bluuebunny/crossref_metadata_2025 huggingface dataset:
Remembering Savva Ivanovich Mamontov
Authors: Vasnetsov Viktor
Published: October 2019

At the memorial service held for Savva Mamontov after his death in 1918, Viktor Vasnetsov delivered his reminiscences by way of a eulogy. He describes his relationship with the impresario, some key moments in the history of the Abramtsevo circle and his own artistic life. In so doing, Vasnetsov conveys the deep affection held by artists in the circle for Mamontov, and hints at some of the reasons why his role was so critical and influential in coaxing the group towards their many artistic successes.
“Dreaming of Russia”
Authors: Davydova Olga
Published: October 2019

The National-Romantic trend in Russian Art Nouveau is characterized by a lyrical approach to the past, including imagery from folklore. This tendency is also identifiable within the global development of Art Nouveau, each country expressing its national identity in highly characteristic forms in design and architecture. Art Nouveau coincided with the zenith of Symbolism and, therefore, transmitted both its universal ideas and the unique creative psychology of the individual artist, who often based personal quest upon local traditions and innate cultural memory. This article analyzes the poetics of this style in Russia. The lyrical and mythological approach towards artistic images, influencing design, form, and meaning, is studied through an examination of the works of artists close to the Abramtsevo circle and the innovative experiments of the World of Art group (1898-1904).
“Parallel” Muftiates as the “Third Force” among Spiritual Administrations of Muslims in Russia
Authors: Bekkin Renat
Published: October 2019

Throughout 230-year long history, from Russian Empire, through Soviet Union to present day Russia the institution of muftiate has continuously served as an instrument of state religious policy in relation to Muslims. Guided by various considerations, the authorities have kept the institution itself but did not allow individual muftiates to create a united all-Russian Muslim centre. The article discusses such a phenomenon of 2010s as “parallel” muftiates, designed to become an alternative, “third force” to the existing federal muftiates. The article hypothesizes that one of the main tasks of “parallel” muftiates is to prevent federal muftiates from uniting.
Diminished amyloid-β uptake by mouse microglia upon treatment with quantum dots, silver or cerium oxide nanoparticles: Nanoparticles and amyloid-β uptake by microglia
Authors: Sikorska K , Grądzka I , Sochanowicz B , Presz A , Męczyńska-Wielgosz S , Brzóska K , Kruszewski MK
Published: October 2019

Alzheimer’s disease (AD) is a chronic neurodegenerative disease leading to progressive dementia in elderly people. The disease is characterized, among others, by formation of amyloid- β (A β) polypeptide plaques in the brain. Although etiology of the disease is not fully understood, recent research suggest that nanomaterials may affect AD development. Here, we described the consequences of exposure of mouse BV-2 microglia to silver nanoparticles (AgNPs, 50 µg/mL), cerium oxide nanoparticles (CeO2NPs, 100 µg/mL), and cadmium telluride quantum dots (CdTeQDs, 3 or 10 µg/mL) in the context of its ability to clear A β plaques. The brain microglial cells play an important role in removing A β plaques from the brain. Cell viability and cycle progression were assessed by trypan blue test and propidium iodide binding, respectively. The uptake of A β and NPs was measured by flow cytometry. Secretion of proinflammatory cytokines was measured with the use of cytometric bead array. A β (0.1 μM) did not affect viability, whereas NPs decreased microglia growth by arresting the cells in G1 phase (CdTeQDs) or in S phase (AgNPs and CeO2NPs) of cell cycle. The uptake of A β was significantly reduced in the presence of AgNPs and CeO2NPs. In addition, the least toxic CeO2NPs induced the release of proinflammatory cytokine, tumor necrosis factor α. In summary, each of the NPs tested affected either the microglia phagocytic activity (AgNPs and CeO2NPs) and/or its viability (AgNPs and CdTeQDs) that may favor the occurrence of AD and accelerate its development.
Effectiveness of a multidisciplinary parent training program for children with developmental disabilities: A single-blind randomized waitlist controlled trial
Authors: Leung Cynthia , Lai Cynthia , Lau Dustin , Leung Shirley , Pin Tamis W
Published: October 2019

This study aimed to evaluate the effectiveness of a multidisciplinary parent training program, Promoting Holistic Development of Young Kids (Poly Kids), using a single-blind randomized waitlist controlled design. The participants included 218 parents of children with developmental disabilities (DD) (intervention group = 107 and waitlist control group = 111). The primary outcomes were child learning, expressive language, fine and gross motor skills (based on individual assessment by respective blinded professionals), and parental reports of child behavior problems. The parents in the intervention group reported significantly lower child behavior problems ( d = .34), higher child task motivation ( d = .63), and lower parenting stress post-intervention ( d = .25), while the children in the intervention group produced significantly more words post-intervention ( d = .82). McNemar test results were significant for movement out of the clinical range in child behavior problems and cognitive skills in the intervention group, but not the control group. The results provided initial evidence on the effectiveness of this train-the-trainer program in supporting families with preschool children with DD in terms of child behavior problems, expressive language, cognitive skills, task motivation, and parenting stress.
Autologous correction in patient induced pluripotent stem cell‐endothelial cells to identify a novel pathogenic mutation of hereditary hemorrhagic telangiectasia
Authors: Zhou Fang , Zhao Xiuli , Liu Xiu , Liu Yanyan , Ma Feng , Liu Bao , Yang Jun
Published: October 2019

