bluuebunny/crossref_metadata_2025_split huggingface dataset:
Relation of CD4+CD25+ Regulatory T Cells to Immune Status in Patients with HIV Infection.
Authors: Tsunemi Sachi , Iwasaki Tsuyoshi , Imado Takehito , Higasa Satoshi , Kakishita Eizo , Okamoto Takahiro , Shirasaka Takuma , Sano Hajime 
Published: October 2019

Abstract
Human immunodeficiency virus (HIV) infection is characterized by marked defects in CD4+ helper T cell (Th) functions that commonly progress to a substantial decline in peripheral CD4+ T cell counts. However, the mechanisms responsible for the loss of Th functions in HIV-infected patients independent of CD4+ T cell counts remains unclear. CD4+CD25+ regulatory T cells (T Reg) are essential for down-regulation of both autoreactive and alloreactive T cells. Therefore, we decided to investigate the role of T Reg in immune status of HIV-infected patients. We examined the expression of cell surface CD25, cytoplasmic IL-4 and cytoplasmic IFN-gamma in peripheral blood CD4+ T cells from both healthy controls (n=9) and HIV-infected patients (n=43). We also compared T Reg functions between the 2 groups. CD4+CD25+ T Reg isolated from both HIV-infected patients and healthy controls strongly expressed CD45RO, HLA-DR, and FoxP3, and suppressed the proliferation of CD4+CD25− T cells, suggesting that CD4+CD25+ T cells from both healthy controls and HIV-infected patients possess phenotypic and functional characteristics of Treg. CD4+CD25high T cells are a subset of circulating CD4+CD25+ T cells in normal humans and exhibit strong in vitro regulatory functions similar to those reported for murine CD4+CD25+ T Reg. We measured the frequency of CD4+CD25high T Reg by analysis of surface CD25 on CD4+ T cells in peripheral blood samples. We also examined Th1 and Th2 frequencies by analysis of cytoplasmic IFN-gamma and IL-4 levels in CD4+ T cells. T Reg from HIV-infected patients with detectable plasma HIV-1 RNA showed a statistically significant increase in CD4+CD25high cell frequency (p<0.05) compared to healthy controls, with T Reg frequencies inversely proportional to CD4+ T cell numbers (p<0.01). However, in HIV-infected patients with undetectable plasma HIV-RNA, frequencies of CD4+CD25high T Reg were not increased and not related to CD4+ T cell numbers. In both HIV-infected patient groups, T Reg frequency was inversely related to Th1 frequency (detectable: p<0.05, undetectable: p<0.001), but positively related to Th2 frequency (detectable: p<0.01, undetectable: p<0.001). Our results indicate that increased frequencies of peripheral blood T Reg were related to disease progression as measured by detectable plasma HIV-1 RNA, decreased peripheral blood CD4+ T cell counts, and polarization toward Th2 immune responses in HIV-infected patients. HIV infection may lead to induction of T reg that inhibit antiviral immune responses, resulting in the progression of the disease. Manipulation of T Reg could help restore antiviral immune responses in HIV infection, and prevent the progression of HIV infection.
CalDAG-GEFI Is a Key Signal Integrator in Platelet Activation and Thrombus Formation.
Authors: Bergmeier Wolfgang , Crittenden Jill R. , Piffath Crystal L. , Wagner Denisa D. , Housman David E. , Graybiel Ann M. 
Published: October 2019

