bluuebunny/crossref_metadata_2025 huggingface dataset:
Health-Related Quality of Life (HRQoL) Measurements in Multiple Myeloma Patients Obtained By EORTC QLQ-C30; A Critical Review of Interpreting HRQoL Data in Longitudinal Studies
Authors: Nielsen Lene Kongsgaard , Jarden Mary , Andersen Christen Lykkegaard , Frederiksen Henrik , Abildgaard Niels
Published: October 2019

Introduction: Multiple myeloma (MM) patients report more symptoms and more severe symptoms than patients with other hematological malignancies, resulting in reduced health-related quality of life (HRQoL). Because survival is improving in MM patients there is an increased need for focus on HRQoL. The aim of this review was to interpret published longitudinal studies in MM patients according to a difference in HRQoL score, which is perceived as beneficial to the patient. Such criteria for minimal important differences (MID) in HRQoL score has been explored by Kvam et al (Eur J Haematol 2010, 84:345-53) and has been defined by a concensus expert panel (Cocks et al, Eur J Cancer 2012, 48:1713-21).

Methods: A literature search was performed May 2016 in PubMed, Embase, PsycINFO and CINAHL. Publications with longitudinal follow-up using the EORTC QLQ-C30 instrument for HRQoL measurement of physical functioning, global quality of life, fatigue or pain were included, and evaluation of mean change from baseline was carried out according to MID as defined by Kvam, and according to minimal, medium and large changes as defined by Cocks.

Results: 11 first-line, 3 consolidation, 2 maintenance, 7 relapse studies, and one population-based study were included. In general, large and medium improvements were reported during first-line treatments. During consolidation with autologous hematopoietic stem cell transplantation, temporary large deteriorations were reported, but equalization occurred with even large improvements compared to baseline reportings. Deterioration or no beneficial change in global quality of life or fatigue are reported by the MM patients during relapse treatment. HRQoL data during maintenance therapy are sparse. For summary of results, see Figure 1.

Conclusions: Guidelines for interpreting HRQoL changes with definitions of MID have been developed, but consensus is missing. Improvements in HRQoL are far more likely during first-line compared to relapse treatment regimens. The background of these findings should be in focus in future studies and HRQoL measurements should be part of maintenance studies in the future.

Disclosures

No relevant conflicts of interest to declare.
Treatment of High Risk Aggressive B Cell Lymphomas with DA EPOCH R- a Retrospective Analysis
Authors: Panny Michael , Troppan Katharina , Neumeister Peter , Nösslinger Thomas , Keil Felix
Published: October 2019

Background: Promising results in patients suffering from high risk DLBCL, Burkitt`s lymphoma (BL), gray-zone lymphoma (GZL) and primary mediastinal B cell lymphoma (PMBCL) treated with DA-EPOCH R have been reported.

In our centres high risk DLBCL - defined as double-hit/double hit score 2 or high/ high-intermediate risk NCCN IPI -, BL, MGZL and PMCL are treated with DA EPOCH R.

Methods: Retrospective analysis of toxicity and efficacy in DA EPOCH R treated patients.

Results: So far 39 previously untreated patients with a median age of 54a (28a - 76a) have been treated with a total of 190 cycles of DA EPOCH R: 16 DLBCL, 9 GZL, 8 PMBCL, 6 BL. 37 Patients have finished treatment, 2 are stilll on treatment.

Targeted ANC < 500/l occurred in 46%, thrombocytopenia < 25.000/l in 20% and anemia <8g/dl in 12,5% of all cycles. Dose escalation was possible in 27 (73%) patients - but only in 5 (38%) of 13 patients > 65a. 15 (63%) of 24 patients aged < 65a received at least dose level 3. Due to peripheral sensory neuropathy, Vincristine had to be dose reduced in 52% of all cycles. Other CTCAE grade III/IV non-hematopoietic toxicities were infrequent and manageable.

After a median follow up of 10 months (range: 1-25) overall survival (OS) rate is 74%. 2 patients in PR were (1 PMBCL, 1 GZL) bridged to allogenic stem cell transplantation , 1 patient in CR had to be switched to a less toxic regimen due to repeated febrile neutropenia after 3 cycles of treatment. In 16 high risk DLBCL patients (8 DHS2/DHL, 8 high/high intermediate NCCN IPI) OS is 80% after a median follow up of 12 month. As predescribed in literature, prognosis of relapsed or refractory patients is poor: 7 out of 8 (4GZL, 2 DLBCL, 1 BL, 1 PMBCL) relapsed/ refractory died. Causes of death were: 2 infectious complications (1 DLBCL HIV associated, 1 GZL) and 5 progressive disease.

Conclusion: Although still preliminary, limited data, DA EPOCH R seems to be a feasible treatment with acceptable toxicity and a promising response rate. Dose escalation is age dependent. Especially in patients with high risk DLBCL DA EPOCH R is an alternative to (insufficient) induction therapy with R CHOP and is challenging more toxic regimens like R ACVBP or R Hyper CVAD.

