bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effort to Protect Totos, the Smallest Tribe in the World.
Authors: Basak Jayasri , Majumdar Deboshree , Pal Nabamita , Paul Sanghita , Mukhopadhyay Soma , Mukhopadhyay Ashis 
Published: October 2019

Abstract
Abstract 5113

Background
Toto is a primitive Indo-Bhutanese tribe residing in a small enclave called Totopara in the Jalpaiguri district of West Bengal, India. Toto culture and language is totally unique to the tribe, and is clearly distinguished from the neighboring Rajbongshis, Koch or the Bhutanese Sharchop tribes. Totos use to do consanguineous marriage and stay in the same locality even after marriage. Totos are becoming extinct since 1950. Total population of Totos are 1377 (smallest tribe of the world) and it is declining day by day. We noticed that their life span is very short (35-40 years). With a motive to investigate the reason behind it we started our work and got the answers of our queries. Almost 50% of them are carrier of beta Thalassemia, which is a genetic disease and are caused by the mutation of beta globin gene. The objectives of our study are,

Materials & Methods
After awareness program, 3-4 ml of blood sample was collected from each Toto along with their written consent. On the spot inexpensive NESTROF (Naked Eye Single Tube Red Cell Osmotic Fragility) test was done. After returning to institute we use to do CBC (Complete Blood Count) followed by HPLC (High Performance Liquid Chromatography). DNA was isolated from those samples detected as carrier or patients using the protocol standardized in our lab. Mutation was detected by ARMS (Amplification Refractory Mutation System)–PCR.

Conclusion
Total individual already screened in the Totopara under the age limit 10- 35 years is 688. Number of Totos and Non-Totos are 628 and 60 respectively. From NESTROFT and CBC result it was revealed that different blood parameters e.g., MCV, MCH, RBC, RDW, Hb% etc of 71% Totos do not match with the normal value and found to be significantly altered. HPLC results show that 51% Totos are carrier of HbE and 20% of them are HbE homozygous. Whereas 97% non-Totos are found to be normal. DNA analysis by ARMS-PCR confirmed the result of HPLC. The percentage of HbE carrier and HbE homozygous is very high among Totos and this is the real cause of their early death. To protect this primitive tribe we have to restrict their marriage between two carriers. For Totos thalassemia carrier detection should be mandatory before marriage.

Disclosures
No relevant conflicts of interest to declare.
Platelet Adhesion Plaques: Anchors for Contraction of the Hemostatic Plug.
Authors: White James G. , Burris Steven M. , Escolar Gines 
Published: October 2019

Abstract
Abstract 5128
Exposure of GPIIb-IIIa and other receptors on the surface of activated platelets, binding of fibrinogen, molding of shape-changed cells into tight aggregates, internal assembly of actin molecules into filaments and movement of talin, an actin-binding protein, to the inner membrane surface provides the framework for clot retraction. However, the direction of contractile force towards the center of large aggregates or clots would lift the hemostatic plug away from the edges of vascular injury. Another mechanism must be present to facilitate the direction of contractile force toward the damaged vessel wall. This may be accomplished by development of adhesion plaques as platelets spread out on the injured vessel. The present study has used scanning (SEM) and transmission electron microscopy (TEM), confocal and immunofluorescence microscopy to detect adhesion plaques developing at sites of contact as platelets spread on surfaces. Rhodamine-phalloidin was used to detect actin filaments, and an anti-talin antibody identified by protein-A gold or Alexa Fluor 488 labeled rabbit anti mouse IgG to demonstrate talin. Normal human platelets were spread on clean glass slides or plastic chambers for intervals of up to 90 min, extracted with Triton X100 or fixed intact then labeled for talin and actin, and prepared for study by the several microscopic techniques. Triton-extracted spread platelets revealed attachment plaques well stained for talin and actin when examined by SEM or TEM. Inmunofluorescence studies of spread platelets stained with rhodamine-phalloidin and antibodies also revealed co-participation of actin filaments and talin in formation of the adhesion plaques. The association of actin and talin remained intact at all intervals for up to 90 min. Clearly adhesion plaques serve as the anchors for contraction and sealing of hemostatic plug to damaged vascular surfaces.

