bluuebunny/crossref_metadata_2025_split huggingface dataset:
Alloimmunisation of Patients Undergoing Bone Marrow Transplantation in the Setting of Leucodepleted Apheresis-Collected Single Donor Platelets.
Authors: Crawford Julie H. , Hughes Annette , Cannell Paul K. , Herrmann Richard 
Published: April 2020

Abstract
Introduction: Immune platelet transfusion refractoriness often leads to requirements for specialised platelet products. Our patients are screened weekly for the presence of platelet antibodies during bone marrow transplantation (BMT) protocols and are routinely issued leucodepleted (LD) packed red cells and apheresis-collected leucodepleted single donor platelets (LDSDP).
Aims: We assessed the alloimmunisation rate in BMT patients, the impact of this on transfusion requirements and addressed the value of continued routine platelet antibody (PAI) screening in the setting of LDSDP.
Results: Routine PAI screens performed on 148 BMT patients over 33-months (January 2002–September 2004) were reviewed; 71 autologous, 55 sibling allogeneic and 22 matched unrelated donor (MUD) BMT patients. Thirteen patients were alloimmunised (antibody specificity anti-HLA (11 patients) and anti-HPA/platelet glycoprotein (2 patients)). The overall alloimmunisation rates were 8.8% (sibling allogeneic 14.5%, MUD 9.1% and autologous 4.2%). Alloimmunisation rates for women were 4.15 times higher than for male patients. Risk factors for secondary alloimmunisation were present in 92.3%. Seven of 9 alloimmunised women had children, 1 of 2 nulliparous women and all 4 male patients had extensive transfusion histories (all 5 were also transfused at other centres where LDSDP were not standard). Alloimmunisation was detected earlier in females, usually on first PAI screen and before transfusion. Alloimmunised patients required specialised platelet products (crossmatched, HLA, directed donations or frozen-thawed SDP). These measures resulted in improved post platelet transfusion increments.
Conclusions: Alloimmunisation continues to occur despite use of leucodepleted transfusion products. Evidence supporting secondary alloimmunisation was present in the great majority of patients with more female patients alloimmunised largely due to their parity. Secondary alloimmunisation is not prevented by transfusion of leucodepleted blood products, however early detection of alloimmunisation assists transfusion decisions. There is indication from this data that platelet antibody could be limited to a PAI screen at presentation (together with HLA typing) in patients who have been transfused or pregnant with a repeat PAI performed following transfusion. Further testing could be delayed until platelet transfusion refractoriness is suspected.
Non-Genotoxic Targeting of p53-Dependent and p53-Independent Signaling Pathways Efficiently Kills Hodgkin/Reed-Sternberg Cells.
Authors: Janz Martin , Stühmer Thorsten , Dörken Bernd , Bargou Ralf 
Published: April 2020

Abstract
Although the majority of patients with classical Hodgkin lymphoma (cHL) can be cured by conventional chemotherapy, a substantial proportion of patients will finally develop treatment-induced secondary malignancies. Therefore, non-genotoxic targeting of major cellular survival pathways could be an interesting novel treatment strategy for patients with cHL. In this study, we focused on the analysis of p53-dependent and p53-independent signaling pathways in Hodgkin/Reed-Sternberg (HRS) cells. To test whether p53 signaling is functional in cHL and whether activation of the p53 pathway is sufficient to kill HRS cells, we employed a recently developed small-molecule antagonist of MDM2, designated nutlin-3a, that disrupts the p53-MDM2 interaction. Nutlin-3a efficiently increased the level of p53 and induced expression of p53 downstream targets in Hodgkin cell lines with wild-type p53, whereas no effects were observed in Hodgkin cell lines that harbor p53 mutations. Activation of the p53 pathway led to strong induction of apoptosis in p53 wild-type Hodgkin cell lines. Knock-down of p53 by RNA interference protected cells from nutlin-induced apoptosis, demonstrating that nutlin-3a exerts its effects strictly through p53. In addition, MDM2 inhibition strongly sensitized HRS cells to cytotoxic drugs, such as doxorubicin, etoposide, or vincristine. In view of the fact that HRS cells are characterized by high constitutive NF-κB activity, we also analyzed the effects of a second non-genotoxic agent, geldanamycin, which is an inhibitor of the HSP90/NF-κB pathway. Titration experiments showed that the pro-apoptotic effects of geldanamycin correlate with the mutation status of IκB proteins, demonstrating strong induction of apoptosis in cell lines with wild-type IκB. Furthermore, Hodgkin cell lines that contain wild-type IκB but lack functional p53 (through mutation or siRNA knock-down) are resistant to nutlin treatment, but still respond to treatment with geldanamycin. This indicates that inhibitors of HSP90 induce apoptosis in HRS cells in a p53-independent manner. Therefore, combined targeting of p53-dependent and p53-independent pathways could be a promising approach to develop highly effective and less genotoxic treatment strategies for patients with cHL.