Abstract
Inside-out activation of platelet integrin αIIbβ3 is a key step in agonist-induced platelet aggregation. Recent studies suggested the involvement of the small GTPase Rap1b in this process as it is highly expressed in platelets and becomes activated during platelet activation. In cell lines, overexpression of the Rap activator CalDAG-GEFI increased αIIbβ3-dependent adhesion, while overexpression of RapGAP, which inactivates Rap1, reduced αIIbβ3 activity. Here we provide evidence that CalDAG-GEFI is an essential component of this pathway in vivo. To generate CalDAG-GEFI knockout mice, we engineered mouse embryonic stem (ES) cells with a deletion that results in a frameshift mutation and a premature stop codon at the position encoding the 37th amino acid of CalDAG-GEFI. These ES cells were then used to derive chimeric mice that yielded germline transmission of the CalDAG-GEFI mutation. Deficiency of CalDAG-GEFI in mutant mice was confirmed by immunohistochemistry and western blot analysis. CalDAG-GEFI−/− platelets showed impaired Rap1b activation and aggregation in response to various agonists, with aggregation being completely blocked when platelets were activated with ADP, thromboxaneA2 analog, or calcium ionophore. Under physiological flow conditions in vitro and in vivo, CalDAG-GEFI-deficient platelets showed normal tethering to basement membrane components but failed to form thrombi. Mice deficient in CalDAG-GEFI were further characterized by a greatly increased bleeding time as well as by a strong protection against collagen-induced pulmonary thrombosis. In summary, we identified CalDAG-GEFI as a key signal integrator in the cascade leading through Rap1 and integrin αIIbβ3 to platelet aggregation and thrombus formation. The fact that CalDAG-GEFI knockout mice are resistant to collagen-induced thrombosis, and do not undergo spontaneous hemorrhaging, suggests that CalDAG-GEFI may be a promising new target for antithrombotic therapy.
Role for Interleukin-6 and Insulin like Growth Factor-I Via PI3-K/Akt Pathway in the Proliferation of CD56-Negative and Positive Multiple Myeloma Cells.
Authors: Sahara Naohi , Takeshita Akihiro , Kobayashi Miki , Shigeno Kazuyuki , Nakamura Satoki , Shinjo Kaori , Naito Kensuke , Otsuki Takemi , Ohnishi Kazunori , Ohno Ryuzo 
Published: October 2019

Abstract
Several studies including ours have suggested that lack of CD56 expression in multiple myeloma (MM) defines a unique patient subset with poorer prognosis. However, the mechanism underlying this aggressive behavior of CD56− MM has not been well elucidated. In this study, we sorted out both CD56− and CD56+ fractions from MM cell lines or patients with MM, and investigated their different responsiveness to interleukin-6 (IL-6) or insulin like growth factor-I (IGF-I), and tried to clarify the course of action in cell cycle distribution. After stained with PE-CD56, CD56− and CD56+ fractions in KMS-21-BM and U-266 cell lines were isolated by the cell sorter, and cultured separately either in the presence or absence of IL-6 (2 ng/ml and 10 ng/ml, respectively). Although CD56− cells in both KMS-21-BM and U-266 cell lines responded significantly to IL-6 (P=0.001 and 0.009, respectively), CD56+ cells did not. Ki-67+ cells in CD56− KMS-21-BM cells, that were significantly fewer than that in CD56+ ones (P=0.0003), increased significantly upon 24-hour incubation with IL-6 (P<0.0001). Western blotting analysis showed that the level of cyclin D1 and p27 protein in CD56− KMS-21-BM cells were up- and down-regulated by IL-6 in a time dependent manner, respectively. IL-6 also brought phosphorylation of Akt (ser473) in the CD56− cells. LY-294002 completely blocked these effects of IL-6. On the other hand, Ki-67+ cells in the CD56+ cells did not respond to IL-6. Although IGF-I did not increase Ki-67+ cells either in the CD56− and CD56+ cells from KMS-21-BM, anti-IGF-I mAb significantly reduced Ki-67+ cells only in the CD56+ cells (P=0.006). IGF-I up-regulated the level of cyclin D1 and phosphorylated Akt in CD56+ KMS-21-BM cells. LY294002 completely blocked these effects of IGF-I. Same results were obtained in the analysis of U-266 cell lines. The MM cells sorted from 17 patients with MM were also examined for CD56 and Ki-67 expression. Four and 13 patients were distributed to the CD56− and CD56+ group, respectively. These MM cells from the patients were cultured with or without IL-6 (10 ng/ml) or IGF-I (500ng/ml) for 24 hours. IL-6 increased the percentage of Ki-67+ cells in the CD56− group more than those in the CD56+ group (P=0.007). Although MM cells did not respond to IL-6, IGF-I significantly increased Ki-67+ cells in the CD56+ group (P=0.005). These results suggest that CD56− and CD56+ MM cells could be stimulated by different cytokines. We here found that CD56− MM cells were proliferated more than CD56+ MM cells in the presence of IL-6, and that this effect of IL-6 was mainly mediated by the activation of PI3-K/Akt pathway. In addition, our results suggest that IGF-I play an important role in the proliferation of CD56+ MM cells via PI3-K/Akt pathway.
Cladribine (CdA) or Fludarabine (F) or High-Dose Intermittent Chlorambucil (Chl) as First-Line Treatment of Symptomatic Chronic Lymphocytic Leukemia? First Interim Analysis of Data from the International Randomized Phase III Trial.
Authors: Karlsson Karin , Strömberg Mats , Jönsson Viggo , Gill Devinder , Hammerström Jens , Wallvik Jonas , Mulligan Stephen , Bradstock Kenneth , Liliemark Jan , Juliusson Gunnar 
Published: October 2019

