bluuebunny/crossref_metadata_2025_split huggingface dataset:
A TACI Mutation in a Patient with Autoimmune Lymphoproliferative Syndrome.
Authors: Campagnoli Maria Francesca , Garelli Emanuela , Baravalle Valentina , Carando Adriana , Alliaudi Carla , Quarello Paola , Ramenghi Ugo 
Published: October 2019

Abstract
The autoimmune lymphoproliferative syndrome (ALPS) is an inherited disorder of lymphocyte apoptosis caused by defects in the Fas apoptotic pathway. It is characterized by the association of chronic lymphoid accumulation and autoimmune manifestations. In a subgroup of patients the disease progresses through antibody deficiency and its clinical and laboratory features overlap those of common variable immunodeficiency (CVID). Some CVID cases are associated with mutations in gene TNFRSF13B, encoding TACI. Aim of this study was to determine whether TNFRSF13B mutations are also associated with ALPS. Methods. Genomic DNA from 31 ALPS patients was extracted from PBMCs after informed consent. Exons and intron-exon boundaries of TNFRSF13B gene were amplified by PCR and sequenced on an ABI PRISM 310 Genetic Analyzer. Results. A T>A transition at nt 275, that determined the non-conservative substitution of Isoleucine with Asparagine (I92N), was found in exon 3 from one patient. The mutation involved the second extracellular cysteine rich domain, which is critical for BAFF binding. It was not found in 100 control chromosomes. The patient showed a classical ALPS phenotype with lymphadenopathy, massive splenomegaly that required splenectomy, hepatomegaly and immune cytopenias (neutropenia and immune thrombocytopenic purpura); he had alopecia and high ANA levels. Serum immunoglobulins were normal (IgG titers at the lower level of the range); lymphocytes showed defective Fas-induced apoptosis. Discussion. TACI mutations might contribute to the ALPS phenotype in some cases. Our data confirm the relationships between ALPS and CVID and provides a possible molecular explanation for overlapping phenotypes.
Adult Langerhans’ Cell Histiocitosis (LCH). A Single Institution Experience.
Authors: Fayad Luis , Cannata Jimena 
Published: October 2019

Abstract
Langerhans’ cell histiocytosis (LCH) is a rare clonal disorder that affects mainly children. Clinical presentation as well as behaviour are heterogeneous and can range from solitary, relatively benign lesions to a fatal disseminated disease. Although the features have been well described in children, they remain poorly defined in adults. We report the clinical characteristics and outcome of 30 adults from a single institution.
Clinical presentation: From 1957 to 2003, 30 patients were diagnosed with LCH at MDA. Median age at presentation was 37 years-old, with 60% of being male. Depending on the organs involved and according to the pediatric classification, 17 cases (57%) had a limited form of LCH (single-system) and 13 cases (43%) had disseminated disease (multisystemic). Patients with single-system LCH had mainly bone (76%) and skin (24%) involvement, which was localized to a single site in 10 cases (58%) and multifocal in the remaining. Patients with limited disease were younger (median age 30 yo). Patients with multisystemic LCH were subdivided into a “high risk” category (5 patients) if lung, liver, spleen, or bone marrow was involved, or into a “low risk” category (8 cases), if none of the risk organs was affected. Six patients had pulmonary involvement, typically as bilateral small nodules although diffuse progressive infiltrates were also present in 2 cases.
Clinical course: Patients with limited disease were mainly treated with steroids, radiation, or immunomodulatory agents while those with multifocal or multi-systemic disease had a more agressive behaviour and were treated with systemic therapy. Patients with pulmonary nodules usually responded to systemic steroids and stop of smoking habit. Median of 3 treatments were given (range 1 to 7). Sixty-one percent of patients had a 30-year overall survival (OS). Single-system LCH had a more indolent course (79% OS at 30 years) than multi-systemic LCH (median OS of 16 years). Risk organ involvement was the most important adverse prognostic factor (median OS of 2.2 years vs. 65% OS at 30 years, p: 0.007).
Figure 1. Global overall survival on LCH patients Figure 1. Global overall survival on LCH patients Figure 2. Overall survival according to the extent of the disease Figure 2. Overall survival according to the extent of the disease
Extended TNF Genotyping Identifies TNF -857T as a Risk Factor for Acute Graft-Versus-Host Disease Following Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Mullighan Charles , Heatley Sue L. , Villalta Natalie R. , Doherty Kathleen V. , Danner Silke , Hahn Uwe , Bradstock Kenneth , Schwarer Anthony P. , Szer Jeffrey , Bardy Peter G. 
Published: October 2019