Disclosures
No relevant conflicts of interest to declare.
Allografts Selectively Photodepleted of GvHD Causing T Cells and Followed by Low-Level Immunosuppression: A Novel Method to Improve Disease Control After HLA-Matched Sibling Transplantations.
Authors: Mielke Stephan , Shenoy Aarthi , Rezvani Katayoun , Yong Agnes S. M. , McIver Zachariah A. , Fellowes Vicki S. , Savani Bipin N. , Kurlander Roger , Khuu Hanh , Boss Carol , Barrett A.John 
Published: October 2019

Abstract
Abstract 515

We established a highly efficient GMP-grade, ex-vivo selective allodepletion process where host-activated donor T cells are eliminated based on their preferential retention of the photosensitizer 4,5-dibromorhodamine 123 (TH9402) and exposure to visible light (Kiadis Pharma, The Netherlands). As relapse of disease largely impairs the overall success of allogeneic stem cell transplantation we aimed to improve this outcome by using selectively T cell depleted allografts in order to reduce post transplant immunosuppression and thereby enhance graft versus malignancy effects. To determine the appropriate level of post transplant immunosuppression we designed a three sequential de-escalation stage trial with grade III-IV acute GvHD as the primary endpoint involving 17 patients per study cohort. Here we report on the first completed study cohort of NIH trial 07-H-0136 where seventeen patients (median age 44 (28-68) years) with hematological malignancies received a CD34-selected (Miltenyi, Germany) stem cell allograft together with 5 × 106/kg selectively depleted donor T cells following an age-adapted, radiation-based preparative regimen (FluCyTBI). Eleven patients had high risk disease (including ALL (Ph+, CR>1), refractory NHL, AML/MDS, AML with chloroma and blast crisis CML). Low-dose cyclosporine was used as sole immunosuppression for 90 days post transplant in the absence of GvHD. At a median follow-up of 385 (119-714) days actuarial probabilities (±SEM) of acute GvHD were 35±12% for grade II-IV and 0% for grade III-IV. Non-relapse mortality (NRM) was low with 17±11%. Overall survival (OS) was 73±12% and relapse-free survival (RFS) was 65±13% with a relapse probability of 21±11% (Figure). A low relapse incidence in a high-risk population suggests functionality of selectively allodepleted T cells. The absence of severe GvHD reflects the efficacy of the allodepletion process. Based on these findings we have initiated recruitment for the next study cohort where post transplant immunosuppression will be limited to 45 days only. Ultimately the aim is to achieve further reduction in immunosuppression in the absence of severe GvHD in order to enhance graft versus malignancy effects and thereby improve the outcome after allogeneic stem cell transplantation especially for patients at high risk for relapse.

Disclosures:
Mielke: Kiadis Pharma, The Netherlands: Research Funding, The current trial is supported under a clinical trial agreement between NHLBI and Kiadis.. Savani:Kiadis Pharma Inc., The Netherlands: Consultancy. Barrett:Kiadis Pharma, The Netherlands: The current trial is supported under a clinical trial agreement between NHLBI and Kiadis..
Comparable Disease-Free and Overall Survival After “Well-Matched” Unrelated Donor and Matched Sibling Donor Transplantation in Acute Myeloid Leukemia with Adverse Risk Karyotype in First Complete Remission: A Report From the Acute Leukemia Working Committee of the Centre for International Blood and Marrow Transplant Research.
Authors: Gupta Vikas , Tallman Martin S. , He Wensheng , Logan Brent , DiPersio John F. , Bunjes Donald W. , Weisdorf Daniel J. 
Published: October 2019