Disclosures

No relevant conflicts of interest to declare.
Correlation of Morphological Dysplasia and Immunophenotypic Features in Myelodysplastic Syndromes
Authors: Velloso Elvira D Rodrigues Pereira , Cosentino Rosana M , Suganuma Liliana Mitie , Barroso Rodrigo de Souza , Bacal Nydia Strachman , Colombini Marjorie P , Silveira Paulo
Published: October 2019

The Myelodysplastic Syndromes (MDS) are a group of clonal hematopoietic stem cell diseases characterized by cytopenias, dysplasia in one or more myeloid cell lines, ineffective hematopoiesis and increased risk for development of acute myeloid leukemia (AML).The pathological hallmark of myelodysplastic syndromes (MDS) is marrow dysplasia, which represents the basis of the World Health Organization (WHO) classification of these disorders. However, morphological diagnosis of MDS may have difficulties, with poor inter-observer agreement.Recently, Della Porta and col.(on behalf of Rete Ematologica Lombarda (REL) Network), described a new morphological score system to define dysplasia, that showed high sensitivity and specificity (>90%) andbetter inter-operator agreement (Leukemia 2015;29:66-75). In addition, new toolsfor diagnostic and prognostic purposes were developed.Flow cytometry (FCM) was addedin standard European Leukemia Net guidelinesasrecommended tool for diagnosis and prognosis in MDS, and a simple MDS-FCM scoring system based on four parameters (mainly CD34 and granulocytic cells) by Ogata and col. showed a good specificity (Haematologica 2009;94:1066-1074).

The aim of this study is to correlate morphological and immunophenotypic features in suspicion of MDS.

The study comprised two steps: the first used for morphological validation, included 33 bone marrow (BM) smears from patients with suspected MDS with or without therapy, reviewed by four experts hematologists blinded to clinical data. Dysplasia was evaluated using WHO 2008 criteria and Della Porta's score: 100 erythroblasts, 100 granulocytic cells and 30 megakaryocytes for dysplasia and 500 nucleated cells for blast cells percentage. The second step included 69 BM samples from patients with suspected MDS (27 with final diagnosis of MDS) and was used to correlate cytology and immunophenotypic findings. The morphological aspects were reviewed by one of the expert hematologists, using the same previous criteria and blinded to clinical data. Inter-operator reproducibility of morphological analyses was assessed by Intraclass Correlation Coefficient (ICC). Acceptable reproducibility was defined as ICC > 0.4. The correlation of minimal morphological criteria with immunophenotypic criteria of Ogata and col. was assessed by Discordance rate and Spearman correlation.

The results are shown in table.

In conclusion our study shows good inter observer reproducitibility, in both used criteria. The use of new criteria of Della Porta to the erythroid series was slightly better, suggesting that can be useful in diagnosing myelodisplasias. Immunophenotypic data had a good agreement with morphologic data in confirmed cases of MDS.

Table 1 Table 1.

Disclosures

No relevant conflicts of interest to declare.
Comparison of the IPSS and IPSS-R in Predicting Survival and Response of Myelodysplastic Syndrome Patients to 5-Azacitidine
Authors: Shapiro Roman M , Lazo-Langner Alejandro
Published: October 2019

Background

The international prognostic risk score (IPSS) is a validated tool in the prognosis of myelodysplastic syndromes (MDS). Patients who fall into the Int-2 and High categories of the IPSS qualify for treatment with 5-azacitidine, a hypomethylating agent that has been shown to provide a survival advantage with these prognostic risk scores. The IPSS was recently updated, with the newer revised IPSS (IPSS-R) risk score taking into account more cytogenetic data than the IPSS. It has not yet been determined whether the IPSS-R provides an advantage over the IPSS in predicting outcome with 5-azacitidine. The objective of this study was to reclassify a cohort of MDS patients at a single academic center using the IPSS-R, and to determine its ability to predict survival and response to 5-azacitidine.

Methods

This study was approved by the institutional board review. A total of 97 patients with a diagnosis of MDS were treated with 5-azacitidine at the London Health Science Center up until 2015. Both the IPSS and IPSS-R could be calculated for 77 patients. Kaplan-Meier survival curves were constructed for the patients categorized based on the IPSS and IPSS-R. The scores were also compared based on objective response rates (ORR = complete response + partial response + hematologic improvement) as defined by the IWG 2006 criteria that were determined for all patients.

Results

All 77 patients included in the study had qualified for 5-azacitidine treatment according to provincial criteria, and had an IPSS score int-2 or high. Upon reclassification using the IPSS-R, 2/77 patients were scored as low, 14/77 patients were scored as intermediate, 24/77 patients were scored as high, and 37/77 patients were scored as very high risk. Kaplan-Meier survival analysis (Figure 1) indicated there was no statistically significant difference between the IPSS-R intermediate, high, and very high survival curves (log-rank p = 0.774). The objective response rate for all 77 patients treated with 5-azacitidine was 23%, with ORR of 21% for IPSS-R intermediate, 17% for IPSS-R high, and 38% for IPSS-R very high.

Conclusions

The IPSS-R is an alternative prognostic risk scoring system that takes into account additional cytogenetic data compared to the IPSS. In our center the IPSS-R has not resulted in prognostic risk categories that are an improvement over the IPSS in predicting survival in response to 5-azacitidine. The IPSS-R very high risk patients had double the objective response rate to 5-azacitidine compared to the intermediate and high risk groups.

Disclosures

Lazo-Langner: Bayer: Honoraria; Pfizer: Honoraria; Daiichi Sankyo: Research Funding.
Impact of Novel Agents on Frequency of Second Salvage Autologous Transplantation in Patients with Multiple Myeloma
Authors: Pathak Prajwol , Unnikrishnan Athira , Zou Fei , Leather Helen , Cogle Christopher R. , Hsu Jack W , Brown Randy A. , Wingard John R. , Norkin Maxim
Published: October 2019

Introduction:

High dose melphalan followed by autologous hematopoietic stem cell transplant (ASCT) is the standard of care in eligible multiple myeloma (MM) patients (pts). It is common practice to collect sufficient stem cells for two transplants, for the first ASCT immediately after induction, and for salvage ASCT following disease progression. We investigated the frequency of salvage second ASCT in the era of novel agents to determine if there has been a change in practice.