Abstract
Extensive evidence supports a role for non-HLA immunogenetic polymorphisms in determining risk and severity of acute GVHD following allogeneic hematopoietic stem cell transplantation (HCT). Tumor necrosis factor (TNF) polymorphisms have been frequently associated with GVHD risk. However, TNF is highly polymorphic and the TNF polymorphism(s) most strongly associated with GVHD remain to be determined. We previously studied three TNF single nucleotide polymorphisms (SNPs; −308, −238 and 488) in a retrospective study of 160 myeloablative HLA-matched related allo-HCTs and found a highly significant association between the intronic TNF 488A allele and risk and severity of acute GVHD and early mortality (Transplantation 2004;27:587). It was unclear if this association was primarily with TNF 488A or secondary to other functionally important TNF alleles in linkage disequilibrium. To clarify the importance of TNF variants in GVHD, we performed extended genotyping of TNF promoter SNPs in the previously described cohort, and in a prospective cohort of myeloablative (n=78) and non-myeloablative (n=56) sibling HLA-matched allo-HCT. Six TNF polymorphisms (−1030, −863, −857, −308, −238, 488) were genotyped in donors and recipients by PCR-SSP. TNF −308/−238/488 haplotypes were directly amplified by PCR, and full TNF haplotypes inferred using Haploview. The incidence of acute GVHD was 53% and 45.8% for myeloablative and non-myeloablative transplants, respectively. Nine TNF haplotypes were identified, and the TNF 488A allele was in strong, but not complete linkage disequilibrium with TNF −857T. TNF −857T was more strongly associated with acute GVHD (OR 11.1) and grade II–IV GVHD (OR 12.3) than 488A (acute GVHD OR 8.6 and grade II–IV GVHD OR 8.9) in the first cohort. These alleles were also associated with acute GVHD in the second cohort, but only in recipients receiving myeloablative conditioning (all acute GVHD, TNF 488A OR 2.9, −857T OR 3.4). Non-significant trends were observed with grade II–IV GVHD, possibly reflecting the reduced frequency of this complication in the later cohort (29% v. 48%). No association or trends were seen in non-myeloablative transplants. Moreover, no associations were identified with the extensively studied −308 or −238 SNPs in either cohort. Together, these data confirm the association between TNF polymorphisms and GVHD in myeloablative transplants, and suggest that −857T, which lies in closer proximity to TNF regulatory regions than 488A, may be the most useful marker for risk prediction. Our results also suggest that the immunogenetic determinants of GVHD vary according to the intensity of conditioning. Further work examining non-myeloablative transplants is required.
Regulation of Phosphatidylserine Exposure by AMP-Dependent Kinase in Human Platelets.
Authors: Verhoeven Arthur J. , Dekkers Dave W.C. , De Cuyper Iris , Gouwerok Eric , De Korte Dirk 
Published: October 2019

