bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fluorescence In Situ Hybridization (FISH) Using a Subtelomeric 11q Probe as a New Diagnostic Tool for Congenital Thrombocytopenia Caused by Deletions in 11q.
Authors: Gadzicki Dorothea , Michaelsen Gudrun , Rudolph Cornelia , Steinemann Doris , Happel Christoph M. , Welte Karl , Klein Christoph , Toennies Holger , Lichter Peter , Kontny Udo , Niemeyer Charlotte Marie , Schlegelberger Brigitte 
Published: April 2020

Abstract
Neonatal thrombocytopenia occurs in about 1% of all newborns. Inherited forms like 11q- or Jacobsen syndrome are rare. However, they may remain undetected with karyotyping because the deleted regions in 11q often involve small subtelomeric regions. Here we report on the detection of deletions in 11q in two newborns with normal routine karyotypes who were shown to carry subtelomeric deletions in 11q by means of fluorescence in situ hybridization (FISH) using a subtelomeric 11q probe (Abbott, Diagnostics, Wiesbaden, Germany). Both children showed thrombocytopenia (18.000/μl and 26.000/μl, respectively) and dysmegakaryopoiesis (absence of normal megakaryocytes and presence of micromegakaryocytes) associated with facial dysmorphism, cardiac defects and psychomotoric retardation. In the second case, the mother and the grandmother also showed mild thrombocytopenia. In both patients, FISH analyses on peripheral blood and bone marrow showed the loss of the telomere-associated region of 11q distal of the MLL gene. In the first patient, the deletion of 11q resulted from an unbalanced complex rearrangement with duplication of 11p. As the source of this chromosomal aberration, a paternal pericentric inversion of chromosome 11 was identified. The partial monosomy 11q and the partial trisomy 11p in the first patient were confirmed by comparative genomic hybridization (CGH) analysis. Array/matrix CGH assisted in determining the breakpoints at 11p15.1 and 11q24.1. No structural aberrations of 11q were found in the mother of the second patient, but further investigations are under way. These findings give further evidence that small subtelomeric deletions of 11q and probably mutations of genes located therein cause thrombocytopenia. Since it can be very difficult to detect these deletions by karyotyping, FISH using a subtelomeric 11q probe seems to be an extremely useful new diagnostic tool. This new method should be applied in children with congenital thrombocytopenia, in particular if they have additional complex dysmorphic features.
Are the Pfa-100® Closure Time and the Skin Bleeding Time Useful in the Diagnostic Work-Up of Patients with Bleeding Diathesis? Correlation with the Severity of Bleeding Symptoms and Sensitivity for Known Defects of Primary Hemostasis.
Authors: Podda GianMarco , Bucciarelli Paolo , Lussana Federico , Cattaneo Marco 
Published: April 2020

Abstract
We compared the performance of PFA-100® closure time (CT) and the skin bleeding time (BT) in the diagnostic work-up of 128 consecutive patients referred to our Centre from June 2002 through June 2003 to be screened for bleeding disorders, due to the presence of bleeding symptoms or the casual finding of abnormal screening tests of hemostasis. The correlation with the severity of the bleeding symptoms and the sensitivity of BT and PFA-100® CT for known defects of primary hemostasis were evaluated. All patients underwent a careful medical interview and were assigned a “bleeding score”, based on the number, type, frequency and severity (development of anemia, need for blood transfusion and/or surgical or medical intervention) of bleeding symptoms. In addition, all patients underwent a first-line screening, which included prothrombin time, activated partial thromboplastin time (APTT), BT and PFA-100® CT (with both the collagen-ADP and the collagen-epinephrine cartridges). The search for Von Willebrand disease (VWD), platelet function disorders (PFD), clotting factor defects and abnormalities of fibrinolysis was performed according to the results of the first line screening tests and the severity and type of bleeding history. Seven (6%) patients had type-1 VWD, 12 (9%) PFD, 29 (23%) defects of clotting factors, 18 (14%) had prolongations of the APTT due to abnormalities that are not associated with bleeding (factor XII deficiency and lupus anticoagulant), while in 63 (49%) all tests gave normal results. The sensitivity of PFA-100® for VWD was 86% (both cartridges), for PFD 75% (collagen-epinephrine) and 8% (collagen-ADP). The sensitivity of BT for VWD was 29%, for PFD 33%. The CT of the PFA-100® CT collagen-epinephrine cartridge was mildly prolonged in 38% of patients with clotting factor deficiency. After dividing the patient population in four quartiles of distribution, according to the severity of the bleeding score (null, low, intermediate and severe), only the CT of the PFA-100® CT collagen-epinephrine cartridge showed a progressive and significant prolongation from the first to the fourth quartile (p=0.04). No association of BT and collagen-ADP CT with the severity of bleeding history was found. In conclusion: 1) CT of the PFA-100® collagen-epinephrine cartridge was significantly associated with the severity of the bleeding history; 2) PFA-100® CT showed a better sensitivity than BT for VWD and PFD; 3) at variance with the BT, PFA-100® could help to discriminate between VWD (prolongation of CT of both cartridges) and PFD (prolongation of CT of the collagen-epinephrine cartridge only).
Ex Vivo Blockade of NF-[dappa]B Signaling in Alloreactive T Cells Prevents Murine Graft-Versus-Host Disease (GVHD).
Authors: O’Shaughnessy Matthew J. , Vogtenhuber Christine , Serody Jonathon S. , Sitcheran Raquel , Baldwin Albert S. , Dang Lenny , Jaffee Bruce , Blazar Bruce R. 
Published: April 2020