Methods:

We retrospectively reviewed medical records of MM pts who had stem cells collected for two ASCTs from 03/1996 to 12/2014. We then compared pts who received their first ASCT prior to January 2007 to those after this time. We excluded pts who received their second transplant as part of a clinical protocol, who received subsequent allogenic transplants, and pts who received 2 ASCT transplants as part of initial therapy. Fisher's exact test was used to assess significance of categorical independent variables and t-test was used to assess significance of continuous variables.

Results:

Forty of 506 pts (7.9%) received salvage second ASCT (Fig 1). Among 195 MM pts who received their first ASCT before 2007, 19 pts (9.7%) received salvage ASCT. After 2007, 21 of 311 MM pts (6.7%) received salvage ASCT (P= 0.24). When comparing salvage ASCT in the two time periods, no significant differences were noted, with the

exception of a slightly lower KPS status (80% vs 70%, P=0.001), and the use of novel agents before the salvage therapy (0/19 (0%) vs 17/21 (81%), p <0.001). Otherwise the clinical characteristics, prior response and response duration, and initial stage were similar. Prior to 2007, 6 of 44 (14%) pts older than 65 years received salvage ASCT, whereas after 2007 only 3 of 110 (3%) patients received salvage ASCT (p = 0.017).

Conclusions:

Outside of clinical trials, the use of salvage second ASCT for relapsed MM is not frequent, and was noted to be decreasing further after 2007. We speculate that this 30% relative reduction in the frequency of salvage ASCT after 2007 is likely explained by the introduction of novel agents such as bortezomib and lenalidomde into the clinical practice, the use of maintenance therapy post-ASCT, and more non-ASCT options for salvage treatment after relapse. Further studies are needed to investigate the optimal timing for salvage ASCT and the patients who will best benefit from such treatment.

Figure 1 Figure 1.

Disclosures

Wingard: Astellas: Consultancy; Gilead: Consultancy; Ansun: Consultancy; Fate Therapeutics: Consultancy; Merck: Consultancy.
Survival Improvement of Patients with Chronic Lymphocytic Leukemia (CLL) in Routine Care. a Retrospective Analysis of All Patients with CLL Who Were Treated in an Oncology Group Practice in Germany Between 1995-2015
Authors: Weide Rudolf , Feiten Stefan , Chakupurakal Geothy , Friesenhahn Vera , Kleboth Kristina , Lutschkin Julia , Thomalla Jörg , van Roye Christoph , Heymanns Jochen , Köppler Hubert
Published: October 2019

Introduction:Progress has been made in the diagnosis and treatment of patients with CLL who receive their treatment within prospective clinical trials. Clinical trials only include patients who fulfill the selection criteria and hence do not reflect outcomes in routine practice. Data with regards to the diagnosis, treatment and survival of patients in routine care reflecting the improvements in survival following implementation of newer therapy strategies and agents are not available.

Methods: A retrospective analysis of all patients with CLL who were treated in an oncology group practice in Germany between 1995-2015. Relevant clinical data concerning diagnosis, treatment and survival were transferred from clinical files into a database and analysed statistically using SPSS.

Results:676 CLL patients with a median age of 67 (35-92) were identified. At initial diagnosis 521 patients (77.1%) were in Binet stage A, 83 (12.3%) Binet stage B and 36 (5.3%) Binet stage C. Due to external diagnosis of 36 patients (5.3%) the stage at initial diagnosis couldn't be retrieved. 361 patients (53.4%) never received any treatment. 315 patients (46.6%) needed therapy with a median of 2 therapy lines (1-13). Regimens most frequently applied were: Bendamustine-containing (68.9%), Rituximab-containing (65.4%), Chlorambucil-containing (55.9%), Bendamustine+Rituximab combinations (48.3%) and Fludarabine-containing (39.4%). Regimens containing recently approved drugs could be found as well. 5.1% received Ibrutinib, 2.9% Obinutuzumab and 2.2% Idelalisib. 17.8% of patients were treated within a clinical trial. 5 and 10 year overall survival was 84.5% and 60.4%. Median overall survival according to Binet stage was 13 years for Binet A, 9 years for Binet B and 9 years for Binet C. Patients who needed therapy had a median overall survival of 11 years (0-42) compared to 17 years (0-27) of patients who never needed any therapy.

Conclusions: 53.4% of CLL-patients never needed any therapy. Patients who needed therapy had a much lower life expectancy compared to patients who never needed therapy. Treatment consisted mainly of Bendamustine, Rituximab, Chlorambucil, Bendamustine+Rituximab combinations and Fludarabine leading to a marked prolongation of survival compared to historical controls and registry data.

Disclosures

No relevant conflicts of interest to declare.
p19INK4d Modulates Human Terminal Erythroid Differentiation By Post-Transcriptionally Regulating GATA1 Expression
Authors: Han Xu , Zhang Jieying , Peng Yuanliang , Chen Huiyong , Chen Xiao , Zhang Yibin , Hu Xiao , Mohandas Narla , An Xiuli , Liu Jing
Published: October 2019