Abstract
Purine analogues are effective treatment for CLL, but despite improved response rates and durations for purine analogues as compared to chlorambucil no survival advantage was documented. Our trial is the first comparing CdA and fludarabine (F) as first-line treatment of symptomatic B-CLL with high dose intermittent chlorambucil (Chl) as control. Patients aged 18–75 years from Scandinavia, Australia and UK were since November 1997 randomised between monthly courses of Chl 10mg/sqm orally day 1–10, F 25mg/sqm iv day 1–5, or CdA 5mg/sqm iv/sc day 1–5. Protocol is seen at www.cck.ki.se/CLL-study/index.html. Randomisation was stratified according to age and stage. Up to six courses were given according to response and toxicity. We now report a scheduled interim analysis of the first included 150 patients; 139 were evaluable and 5 excluded for not fulfilling eligibility criteria and 6 for incomplete data reporting. There was no significant difference in age, Binet stage or time from diagnosis to inclusion between the groups. Median age was 63 years and median time from diagnosis to inclusion was 16 months. Overall responses according to NCI criteria and intention to treat were documented in 57%/67%/74% of patients in the Chl/F/CdA arms, respectively (Chl vs CdA, p=0.06, Chi-test). The number of CR/nPR/PR in the different treatment groups were for Chl 2/3/22 (n=47), for F 0/2/28 (n=45), and for CdA 2/3/30 (n=47). NCI grade III and IV platelet toxicity was seen in 25% and 20% for Chl, 24% and 5% for F, and 36% and 11% for CdA. NCI grade III–V infections were seen in 25%/28%/30%, respectively. One F patient died in septic shock from E coli, and two CdA patients died from RS virus infection and aspergillosis, respectively; none of them responded to CLL-treatment. Autoimmune hemolysis was seen only with purine analogues, 4 during F and 3 during CdA treatment. We conclude that toxicity was feasible, and infections did not differ between arms. So far there is no significant difference in response rates between the therapies, but CdA seems at least as good as F. Inclusion is currently ongoing but will close during 2004, and a full evaluation is scheduled thereafter.
Evaluation of ADAMTS-13 Activity and VWF:Ag Levels in Patients with Ischemic Stroke.
Authors: Krause Manuela , Boehm Martina , Stier-Brueck Ingrid , Scharrer Inge 
Published: October 2019

Abstract
Background: The role of von Willebrand factor cleaving protease (ADAMTS-13) in the pathogenesis of ischemic stroke is still undefined. ADAMTS-13 cleaves ultra-large multimers of von Willebrand factor (ULVWF), and is believed to regulate the size thus the activity of VWF, especially on sites of high shear stress. By assuming the hypothesis, that a deficiency of ADAMTS-13 influences the thrombotic tendency such as intravascular aggregation and platelet thrombus formation, we analysed adult patients with onset of cerebral ischemia.
Methods and Patients: 158 unrelated patients (female:89 /male:69) with an objectively confirmed ischemic stroke (n=126) or TIA (n=32), median age at first onset:42 years (range:17–71 years), and 82 healthy subjects, median age:28 years (range:16–52 years), were studied. In our study group we analysed on the one hand 95 patients (pts) with early onset of stroke/TIA <45 years (median age:32 years), and on the other hand 63 elderly pts >45 years (median age:54 years). None of the pts enrolled had overt evidence of autoimmune or malignancy disease. ADAMTS13 was measured with an assay based on the positive correlation between multimeric size and Ristocetin Cofactor activity of the VWF.
Results: In our population the ADAMTS-13 activity was in median 95% (range= 40–185%) significantly higher than 91% (range=52–150%) among controls (p=0.002; OR 0.54). In the subgroup analysis we found no statistical differences in the mean ADAMTS-13 activities among pts with early onser (99% vs 91%; p=0.76, OR 0.93), and among pts with onset at age 45 years as compared to healthy controls (88% vs 91%; p=0.30, OR 1.25). Furthermore, we found statistical differences in the mean VWF:Ag levels among all pts as compared to healthy controls (135% vs 110%; p=0.01, OR 0.63), and in the elderly pts (140% vs 110%; p=0.01, OR 1.63). The young pts showed no statistical differences in the mean VWF:Ag levels compared with the control group (127% vs 110%; p=0.98, OR 0.99).
Conclusion: The results from this study show that higher levels of ADAMTS-13 activity and VWF:Ag as compared to healthy controls are associated with ischemic stroke/TIA. Only in the elderly pts we observed a tendency to a negative association between ADAMTS-13 and VWF (lower ADAMTS-13 levels and higher levels of VWF:Ag). It may be, that higher ADAMTS-13 levels have a relevance to prevent thrombotic events.
Further studies are needed to confirm whether the relationship between VWF and ADAMTS-13 plays an additional role in the pathogenesis on the onset of ischemic stroke.
Routine Prophylactic Platelet Transfusions Are Not Necessary for Patients in Clinically Stable Condition after Autologous Peripheral Stem Cell Transplantation(ASCT): Updated Results in 106 Patients.
Authors: Schaefer-Eckart Kerstin , Frank Markus , Wilhelm Martin , Wandt Hannes 
Published: October 2019