Abstract
A failure of IL-2 transcription has been associated with tolerance induction. We hypothesized that inhibition of the NF-κB pathway in alloreactive T-cells, which is critical for IL-2 transcription, could lead to alloantigen-specific hyporesponsiveness and prevention of GVHD. PS1145, a potent inhibitor of IκB kinase, and hence NF-κB activation, was added to an MLR culture consisting of CD4+ T-cells and MHC class II-disparate stimulators. Inhibition of NF-κB activity was verified by EMSA and confocal microscopy. Global inhibition of cytokine production and T-cell hyporesponsiveness was observed which persisted after washing T-cells and re-exposure to alloantigen. Responses to non-specific mitogens remained largely intact and alloantigen hyporesponsiveness was reversed by exogenous IL-2. Treatment of T cells and stimulator cells with PS1145 was required for maximal effect. Depletion of CD4+CD25+ cells from the MLR indicated that these cells were not required for tolerance induction in this system. Using an MLR system containing alloreactive and non-alloreactive transgenic T cells indicated that PS1145 treatment increased the rate of T-cell apoptosis selectively in alloreactive cells. Data from each of 4 experiments showed that GVHD in recipients of ex vivo PS1145 treated cells was profoundly inhibited, whereas CD4+ T-cells recovered from a vehicle-treated 7-day MLR were uniformly fatal upon adoptive transfer into sublethally irradiated MHC class II-disparate recipients. Studies addressing non-alloreactive in vivo responses of PS1145 treated T cells will also be presented. Our studies indicate that the NF-κB pathway is a critical regulator of productive alloresponses and provide a novel ex vivo approach to induce alloantigen-specific tolerance as a means of preventing GVHD.
Epitope Mapping during FVIII Inhibitor Elimination with Rituximab Revealed Conformational Epitopes on FVIII and Identifies Potentially Small Molecules for Novel Therapies.
Authors: Königs Christoph , Kessel Christoph , Linde Richard , Klingebiel Thomas , Kreuz Wolfhart 
Published: April 2020