Erythropoiesis is a process during which hematopoietic stem cell (HSCs) are first committed to erythroid progenitors, which subsequently undergo terminal erythroid differentiation to produce mature red blood cells. During terminal erythroid differentiation, proerythroblasts undergo 4-5 mitoses to sequentially generate basophilic erythroblasts, polychromatic erythroblasts and orthochromatic erythroblasts that expel their nuclei to produce enucleated reticulocytes. Terminal erythropoiesis is a tightly regulated process. The most well studied regulatory mechanisms include EPO/EPOR mediated signal transduction and transcription factors among which GATA1 plays critical role. Terminal erythroid differentiation is also tightly coordinated with cell cycle exit, which is regulated by cyclins, cyclin-dependent kinases and cyclin-dependent kinase inhibitors (CDKI), yet their roles in erythropoiesis remain largely undefined. Our RNA-seq of human terminal erythroid differentiation shows that of seven CDKI members, only three of them, p18INK4c, p19INK4d and p27KIP1, are abundantly expressed in erythroid cells and their expressions are significantly upregulated in late stage erythroblasts, which were further confirmed by western blotting analysis. In contrast to demonstrated roles of p18INK4c and p27KIP1 in terminal erythroid differentiation, the function of p19INK4d this process has not been studied. To explore the role of p19INK4d during human erythropoiesis, we employed a shRNA-mediated knockdown approach in CD34+ cells and found that p19INK4d knockdown delayed erythroid differentiation, inhibited cell growth, led to increased apoptosis and generation of abnormally nucleated late stage erythroblasts. Unexpectedly, p19INK4d knockdown did not affect cell cycle. Rather it led to decreased GATA1 protein levels. Importantly, the differentiation and nucleus defects were rescued by ectopic expression of GATA1. As GATA1 protein is protected by nuclear HSP70, to explore the mechanism for the decreased GATA1 protein levels, we examined the effects of p19INK4d knockdown on HSP70 and found p19INK4d knockdown led to decreased nuclear localization of HSP70 due to reduced ERK activation. Further biochemical analysis revealed that p19INK4d directly binds to Ras kinase inhibitor PEBP1 and that p19INK4d knockdown increased the expression of PEBP1 which in turn led to reduced ERK activation. These results demonstrate that p19INK4d maintains GATA1 protein levels through PEBP1-pERK-HSP70-GATA1 pathway. Our findings identify previously unknown and unexpected roles for p19INK4d in human terminal erythroid differentiation via a novel pathway.

Disclosures

No relevant conflicts of interest to declare.
Single-Agent Ibrutinib in Recurrent/Refractory Central Nervous System Lymphoma
Authors: Grommes Christian , Pastore Alessandro , Gavrilovic Igor , Kaley Thomas , Nolan Craig , Omuro Antonio M. , Wolfe Julia , Pentsova Elena , Hatzoglou Vaois , Mellinghoff Ingo , DeAngelis Lisa
Published: October 2019

BACKGROUND: Primary Central Nervous System Lymphoma (PCNSL) is an aggressive primary brain tumor with median progression free survival (PFS) after upfront methotrexate-based chemotherapy of 2-3 years. Outcome and treatment options are poor for recurrent/refractory (r/r) disease. Response rates (ORR) range between 30-60% with a PFS of 2-5 months. Ibrutinib has shown promising clinical response in Mantel cell lymphoma, CLL, and Waldenström. This trial investigates Ibrutinib in patients with r/r PCNSL and SCNSL.

METHODS: Eligible patients had r/r PCNSL or Secondary CNS Lymphoma (SCNSL), age≥18, ECOG≤2, normal end-organ function, and unrestricted number of CNS directed prior therapies. In patients with SCNSL disease, systemic disease needed to be absent.

RESULTS: Twenty patients were enrolled (3 at 560 mg; 17 at 840 mg). Median age was 69 (range 21-85); 12 were women. Median ECOG was 1 (0: 2, 1: 12, 2: 6). 65% had PCNSL and 35% SCNSL; 70% had recurrent disease. Eleven had parenchymal disease, 3 isolated cerebrospinal fluid (CSF) involvement and 6 both. Five grade 4 adverse events were observed in 4 patients (lymphopenia (2), sepsis (1), neutropenia (2)). Ten patients developed grade 3 toxicities, including lymphopenia in 3 patients, thrombocytopenia in 2, hyperglycemia in 2, lung infection in 2, neutropenia in 1, urinary tract infection in 1, colitis in 1, and fungal encephalitis in 1. The most common toxicities were hyperglycemia, anemia, and thrombocytopenia. After a median follow-up of 193 days, 19/20 patients were evaluated for response: 8 CR, 7 PR, 1 SD and 3 PD; 75% (15/20) ORR. The median PFS is 7.29 months (95% CI: 3.80-15.43 months (longest: 15.3 months)). The mean Ibrutinib concentration in the CSF 2h post administration at day 1 and 29 is 1.75 ng/mL (3.97 nM) and 2.51 ng/mL (5.6 nM) which is above the IC50 (1nM) required in vitro to reduce growth of lymphoma cells.An additional treatment arm has been added to the trial which will evaluate adverse events of the combination of ibrutinib and high-dose methotrexate chemotherapy. Enrollment into the combination arm is ongoing and updates will be presented at the meeting.

CONCLUSION: Patients with CNS lymphoma tolerate Ibrutinib with manageable adverse events. Drug concentrations in CSF are higher at steady state (day 29) and meaningful CSF concentrations are reached. Clinical response was seen in 75% of CNS lymphoma patients. A combination arm will assess the adverse events of ibrutinib in combination with high-dose methotrexate chemotherapy.

Disclosures

No relevant conflicts of interest to declare.
Gender Differences in the Development of CMR Abnormalities and Cardiac Complications: A Multicentric Prospective Study in a Large Cohort of Thalassemia Major Patients
Authors: Pepe Alessia , Maffei Silvia , Pistoia Laura , Ciancio Angela , Palazzi Giovanni , Argento Crocetta , De Franceschi Lucia , Mangione Maurizio , Vallone Antonino , Missere Massimiliano , Positano Vincenzo , Meloni Antonella
Published: October 2019

Introduction. We aimed to prospectively assess if the male gender was associated with an higher risk of progressive cardiac iron accumulation, development of biventricular dysfunction and myocardial fibrosis assessed by CMR, and development of cardiac complications including heart failure (HF), arrhythmias and pulmonary hypertension (PH).