Hereditary hemorrhagic telangiectasia is a rare disease with autosomal dominant inheritance. More than 80% hereditary hemorrhagic telangiectasia patients carry heterozygous mutations of Endoglin or Activin receptor‐like kinase‐1 genes. Endoglin plays important roles in vasculogenesis and human vascular disease. In this report, we found a novel missense mutation (c.88T > C) of Endoglin gene in a hereditary hemorrhagic telangiectasia 1 patient. Induced pluripotent stem cells of the patient were generated and differentiated into endothelial cells. The hereditary hemorrhagic telangiectasia‐induced pluripotent stem cells have reduced differentiation potential toward vascular endothelial cells and defective angiogenesis with impaired tube formation. Endoplasmic reticulum retention of the mutant Endoglin (Cys30Arg, C30R) causes less functional protein trafficking to cell surface, which contributes to the pathogenesis of hereditary hemorrhagic telangiectasia. Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 genetic correction of the c.88T > C mutation in induced pluripotent stem cells revealed that C30R mutation of Endoglin affects bone morphogenetic protein 9 downstream signaling. By establishing a human induced pluripotent stem cell from hereditary hemorrhagic telangiectasia patient peripheral blood mononuclear cells and autologous correction on mutant hereditary hemorrhagic telangiectasia‐induced pluripotent stem cells, we were able to identify a new disease‐causing mutation, which facilitates us to understand the roles of Endoglin in vascular development and pathogenesis of related vascular diseases.
Maestro tissue factor reaches new hEIGHT
Authors: Pryzdial Edward L. G.
Published: October 2019

In this issue of Blood, Kamikubo et al broaden the already diverse tissue factor (TF) repertoire to include activation of anti–hemophilia A clotting factor VIII (FVIII), which amplifies coagulation initiation.1
AML: exposed and exploited?
Authors: Auletta Jeffery J.
Published: October 2019

In this issue of Blood, Gillissen and colleagues characterize donor-derived cytotoxic antibodies, isolated from allogeneic hematopoietic cell transplant (HSCT) patients with acute myelogenous leukemia (AML) in sustained remission, that targeted the spliceosome U5 snRNP200 complex expressed on the cell membrane of AML blasts. Mechanistically, in vitro antibody-dependent cytotoxicity did not cause leukemia cell apoptosis, but rather destabilization of the cell membrane cytoskeleton and subsequent pore formation, resulting in cellular swelling and extravasation of intracellular contents (oncosis). In addition, in vivo reduction in AML burden using a U5 snRNP200–specific antibody was demonstrated in a murine SCID xenograft model. Collectively, the authors’ work suggests a potential role for donor-derived antibodies in mediating graft-versus-leukemia (GVL) activity following allogeneic HSCT.1
The delectability of platelets to a phagocyte
Authors: Kickler Thomas S.
Published: October 2019

In this issue of Blood, Saris et al show that some HLA antigens, namely, B8, B12, and B35, vary in expression on the platelets of some individuals and that this is a constant variant in these people.1
PMBCL: a molecular diagnosis?
Authors: Lim Megan S.
Published: October 2019

In this issue of Blood, Mottok et al1 demonstrate the utility of a molecular assay that assesses the expression of 58 genes to distinguish primary mediastinal B-cell lymphoma (PMBCL) from diffuse large B-cell lymphoma (DLBCL) by using routinely available formalin-fixed paraffin-embedded tissue (FFPET) biopsies. The results could improve diagnostic accuracy for patients with PMBCL and may have important implications for clinical trial selection and interpretation of clinical outcomes for patients with this rare form of lymphoma.
Hemoglobin E: a balanced polymorphism protective against high parasitemias and thus severe P falciparum malaria
Authors: Chotivanich Kesinee , Udomsangpetch Rachanee , Pattanapanyasat Kovit , Chierakul Wirongrong , Simpson Julie , Looareesuwan Sornchai , White Nicholas
Published: October 2019

Hemoglobin E is very common in parts of Southeast Asia. The possible malaria protective effects of this and other inherited hemoglobin abnormalities prevalent in Thailand were assessed in a mixed erythrocyte invasion assay. In vitro, starting at 1% parasitemia,Plasmodium falciparum preferentially invaded normal (HbAA) compared to abnormal hemoglobin (HbH, AE, EE, HCS, β-thalassemia E) red cells (HRBCs). The median (range) ratio of parasitization of HRBCs (n = 109) compared to the controls of different major blood groups was 0.40 (0.08, 0.98), less than half that of the normal red cells (NRBCs) compared to their controls 0.88 (0.53, 1.4;P = .001). The median (range) parasitemia in the HRBCs was 2% (0.1%-9%) compared to 5.2% (1.2%-16.3%) in the NRBCs (P = .001). The proportion of the RBC population that is susceptible to malaria parasite invasion can be described by a selectivity index (SI; observed number of multiply invaded RBCs/number predicted). The heterozygote AE cells differed markedly from all the other cells tested with invasion restricted to approximately 25% of the RBCs; the median (range) SI was 3.8 (1-15) compared with 0.75 (0.1-0.9) for EE RBCs (P < .01). Despite their microcytosis, AE cells are functionally relatively normal in contrast to the RBCs from the other hemoglobinopathies studied. These findings suggest that HbAE erythrocytes have an unidentified membrane abnormality that renders the majority of the RBC population relatively resistant to invasion by P falciparum. This would not protect from uncomplicated malaria infections but would prevent the development of heavy parasite burdens and is consistent with the “Haldane” hypothesis of heterozygote protection against severe malaria for hemoglobin E.
Kit signaling inhibits the sphingomyelin-ceramide pathway through PLCγ1: implication in stem cell factor radioprotective effect
Authors: Maddens Stéphane , Charruyer Alexandra , Plo Isabelle , Dubreuil Patrice , Berger Stuart , Salles Bernard , Laurent Guy , Jaffrézou Jean-Pierre
Published: October 2019