BIC and miR-155 Are Highly Expressed in Hodgkin, Primary Mediastinal and Diffuse Large B Cell Lymphomas.
Authors: Kluiver Joost , Poppema Sibrand , de Jong Debora , Blokzijl Tjasso , Harms Geert , Jacobs Susan , Kroesen Bart-Jan , van den Berg Anke 
Published: April 2020

Abstract
In a previous study we demonstrated high expression of the non-coding BIC gene in the vast majority of Hodgkin’s lymphomas (HLs). It is now known that BIC is a primary microRNA (pri-miRNA) that can be processed to miRNA-155. We analyzed HL and NHL cell lines and tissue samples to determine miR-155 expression levels. High levels of miR-155 could be demonstrated in all HL tissue samples and most cell lines. Most NHL subtypes were negative for BIC as determined by RNA-ISH. However, 8/8 cases of primary mediastinal B cell lymphoma (PMBL) showed BIC staining in 12% of the tumor cells. QRT-PCR analysis confirmed high relative BIC levels (35). 12/18 cases of DLBCL showed a variable BIC staining with on average 7% of positive cells. Interestingly, activated B cell-like (ABC-like) DLBCL showed on average 12% of tumor cells positive for BIC while germinal centre B cell-like (GCB-like) DLBCL only showed on average 4% of cells positive for BIC. Differences in BIC expression levels were confirmed by qRT-PCR analysis revealing average BIC levels of 22 for ABC-like and 12 for GCB-like DLBCL. Northern blot analysis showed expression of miR-155 in all DLBCL and PMBL derived cell lines and tissue samples analyzed. In summary, we demonstrate expression of pri-miRNA BIC and its derivative miR-155 in HL, PMBL and DLBCL. In DLBCL, BIC expression is most pronounced in the ABC-like phenotype, possibly as a result of NF-κB activation.
Bone Marrow Mast Cells Are Significantly Increased in Patients with Waldenstrom’s Macroglobulinemia, and Their Number Following Therapeutic Intervention Is Dependent on Extent of Response.
Authors: Santos Daniel Ditzel , Chemaly Mariana Z.A. , Tournilhac Olivier , OConnor Kelly E. , Hatjiharissi Evdoxia , Leleu Xavier , Xu Lian , Branagan Andrew R. , Manning Robert J. , Patterson Christopher , Ho Allen W. , Hunter Zachary R. , You James M. , Tai Yu-Tzu , Kutok Jeffery L. , Anderson Kenneth C. , Munshi Nikhil , Treon Steven P. 
Published: April 2020

Abstract
INTRODUCTION: In previous studies, we demonstrated that mast cells (MCs) may support the growth and survival of lymphoplasmacytic cells (LPC) in WM patients through expression of several ligands including CD40L, B-LYS, PDGFa and VEGF. While increased numbers of MC, as well as their association with LPC in the bone marrow (BM) of patients with WM has previously been reported, their enumeration before and following therapeutic intervention remains to be clarified.