Methods. We considered 1711 TM patients (899 females, 31.09±9.08 years), consecutively enrolled in the Myocardial Iron Overload in Thalassemia (MIOT) Network. Myocardial iron overload was assessed by the multislice multiecho T2* technique. Biventricular function was quantified by cine images. Late gadolinium enhancement (LGE) images were acquired to detect myocardial fibrosis.

Results. Although having a similar risk of accumulating iron, males showed a significant higher risk of developing cardiac dysfunction, heart failure, arrhythmias and cardiac complications globally considered (Table 1). Figure 1 shows the Kaplan-Meier curves for the outcomes for which the male sex was a significant prognosticator. Until 20-30 years of follow-up the two lines (male and female sex) were almost overlapping while after they clearly diverged. So, patients were divided in two groups based on the follow-up duration. A significant gender-specific difference in the frequency of ventricular dysfunction and cardiac complications appeared for patients followed for at least 20 years. So, two subgroups of patients were identified: patients followed for less than 20 years and patients followed for more than 20 years. In the first subgroup males and females had a comparable risk of developing cardiac iron overload, ventricular dysfunction and cardiac complications. Conversely, if a follow-up longer than 20 years was considered, males exhibited a significant higher risk of having ventricular dysfunction, heart failure, arrhythmias, and cardiac complications.

Conclusion. Females seem to tolerate iron toxicity better, possibly as an effect of reduced sensitivity to chronic oxidative stress. According to the International Guidelines, TM patients should perform a complete cardiac evaluation every year. Our study suggested that in females older than 20 years the follow-up may be performed every 24 months, thus reducing health care costs.

Table 1 Table 1. Figure 1 Figure 1.

Disclosures

Pepe: Chiesi Farmaceutici and ApoPharma Inc.: Other: Alessia Pepe is the PI of the MIOT project, that receives no profit support from Chiesi Farmaceutici S.p.A. and ApoPharma Inc.. De Franceschi:F. Hoffmann-La Roche Ltd, Basel, Switzerland: Research Funding.
Organization and Regulation of the Human Genome
Authors: Ren Bing
Published: October 2019

The 3-dimensional (3D) chromatin organization plays a critical role in gene regulation. Great strides have been made recently to characterize and identify cis regulatory elements from epigenome profiles in different cell types and tissues, but efforts have just begun to functionally characterize these long-range control elements. Mapping interactions between enhancers and promoters, and understanding how the 3D landscape of the genome constrains such interactions is fundamental to our understanding of genome function. I will present recent findings related to 3D genome organization in mammalian cells, with a particular focus on how chromatin organization contributes to transcriptional regulation. I will describe higher-order organizational features that are observed at the level of both the whole chromosome and individual loci. I will highlight changes in genome organization that occur during the course of differentiation, and discuss the functional relationship between chromatin architecture and gene regulation. Taken together, mounting evidence now shows that the genome organization plays an essential role in orchestrating the lineage-specific gene expression programs through modulating long- range interactions between enhancers and target genes.

Disclosures

Ren: Arima Genomics, Inc.: Equity Ownership, Patents & Royalties; Eli Lilly: Employment.
New Concepts in Genome Regulation
Authors: Dean Ann , Lee Jongjoo , Dale Ryan , Krivega Ivan
Published: October 2019

Manipulating gene regulation to favor gamma-globin transcription over beta-globin transcription has been a goal of research in erythropoiesis for decades because of its relevance to amelioration of the pathophysiology of sickle cell disease and beta-thalassemia. A fundamental unanswered question in biology is how the unique pattern of gene expression, the transcriptome, of the many different individual mammalian cell types arises from the same genome blueprint and changes during development and differentiation. There is a growing appreciation that genome organization and the folding of chromosomes is a key determinant of gene transcription. Within this framework, enhancers function to increase the transcription of target genes over long linear distances. To accomplish this, enhancers engage in close physical contact with target promoters through chromosome folding, or looping. These long range interactions are orchestrated by cell type specific proteins and protein complexes that bind to enhancers and promoters and stabilize their interaction with each other. We have been studying LDB1, a member of an erythroid protein complex containing GATA1, TAL1 and LMO2. The LDB1complex activates erythroid genes through occupancy of virtually all erythroid enhancers. LDB1engages in homo- and heterotypic interactions with proteins occupying the promoters of erythroid genes to bring them into proximity with their enhancers. We find that enhancer long range looping activity can be redirected. Both targeting of the beta-globin locus control region to the gamma-globin gene in adult erythroid cells by the tethering of LDB1 or epigenetic unmasking of a silenced gamma-globin gene lead to increased locus control region (LCR)/gamma-globin contact frequency and reduced LCR/beta-globin contact. The outcome of these manipulations is robust, pan-cellular gamma-globin transcription activation with a concomitant reduction in beta-globin transcription. These examples suggest that chromosome looping can be considered a therapeutic target for gene activation or gene silencing to ameliorate genetic diseases such as sickle cell disease and beta-thalassemia.