Previous studies demonstrated that Kit activation confers radioprotection. However, the mechanism by which Kit signaling interferes with cellular response to ionizing radiation (IR) has not been firmly established. Based on the role of the sphingomyelin (SM) cycle apoptotic pathway in IR-induced apoptosis, we hypothesized that one of the Kit signaling components might inhibit IR-induced ceramide production or ceramide-induced apoptosis. Results show that, in both Ba/F3 and 32D murine cell lines transfected with wild-type c-kit, stem cell factor (SCF) stimulation resulted in a significant reduction of IR-induced apoptosis and cytotoxicity, whereas DNA repair remained unaffected. Moreover, SCF stimulation inhibited IR-induced neutral sphingomyelinase (N-SMase) stimulation and ceramide production. The SCF inhibitory effect on SM cycle was not influenced by wortmannin, a phosphoinositide-3 kinase (PI3K) inhibitor. The SCF protective effect was maintained in 32D-KitYF719 cells in which the PI3K/Akt signaling pathway is abolished due to mutation in Kit docking site for PI3K. In contrast, phospholipase C γ (PLCγ) inhibition by U73122 totally restored IR-induced N-SMase stimulation, ceramide production, and apoptosis in Kit-activated cells. Moreover, SCF did not protect 32D-KitYF728 cells (lacking a functional docking site for PLCγ1), from IR-induced SM cycle. Finally, SCF-induced radioprotection of human CD34+ bone marrow cells was also inhibited by U73122. Altogether, these results suggest that SCF radioprotection is due to PLCγ1-dependent negative regulation of IR-induced N-SMase stimulation. Beyond the scope of Kit-expressing cells, it suggests that PLCγ1 status could greatly influence the post-DNA damage cellular response to IR, and perhaps, to other genotoxic agents.
Deterministic regulation of hematopoietic stem cell self-renewal and differentiation
Authors: Müller-Sieburg Christa E. , Cho Rebecca H. , Thoman Marilyn , Adkins Becky , Sieburg Hans B.
Published: October 2019

Most current theories assume that self-renewal and differentiation of hematolymphoid stem cells (HSCs) is randomly regulated by intrinsic and environmental influences. A direct corollary of these tenets is that self-renewal will continuously generate functionally heterogeneous daughter HSCs. Decisions about self-renewal versus commitment are made by individual, single HSCs and, thus, require examination on the clonal level. We followed the behavior of individual, clonally derived HSCs through long-term, serial repopulation experiments. These studies showed that daughter HSCs derived from individual clones were remarkably similar to each other in the extent and kinetics of repopulation. Moreover, daughter HSCs within a clone showed equivalent contributions to the myeloid or lymphoid lineages. Lineage contribution could be followed because of the discovery of a new subset of HSCs that gave rise stably to skewed ratios of myeloid and lymphoid cells. Overall, the data argue that self-renewal does not contribute to the heterogeneity of the adult HSC compartment. Rather, all HSCs in a clone follow a predetermined fate, consistent with the generation-age hypothesis. By extension, this suggests that the self-renewal and differentiation behavior of HSCs in adult bone marrow is more predetermined than previously thought.
Native human blood dendritic cells as potent effectors in antibody-dependent cellular cytotoxicity
Authors: Schmitz Marc , Zhao Senming , Schäkel Knut , Bornhäuser Martin , Ockert Detlef , Rieber Ernst Peter
Published: October 2019

Functional studies on native human dendritic cells (DCs) are hampered by technical difficulties in preparing fresh DCs. Recently, with the help of the monoclonal antibody M-DC8, we succeeded in isolating a major subpopulation of human blood DCs by a one-step immunomagnetic separation procedure. These cells strongly express FcγRIII (CD16) and FcγRII (CD32) and are quite efficient in the antigen-specific activation of naive T cells. Because some Fcγ receptor-bearing cell types are known as effector cells in antibody-dependent cellular cytotoxicity (ADCC), we investigated whether M-DC8+ DCs are capable of effectuating ADCC. In this report we show that freshly prepared M-DC8+ DCs efficiently mediate tumor-directed ADCC and that both types of Fcγ receptors as well as tumor necrosis factor α essentially contribute to the cytotoxic activity. The results provide evidence that, in addition to their pivotal role in primary T-cell activation, a subset of blood DCs displays efficient cytotoxicity in ADCC.
High-dose immunosuppressive therapy for severe systemic sclerosis: initial outcomes
Authors: McSweeney Peter A. , Nash Richard A. , Sullivan Keith M. , Storek Jan , Crofford Leslie J. , Dansey Roger , Mayes Maureen D. , McDonagh Kevin T. , Nelson J. Lee , Gooley Theodore A. , Holmberg Leona A. , Chen C. S. , Wener Mark H. , Ryan Katherine , Sunderhaus Julie , Russell Ken , Rambharose John , Storb Rainer , Furst Daniel E.
Published: October 2019