METHODOLOGY: We therefore enumerated MC using tryptase staining in whole sections of BM trephine biopsies before and after therapy for 19 patients (median age 59; range 49–85 years) with the consensus panel diagnosis of WM who received rituximab (n=4), rituximab plus fludarabine (n=14) or bortezomib (n=1). Biopsies from 7 non-WM, age matched patients with normal trilineage myelopoiesis and who demonstrated no evidence of disease were used as controls.
RESULTS: At baseline, an increased number of BMMC was observed for WM patients (median 49/mm2; range 0.4–149.5/mm2) versus control patients (median 14/mm2, range 3.8–31.4/mm2); p=0.003. Among non-responding patients, the median number of BMMC rose from 42.76/mm2 (range 23.3–134.6/mm2) to 72.59/mm2 (range 59.1–77.9/mm2, n=4), while it remained stable for minor responders 14.9/mm2 (range 0.4–26.6/mm2) to 15.8/mm2 (range 11.9–44.8/mm2, n=3), and decreased for major responders (CR and PR) from 56.6/mm2 (range 7.1–149.5/mm2) to 34.13/mm2 (range 3.3–110.9/mm2, n=12).
CONCLUSION: These preliminary studies demonstrate that BMMC are significantly increased in patients with WM, and their number following therapeutic intervention is dependent on extent of response.
No Association between Chlamydia Psittaci and Ocular Adnexal Lymphoma in North American Samples.
Authors: Zhang Grace S. , Winter Jane N. , Variakojis Daina , Reich Steven , Lissner Gary S. , Bryar Paul , Regner Mary Ann , Mangold Kathy , Kaul Karen 
Published: April 2020

Abstract
Previous studies linked infection by Chlamydia psittaci to ocular adnexal lymphomas in Italian patients (Ferreri et al., JNCI 2004). Patients treated with doxycycline to eradicate the organism have demonstrated disease regression (Ferreri et al., J. Clin Oncol, 2005). To assess whether Chlamydia psittaci is associated with ocular adnexal lymphomas in a North American population, we studied 24 ocular adnexal lymphoma biopsy specimens from adults, 16 control lymphoma specimens from patients with systemic lymphomas not involving the ocular adnexa and five benign control ocular tissue samples. There were fifteen MALT, five diffuse, large B-cell, three follicular, and one mantle cell lymphoma specimens. The presence of Chlamydia psittaci DNA was investigated by PCR in each group. DNA was isolated from sections of formalin-fixed, paraffin-embedded tissue. Two different assays were utilized: 1) a previously published conventional PCR/gel based assay targeting a 111 bp fragment of the 16S gene [Madico, J. Clin Micro, 2000] and 2) a realtime PCR assay amplifying a 148 bp portion of the 16S gene with detection via a specific fluorescent probe. Amplification was carried out to 60 cycles. Positive controls consisted of isolated DNA from Chlamydia psittaci strain VS1, CP3, and FP. A human DNA internal control was utilized to assess DNA quality and amplification success. Both assays detected Chlamydia psittaci DNA from isolated strains. All patient samples in all categories yielded negative results using both assays. Thus, we could not confirm that Chlamydia psittaci is associated with ocular adnexal lymphomas in our patient population. Differences in the prevalence of Chlamydia psittaci infection in North America and Europe or technical differences in application of the assays may underlie the differences in our findings when compared to the Italian series. Confirmation with larger numbers of specimens is required.
Quantification of ADAMTS-13 Activity and Antigen from the Same Sample by a Newly Developed Combi-Actibind Assay (TECHNOZYM® ADAMTS-13).