Disclosures

No relevant conflicts of interest to declare.
The Hemoglobin D Syndromes
Authors: CHERNOFF AMOZ I.
Published: October 2019

The clinical and hematologic manifestation of the Hgb D trait and homozygous Hgb D disease have been presented. Family studies suggest that Hgb D is transmitted in a manner identical to that of Hgb A.
Effect of Indole Compounds on Vitamin B12 Utilization
Authors: DREXLER JERRY
Published: October 2019

Indole and indoleacetic acid are shown to inhibit the growth of a vitamin B12 dependent microorganism, a mutant strain of Escherichia coli. It is suggested that the mechanism of that inhibition may be a competitive inhibition of some enzymatic system necessary for the utilization of vitamin B12.
Studies on Lymphocytic Function
Authors: BRAUNSTEINER H. , PAERTAN J. , THUMB N.
Published: October 2019

Cellular reactions in inflammed areas were observed on coverslips. After an initial granulomonocytic phase a massive infiltration by lymphocytes took place. These lymphocytes rapidly hypertrophy and become "mononuclear," "epitheloid" and even giant cells. By injecting 15 to 30 millions of these cells from tularin- (tuberculin) positive donors to negative recipients, allergy of the delayed type is specifically transferred as with lymphocytes. The function of the lymphocyte is discussed on the basis of these findings.
Clinical Evaluation of 6-Chloropurine in Leukemia of Adults
Authors: ELLISON ROSE RUTH , KARNOFSKY DAVID A. , BURCHENAL JOSEPH H.
Published: October 2019

1. 6-Chloropurine, an analogue of 6-mercaptopurine, has been shown to produce complete remissions in three and partial remissions in eight of 30 adults with acute leukemia. No beneficial effects were obtained in four additional patients known to be resistant to 6-mercaptopurine.

2. Three of four patients with chronic granulocytic leukemia responded to 6-chloropurine.

3. Slight objective improvement was noted in patients with lymphomas but only at the point of hematologic toxicity.

4. A high incidence of jaundice was noted, but it is difficult to relate this definitely to the therapy.

5. While 6-chloropurine appears to be slightly superior to 6-mercaptopurine in acute leukemia in adults, the pattern of response would indicate that these agents have a similar mechanism and range of therapeutic activity.
Familial Neonatal Neutropenia with Maternal Leukocyte Antibodies
Authors: BRAUN E. H. , BUCKWOLD A. E. , EMSON H. E. , RUSSELL A. V.
Published: October 2019

The history of a family with three cases of neonatal neutropenia is reported. During the mother’s last pregnancy leukocyte antibody titers were found to be rising, culminating in a titer of 1:512 at delivery, when leukolysins and passive transfer of antibody to the fetus in the cord blood were also demonstrated. This pregnancy resulted in an infant with a severe degree of neutropenia during its neonatal period. The original sensitization was attributed to prior transfusion with the husband’s whole fresh blood. The whole process can be compared with the sensitization produced by the Rh factors.
HEMOPOIESIS IN RIBOFLAVIN-DEFICIENT RATS
Authors: ENDICOTT K. M. , KORNBERG ARTHUR , OTT MAURINE
Published: October 2019

Leukopenia, granulocytopenia, and, occasionally, anemia develop in rats fed a purified diet deficient in riboflavin. Folic acid (L. casei factor) corrects the leukopenia and granulocytopenia. Riboflavin will prevent all the dyscrasias but will correct only the anemia.

The bone marrow in granulocytopenic rats is hypoplastic and is almost completely depleted of cells of the granulocytic series. Cells of the erythroid series are decreased in number. The myelogram of rats made folic acid deficient by the inclusion of sulfasuxidine in a purified diet resembles this picture and in both cases the response of the marrow and the blood to folic acid therapy is similar.

The bone marrow in riboflavin-deficient rats having both granulocytopenia and anemia is depleted of granulocytic cells but shows an erythroid hyperplasia. This myelogram differs from that seen in sulfasuxidine-induced folic acid deficiency anemia and granulocytopenia in which there is erythroid hypoplasia. The two anemias differ further in that the folic acid deficiency anemia responds to folic acid therapy whereas the riboflavin deficiency anemia responds to riboflavin therapy but not to folic acid therapy.
FURTHER OBSERVATIONS ON THE SPECIFICITY OF THE FOLIC ACID MOLECULE
Authors: SPIES TOM D. , LOPEZ GUILLERMO GARCIA , STONE ROBERT E. , MILANES FERNANDO , BRANDENBURG ROBERT O. , ARAMBURU TOMAS
Published: October 2019

Methyl folic acid,2 N-(4-(4-quinazoline) amino) benzoyl)-glutamic acid, the Mg salt of formyl pteroyl glutamic acid, the Mg salt of formyl pteroic acid, pteroyl aspartic acid, oxyfolic acid and oxypteroic acid have been studied as to their effect on blood regeneration in selected cases of Addisonian pernicious anemia, nutritional macrocytic anemia and tropical sprue. In the amounts administered, only the Mg salt of formyl pteroyl glutamic acid was effective in producing reticulocytosis and an increase in red blood cells, hemoglobin, white blood cells and platelets, and it was not as effective per unit of weight as was folic acid per se. Presumably this compound is slowly changed into folic acid in the body. It is of special interest that the Mg salt of formyl pteroic acid (Streptococcus lactis factor) was negative in producing hemopoiesis. These observations show the very great specificity of the folic acid molecule.
THE ACTIVATION OF PROTHROMBIN, WITH SPECIAL REFERENCE TO "THROMBOPLASTIC ENZYME" (TRYPTASE)
Authors: FERGUSON JOHN H. , TRAVIS BURTON L. , GERHEIM EARL B.
Published: October 2019

A number of high-potency purified prothrombin preparations,61 in 0.2-0.5 per cent solution in borate buffer (pH = 7.7), maintain for days or weeks a stable thrombin-forming ability, whether (a) with buffer alone, (b) with brain thromboplastin suspension, (c) with CaCl2. Nevertheless, they all contain a trace of thrombin and continue to activate "spontaneously" at a very slow rate. Optimal addition of Ca-salt somewhat accelerates this and usually leads to maximal (complete) thrombin formation in 2-11 days at room temperature. Except in a few cases, where ionized Ca++ is demonstrable, thromboplastin, alone, is without effect, but added with calcium, it completes the activation in a matter of minutes or hours depending principally upon the concentration used.