Systemic sclerosis (SSc) is a multisystem disease of presumed autoimmune pathogenesis for which no proven effective treatment exists. High-dose immunosuppressive therapy (HDIT) has been proposed as an investigational treatment for severe autoimmune diseases. Nineteen patients with poor-prognosis SSc underwent HDIT. The median age was 40 years (range, 23-61 years), the median modified Rodnan skin score (a measure of dermal sclerosis) was 31, and the median DLCO was 57%. Conditioning therapy involved 800 cGy total body irradiation (TBI) (± lung shielding to approximately 200 cGy), 120 mg/kg cyclophosphamide, and 90 mg/kg equine antithymocyte globulin. CD34-selected granulocyte–colony-stimulating factor–mobilized autologous blood stem cells provided hematopoietic rescue. With median follow-up at 14.7 months, the Kaplan-Meier estimated 2-year survival rate was 79%. Three patients died of treatment complications and one of disease progression. Two of the first 8 patients had fatal regimen-related pulmonary injury, a complication not found among 11 subsequent patients who received lung shielding for TBI. Overall, internal organ functions were stable to slightly worse after HDIT, and 4 patients had progressive or nonresponsive disease. As measured by modified Rodnan skin scores and modified health assessment questionnaire disability index (mHAQ-DI) scores, significant disease responses occurred in 12 of 12 patients evaluated at 1 year after HDIT. In conclusion, though important treatment-related toxicities occurred after HDIT for SSc, modifications of initial approaches appear to reduce treatment risks. Responses in skin and mHAQ-DI scores exceed those reported with other therapies, suggesting that HDIT is a promising new therapy for SSc that should be evaluated in prospective randomized studies.
Phase 2 trial of infusional cyclophosphamide, doxorubicin, and etoposide in patients with poor-prognosis, intermediate-grade non-Hodgkin lymphoma: an Eastern Cooperative Oncology Group trial (E3493)
Authors: Sparano Joseph A. , Weller Edie , Nazeer Tipu , Habermann Thomas , Traynor Ann E. , Manalo Jane , Cassileth Peter
Published: October 2019

Preclinical and clinical evidence suggest a potential advantage for infusional therapy in lymphoma. Sixty-two analyzable patients with predominantly intermediate-grade non-Hodgkin lymphoma received cyclophosphamide (200 mg/m2 per day), doxorubicin (12.5 mg/m2 per day), and etoposide (60 mg/m2per day) (CDE) by continuous intravenous infusion for 4 days (96 hours) every 3 weeks for a maximum of 8 cycles. By the age-adjusted International Prognostic Index (IPI), 42% were at high risk and 58% were at high-intermediate risk. Complete response (CR) occurred in 30 (48%) patients (95% confidence interval [CI], 35%, 64%), and partial response occurred in 16 (26%) patients, yielding an overall response rate of 74% (95% CI, 62%, 84%). Failure-free survival (FFS) rates at 1 and 2 years were 55% (95% CI, 43%, 67%) and 50% (95% CI, 38%, 62%), respectively. When comparing the outcome for 62 patients receiving infusional CDE with historical data derived from 927 IPI-matched lymphoma patients using a Cox proportional hazards model, there was a nonsignificant trend favoring CDE in FFS (P = .12) and overall survival (P = .09). Severe or life-threatening toxicity included neutropenia (68%), anemia (57%), thrombocytopenia (44%), and infection (24%). Two patients (3%) died of treatment-related infectious complications. The primary end point of improving 1-year FFS from 55% to 70% was not achieved with infusional CDE given as initial therapy in patients with poor-risk intermediate-grade lymphoma. It is unlikely that infusional therapy as used in this study produces a 25% or greater relative improvement in FFS compared with standard therapy.
Developmental kinetics and lifespan of dendritic cells in mouse lymphoid organs
Authors: Kamath Arun T. , Henri Sandrine , Battye Frank , Tough David F. , Shortman Ken
Published: October 2019

The labeling kinetics of 5 dendritic cell (DC) subtypes within the lymphoid organs of healthy laboratory mice during continuous administration of bromodeoxyuridine (BrdU) was determined to investigate developmental relationships and determine turnover rates. Individual DC subtypes behaved as products of separate developmental streams, at least as far back as their dividing precursors. The rate of labeling varied with the lymphoid organ and the DC subtype. Labeling was faster overall in spleen and mesenteric lymph nodes (LNs) and slower in thymus and skin-draining LNs. The CD8+ DC subtype displayed the most rapid turnover, with a uniformly short (3-day) lifespan in spleen but with distinct short-lived and longer-lived subgroups in thymus. All the skin-derived DCs in LNs showed delayed and slow BrdU labeling, indicating a long overall lifespan; however, this was shown to reflect a long residence time in skin rather than a long-duration presenting antigen in the draining LN. Epidermal-derived Langerhans DCs displayed longer BrdU labeling lag and slower overall turnover than the dermal-derived DCs, and the movement of fluorescent Langerhans DC from skin to LN was slower than that of dermal DCs following skin painting with a fluorescent dye. However, once they arrived in lymphoid organs, all DCs present in healthy, uninfected mice displayed a rapid turnover, and this turnover was even faster after antigenic or microbial product stimulation.
Glucocorticoids and rituximab in vitro: synergistic direct antiproliferative and apoptotic effects
Authors: Rose Andrea L. , Smith Barbara E. , Maloney David G.
Published: October 2019