Authors: Geiter Sabine , Colle Sebastién , Kyrle Paul A. , Scheiflinger Fritz , Binder Bernd R. , Vetr Helga 
Published: April 2020

Abstract
Defects in activity and/or antigen levels of ADAMTS-13, the von Willebrand Factor (vWF) cleaving protease, are viewed as the main cause inducing the microvascular thrombotic disorder TTP (thrombotic thrombocytopenic purpura) that is in more than 90% of cases fatal if not treated early and appropriately. Malfunction of ADAMTS-13 with respect to cleave multimeric vWF can be caused by auto-antibodies directed against ADAMTS-13, by decreased presence of ADAMTS-13 in the circulation or by defective activity of ADAMTS-13. Therefore rapid and reliable diagnosis of ADAMTS-13 parameters is a clinical need. We present here a new assay for quantification of ADAMTS-13 activity and antigen levels based on ELISA format and thus suitable for fast and routine measurements.
The method is based on the Actibind® technology, using a monoclonal antibody to capture ADAMTS-13 from plasma to a microtiter plate. Activity is then assayed by a fluorescent substrate. After removal of the substrate solution, antigen can be assayed by standard ELISA technology using a polyclonal antibody followed by a peroxidase labelled secondary antibody. Thus both parameters can be measured from the same sample consequently in the same assay allowing calculation of specific activities of ADAMTS-13.
Dilutions of normal plasma (pool of 100 normal donors) were used as calibrators. The standard curve for the activity assay comprises a range from 3.7% to 100 % of normal plasma activity, thus samples of severe TTP (definition: activity below 5%) can be detected with sufficient sensitivity. 42 samples from TTP patients were tested in this new activity/antigen assay as well as in the TECHNOZYM ADAMTS-13 INH assay that allows determination of anti ADAMTS-13 auto antibodies. Using these two assays a significant negative correlation was found between ADAMTS-13 activity and the presence of autoantibodies, 82% of the samples with extremely low ADAMTS-13 activity (&lt;5%) had high levels of autoantibodies. A less stringent positive correlation was found between ADAMTS-13 antigen and ADAMTS-13 activity. Out of 30 samples with low ADAMTS-13 activity (&lt; 25%) 15 samples had antigen levels less than 50% of normal. Out of the 42 samples from TTP patients, 3 samples were found without auto antibodies but still low activity. 2 of these samples also had low antigen levels. Moreover, the TTP samples had significantly higher levels of vWF:CBA activity (p&lt;0.05) consistent with the presence of high molecular weight vWF.
This combined new assay clearly adds to the diagnostic possibilities to differentiate patients with TTP with respect to defects in ADAMTS-13 function and antigen concentration. In addition, the results of this assay correlate with the clinical picture and the decreased functional activity of ADAMTS-13 leading to increased levels of vWF multimers and in turn platelet consumption.
A Pathophysiological Role for ADAMTS13 in Thrombotic Thrombocytopenic Purpura (TTP) without Severe ADAMTS13 Deficiency.
Authors: Hovinga Johanna A. Kremer , George James N. , Vesely Sara K. , Terrell Deirdre R. , Budde Ulrich , Sulzer Irmela , Lammle Bernhard 
Published: April 2020

Abstract
Background: A severe ADAMTS13 deficiency (&lt;5% of the normal) is a specific finding for an acute thrombotic microangiopathy commonly labeled as idiopathic TTP. However, the sensitivity of this finding for the clinical diagnosis of idiopathic TTP is only ~60% (range 33–100% in different studies) and the pathophysiology underlying idiopathic TTP in the absence of severe ADAMTS13 deficiency has not yet been elucidated.
Methods: 74 consecutive patients with acute acquired idiopathic TTP-HUS from the Oklahoma TTP-HUS registry (partly published by Vesely et al. [Blood2003;102:60–8]) were included into this study. For laboratory investigation, serum samples had been withdrawn before initiation of any form of treatment. ADAMTS13 activity was determined by quantitative immunoblotting, ADAMTS13 autoantibodies were evaluated by a routine inhibitor screening test and the TECHNOZYM ADAMTS13 INH ELISA, and patients’ VWF multimers were analyzed by SDS-agarose electrophoresis to assess endogenous VWF proteolysis arisen in vivo.
