=0.031) for extensive pq cGVHD, and elevated ALP (p=0.001) for overall pq cGVHD were independent risk factors. The prediction model of subsequent pq cGVHD validated these risk factors with respect to the incidences of overall pq cGVHD (48.6% versus 91.9% for no risk factor versus 1~3 risk factor(s)) and of extensive pq cGVHD (34.2% versus 59.6% versus 92.2% for no / 1 / 2~3 risk factors). HLA-disparity and stem cell source did not influence on the development of pq cGVHD in this cohort. The GSS and probability of systemic immunosuppressive treatment at 2 year after diagnosis of cGVHD was estimated as 55.9% and 51.9%. The GSS was significantly associated with the performance status (p=0.004) and lymphocytopenia (≤ 1,000/μl, p=0.022) at diagnosis of cGVHD by Cox’s proportional hazard model.
Conclusion: Severe aGVHD, PTF, lymphocytopenia and elevated ALP may be useful predictive factors for the development of pq cGVHD in a cohort of patients who experienced a GVHD after allogeneic SCT.
Figure. The significance of the predictive model for the development of overall (A) or extensive chronic graft-versus-host disease (GVHD; B) after occurence of acute GVHD Figure. The significance of the predictive model for the development of overall (A) or extensive chronic graft-versus-host disease (GVHD; B) after occurence of acute GVHD
Allogeneic-SCT in Patients with Hematological Malignancies Aged 60 Years or Older: Importance of HLA-Matching.
Authors: Schleuning Michael , Judith Dirk , Kaltenhaeuser Joerg , Baurmann Herrad , Schwerdtfeger Rainer 
Published: April 2020

Abstract
Despite the introduction of toxicity-reduced conditioning there is still much debate if patients aged 60 years or more should be considered as candidates for allogeneic stem cell transplantation (allo-SCT) and the experience with this patient group remains limited. This report summarizes our cumulative experience in a cohort of 31 consecutive elderly patients (age 60–69, median 61 years) with hematological malignancies treated with, depending on the underlying disease, a variety of toxicity-reduced conditioning regimens followed by allo-SCT. All but four patients received mobilized peripheral blood stem cells as graft and 12 patients had a sibling and 19 an unrelated donor (MUD), respectively. In the majority of patients the underlying disease was acute leukemia or progressive MDS (n=2) with only 25% being in complete remission at the time of transplant. The remaining 8 patients suffered from either chronic leukemia (CML, n=3; CLL, n=1), osteomyelofibrosis (n=3) and multiple myeloma (n=1). For this patient cohort the conditioning regimens applied were all myeloablative as evidenced by full donor chimerism at day +30 in all patients. The time to recovery of absolute neutrophil count &gt;/=0.5 x 10(9)/l was 10-34 days (median 19 days). Despite the high relapse risk in the majority of patients we observed only 4 deaths because of recurrent leukemia. Severe acute GVHD (grade 3 and 4) occurred in 26% and the non relapse mortality was 39% with 5 patients dying from GVHD, 6 from infection and one from toxicity. With a median follow-up of 16 months (range 2 – 66m) for surviving patients the Kaplan-Meyer procedure estimates a 39% probability of survival at 4 y after transplantation. Interestingly, there is no difference what so ever in survival if patients had an identical sibling donor or a MUD. Patients with acute leukaemia had a trend to inferior survival as compared to the others; however, the remission status of these patients had no impact on survival. The most important risk factor for inferior survival was an incomplete HLA-match (8/8). None of the patients who received a mismatched graft survived more than 13 months, whereas in fully HLA-matched transplantations 56% of patients are currently alive and the Kaplan-Meier estimate for survival is 49% at 4 years. Our data support the notion that toxicity-reduced conditioning followed by allo-SCT may be still myeloablative in the elderly, nevertheless it can safely be applied and has excellent anti-leukemic efficacy. However, further efforts are needed for control of GVHD and infectious complications, especially in this high-risk population. A first step in that direction would be, to only accept fully HLA-matched donors.
Analysis of TREC (T Cell Receptor Excision Circles) Levels in CD94 Expressing CD8 T Cells in Chronic GVHD.
Authors: Iwao Noriaki , Tanaka Junji , Kato Naoko , Kondo Takeshi , Miura Yoko , Toubai Tomomi , Shigematsu Akio , Kondo Keiichi , Ohta Shuichi , Asaka Masahiro , Imamura Masahiro 
Published: April 2020

Abstract
CD94 is one of the C-type lectin family members, forms a heterodimer with NKG2 gene family, and CD94 /NKG2A are inhibitory receptors. Not only NK cells but a subset of T cells express CD94/NKG2A, and previously we revealed the proportion of CD94/NKG2A expressing CD8 T cells were higher in patients with chronic graft versus host disease (GVHD), and CD94 expressing T cells have suppressive effects on mixed lymphocyte culture(MLC). We focus on CD94 positive T cell during T cell reconstitution after allogeneic hematopietic stem cell transplantation (allo-HSCT). T cell receptor excision circles (TREC) are suggested to be a useful marker of recent thymic output. In this study, we attempt to study TREC-containing CD8 T cell subset expressing CD94, and to examine the relation of TREC DNA level in CD94 expressing CD8 T cell and GVHD. We analyzed peripheral blood mononuclear cells (PBMCs) isolated from 24 patients (82 samples) undergone allo-HSCT including 15 patients with bone marrow transplantation and 9 patients with non-myeloablative stem cell transplantation. Informed consent was obtained from all patients. CD4 positive T cells were separated from PBMCs by magnetic cell sorting, and CD4 negative cell population was divided into CD94 positive CD8 T cells and CD94 negative CD8 T cells by fluorescence activated cell sorter. Genomic DNA was extracted from these separated T cell subsets. TREC DNA copy numbers per 105 isolated T cells (TREC level) were quantified by real time PCR. We investigated TREC levels in clinical status with pre-allo-HSCT, no episodes of GVHD or before manifestation of GVHD (No GVHD), chronic GVHD on disease (C-GVHD), and no symptoms and remission status of GVHD after immunosuppressive therapy (R-GVHD). Statical analyses were carried out by Mann-Whitney U test. There were no significant differences in TREC level of sorted CD4 positive T cells in C-GVHD compared with No GVHD (p=0.75) and R-GVHD (p=0.61), and also CD94 negative CD8 T cells in C-GVHD compared with No GVHD (p=0.79) and R-GVHD (p=0.20). On the other hand, TREC level of CD94 positive CD8 T cells in C-GVHD decreased in comparison with No GVHD (p=0.015) and R-GVHD (p=0.0019). The reduction of TREC level is thought to be induced not only by low thymic output but also by dilution of TREC concentration due to peripheral T cell expansion without duplications of TREC. These results may suggest that CD94 positive T cells play a role in modulation of GVHD, and proliferate during chronic GVHD with dilution of TREC in CD94 positive CD8 T cells. It is suggested that TREC level of CD94 expressing CD8 T cells may be useful markers of chronic GVHD.