Rituximab, a chimeric human immunoglobulin G1(IgG1) anti-CD20 monoclonal antibody has been shown to mediate cytotoxicity in malignant B cells via several mechanisms in vitro. These include direct antiproliferative and apoptotic effects, complement-dependent cytotoxicity (CDC), and antibody-dependent cell-mediated cytotoxicity (ADCC). Glucocorticoids (GCs) are often administered in conjunction with rituximab in chemotherapeutic regimens or as premedication to reduce infusion-related symptoms. The effects of GCs on CDC and ADCC, and the direct apoptotic and antiproliferative effects of rituximab are unknown. Therefore, we evaluated these mechanisms in 9 B-cell non-Hodgkin lymphoma (B-NHL) cell lines using rituximab and GCs. Rituximab and dexamethasone induced synergistic growth inhibition in 6 B-NHL cell lines. Dexamethasone and rituximab induced significant G1 arrest in 9 of 9 cell lines. The combination of rituximab and dexamethasone resulted in supra-additive increases in phosphatidylserine exposure and hypodiploid DNA content in 5 and 3 B-NHL cell lines, respectively. CDC and ADCC were neither impaired nor enhanced when dexamethasone and rituximab were administered concurrently. However, preincubation of both effector and tumor cells with dexamethasone reduced specific lysis in ADCC assays in 4 B-NHL cell lines. Preincubation of tumor cell lines with dexamethasone significantly increased cell sensitivity to CDC in 3 B-NHL cell lines. We conclude that the addition of dexamethasone to rituximab results in supra-additive cytotoxicity with respect to its direct antiproliferative and apoptotic effects, induces a cell-dependent increased sensitivity to rituximab-induced CDC, and has minimal negative impact on ADCC when used simultaneously with rituximab.
Protection of CLL B cells by a follicular dendritic cell line is dependent on induction of Mcl-1
Authors: Pedersen Irene M. , Kitada Shinichi , Leoni Lorenzo M. , Zapata Juan M. , Karras James G. , Tsukada Nobuhiro , Kipps Thomas J. , Choi Yong Sung , Bennett Frank , Reed John C.
Published: October 2019

Chronic lymphocytic leukemia (CLL) B cells have defects in apoptosis pathways and therefore accumulate in vivo. However, when removed from the patient and cultured in vitro, these malignant cells rapidly undergo apoptosis. Recent studies suggest that leukemia cell survival is influenced by interactions with nonleukemia cells in the microenvironment of lymph nodes, marrow, and other tissues. To model such cell-cell interactions in vitro, we cultured freshly isolated CLL B cells with a follicular dendritic cell line, HK. CLL B cells cocultured with HK cells were protected from apoptosis, either spontaneous or induced by treatment with anticancer drugs. Protection against spontaneous apoptosis could also be induced by coculturing the CLL B cells with normal dendritic cells (DCs) or with a CD40-ligand (CD154)–expressing fibroblast cell line. Examination of the expression of several apoptosis-regulatory proteins revealed that coculture with HK cells or DCs induced up-regulation of the antiapoptotic Bcl-2 family protein Mcl-1 in CLL B cells, whereas CD40 ligation increased expression of Bcl-XL. Cell-cell contact was required for HK-induced protection, and introducing neutralizing antibodies against various adhesion molecules showed that CD44 was involved in HK-mediated survival, whereas CD40, intercellular adhesion molecule–1 (ICAM-1) and vascular cell adhesion molecule–1 (VCAM-1) were not. Anti-CD44 antibodies also blocked Mcl-1 induction by HK cells. Mcl-1 antisense oligonucleotides reduced leukemia cell expression of Mcl-1, and significantly suppressed HK-induced protection against apoptosis, whereas control oligonucleotides had no effect. Thus, HK cells protect CLL B cells against apoptosis, at least in part through a CD44-dependent mechanism involving up-regulation of Mcl-1, and this mechanism is distinct from that achieved by CD40 ligation. Consequently, the particular antiapoptotic proteins important for CLL survival may vary depending on the microenvironment.
Antiphospholipid Syndrome in Patients with Systemic Lupus Erythematosus Treated by Autologous Hematopoietic Stem Cell Transplantation.
Authors: Statkute Laisvyde , Burt Richard K. , Oyama Yu , Verda Larissa , Krosnjar Nela , Traynor Ann E.
Published: October 2019

The association of antiphospholipid antibodies (APLA) with either vascular thrombosis or fetal morbidity is known as the antiphospholipid syndrome (APS). Since systemic lupus erythematosus (SLE) is frequently associated with APS, we analyzed 44 patients with severe and refractory SLE involved in a larger study of autologous hematopoietic stem cell transplantation (HSCT) utilizing cyclophosphamide and antithymocyte globulin (ATG) conditioning to determine the efficacy of this treatment in eliminating APLA and preventing recurrent thrombi. The prevalence of SLE-related APS in our patient population was 63% (45% met Sapporo criteria for definite APS, and 18% had history of APLA positivity with associated clinical APS features). Nineteen patients with APS had lupus anticoagulant (LA) and/or high titers of anticardiolipin antibodies (ACLA) at the entry into study. Ten patients were positive for LA. Fourteen patients showed elevated titers of ACLA, IgG and/or IgM. When followed for up to 30 months after autologous HSCT, 70% of initially LA-positive patients became and remained negative for anticoagulant. 79% demonstrated normalization of ACLA titers during maximum of 28 months follow-up. 64% of patients refractory to chronic anticoagulation regimen were able to discontinue from anticoagulation therapy at average of 6 months post-transplant and remained thrombotic events-free and in complete SLE remission for up to 78 months after HSCT. There was no treatment-related mortality or severe adverse events, including stem cell engrafment-related hematological complications or overhelming bacterial, fungal or viral infection, in any of the 28 SLE-related APS patient. Autologous hematopoietic stem cell transplantation may be performed safely and appears to be effective therapy for eliminating SLE-related APLA and preventing further thrombotic events.
PKC412, a FLT3 Inhibitor, Effectively Induces Apoptosis of Leukemia Cells with 11q23 Translocation Particularly Having D835 Mutation.
Authors: Takahashi Kazuya , Goi Kumiko , Furuichi Yoshiyuki , Inukai Takeshi , Sugita Kanji , Nakazawa Shinpei
Published: October 2019