Abstract
Abstract 526

In patients with acute myeloid leukemia (AML) in first complete remission (CR1), the indications for matched sibling donor (MSD) transplants and unrelated donor (URD) haemopoietic stem cell transplantation (HSCT) are different. We sought to determine the prognostic impact of donor type on the outcomes of AML with adverse risk karyotype in CR1, a high-risk AML population considered as a standard indication for MSD and URD HSCT. We evaluated the outcomes of 584 patients undergoing allogeneic HSCT for AML with adverse risk karyotype in CR1 between 1995 and 2006, reported to the CIBMTR. Adverse risk karyotype was defined according to SWOG/ECOG classification. Cytogenetics abnormalities were further classified as: complex karyotype (3 or more abnormalities), 32%; and Non-complex divided as abnormal chromosome 7, 25%; chromosome 5, 9%; MLL gene rearrangements, 18%; t (6;9), 5%; and others, 10%. 226 patients underwent MSD and 358 URD. URD were classified based on high resolution typing as:” well matched” [n=254 (71%)] with no known disparity at HLA A, B, C, DRB1; and, “partially matched” [n=104 (29%)] with one locus known or likely mismatched. Previous MDS was present in 19% and 14% had therapy-induced (t-AML). Conditioning regimens were myeloablative and reduced intensity in 74% and 26%, respectively. At 3 years treatment-related mortality (TRM) incidence was 28% (95% CI 24-31); relapse 36%(32-40); disease-free survival (DFS) 36%(32-41) and overall survival (OS) 39%(35-44). Multivariate analyses are summarized in the table. “Well matched” URD and MSD yielded similar DFS and OS, while outcomes were significantly inferior for “partially matched” URD. Cytogenetically defined subsets had similar outcomes. Evaluated as a time-dependent covariate, chronic GVHD had a significantly lower risk of relapse (RR 0.68, p=0.046), while acute GVHD had no effect (RR 0.99, p=0.96). “Well matched” URD and MSD lead to similar DFS and OS in AML CR1patients with adverse risk karyotype. The pool of patients who may benefit from graft-vs-leukemia effect generated with allogeneic HSCT may be considerably expanded with “well-matched” URD HSCT. If a suitable MSD is not availabel, “well-matched” URD should be strongly considered where a MSD HSCT would otherwise be undertaken.

Disclosures:
No relevant conflicts of interest to declare.
Allogeneic Hematopoietic Cell Transplantation Can Cure Some Patients with Acute Leukemia in Relapse or Primary Induction Failure: A CIBMTR Study.
Authors: Duval Michel , He Wensheng , Klein John P. , Tallman Martin S. , DiPersio John F. , Bunjes Donald W. , Weisdorf Daniel J. 
Published: October 2019

Abstract
Abstract 528
Acute leukemia refractory to chemotherapy is uniformly fatal without hematopoietic cell transplantation (HCT). However, the benefit of transplantation for patients not in complete remission (CR) is controversial.

PATIENTS AND METHODS:
2,255 patients transplanted (at 252 centers in 38 countries) with an allogeneic donor after myeloablative conditioning regimen between 1995 and 2004 for acute leukemia in relapse or with primary induction failure were reported to the Center for International Blood and Marrow Transplant Research (CIBMTR). 1,673 had acute myeloid leukemia (AML) and 582 acute lymphoblastic leukemia (ALL). Median age was 38 and 29 years for AML and ALL patients, respectively. Presence of circulating blasts, >25% marrow blasts, Karnofsky score < 90 %, or transplant in first relapse was observed in nearly half of the patients (see Table).

RESULTS:
100 day mortality was 39% in AML and 41% in ALL. The median follow-up of survivors was 5 years. Overall survival (OS) was 19% (95% confidence interval (CI) 17-21) for AML and 16% (13-20) for ALL patients. For AML patients, five adverse pre-transplant variables significantly impacted OS : first CR duration < 6 months, blasts in the blood, donor other than HLA-identical sibling or partially matched unrelated, Karnofsky score < 90%, poor-risk cytogenetics. 106 AML patients had none of these high risk criteria, with a 3 years OS of 44% (35-54). For ALL patients, survival was worse with either: 1st refractory or ' 2nd relapse, ≥ 25% marrow blasts, CMV seropositive recipient, age ≥10 years. 8 ALL patients had no high risk criteria, with a 3 years OS of 85% (53-100). Grade 3-4 acute graft-vs-host disease (GVHD) occurred in 23% of AML and 27% of ALL patients. Chronic GVHD developed in 27%. Leukemia was the cause in 42 % of AML and 37% of ALL patients' deaths.