Abstract
Introduction: One of the properties of platelets is the ability, upon activation, to expose phosphatidylserine (PS), providing a negative surface necessary for the docking of the blood coagulation complexes. During storage, PS exposure should be kept to a minimum to achieve optimal transfusion recoveries. Many events underlying PS exposure are still unknown despite intensive research over the last 20 years. The involvement of a phosphorylation event has been suggested and we investigated the involvement of AMP-dependent kinase (AMPK) as a candidate involved in the cascade leading to PS exposure.
Methods: Platelets were washed free of plasma by three consecutive wash steps in substrate-free medium, preventing aggregation during the first two steps by acidification of the platelets with acid citrate dextrose (ACD). Platelets were subsequently incubated with different metabolic substrates for 18 h. The nucleotide levels in the cytosolic fractions of the platelets were analyzed by HPLC, PS exposure via flow cytometry using AnnexinV-FITC as label and phosphorylated AMPK (reflecting activation of AMPK) was determined by immunoblotting using antibodies against phosphorylated AMPK (P-AMPK).
Results: When glucose was present, hardly any phosphorylation of AMPK was observed. Under substrate-deprived conditions, AMPK becomes highly phosphorylated. In the presence of mitochondrial substrates (pyruvate, acetate) AMPK phosphorylation was clearly higher than with glucose. The analysis of the nucleotide profiles showed a good correlation between phosphorylated AMPK and AMP/ATP ratios. Interestingly, PS exposure showed a strong correlation with AMP/ATP ratios and the phosphorylation of AMPK. Inhibition of LKB1, the upstream kinase of AMPK, with 5-iodotubericidin partially suppressed AMPK phosphorylation and PS exposure in response to substrate deprivation.
Conclusion: In human platelets, activation of AMPK induced by metabolic variation strongly correlates with PS exposure, suggesting a role for AMPK in the induction of PS exposure.
Definition of the Apoptotic Pathway Induced in Proliferating Endothelial Cells by Cleaved High Molecular Weight Kininogen.
Authors: Sun Danyu , McCrae Keith R. 
Published: October 2019

Abstract
Previous studies from our laboratory and others have demonstrated that cleaved high molecular weight kininogen (HKa) induces selective apoptosis of proliferating endothelial cells and inhibits angiogenesis. Recently we reported that the induction of endothelial cell apoptosis by HKa was matrix dependent and requires the generation of reactive oxygen species. To further define the mechanisms of HKa-induced endothelial cell apoptosis, we have investigated the pathway through which apoptosis occurs. HKa-induced endothelial cell apoptosis was caspase-dependent, as demonstrated by the cleavage of pro-caspase 3 and the inhibition of apoptosis by the broad spectrum caspase inhibitor z-VAD-fmk. Activation of the intrinsic pathway of apoptosis was also confirmed by demonstrating cleavage of pro-caspase 9 as well as the release of cytochrome C from mitochondria. These results are consistent with our previous work, as apoptosis induced through oxidative stress is a well known activator of the intrinsic apoptotic pathway. Given the resistance to HKa-induced endothelial cell apoptosis conferred by culture of endothelial cells on collagen I or IV, we questioned whether MMPs may be involved in this process. Interestingly, we observed that the broad spectrum MMP inhibitor GM6001 completely protected endothelial cells from HKa-induced apoptosis. Moreover, we observed that HKa-induced apoptosis was associated with PECAM-1 cleavage within 1 hour of exposure, and that inhibition of apoptosis by either GM6001 or z-VAD-fmk also inhibited the cleavage of PECAM-1. These findings are likely to be relevant to the apoptotic pathway, as intact PECAM-1 has been previously reported to inhibit cytochrome c release after exposure of cells to cytotoxic stimuli, while a cleaved cytoplasmic domain of PECAM-1 enhances the susceptibility of cells to apoptosis. In summary, our studies demonstrate that the antiangiogenic activity of HKa may be critically dependent on MMP activation, which in turn leads to ROS generation and caspase activation. The subsequent cleavage of PECAM-1, presumably by caspases, may be another important event in HKa-induced apoptosis.
Analysis of Causes and Outcome of Thrombocytopenia as a Presenting Symptom in a Single Institution.
Authors: Kraemer Doris M. , Bauer Barbara , Rath Joerg C. , Einsele Hermann 
Published: October 2019