About 30% cases of acute myeloid leukemia (AML) show FLT3 mutations such as internal tandem duplications within the juxtamembrane domain and point mutations within the kinase domain (D835 et al), which result in constitutive activation of FLT3 kinase activity. Although it was recently shown that acute lymphoblastic leukemia (ALL) with 11q23 translocation, often found in infantile leukemia with very poor prognosis, express FLT3 at high levels, few studies have been performed regarding the biological implication of FLT3 in leukemia with 11q23 translocation. We recently found that 2 of 6 cell lines with 11q23 translocation had D835 mutation in FLT3 (KOCL-33 with 11; 19 translocation and B-precursor phenotype, and KOCL-48 with 4; 11 translocation and monocytoid phenotype). In the present study, we evaluated the in vitro inhibitory effects of PKC412 (a kinase inhibitor of FLT3) using leukemic cell lines with or without D835 mutation. 3H-thymidine uptakes were inhibited by PKC412 at 50–200 nM very markedly in two 11q23-cell lines with D835 mutation (>90% inhibition), moderately (50–80% inhibition) in 11q23-cell lines without FLT3 mutation (n=5), but only modestly (<50% inhibition) in other B-precursor cell lines without FLT3 mutation (n=5). Two 11q23-cell lines with D835 mutation were effectively induced into apoptosis by PKC412, which was preceded by a prompt disappearance of constitutive phosphorylation of STAT5 before PKC412 treatment. The RNase protection assay revealed a decrease in mRNA of Bfl-1 which is known to be up-regulated by the NFkB activity, but showed no changes in mRNA of apoptosis-related molecules such as Bcl-2, Bcl-xl, Bad, Fas, FasL, and TRAIL. These results suggest that PKC412 is a potentially useful agent for the treatment of leukemia with 11q23 translocation, particularly having FLT3 mutation.
The Expression of Human Leukocyte Formin Protein, a New Protein That Associates with AKT, Is Correlated with the Expression of ZAP-70 in CLL.
Authors: Favaro Patricia M.B. , Medina Samuel S. , Traina Fabiola , Oliveira Gislaine B. , Metze Irene L. , Costa Fernando F. , Saad Sara T.O.
Published: October 2019

Recently, we have cloned a new human gene (GenBank Accession No. AY278319), belonging to the formin family. This new gene, called for us human leukocyte formin, presents the common domains found in formin-related proteins: FH1, FH2 and FH3 domains. Western Blotting analysis has demonstrated that the protein encoded by this gene is overexpressed in lymphoid malignancies and cancer cell lines. Based on this pattern of expression, our objective was to investigate the expression of human leukocyte formin protein, by Western blotting analysis, in mononuclear cells from chronic lymphocytic leukemia (CLL) patients, isolated on a Ficoll-Hypaque gradient. We studied 18 CLL patients with median age of 65 y.o. (range, 45 to 86) out of treatment for at least three months (Rai 0 n=8; Rai 1 n=6; Rai 2 n=1; Rai3 n=1; Rai 4 n=2). As normal control we used 6 blood donors. Our data showed an overexpression of the human leukocyte formin in the CLL group when compared with the control group (p= 0.0354), as well as a positive correlation of this protein and ZAP-70 in the CLL group (Spearman test, p= 0.0107). The expression of ZAP-70 has been associated with rapid progression and poor survival and can be used as a prognostic marker. Previously we had described that human leukocyte formin protein associates with Akt, a critical survival regulator in many different cell types. The association was observed in a protein extract of Jurkat cell line and in peripheral blood leukocytes from CLL patients. In an attempt to confirm the association between Akt and human leukocyte formin, we performed cotransfections in 293 cells using an expression vector pEGFP containing FH2 or FH3 domains, and an expression vector of pCMV-HA containing the full length of Akt. The results showed that both FH2 and FH3 domains are involved in the association with Akt. The correlation of human leukocyte formin and ZAP-70 expression, and the association of human leukocyte formin protein with Akt suggest that this new protein may be involved in the signaling pathway of leukemia cell survival and is possibly a new prognostic marker.
PCR-Based Liposomal Amphotericin B Treatment Following Allogeneic Stem Cell Transplantation Is a Safe Treatment Strategy: Preliminary Results of a Prospective Study.
Authors: Hebart Holger , Klingspor L. , Klingebiehl Thomas , Loeffler Juergen , Tollemar J. , Ljungman Per , Wandt Hannes , Dornbusch H.J. , Schumacher Ulrike , Meisner Christoph , Engel C. , Kanz Lothar , Ringden Ole , Einsele Hermann
Published: October 2019

A prospective randomized multi-center trial was initiated comparing PCR-based preemptive and empirical antifungal therapy with AmBisome in patients after allogeneic stem cell transplantation.

A total of 408 patients were randomized at the time of transplant to the antifungal treatment strategy. PCR screening was planned twice weekly during stay in hospital and once weekly after discharge until day 100 post-transplantation. Antifungal prophylaxis consisted of up to 200 mg fluconazole/day. Antifungal therapy was initiated in the PCR group in PCR+ patients with signs of infection and patients with 2 consecutive positive PCR results, and in the empirical treatment group (group B) after 120 hours of febrile neutropenia not responding to broad-spectrum antibacterial therapy. Antifungal treatment consisted of AmBisome at 3 mg/kg for 3 days (loading dose) followed by AmBisome at 1 mg/kg in clinically stable patients, AmBisome at 3 mg/kg was continued in clinically unstable patients. Treatment was stopped according to pre-defined study rules.