Abstract
Hemophilia A is an X-linked bleeding disorder. Mutations in the Factor VIII (FVIII) gene result in reduced or non-functional expression of FVIII. Patients are commonly treated by substitution of recombinant (rFVIII) or plasma derived (pdFVIII) FVIII preparations. However, a significant number of patients treated develop an antibody response (inhibitors) against substituted FVIII. Various Immune Tolerance Therapies (ITTs) are applied to overcome the FVIII specific immune response.
We recently reported on a novel approach to eliminate the FVIII specific immune response by several rounds of rituximab (Mabthera®) infusions after several failed ITT approaches.
In this study, phage displayed random peptide libraries were screened with sera from a patient before, under and after rituximab treatment. After several rounds of biopanning more than 100 phage clones specific for FVIII-inhibitors were identified and sequenced. One amino acid motif - EVPN - was most common in isolated clones. Computer based analysis identified a conformational epitope in the A2 domain of FVIII. The clones specifically inhibit binding of inhibitor to FVIII. Also sera from other inhibitor patients cross reacts with isolated phage clones. Inhibitor sera also bind to synthetic peptides, which sequences correspond to the peptide insert on isolated phage. Functional studies to target inhibitor specific B cells with isolated peptides are currently being performed.
Specific ligands for FVIII inhibitors were identified. The epitope was mapped to the A2 domain. When inhibitor relapsed after Rituximab treatment, the same epitope was recognized. Small peptides representing the epitope on the phage (mimotopes) bound to inhibitor positive sera and even cross-reactivity with heterologous sera was observed. The inhibitor specific peptide ligands may serve as useful tools to develop novel approaches for inhibitor elimination therapies.
Factor V G1691A and Factor II G20210A Mutations and Clinical Expression of Severe Hemophilia A (&lt; 2%) in Children: Impact on Bleeding Episodes and Joint Damage.
Authors: Nowak-Gottl Ulrike , Escuriola Carmen , Schobess Rosemarie , Duering Christine , Bidlingmaier Christoph , Pollmann Hartmut , Horneff Sylvia , Kurnik Karin 
Published: April 2020

Abstract
It has been recently shown that the first bleeding onset in children with severe hemophilia A (HA) carrying prothrombotic risk factors is significantly later in life than in non-carriers.1 The present multicenter study was performed to determine whether the factor (F) V G1691A or the F II G20210A are associated with decreased annual bleeding episodes (ABE) in 106 pediatric PUP patients with severe HA (Intron 22 58.6%) consecutively admitted to German pediatric hemophilia treatment centers. Treatment was initiated according to the frequency of bleedings, and most patients received on demand therapy with a switch over to prophylactic therapy 3x/week (40–60 IU/kgKG factor VIIII concentrate) when more than three bleedings (range 2–6) had occurred into the same joint (n=49). Prospective median(range) patient follow-up was 14(4–35) years. Heterozygosity of the FV mutation was found in 8 subjects, homozygosity in one, and 5 children carried the FII mutation once combined with protein C-deficiency. Carriers of the FV and FII mutations had significantly fewer ABE than non-carriers (p=0.004). 66 of 106 PUP patients developed at least one target joint with a median(range) Pettersson score of 1(0–12) available in 57 patients clearly dependent on age (p=0.039) as well as ABE (p=0.037). The “Nuss” joint score available in 33 subjects highly correlated with the Pettersson score (p=0.007). Data presented here give evidence that the clinical expression of severe HA in children is influenced by the co-expression of the FV and FII mutation.
Slug Plays an Essential Role in the Radioprotection of Hematopoietic Progenitors In Vivo by Antagonizing p53-Mediated Apoptotic Pathways.
Authors: Wu Wen-Shu , Xu Dong , Heinrichs Stefan , Look A. Thomas 
Published: April 2020

Abstract
An antiapoptotic role for Slug/Snail in mammals was suggested by studies in C. elegans, where CES-1/Scratch, a member of the Slug/Snail superfamily, was found to control the apoptotic death of NSM sister neurons by acting as a transcriptional repressor of EGL-1, a BH3-only proapoptotic protein. Identification of Slug as the target gene of the E2A-HLF oncoprotein in human pro-B leukemia cells led us to demonstrate its antiapoptotic function in IL-3-dependent murine pro-B cells. In contrast to its aberrant expression in pro-B leukemia cells, endogenous Slug is normally expressed in both LT-HSC and ST-HSC, as well as committed progenitors of the myeloid series, but not in pro-B and pro-T cells, implying its function in myelopoiesis. Using Slug−/− mice produced in our laboratory, we showed that these knockouts are much more radiosensitive than Slug+/− and wild-type mice, and that apoptotic cells increase significantly in the hematopoietic progenitor cells of Slug−/− mice as compared to wild-type mice following γ-irradiation, indicating a radioprotective function in vivo. We showed here that although the development of myeloid progenitors is not impaired under steady-state conditions, their repopulation is incomplete γ-irradiated in in Slug−/− mice. We demonstrate further the radiation-induced death of Slug−/− mice is exclusively a result of bone marrow failure with no apparent contribution from systemic injures to other tissues. By two-way bone marrow transplantation, we provide firm evidence that Slug protects mice from γ-irradiation-induced death in a cell-autonomous manner. Interestingly, regenerative capacity of hematopoietic stem cells (HSC) was retained in irradiated Slug−/− mice, which could be rescued by wild-type bone marrow cells after irradiation, indicating that Slug exerts its radioprotective function in myeloid progenitors rather than HSCs. Furthermore, we establish that Slug radioprotects mice by antagonizing downstream of the p53-mediated apoptotic signaling through inhibition of the p53-resposive proapoptotic gene Puma, leading in turn to inhibition of the mitochondria-dependent apoptotic pathway activated by γ-irradiation in myeloid progenitors. More interestingly, we observed that Slug is inducible by γ-irradiation in a p53-dependent manner. Together, our findings implicate a novel Slug-mediated feedback mechanism by which p53 control programmed cell death in myeloid progenitor cells in vivo in response to γ-irradiation.
Treatment of Anemia with Aranesp® (Darbepoetin-alfa) Given Once Every Two Weeks in HIV Seropositive Patients.
Authors: Tulpule Anil , Dharmapala Dharshika , Burian Pamela , Espina Byron M. , Jayne Gerald , George Shelly , Levine Alexandra M. 
Published: April 2020