Abstract
Patients suffering from thrombocytopenia admitted to the emergency room during nights and weekends might be a problem, depending from the availability of a haematologist on duty. We evaluated retrospectively the medical history of all patients entering our department receiving a bone marrow biopsy due to severe thrombocytopenia in the years from 2000–2003. We analyzed 441 patients admitted due to thrombocytopenia. 146 patients had a platelet count < 50,000/μl. 69 of these patients suffered from severe thrombocytopenia < 20,000/μl.
All patients entering the ER during daytime received a bone marrow biopsy during the first three hours post admission. If patients were admitted during the night, they received a biopsy the following morning. Bone marrow smears were stained according to Pappenheim and evaluated as soon as they were dry. Diagnoses were as the following: acute leukaemia 54; immune thrombocytopenia 31; aplastic anaemia/toxic bone marrow failure/myelodysplastic syndrome 36; Non Hodgkin’s Lymphoma 7; carcinosis of bone marrow 8; myeloproliferative syndrome 3; viral infection (Hanta, HIV) 2; sepsis 1; hypersplenism 2; thrombotic thrombocytopenic purpura 1; heparin induced thrombocytopenia type II 1.
During their time in the hospital only one patient of the 146 patients developed severe bleeding with the need of transfusion of blood packs. The 73 y old male patient with primary diagnosis of rectal carcinoma and 23,000 platelets at admission was diagnosed with secondary acute myeloid leukaemia. He showed lower gastrointestinal bleeding since rectal carcinoma could not be treated by surgery due to the leukaemia.
In summary 1 out of 146 patients with severe thrombocytopenia showed clinical bleedings (<1%), indicating this as a very rare event. Due to the fact that only 31 out of 146 patients (or 20 of 69 patients with platelets < 20,000/μl) with severe thrombocytopenia suffered from immune mediated thrombocytopenia immediate consultation of the haematologist is essential to allow rapid diagnosis and adequate treatment strategies in patients presenting with thrombocytopenia in the emergency department.
Three Novel ADAMTS-13 Gene Mutations in Thrombotic Thrombocytopenic Purpura (TTP).
Authors: Kim Nam Keun , Jang Moon Ju , Kim Sun , Lee Sun Ju , Park Hye Won , Oh Doyeun , Kang Myung Seo 
Published: October 2019

Abstract
ADAMTS-13 is an enzyme which cleave von Willebrand factor (VWF) to prevent excessive platelet thrombus formation. Mutation of ADAMTS-13 is associated with congenital thrombotic thrombocytopenic purpura (TTP). In this study, we report three novel missense mutations of ADAMTS-13 gene in TTP families. Genetic analysis of ADAMTS-13 gene was performed in 6 TTP families. Three novel ADAMTS-13 gene mutations (2708C>T [S903L], 3650T>C [I1217T] and 3941C>T [S1314L]) detected in 3 families. They were all found with heterozygous genotype in exon 21, 26 and 28, respectively. The 3650T>C mutation was found at exon 26 in the patient and his mother. A heterozygous guanine to adenine substitution was found at 5′ splicing site of intron 3(IVS+ 1) in the patient, his brother and his father. The plasma ADAMTS-13 activities of the patient, father, mother, and brother were less than 3%, 56%, 55% and 62% respectively. ADAMTS-13 inhibitors were not detected in all family members. 3941C>T was found at exon 28 in the patient and her father. The plasma ADAMTS-13 activities of the patient, father, and mother were 96%, 83%, and 83% respectively.
Anti-D Immunglobulin in the Treatment of Childhood Chronic Idiopathic Thrombocytopenic Purpura.
Authors: Turkkan Emine , Pekun Fugen 
Published: October 2019