CONCLUSIONS:
1. HCT can cure some patients with acute leukemia in relapse or with PIF, particularly those lacking defined high risk features. 2. These pre-transplant variables should be considered when deciding whether or not to offer HCT to such patients.

Disclosures:
No relevant conflicts of interest to declare.
Total Therapy 3 Successor (TT3-S) Protocol (2006-66)–Confirming Results Reported for Initial TT3 (TT3-I) Protocol (2003-33) for Newly Diagnosed Multiple Myeloma (MM).
Authors: Nair Bijay , Alsayed Yazan , Waheed Sarah , Anaissie Elias , van Rhee Frits , Szymonifka Jackie , Hoering Antje , Crowley John , Barlogie Bart 
Published: October 2019

Abstract
Abstract 531

Introduction:
TT3-I imparted high CR rates and superior overall survival (OS) and event-free survival (EFS). Bortezomib pharmaco-genomic studies, performed concurrently, revealed that gene expression profiling (GEP) changes 48hr after a bortezomib test-dose had major prognostic significance, which we wished to validate in TT3-S. The purpose of this work is to compare clinical outcomes among the 303 TT3-I and 177 TT3-S patients.

Methods:
Our MM data base was scrutinized for patient characteristics and outcomes of the 2 cohorts in terms of CR rates and duration, as well as OS and EFS, in the context of major prognostic variables (cytogenetic abnormalities (CA), B2M, LDH, GEP risk).

Results:
Patients in the 2 studies were comparable except for a higher incidence of several adverse features in TT3-S: albumin <3.5g/dL in 45% v 25% (p<0.001), B2M >=3.5mg/L in 57% v 45% (p=0.012) and GEP-defined high-risk in 22% v 15% (p=0.035). Outcome comparisons for TT3-S v TT3-I were almost identical: 3-year estimates of OS 80% v 82%, EFS 81% v 77%, sustained CR 61% v 62%; in GEP low-risk/no CA subgroup: OS 94% v 91%, EFS 90% v 87%, sustained CR 98% v 94% (Figure 1a); GEP low-risk/CA subgroup: OS 81% v 79%, EFS 81% v 74%, sustained CR 86% v 90%; GEP high-risk/no CA subgroup: OS 70% v 82%, EFS 75% v 46%, sustained CR 75% v 30%; GEP high-risk/CA subgroup: OS 48% v 41%, EFS 52% v 38%, sustained CR 60% v 55% (Figure 1b). None of the Kaplan-Meier plots for these GEP-risk/CA constellations were different between TT3-S and TT3-I.

Conclusions:
We have validated the remarkable success of the TT3 principle in low-risk MM, whereas high-risk disease deserves novel strategies as being pursued in TT5 with dose-dense and less dose-intense therapies.

Disclosures:
van Rhee: Genzyme Corporation: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.
Long-Term Survival Analysis of the North American Intergroup Study C9011 Comparing Fludarabine (F) and Chlorambucil (C) in Previously Untreated Patients with Chronic Lymphocytic Leukemia (CLL).
Authors: Rai Kanti R. , Peterson Bercedis L. , Appelbaum Frederick R. , Tallman Martin S. , Belch Andrew , Morrison Vicki A. , Larson Richard A. 
Published: October 2019