Results: An ADAMTS13 activity &lt;5% was found in 22 (30%), an ADAMTS13 activity &lt;10% in 28 (38%) patients. The ADAMTS13 INH ELISA gave positive results (≥15 arbitrary antibody U/ml) in 48 (65%) patients as well as in 15/113 (13.3%) controls. Setting a higher threshold of 35 arbitrary antibody U/ml (high titer), 33/74 (44.5%) patients and 3 (2.7%) controls showed positive results for autoantibodies to ADAMTS13. Out of these 33 patients, 28 had a positive inhibitor screening test, 24 had an ADAMTS13 activity &lt;10%, and 25 displayed impaired or severely impaired VWF proteolysis on multimer analysis. In the intermediate antibody titer group (15-&lt;35 U/ml) only 2/15 patients had an ADAMTS13 activity &lt;10%, and 14/15 (93%) displayed normal or even increased VWF proteolysis. Of the remaining 26 (35%) patients without detectable ADAMTS13 autoantibodies, 2 (8%) had an ADAMTS13 activity &lt;10%, and 22 (85%) had normal or even increased VWF proteolysis.
Conclusions: Most patients suffering from acute idiopathic TTP-HUS and high titers of ADAMTS13 autoantibodies had impaired VWF proteolysis in line with severely decreased ADAMTS13 activity. Patients without ADAMTS13 antibodies as well as the majority of patients with intermediate antibody titers demonstrated normal or even increased VWF proteolysis. (Severely) impaired proteolysis of endogenous VWF was observed in 3 patients with high titer anti-ADAMTS13 antibodies and normal ADAMTS13 activity (50–100%). Thus, our data suggest a pathophysiological role of ADAMTS13 in at least some patients despite normal ADAMTS13 activity by standard assay.
The Effect of Tumor Necrosis Factor (TNFα) Inhibition on Platelet Counts in Adults and Children with Persistent ITP.
Authors: Tarantino Michael D. , Stephens Dana M. , Imfeld Karen L. , Young Guy , Williams Shirley A. , Lucas Yvonne L. , Nugent Diane J. 
Published: April 2020

Abstract
Immune Thrombocytopenic Purpura (ITP) is hematologic disorder caused by autoimmune opsonization and premature destruction of platelets. Approximately 20% of children and the majority of adults will have a chronic course of ITP with antibody-mediated destruction of platelets and the inability to clear immune complexes. Pro-inflammatory cytokines IL-2, TNF-α and IFN-γ are secreted following a Th1 response and are elevated in patients with chronic ITP. Recent treatments for chronic ITP in adults and children have focused on quelling dysregulated T and B lymphocyte activity. We conducted a pilot clinical trial of etanercept, a fusion peptibody composed of the soluble TNF-α receptor and the Fc portion of Ig. Hematologic and immunologic responses were measured before, during (blood counts only) and after a 12-week regimen of 0.4mg/kg/dose (max dose 25 mg) of subcutaneous etanercept, given twice weekly. Only fixed-dose prednisone or danazol were allowed as concomitant ITP medications. Cytokine concentrations were measured pre- and post-treatment from plasma and cultured lymphocytes (unstimulated or PHA- [2.5ng/ml] stimulated). Sixteen patients (9 male, 7 female), with chronic ITP (8 post-splenectomy) an initial platelet concentration &lt; 30 × 109/L were enrolled with data evaluable for 15. The mean age of the patients was 31 years (median 32, min/max 7/52 years). One patient withdrew due to dizziness thought related to the study drug. One patient developed autoimmune hemolytic anemia, was diagnosed with Evans syndrome at treatment week 3 and withdrew from the study. Otherwise, no severe or serious related adverse events were observed. Two patients had a complete platelet response (CR, plt count &gt; 150 × 109/L at least 6 weeks post treatment), 6 had a partial response (PR, plt count &gt; 30 × 109/L and double baseline plt count), one had a minimal response (MR, plt count &gt; 30 × 109/L) and 5 had no platelet response (NR). Two of the patients with PR had a platelet count rise that lasted at least until the end of the 8 week post-treatment observation period. The platelet response rate in nonsplenectomized and splenectomized patients was 67% and 28%, respectively, p = 0.1. Concentrations of IL-1β, IL-4, IL-6, IL-10 IFN-γ and TGF-β, before and at the end of the 12-week treatment course, showed marked inter-patient variation and did not predict the platelet count outcome. Both patients that had a CR had a reduction in PHA-stimulated TNF-α of 33 and 30%, respectively. Others with NR to treatment had similar declines in TNF-α, however. Although the precise mechanism of action is unclear, etanercept treatment led to at least a partial platelet response in 50% of patients with persistent ITP and was well-tolerated.