Abstract
We present our extended experience with an only therapeutic platelet transfusion strategy in patients after autologous peripheral stem cell transplantation(ASCT). Clinically stable patients(fever < 38,5°Celsius, no local infections, no sepsis syndrome) received single donor apheresis platelet transfusions only in the case of bleeding WHO ≥ II°, while prophylactic platelet transfusions were given to clinically instable patients if the morning platelet count was < 10/nl. In a first analysis after 50 patients we have shown that this strategy was safe, with no bleeding greater than WHO II°. In a retrospective matched-pair analysis the total number of platelet units transfused was reduced to 50% compared to our former strategy with routine platelet transfusions given when the morning platelet count was below 10/nl. (ASH 2002).
We now analysed 106 patients with a total number of 140 ASCTs. Median age was 54 years(19–70): The diagnoses were acute leukemia(17), lymphoma(34), solid tumors(9) and multiple myeloma(46). The conditioning regimens corresponded to standard protocols.
Median days of thrombocytopenia < 20/nl and 10/nl were 6(0–92) and 3(0–62) respectively, with a total number of days with thrombocytopenia <20/nl and <10/nl of 989 and 508. Hemorrhages WHO I° and II° was observed in only 49 out of 140(35%) ASCTs. We observed no bleeding greater than WHO II°. The median number of platelet units was 1(0–18). 48 out of 140(34%) transplantations could be performed without platelet transfusions. In multiple myeloma this percentage was even higher: 32/68(47%). The indications for prophylactic transfusions were mainly FUO(21/61 – 34%) and mucositis with or without fever(19/61–31%). Considering age below or above the median age of 54 years or different diagnoses, there was no difference in days with platelets <10/nl, <20/nl, bleeding complications or median number of platelet units transfused. The total number of 234 transfusions in these 140 transplantations could have been even further reduced, because 15%(36/234) of the transfusions were given without a clear indication regarding the study regimen, because of a learning effect with this new strategy. This new strategy has shown to be very safe and prophylactic platetelet tansfusions are probably not necessary in clinically stable patients with fever as the only sign of an infection.
We are just starting a multicenter randomised study comparing this new strategy with the former strategy of routine prophylactic platelet transfusion.
Accessing the Erythrocyte Cytoplasm: A Method for Manipulating the Intracellular Environment of Erythrocytes for the Study of Malaria Invasion, Trafficking and Growth.
Authors: Murphy Sean C. , Bhattacharjee Souvik , Harrison Travis , Haldar Kasturi 
Published: October 2019

Abstract
Invasion of erythrocytes by malaria parasites requires participation of both parasite ligands and host determinants. Further recent studies show that erythrocyte G protein signaling regulates malarial infection. Many of the Gs-associated signaling components reside on the cytoplasmic leaflet of the erythrocyte plasma membrane, rendering them inaccessible to most extracellular probes. Since erythrocytes are enucleated and terminally differentiated, they cannot be transfected to express exogenous transgenes. We have modified methods of hypotonic lysis and isotonic resealing to generate loaded erythrocyte ghosts that can be efficiently infected by Plasmodium falciparum and sustain normal levels of intraerythrocytic parasite growth and replication. Further, we show that these ghosts can be filled with various membrane-impermeable peptides or other proteinaceous cargoes for studying signaling and transport events inside the erythrocyte. Resealed ghosts morphologically resemble normal erythrocytes, albeit with reduced hemoglobin content. Other measures of erythrocyte function and malarial infection are being investigated. Studies will be presented on the use of ‘reconstituted’ erythrocytes in elucidating mechanisms parasite invasion as well as parasite protein trafficking in erythrocytes infected by P. falciparum.
The Venezuelan Type of Nondeletional Hereditary Persistence of Fetal Hemoglobin Is Due C →T Mutation at Nucleotide -211 of the Aγ Globin Gene.
Authors: Arends Anabel , Urquiola Martha Bravo , Montilla Silvia , Lic Gloria García , De L. Dalia Velásquez , Alvarez Maritza , Gerard Natalie , Krishnamoorthy Rajagapol 
Published: October 2019