The two arms were well balanced with regard to age, gender, donor type, underlying disease and stage, and conditioning regimens applied. Out of 403 evaluable patients 196 were randomized to group A and 207 to group B. AmBisome therapy was initiated in 109 patients in group A and in 76 patients in group B (P<0.05). Eleven patients in group A and 16 patients in group B developed proven invasive fungal infections (IFI), overall mortality at 100 and 180 days was not different. Survival curves demonstrated a lower mortality until day 30 post-transplantation for patients receiving PCR-based antifungal therapy with AmBisome (deaths in group A, n=4; group B, n=13; P=0.03). During this early time period when regular PCR-screening was performed, only 1 patient in group A (candidiasis) and 5 patients in group B (invasive aspergillosis, n=1; candidiasis, n=4) succumbed to IFI (P=0.1).

This study prospectively compared PCR-based versus empirical antifungal therapy with AmBisome after allogeneic SCT. A reduction in early mortality and a trend for a lower rate of proven invasive fungal infections was documented until day 30 post-transplantation indicating that close fungal PCR monitoring allows to stratify patients at high risk for the subsequent development of invasive fungal infections. Survival at day 100 and 180 post-transplantation was not different.
Effect of Polymorphisms in Folate Related Genes on Side Effects of High Dose MTX Treatment in Childhood Acute Lymphoblastic Leukemia.
Authors: Tissing Wim , Huang Lisha , de Jonge Robert , Pieters Rob
Published: October 2019

Introduction. High dose (2 – 3 g/m2) methotrexate (MTX, an anti-folate) is used in the treatment of pediatric acute lymphoblastic leukemia (ALL) with side effects in a considerable number of children. We studied the prevalence of MTX related side effects and a possible relation with common polymorphisms in folate metabolism related genes in the treatment of childhood ALL.

Methods. In this retrospective study, DNA was isolated from leukemic samples of 81 children with ALL. High risk (HR) ALL patients were treated with 4 courses of 3 g/m2, non-high risk (NHR) patients with 3 times 2g/m2. The following effects were monitored: serum MTX levels, transfusion need, liver toxicity, skin toxicity and hematological toxicity. Common gene mutations in methylenetetrahydrofolate reductase (MTHFR 677C>T and MTHFR 1298A>C), methionine synthase (MS 2756A>G), methionine synthase reductase (MTRR 66A>G), serine hydroxymethyl transferase (SHMT 1420C>T), methylenetetrahydrofolate dehydrogenase (MTHFD1 1958G>A), thymidylate synthase (TS 2R3R) and reduced folate carrier (RFC 80G>A) were detected by PCR-RFLP.

Results. 57% of the patients had prolonged increased serum levels of MTX in at least 1 course. 59% patients had the MTX courses at the scheduled time, 35% had one course postponed and 6% 2 courses. Mucositis was observed in 22% patients after 1 out of 3 courses, 19% after 2 and 20% after 3 courses. 28% patients had 1 transfusion, 6% had 2 and 1% had 3 transfusions during the MTX courses. Transaminases were elevated 2 times above the normal upper level in 25% of the patients. Skin rash was observed in 32% patients.

Using Mann Whitney analysis we found a significant relation between polymorphisms of the MTHFR 1298A>C and for the MTRR 66A>G genes and side effects. The MTHFR 1298A>C polymorphism was related to less transfusions (p<0.05). Recovery of WBC count in HR ALL patients tended to be slower in patients with a MTHFR 1298AA genotype (P=0.053). HR patients with a MTRR 66AG genotype had a slower recovery of platelet count (p=0.004)

Conclusion. Common polymorphisms in the MTHFR (677C>T), MS, SHMT, TS, RFC, and MTHFD1 genes are not related to the occurrence of side effects of MTX treatment in childhood ALL. Polymorphisms in the MTHFR (1298A>C) and MTRR (66A>G) gene are related to hematological toxicity.
B-Aggressive Lymphoma Gene (BAL) Is a Risk-Related, γIFN-Inducible Gene That Is Expressed in Primary Diffuse Large B-Cell Lymphomas with a Brisk Host Inflammatory Response.
Authors: Juszczynski Przemyslaw , Kutok Jeffery L. , Aguiar Ricardo C.T. , Mitra Joydeep , Shipp Margaret A.
Published: October 2019

Although diffuse large B-cell lymphoma (DLBCL) is potentially curable with current therapy, a significant number of DLBCL patients still die of their disease. In a broad-based screen for genes and pathways associated with poor DLBCL outcome, we previously identified a novel risk-related gene, termed BAL (B-aggressive lymphoma). Thereafter, we cloned and characterized a major BAL partner protein, BBAP (B-aggressive lymphoma and BAL binding partner), described the nuclear trafficking patterns of both proteins and demonstrated that BAL functions as a modulator of transcription. In the current study, we characterized BAL expression in normal tonsil and primary DLBCLs and defined BAL regulation and potential functions in both cell types. Immunohistochemical staining of normal tonsil revealed that BAL was expressed by small numbers of germinal center (GC) cells and interfollicular cells with the morphologic features of centroblasts (with the GC) and immunoblasts (within the interfollicular areas). In primary DLBCLs, BAL was expressed by the malignant B cells. To gain insights regarding the regulation of BAL and BBAP expression in DLBCLs, we analyzed a series of 176 primary tumor biopsies transcriptionally profiled an U133A/B Affymetrix microarrays. BAL/BBAP high-expressing DLBCLs had evidence of a brisk host immune response, including increased expression of T/NK-cell receptor and activation pathway components, complement cascade members, macrophage/dendritic cells markers and γIFN-induced transcripts, raising the possibility that BAL was induced by γIFN. Consistent with this hypothesis, γIFN treatment of DLBCL cell lines with low basal levels of BAL markedly increased BAL expression. In silico analysis revealed that the BAL and BBAP genes are located in 3q21 in head-to-head orientation and share the same CpG-related promoter. This shared promoter contains a conserved γIFN-responsive module composed of IRF and STAT binding elements. The BAL/BBAP bidirectional promoter was cloned into a pGL3 luciferase promoterless reporter vector and found to increase luciferase activity > 20-fold following γIFN stimulation. To gain further insights into regulation of BAL transcription, mutant versions of BAL promoter were generated and cloned. Luciferase assays demonstrated that the IRF binding site was necessary for γIFN-induced BAL transcription, whereas the STAT1 binding site had an accessory role. Taken together, these results demonstrate that BAL and BBAP genes are transcriptionally activated by γIFN in DLBCLs with features of a brisk host immune response including γIFN signaling. Preliminary studies suggest that BAL limits the efficacy of the observed host inflammatory response.
Response of the Thrombocytopenia Associated with Dengue Hemorrhagic Fever to WinRho® SDF: A Case Series.
Authors: De Castro Reynaldo Angelo C. , Dixit Jitendra R. , Genereux Maurice G.
Published: October 2019