Abstract
Chronic idiopathic (immune) thrombocytopenic purpura (ITP) develops in approximately 20% of children with acute ITP. The therapies for chronic refractory ITP include splenectomy, steroids, intravenous immunoglobulin (Ig), anti-D Ig, monoclonal antibodies, danazol, chemotherapy, plasma exchange, and others. We report five 5 RhD-positive, pediatric,, non-splenectomized patients with chronic immune thrombocytopenic purpura (ITP), treated with anti-D. Patients were refractory to steroids and high dose intravenous immunoglobulin (IVIG). The mean age of the patients was 10.5 years, duration of thrombocytopenia was more than 6 months, and the median platelet count was 6x 10(9)/l. Patients received a single intravenous dose of 75 microg/kg anti-D, and were monitored for response to treatment and adverse events. Complete blood cell and renal function tests were assessed at baseline and 1, 7, 14 and 28 days after infusion. By 24 hours after treatment two of patients had achieved a platelet count>20 x 10(9)/l. Mild headache, fever, or chills occurred in one patients and ameliorated with prednisone and acetaminophen premedication. None of the patients had more than 0.5 gm/dl fall in the hemoglobin level and renal dysfunction following anti-D infusion by day 1,7, 14, 28. A single 75 microg/kg dose of anti-D may increase the platelet count in some children with chronic resistant immune thrombocytopenic purpura with an acceptable safety profile. It is a valuable, safe, well tolerated, and cheap alternative to conventional chronic ITP treatments in selected non-splenectomized patients.
Treatment of Thrombotic Thrombocytopenic Purpura with Heparin, Vincristine and Dexamethasone.
Authors: Wang Jinghua , Liu Na , Liu Fang , Ruan Changgeng , Liu Juan , Cen Xi , Li Xiaoyun , Dai Haibin , Guo Bo , Su Yanhua , Zhang Yanqing , Zhang Jian , Guan Yue , Wang Hongmei 
Published: October 2019

Abstract
Thrombotic thrombocytopenic purpura(TTP) is a serious, low morbidity and high mortality disease, which can simultaneously affect multiple systems in the patients’ body. In the event that the patients cannot be treated by plasma exchange(PE), mortality will be 95–100%(1). Between September, 2000 and May, 2003, thirteen patients with TTP were treated mainly by heparin, vincristine, dexamethasone, six of whom have acceptted one or two PE. The results were excellent. Twelve of the thirteen patients survived. One patient was dead. The ADAMTS13 activity was measured in 10 patients using a Residual-Collagen Binding Assay(R-CBA).
Bortezomib-Induced Down-Regulation of KIT Is Mediated by Inhibition of Sp1 and NF-kB in AML1/ETO-Positive Cells.
Authors: Liu Shujun , Pang Jiuxia , Yu Jianhua , Liu Zhongfa , Huynh Lenguyen , Kearney Jessica , Paschka Peter , Klisovic Rebecca B. , Baiocchi Robert , Blum William , Perrotti Danilo , Marcucci Guido 
Published: October 2019