Abstract
Abstract 536
Long-term outcomes following novel therapies for CLL have rarely been reported. Between 10/90 and 12/94, 509 eligible, untreated patients (pts) with symptomatic CLL were enrolled by 4 cooperative groups onto study C9011; 179 were randomized to F, 193 to C, and 137 to F+C. After slightly more than 5 years median follow up, with the time of last follow-up in June 1999, we reported in 2000 (NEJM 343:1750) that F provided significantly higher response rates and longer remission duration and progression-free survival (PFS) than C (p<0.001 for all 3 endpoints). The combination arm with F+C was stopped early because of high morbidity and mortality. There was no difference in overall survival (OS) among the 3 groups. Nearly 10 years have now elapsed since this report. Therefore, with the time of last follow-up in January 2009, we analyzed the long term outcomes of pts enrolled on the study. PFS was defined as the time between randomization and the occurrence of progressive disease or death due to any cause.

Results:
Of the 509 pts, 85% have now died; among pts on the F and C arms, 92% have progressed. We found that F treatment resulted in significantly longer PFS than did C (p < 0.001), with notable differences in PFS at 2, 3, and 4 years (Table). While the F and C arms had the same OS during the initial 5 years following randomization, our current analysis with longer follow-up shows that pts treated on the F arm had better survival than did those on the C arm during the ensuing years (Figure). The p-values for this difference are 0.04 (unadjusted for covariates) and 0.07 (covariate-adjusted). The emergence of improved survival following initial F treatment, appearing only after 5-6 years, is an unexpected and noteworthy finding. Reporting second malignancies was required on this study. There were 27 epithelial cancers reported (9 on F, 11 on C, 7 on F+C), involving colon, lung, breast, prostate, pancreas, liver, bladder and skin (6 squamous, 2 melanoma). Seven therapy-related myeloid neoplasms (t-MN) were reported; 6 were on F+C; 1 on F. Richter's transformation to non-Hodgkin lymphoma was reported in 34 pts; prolymphocytic leukemia occurred in 10; Hodgkin lymphoma in 6; myeloma in 2; hairy cell leukemia in 1. These cases were distributed with 18 on F, 18 on C, and 17 on F+C. Thus, the overall incidence of second malignancies reported was 17%.

Conclusion:
Initial treatment with F provides better long-term outcomes than initial treatment with C. Second malignancies are common, but the overall incidence is not increased on the F-containing treatment arms except for t-MN.

Disclosures:
No relevant conflicts of interest to declare.
Ectopic Human Mesenchymal Stem Cell-Coated Scaffolds Create An In Vivo Leukemia Niche in NOD/SCID Mice.
Authors: Vaiselbuh Sarah Rivkah , Edelman Morris , Lipton Jeffrey Michael , Liu Johnson M. 
Published: October 2019

Abstract
Abstract 559

Introduction:
Different cellular components of the normal hematopoietic niche have been identified. However, the niche for malignant hematopoiesis remains to be elucidated. Recent work of other groups has suggested that hematopoietic stem cells (HSC) within the bone marrow anchor themselves in place by attaching to osteoblasts and/or vascular sinusoid endothelial cells. We have recently identified mesenchymal stem cells (MSC) as niche-maker cells and found a crucial role of the SDF-1/CXCR4 axis in this process. Stromal Derived Factor-1 (SDF-1/CXCL12) regulates stem cell trafficking and the cell cycle via its receptor CXCR4.

Methods:
Polyurethane scaffolds, coated in vitro with human bone marrow MSC, were implanted subcutaneously in non-irradiated NOD/SCID mice. CD34+ HSC or primary AML cells (from a leukapheresis product) were injected either in situ or retro-orbitally in the mice and analyzed for engraftment. The mice were treated twice per week with in situ injections of SDF-1, AMD3100 (a CXCR4 antagonist) or PBS (control). After 2 to 4 weeks, the scaffolds were processed and evaluated for cell survival in the mesenchymal niche by immunohistochemistry.