Experimental analysis of the activation process stresses the participation of (1) Ca-ions, (2) thromboplastic P-lipid factor (cephalin), (3) plasma and tissue tryptase (proteolytic enzyme). Each of these factors is studied in detail with reference to mode of action, optimal concentration, side-effects, and relation to inhibitors.

Inhibition of prothrombin activation may be considered under the following heads: (1) "decalcifying" agents (e.g., oxalates, citrates, etc.), which (a) depress Ca-ionization and thus prevent thrombin formation, and (b), under special circumstances, reverse the process of activation; (2) "antithromboplastic" (? anticephalin) agents, (e.g., heparin, and probably soybean trypsin-inhibitor, to some extent); (3) "antitryptase" agents (e.g., crystalline trypsin-inhibitors from pancreas and soybean), which inhibit the thromboplastic enzyme (accessory factor). Excess Ca++ slows rate of thrombin formation.

The evidence suggests that thrombin formation proceeds via an "intermediary" calcium-prothrombin-cephalin (thromboplastic phosphatide) complex or compound. The amounts of (a) thromboplastic P-lipid (cephalin or "thromboplastin") and (b) Ca++ determine both the rate of activation and the final thrombin yield. However, the ultimate ("ripe") thrombin owes none of its activity to the presence of any calcium or phospholipid.

The three types of activator (Ca, thromboplastin, and thromboplastic enzyme) occur as "trace impurities" in prothrombin preparations, but Seegers’ most purified materials are tryptase-free. Tryptase (and trypsin) are "thromboplastic" only in the presence of adequate calcium and phospholipid factors, which may, however, be "mobilized" from protein combination, including prothrombin. In this way the two basic activators are, in a sense, "catalyzed" in their prothrombin-activating reactions.

The significance of proteolytic actions (fibrinolysis, fibrinogenolysis, prothrombinolysis, and sometimes thrombinolysis) by natural tryptase of plasma and tissue origin is investigated and discussed in relation to the broader aspects of the blood-coagulation problem.
ADULT GAUCHER’S DISEASE, WITH SPECIAL REFERENCE TO THE VARIATIONS IN ITS CLINICAL COURSE AND THE VALUE OF STERNAL PUNCTURE AS AN AID TO ITS DIAGNOSIS
Authors: GROEN J. , GARRER A. H.
Published: October 2019

The authors present nine cases of Gaucher’s disease in which the diagnosis, suspected on clinical grounds, was definitely established by the detection of the Gaucher cells in the smear of the sternal marrow. The authors review the varieties of the clinical picture in this disease. They discuss the importance of sternal puncture with special reference to the possibility of establishing the diagnosis in subclinical cases.
THE FREQUENCY OF MEGAKARYOCYTES IN AUTOPSY SECTIONS
Authors: BRILL ROBERT , HALPERN M. M.
Published: October 2019

1. Megakaryocytes can be demonstrated with great frequency in the viscera.

2. Megakaryocytes were present in the lungs in all of 50 autopsies studied.

3. They were next most frequently found in the spleen.

4. The simultaneous occurrence in the various organs, in the absence of extra-medullary hematopoiesis, is roughly related to the concentration in the lungs.

5. Under normal conditions megakaryocytes in small numbers circulate in the blood stream.
EXOGENOUS HEMOCHROMATOSIS RESULTING FROM BLOOD TRANSFUSIONS
Authors: SCHWARTZ STEVEN O. , BLUMENTHAL SUNOLL A.
Published: October 2019

1. Five patients who died as a result of a variety of diseases, all characterized by severe anemia for which numerous transfusions had been given, and all of whom developed features of hemochromatosis are presented.

2. Eight similar cases found in the literature are summarized.

3. It is postulated that the hemochromatosis developing in these patients is the end result of the deposition and subsequently irritating action of the excess amounts of iron in the parenchymatous tissues.

4. The underlying anemia and the not infrequent transfusion reactions are thought to act as predisposing factors for the development of exogenous hemochromatosis.

5. The name Exogenous Hemochromatosis is proposed for this syndrome.

6. The clinical similarities and dissimilarities and the differences in pathogenesis between exogenous and endogenous hemochromatosis are discussed.
THE PLACE OF THE SPLEEN IN THE ENDOCRINE SYSTEM
Authors: DOWNS ARDREY W.
Published: October 2019

While the evidence for an endocrine function of the spleen is meagre and the exact nature of this action is not clear it does seem to be fairly well established that the spleen must be considered as an important part of the mechanism whereby a normal corpuscle, and possibly platelet, content of the circulating blood is maintained.

This regulation seems to be due to the production by the spleen of one or more hormones.

These hormones appear to affect the processes of the bone marrow and also may act upon the corpuscles in the circulating blood.

The conception of hyperactivity of the spleen in the human being seems to be firmly established but little is known of the clinical appearance of its hypoactivity.
PATHOLOGY OF THE RUPTURED SPLEEN IN ACUTE VIVAX MALARIA
Authors: LUBITZ JOSEPH M.
Published: October 2019

1. Opportunity to study the pathologic changes in ruptured spleen of acute vivax malaria was afforded by splenectomy.

2. Subcapsular hematoma precedes rupture.

3. The capsular tear is an obvious consequence of the changes in the spleen occurring in acute malaria. Small hemorrhages occur in the vicinity of the capsule or deep in the tissues.