Abstract
Background: Anemia is common in patients (pts) infected with the Human Immunodeficiency Virus (HIV). In these pts, anemia is associated with diminished quality of life (QoL) and is an independent prognostic factor for mortality. Aranesp® (darbepoetin-alfa) is a recombinant erythropoiesis-stimulating protein with extended serum half-life allowing for less frequent dosing than conventional recombinant erythropoietin. We studied Aranesp® in anemic HIV-positive pts to evaluate safety, hemoglobin (Hgb) response, and quality of life (QoL).
Methods: Entry criteria included HIV infection, Hgb &lt; 11.5 g/dL, and adequate renal and liver function. Patients with anemia due to bleeding, malignancy, B12, folate, or iron deficiency were excluded. Aranesp® 3.0 mg/kg was given sc every 2 weeks (Q2W) for 24 weeks. Treatment response was defined as Hgb increase of ≥ 1.5 g/dL from baseline sustained for ≥ 4 weeks duration at any time during the study. If no response by wk 8, Aranesp® was escalated to 5.0 μg/kg.
Results: Thirteen female and 10 male pts have been enrolled to date. Median age was 43 years (range 27–67), median entry Hgb was 10.6 g/dL (range 8.6–11.5), and median CD4 count was 197 cells/mm3 (range 29–821). Nine (39%) pts had a prior clinical AIDS-defining illness or opportunistic infection, 19 (87%) pts received concurrent HAART therapy; 11 (58%) were receiving a zidovudine- containing regimen with either combivir (n=6) or trizivir (n=5). Aranesp® was well tolerated. The only treatment-associated side effect was mild pain at the injection site. Pts who received ≥ 8 wks of Aranesp® were analyzed (n=20). 17 of the 20 pts (85%) had a Hgb response (median duration 12+ weeks [range 4+ to 22+]). One of these pts required a dose escalation. Evaluation of change in QoL using a Linear Analog Scale in 20 evaluable patients showed a mean improvement in energy level (+23.2 mm), activity level (+19.8 mm) and overall QoL (+15.5 mm) over the first 8 wks of treatment.
Conclusions: In HIV-infected pts with anemia, Aranesp® at a dose of 3.0 mg/kg Q2W is well tolerated, is associated with a Hgb response in the majority of pts, effectively maintains Hgb levels, and improves QoL. Further work is on-going.
Akt Regulates the Activation-Induced Apoptosis of T Cells Mediated by HIV-1 gp120.
Authors: Anand Appakkudal R. , Ganju Ramesh K. 
Published: April 2020