Results:
We created in vitro MSC-coated scaffolds that retained inoculated AML cells in the presence of SDF-1, while AML cells seeded on empty scaffolds were not retained. In vivo in NOD/SCID mice, the MSC-coated scaffolds, in the presence of SDF-1 enabled homing of both in situ injected normal CD34+ HSC and retroorbital- or in situ injected primary human AML cells. The scaffolds were vascularized and showed osteoclasts and adipocytes present, suggestive of an ectopic human bone marrow microenvironment in the murine host. Finally, the SDF-1-treated scaffolds showed proliferation of the MSC stromal layer with multiple adherent AML cells, while in the AMD3100-treated scaffolds the stromal lining was thin and disrupted at several points, leaving AML cells free floating in proximity. The PBS-treated control-scaffold showed a thin single cell MSC stromal layer without disruption, with few AML cells attached.

Conclusion:
The preliminary data of this functional ectopic human microenvironment in NOD/SCID mice suggest that AMD3100 (a CXCR4 antagonist) can disrupt the stem cell niche by modulation of the mesenchymal stromal. Further studies are needed to define the role of mesenchymal stem cells in maintaining the hematopoietic/leukemic stem cell niche in vivo.

In Vivo Leukemia Stem Cell Niche:
(A) Empty polyurethane scaffold. (B)Vascularization in SQ implanted MSC-coated scaffold (s) niche in NOD/SCID mice. (C) DAB Peroxidase (brown) human CD45 positive nests of AML cells (arrows) 1 week after direct in situ AML injection. (D) Human CD45 positive myeloid cells adhere to MSC in vivo (arrows).

Disclosures:
No relevant conflicts of interest to declare.
Expression of Mir-196b Is Not Exclusively MLL-Driven but Especially Linked to Activation of HOXA Genes in Children with Acute Lymphoblastic Leukemia.
Authors: Schotte Diana , Lange-Turenhout Ellen , Stumpel Dominique JPM , Stam Ronald W. , Buijs-Gladdines Jessica , Meijerink Jules P.P. , Pieters Rob , Den Boer Monique L. 
Published: October 2019

Abstract
Abstract 580
MicroRNAs (miRNAs) play important roles in diverse biological processes including hematopoiesis. As a consequence, aberrant expression of miRNAs may contribute to hematopoietic malignancies. It has been reported that miR-196b is transcriptionally activated by MLL and MLL-fusion genes and is therefore highly expressed in MLL-rearranged leukemia. In order to investigate whether high expression levels of miR-196b are restricted to MLL-rearranged leukemia cases, we measured the expression in samples of 72 selected pediatric acute lymphoblastic leukemia (ALL) cases i.e. MLL-rearranged and non-MLL-rearranged precursor B-ALL and T-ALL patients. MiR-196b was highly expressed in 9/12 MLL-rearranged precursor B-ALL patients, but also in 14/22 T-ALL patients. In particular, 100% of T-ALL cases carrying CALM-AF10 (n=5), MLL-AF6 (n=2), SET-NUP214 (n=3) and an inversion of chromosome 7 (n=1) showed high expression levels of miR-196b comparable to the high levels found in MLL-rearranged ALL. Like MLL-rearrangements, these genetic abnormalities have been functionally linked with upregulation of HOXA cluster genes. MiR-196b expression levels in these patients were strongly correlated with the expression of HOXA family but not with HOXB and HOXC cluster genes (Rs ≥ 0.7, P≤0.003). Since miR-196b is located between HOXA9 and HOXA10 on chromosome 7, our data suggest co-activation of miR-196b and HOXA family genes in pediatric ALL. In parallel to the high expression level of miR-196b we found decreased methylation at CpG islands located 5' of miR-196b in MLL-rearranged cases compared to normal bone marrow cells, which suggests an epigenetic origin for the high expression level of miR-196b in these patients. Despite the fact that MLL-rearranged ALL patients often respond poorly to prednisolone and L-asparaginase, upregulation of miR-196b was not indicative for the resistance to these drugs in pediatric ALL. In conclusion, high-level expression of miR-196b is not only MLL-driven, but can also be found in other types of leukemia that display aberrant activation of HOXA genes.

Disclosures:
No relevant conflicts of interest to declare.