4. A rapidly enlarging spleen with underlying vascular alteration predisposes to rupture. Minimal trauma may be a precipitating element.

5. Diffuse cellular hyperplasia, subintimal and adventitial leukopoiesis, dilated sinuses and occasional thrombosis and infarction constitute the characteristic pattern in malaria.

6. Changes in the left lung base may serve as a diagnostic aid in cases in which the diagnosis of splenic rupture is considered.
Elevated adenosine deaminase and purine nucleoside phosphorylase activity in peripheral blood null lymphocytes from patients with acquired immune deficiency syndrome
Authors: Murray JL , Loftin KC , Munn CG , Reuben JM , Mansell PW , Hersh EM
Published: October 2019

The purine metabolic enzymes adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) are important in lymphocyte differentiation, and genetic deficiencies of either enzyme have been associated with hereditary immunodeficiency states. Both ADA and PNP activity were measured in null cell-enriched and T cell-enriched peripheral blood lymphocytes from 16 patients with the acquired immune deficiency syndrome (AIDS), seven patients with the AIDS-related symptom complex (ARC), and seven asymptomatic homosexuals. ADA activity in nmol/10(6) lymphocytes/h was significantly elevated in null lymphocytes from AIDS (161 +/- 12) as compared with 23 healthy heterosexual controls (127 +/- 8;P less than .025). PNP activity was also significantly increased in null lymphocytes from AIDS patients (96 +/- 10;P less than .005) as well as those from ARC patients (84 +/- 11:P less than .025) relative to controls (61 +/- 5). No significant differences in enzyme activity were noted in T cell-enriched cells in any group. Along with elevated enzyme activity, AIDS patients had small yet significant increases in the percentages of HLA-DR (P less than .025), terminal deoxynucleotidyl transferase (TdT) (P less than .0001), and peanut agglutinin receptor (P less than .0001) positive lymphocytes in the null fraction compared with controls. TdT-positive cells appeared morphologically as large lymphoblasts with irregular nuclei. The data imply that the cellular immune deficiency in AIDS is not a result of deficiencies in lymphocyte ADA or PNP activity, but is more likely associated with an increase in an immature and/or activated lymphocyte subset.
Neutrophil-specific granule deficiency includes eosinophils
Authors: Rosenberg HF , Gallin JI
Published: October 2019

Neutrophil-specific granule deficiency is a disorder of leukocyte maturation associated with decreased levels of mRNA for a distinct subset of granule proteins. Our work indicates that this disorder, previously thought to be limited to the neutrophil lineage, can also include eosinophils. Immunofluorescence staining led to the discovery of a small but distinct population of peripheral white blood cells containing eosinophil peroxidase (EPO). Unlike normal eosinophils, these EPO+ cells do not have large, eosin-staining cytoplasmic granules, and are indistinguishable from granule-deficient neutrophils by light microscopy. The EPO+ cell lineage did resemble the normal eosinophil lineage in its ability to respond dramatically to granulocyte-macrophage colony-stimulating factor (GM-CSF); the size of the EPO+ peripheral cell population increased approximately 70-fold over baseline in response to GM-CSF administration. The EPO+ cells contained eosinophil Charcot-Leyden crystal protein, but were deficient in three eosinophil-specific granule proteins; neither eosinophil cationic protein, eosinophil-derived neurotoxin, nor major basic protein could be detected in these EPO+ cells, despite the presence of mRNA transcripts for each of the three absent proteins.
Activation of phospholipase D by interleukin-8 in human neutrophils
Authors: Sozzani S , Agwu DE , Ellenburg MD , Locati M , Rieppi M , Rojas A , Mantovani A , McPhail LC
Published: October 2019

Interleukin 8 (IL-8), a member of the C-X-C branch of the chemokine superfamily, stimulated the breakdown of 1-O-[3H]alkyl-2-acyl-sn- glycero-3-phosphocholine ([3H]EAPC) and the formation of 1-O-[3H]alkyl- 2-acyl-phosphatidic acid ([3H]-EAPA) in human polymorphonuclear leukocytes (PMN) in the presence of cytochalasin B. In addition, the mass of diradyl-PA was increased with similar kinetics. In the presence of ethanol, 1-O-[3H]alkyl-2-acyl-phosphatidylethanol ([3H]EAPEt) was formed at the expense of [3H]EAPA formation, indicating the activation of phospholipase D by the cytokine. The effect was time- and concentration-dependent, reaching a plateau at 30 seconds with the maximally activating concentration of 120 nmol/L IL-8. Preincubation of cells with 1 microgram/mL Bordetella pertussis toxin inhibited the breakdown of [3H]EAPC and [3H]EAPA formation, indicating a role for a pertussis toxin-sensitive guanosine triphosphate-binding protein. Formation of phosphatidic acid (PA) correlated with activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, the oxidative burst enzyme, with both events occurring in the same concentration range. Inhibition of PA formation, by the presence of ethanol, also inhibited the oxidative burst stimulation by IL-8. Pretreatment of PMN with 10 nmol/L platelet-activating factor potentiated both [3H]EAPA accumulation and activation of NADPD oxidase by IL-8. Collectively, these data show that IL-8 stimulates the metabolism of choline-containing phosphoglycerides in human PMN and support a role for PA in the signaling mechanisms used by IL-8 to stimulate PMN function.