Background: Dengue Fever (DF) illness caused by any of the four serotypes of dengue virus (flavivirus) is currently the most important mosquito borne viral disease in the tropical areas of the world. Dengue hemorrhagic fever (DHF) is a potentially lethal complication of DF.

An estimated 500,000 cases of DHF require hospitalisation each year, of which a very large proportion are children. The case fatality rate varies from 1 to 5% and can exceed 20% without proper treatment.

Thrombocytopenia in DHF: Thrombocytopenia is a constant finding in DHF/Dengue Shock Syndrome (DSS). The platelet count usually decreases to below 100,000 /mm3, 1–2 days before defervescence and remains low for 3–5 days in most cases. The cause of thrombocytopenia is not clearly understood and may be due to decreased platelet production, increased peripheral destruction, or both.

Method: We report a case series in 19 patients from different hospitals around Manila. Pediatric patients who fulfilled the WHO criteria of DHF: high fever, bleeding, positive tourniquet test, moderate to severe thrombocytopenia (<100,000 /mm3) and hemoconcentration (Hematocrit increase> 20%) were included in the case study after voluntary, written, informed consent was provided by parent or proxy.

All subjects meeting the eligibility criteria received a single dose of WinRho® SDF which was administered at a dose of 20 μg/kg over 15 minutes. WinRho® SDF was administered when the platelet count dropped below 100,000 /mm3. Platelet counts were measured every 24 hours.

Findings: All 19 patients showed an increase in the platelet count after administration of WinRho® SDF. Figure 1 summarizes the average platelet count for the 19 patients before and after treatment with WinRho® SDF. Day 0 indicates the date of administration of WinRho® SDF. 48 hours after administration of WinRho® SDF, the platelet count returned to normal (> 150,000) in all patients. All 19 patients also showed clinical improvement and were discharged from hospital within 96 hours. There were no serious adverse events observed in any patient.

Fig 1: Change in average platelet count before and after treatment with WinRho® SDF. Fig 1:. Change in average platelet count before and after treatment with WinRho® SDF.

Conclusion:

All 19 patients showed a rapid increase in the platelet count after administration of WinRho® SDF. 48 hours after administration of WinRho® SDF, the platelet count returned to normal in all patients. All the patients showed clinical improvement and were discharged from hospital within 96 hours.

WinRho® SDF is currently licensed to treat immune thrombocytopenia purpura (ITP) in pediatric or adult patients. Unlike other disease states which may produce ITP, the thrombocytopenia associated with DF has an acute onset and a high mortality rate if untreated. The response observed in these patients demonstrates that the thrombocytopenia associated with DF responds rapidly to treatment with WinRho® SDF.
In Vivo Expansion of Transduced Human Erythroid Cells Using an Mpl-Based Cell Growth Switch.
Authors: Nagasawa Yasuo , Wood Brent , Nourigat Cynthia L. , Papayannopoulou Thalia , Blau C. Anthony
Published: October 2019

Cell growth switches are engineered signaling molecules that can trigger cell growth in response to artificial ligands such as chemical inducers of dimerization (CIDs). We have previously shown that a cell growth switch comprised of the intracellular portion of the thrombopoietin receptor, Mpl, allows for the CID-dependent, in vivo expansion of genetically modified primary hematopoietic cells in mouse and dog models. Here we report the application of this approach to the in vivo expansion of genetically modified primary human hematopoietic cells using an immune deficient mouse model. A lentivirus vector encoding a CID-activatible deriviative of Mpl (F36VMpl) and a green fluorescent protein (GFP) reporter was used to transduce human cord blood CD34+ cells. Transduced human cord blood CD34+ cells expanded 347–495 fold in cultures containing no added growth factors and 100 nM of AP20187. We proceeded to test CID-responsiveness following transplantation into NOD-SCID-beta 2 microglobulin null mice. Since significant human red cell engraftment persists for only a few weeks post transplantation in this model, mice were evaluated at 3 weeks post transplantation, following a 2 week course of treatment with either CID (AP20187 10 mg/kg/day) or control vehicle alone (without CID). In 2 experiments totalling 42 mice, CID-administration resulted in a significant rise in GFP positive human cells, with the predominant response occuring among human erythroid cells (exp.1: 4.0 fold, P = 0.0003, exp.2: 12.7 fold, P = 0.038). An increase in transduced human erythroid progenitor cells was observed in the spleens (but not the femurs) of CID treated mice. Effects on other hematopoietic lineages were minor and variable. The effect of CID treatment was no longer evident five weeks after the last dose. The restriction of CID-induced cell growth to the erythroid lineage may make this approach well suited for gene therapy applications in sickle cell anemia and beta thalassemia.