Abstract
Hereditary Persistence of Fetal Hemoglobin (HPFH) is a clinically benign condition characterized by continuous synthesis of fetal hemoglobin (Hb F) in adult life without major related hematological changes. The nondeletional HPFH is characterized by hemoglobin F level between 5% and 30%, in heterozygotes state. This condition has been observed among Blacks, Italians, Greeks, English and Chinese. Two different forms are known, characterized by the type of chain involve, either Gγ or Aγ. Several point mutations in the promoter region of the Aγ globin gene have been identified in ndHPFH: −202, −198, −196, −195, −117, −114, deletion 4pb (−222 −225) and −158.
The propositus is a 36 year old black female with a moderate increased level of Hb F, also her sister and her sibling had the same condition. She was discovered during studies due to the presence of splenomegaly and gastroesophageal reflux. The Hbs A, A2 and F were eluted and quantified by cation exchange high-performance liquid chromatography (HPLC-CE), using Beta Tal short program, Variant BioRad (Hercules, California, USA). DNA was isolated from peripheral blood leukocytes by a salting-out extraction procedure. The Aγ and Gγ promoters globins gene were amplified independently by PCR and the presence of the HPFH mutation was determined by denaturing gradient gel electrophoresis technique (DGGE) as previously described Gottardi et al., (1992). The Aγ promoter globin gene was sequenced in three members of the family using the BigDyeTM Terminator Cycler Sequencing Kit and the ABI PRISMTM 310 Genetic Analyzer (PE Applied Biosystems, Foster City, CQ, USA).
Her hematological studies revealed Hb: 14.2 g/dl, Hcto: 41.1 %, MCV: 94.4 fl, MCHC: 34.5 fl, Hb A: 81.9 %, Hb A2: 2.1 % and Hb F: 6.3 %. The sibling hematological finding was Hb: 13.1 g/dl, Hcto: 41.8 %, MCV: 83.7 fl, MCHC: 31.3 fl, Hb A: 83.2 %, Hb A2: 3.2 % and Hb F: 5.1. The sister was Hb: 14.3 g/dl, Hcto: 46.4 %, MCV: 105.9 fl, MCHC: 30.8 fl, Hb A: 83.7 %, Hb A2: 3.3 % and Hb F: 2.7 %. No spleen enlargement was seen in the other two affected members of the family. The DGGE results in the Aγ promoter, suggested the presence of a non-deletion form of HPFH in heterozygotes state. The Aγ promoter region revealed the presence of a substitution C→T at position −211. This mutation does not modify any restriction site.
To our knowledge the −211 (C→T) mutation in the Aγ promoter have not been reported previously. Since modulation of g-globin gene expression in adults could provide a therapeutic intervention in hemoglobinopathy, the study of the molecular mechanisms generating HPFH is medically relevant.
Erythropoietin (EPO) Plasma Levels Reflect the Course of Clinical Vaso-Occlusive Complications in Sickle Cell Disease.
Authors: Duits Ashley J. , Akin Sakir , Ozveren Erkan , Mac Gillavry Melvin R. , Brandjes Dees P. , Schnog John-John B. 
Published: October 2019

Abstract
Erythropoietin (EPO) has an important role in both erythropoieses and angiogenesis. EPO levels are elevated in sickle cell patients due to severe anemia, but tissue hypoxia, due to for example vaso-occlusion, may also contribute to EPO elevation. In this study we sequentially determined EPO plasma levels in HbSS sickle cell patients (n=14, 39 samples) presenting with vaso-occlusive complications at our emergency ward and the period thereafter. Patients were also requested to fill out a pain-intensity score during these events. Our results show a striking correlation between the course of vaso-occlusive events and plasma EPO levels. Vaso-occlusive crises were accompanied by impressive increments of plasma EPO levels with quick reduction of plasma EPO levels upon crisis amelioration. Changes in plasma EPO levels also closely reflected changes in pain intensity. The results of this study warrant further analysis of the potential role of EPO in the pathophysiology of sickle cell disease related complications, as well as the applicability of employing EPO levels as a marker of vaso-occlusive disease activity.