Abstract
Multiple mechanisms contribute to the loss of CD4+ T cells in HIV-1 infected individuals. Activation-induced apoptosis of bystander T cells mediated by HIV-1 gp120 is one of the critical mechanisms leading to T cell loss in AIDS. Clinical studies have shown that T cells in the lymph nodes of HIV-1 infected individuals undergo activation-induced apoptosis. In the present study, we used a model where T cells undergo apoptosis after HIV-1 gp120/CD4 cross-linking in conjunction with CD3/T cell antigen receptor activation. We have shown that treatment with HIV-1 gp120 (10 nM) and anti-gp120 MoAb induces approximately 20–25% apoptosis in Jurkat T cells in the presence of immobilized anti-CD3 antibody. However, the molecular mechanism by which HIV-1 gp120 mediates the apoptosis of T cells is still unclear. We have also examined the role of Akt/Protein kinase B in HIV-1 gp120-induced apoptosis. Akt is a cell survival molecule that has been shown to block cell death. We observed a decrease in Akt phosphorylation upon gp120 treatment of Jurkat T cells and peripheral blood mononuclear cells (PBMCs). In contrast, only CD3 stimulation was shown to increase the phosphorylation of Akt. To further confirm the role of Akt in gp120-induced apoptosis, Jurkat T cells were transfected with HA epitope-tagged wild type Akt, dominant-negative Akt that lacks kinase activity, or with a control vector. The transfected cells were treated with gp120 and apoptosis was evaluated by Annexin-PI staining. The T cells expressing wild type Akt showed reduced gp120 apoptosis as compared to the vector control-expressing cells. Conversely, expression of a dominant-negative mutant of Akt accelerated cell death as compared to the vector control. We then further assessed the role of upstream regulators of Akt, such as PI-3 kinase. In this regard, we have shown that inhibition of PI-3 kinase leads to enhanced gp120-induced apoptosis. At present, we are elucidating downstream effectors of the Akt pathway. Taken together, these studies suggest that Akt plays a key role in HIV-1 gp120-induced apoptosis, and that identification of Akt-mediated signaling pathways may provide novel therapeutic targets to combat immune deficiency in AIDS.
Stanford V (SV) Regimen Versus ABVD for the Treatment of Advanced Hodgkin Lymphoma (HL): Results of a UK NCRI/LTO Randomised Phase II Trial.
Authors: Johnson Peter , Hoskin Peter , Horwich Alan , Hancock Barry , Smith Paul , Stevens Lindsey , Qian Wendy , Sweetenham John William 
Published: April 2020

Abstract
Background The ABVD regimen (doxorubicin, bleomycin, vinblastine, dacarbazine) is widely accepted as standard treatment for patients with advanced HL. There are potential long term cardiac and pulmonary toxicites, particularly in combination with mediastinal radiotherapy. The SV regimen comprises brief duration combination chemotherapy, with relatively low cumulative doses of alkylating agents, doxorubicin and bleomycin, followed by consolidation radiotherapy to sites of bulk disease. High response and disease free survival rates are reported in single institution and multi-centre phase II studies. This randomized phase II trial comparing SV with ABVD was designed to determine 1) the feasibility of the SV regimen; 2) response rate compared with ABVD; and 3) acute toxicity of SV.
Methods Patients with previously untreated advanced HL were randomised to receive SV chemotherapy for 12 weeks or 6 to 8 cycles of ABVD. All patients with bulky disease (mediastinal disease &gt; 1/3 of CTR, nodal masses &gt;5cm, macroscopic splenic nodules on CT scan) received involved field radiotherapy (IFRT). ABVD (days 1 & 15: doxorubicin 25mg/m2, bleomycin 10units/m2, vinblastine 6mg/m2, dacarbazine 375mg/m2), q28. SV (mustine 6mg/m2, wks 1,5,9, doxorubicin 25mg/m2, wks 1,3,5,7,9,11, vinblastine 6mg/m2 wks 1,3,5,7,9,11, prednisone 40mg/m2 alternate days, vincristine 1.4mg/m2 wks 2,4,6,8,10,12, bleomycin 5iu/m2 wks 2,4,6,8,10, 12, etoposide 60 mg/m2 wks 3,7,11).
Results From Mar 1998 to Jan 2002, 150 (74 SV, 76 ABVD) patients were randomised from 15 UK centers. Pre-treatment characteristics were balanced between the two groups: median (range) age 33 (18–67), male 57%, stage II 43%, III 27%, IV 29%, 76% with B symptoms. 97% SV patients completed 12 weeks treatment, 88% patients received 6 or more cycles of ABVD. In both groups, 75% patients received IFRT. Grade 3 or 4 toxicity was rare. 49% SV and 79% ABVD patients required G-CSF. Overall response rate was 86% in SV and 88% in ABVD. With median follow-up of 42 months, 2-year overall survival was 95%, (95% CI=92%–98%). No treatment related death occurred.
Conclusion Stanford V is a potentially less toxic alternative to ABVD. Phase III trial evaluation is underway in the UK.