Abstract
Activating mutations of KIT, encoding a type III receptor tyrosine kinase, are frequently detected in core binding factor AML (i.e., AML1/ETO and CBFB/MYH11 AML), promote cell survival and proliferation of leukemic cells and predict poor outcome. Kinase inhibitors (e.g., imatinib or PKC-412) have been shown to block constitutively activated KIT. However, novel therapeutic approaches that target mutated KIT are necessary, since resistance to these agents can be predicted in a substantial proportion of patients in these subgroups of AML. We observed that expression levels of KIT in AML1/ETO-positive Kasumi-1 cells are more than 25 fold higher than in AML1/ETO-negative AML cell lines (i.e., THP-1, K562, and MV4–11). We also found that in Kasumi-1 cells, bortezomib (Velcade), a proteasome inhibitor already used in the clinic, induces time- (60nM for 0, 1, 3, 6 12 and 24hr) and dose- (0, 1, 6, 20, 60 and 100nM for 24hr) dependent down-regulation (>90%) of total KIT expression. Noticeably, dephosphorylation of KIT occurred 6hr before reduction of the total KIT level after bortezomib exposure. We also found >50% down-regulation of KIT expression in patients’ primary blasts treated with 60nM bortezomib for 24hr and in Kasumi-1 cells treated with the proteasome inhibitor MG132 (300nM for 24hr). Down-regulation of KIT appeared to be associated with inhibition of NF-kB and Sp1, which is necessary for regulation of the KIT promoter activity by the SCL complex. In fact, treatment with bortezomib inactivated NF-kB and decreased transcription of Sp1 in Kasumi-1 cells. Furthermore, exposure to the NF-kB or Sp1 inhibitors parthenolide (30μM for 24hr) and mithramycin (100ng/ml for 24hr), respectively, resulted in a dose-dependent decrease in KIT expression in Kasumi-1 cells. When cells were treated with bortezomib (20nM) in combination with mithramycin (30ng/ml for 24hr), we observed synergy in down-regulation of KIT RNA and KIT protein. This correlated with growth arrest and increased cell death. Interestingly, the magnitude of these effects was higher when Kasumi-1 cells were pretreated with mithramycin for 24hr before being exposed to bortezomib. Taken all together, our data suggest that bortezomib downregulates KIT expression and might also inhibit KIT phosphorylation and should be considered in future therapeutic strategies targeting AML subgroups harboring mutated KIT.
Cell Penetrating Activity of Bax Inhibiting Peptides (BIPs) and Its Utilization for Protein Transduction.
Authors: Gomez Jose A. , Gama Vivian , Distelhorst Clark , Matsuyama Shigemi 
Published: October 2019

Abstract
The cellular membrane has a strict regulation for molecular trafficking, and there is a strong need for the development of new technology for drug delivery into cells with minimum toxicity. Previously, we developed the cell permeable penta-peptides designed from Bax binding domain of Ku70 that are called “Bax Inhibiting Peptides (BIPs)” (i.e. VPMLK, VPTLK, VPALR). Among BIPs, VPTLK showed the best cell penetrating activity. We also found that one of the scrambled control mutant peptides (i.e. KLPVM) shows a strong cell permeable activity without an activity to bind and inhibit Bax-mediated cell death. We named these cell permeable peptides: BIP-derived cell permeable penta-peptides (BCPs). In this study, we determined the mechanism of membrane penetration by these penta-peptides. Human umbilical endothelial cells (HUVEC), Dami cells (human megakaryocytic cell line), and HeLa cells were used for the examination of cell entry of BCPs. Fluorescent dye (Fluorescein) was conjugated to the N-terminus of BCPs, and the cell entry of BCPs was monitored by FACS and confocal micropscope. BCP entered the cells in a dose dependent manner (10 uM-1.6 mM range was tested), and it did not show any significant toxicity even at 1.6 mM. The entry of BCP into the cells was detected within 15 minutes at 37C, and it reaches maximum approximately at 6 hrs of incubation. We found that BCPs entered the cells even at 0 or 4 C, suggesting that energy-independent mechanism is involved in the cell penetration of BCPs. We also tested whether BCPs has an ability to deliver a cargo protein (protein transduction activity) such as 30 kDa Green Fluorescent Protein across the cellular membrane. We used VPTLK and KLPVM for the examination of GFP delivery from the medium into the cell. Recombinant GFP proteins with and without VPTLK or KLPVM on the C-teminus were generated. The delivery of GFP was detected at a concentration as low as 20 nM within 6 hours of incubation at 37C. GFP without BCP was not imported into the cell, indicating that BCP has a cargo delivery activity (protein transduction). There are several bioactive proteins that can become effective therapeutics if they can be delivered into the cell. To be noted, BCPs are less toxic than the previously examined CPPs that had protein transduction activity, such as TAT peptide designed from the human immunodeficiency virus (HIV). BCPs did not show significant toxicity to bone marrow stem cells and endothelial cells, indicating that BCPs are effective tools for drug delivery in stem cells, and vasculature cells. BCPs may become a useful tool to deliver bioactive molecules into the cells, to protect cells from damage, to regulate cellular activity, and to control differentiation.