An Unusual Form of Immune Thrombocytopenic Purpura Characertized by a Platelet Activating IgG Antibody.
Authors: Boyle Soames , Alexy Tamas , Rhochanda Leanne , Liebman Howard A. 
Published: April 2020

Abstract
Autoimmune thrombocytopenia (ITP) is classically a disorder of accelerated platelet clearance and ineffective platelet production due to antibodies directed against common platelet membrane glycoproteins. The most common clinical manifestation of severe thrombocytopenia is bleeding. We report an unusual case of ITP in a 49 y.o. female with a 4 year history of thrombocytopenia, venous and arterial thrombosis secondary to a platelet activating IgG antibody.
The patient was referred to one of the authors (HAL) for a second opinion with a diagnosis of ITP and a history of venous and arterial thrombosis. On presentation the patient was noted to have a platelet count of 34 × 109/L, a decreased fibrinogen of 128 mg/dL and D-dimers of greater than 4000 ng/ml. A diagnosis of chronic DIC was made and the patient was treated with therapeutic doses of LMW heparin resulting in an increase in her fibrinogen to 260 mg/dL, reduction in her D-dimers to &lt;500 ng/ml and an increase in her platelet count to 118 × 109/L. An extensive clinical and laboratory evaluation, included screening for occult malignancy, cavernous hemangioma, antiphospholipid antibodies, dysfibrinogenemia, inhibitory antibodies against protein C and S, failed to disclose an etiology for her chronic consumptive coagulopathy. After 2 months of LMWH therapy the patient had a progressive drop in her platelet count to &lt;30 × 109/L without evidence of recurrent coagulopathy. The patient’s thrombocytopenia was refractory to corticosteroids, Rituxan, azathioprine, cyclosporine, but responsive to IVIG. When the patients LMW heparin was stopped, the patient had evidence of a recurrence of her DIC as defined by an increase in her D-dimer and decreased fibrinogen. The patient underwent an open splenectomy which was complicated by portal vein and superior mesenteric vein thrombosis requiring catheter directed fibrinolysis and reinstitution of LMW heparin therapy. Repeated ELISA and functional assays for heparin induced thrombocytopenia were negative. However, the patient’s heat inactivated plasma and serum were shown to induce spontaneous aggregation. Patient IgG was subsequently isolated by staph A chromatography and found to be a potent inducer of platelet aggregation. Platelet specific IgG was further isolated from the patient’s IgG by affinity chromatography on a column of solubilized platelet membrane proteins. The affinity isolated antibody was then analyzed by the Western blotting using solubilized platelet membranes. Patient antibody was found to react with a 50 kD platelet membrane protein which has yet to be fully characterized.
At the present time the patient remains on LMW heparin, but requires frequent infusions of IVIG to maintain platelet counts above 30 × 109/L. We are aware of only one other reported case of a spontaneous platelet activating antibody and no other case characterized by chronic DIC.