bluuebunny/crossref_metadata_2025 huggingface dataset:
Risk of Ischemic Stroke Associated With Functional Thrombin-Activatable Fibrinolysis Inhibitor Plasma Levels
Authors: Santamaría A. , Oliver A. , Borrell M. , Mateo J. , Belvis R. , Martí-Fábregas J. , Ortín; R. , Tirado I. , Souto J.C. , Fontcuberta J.
Published: September 2003

Background and Purpose—

Recently, a novel procarboxypeptidase B–like proenzyme, called thrombin-activatable fibrinolysis inhibitor (TAFI), has been described. It plays an important role in the delicate balance between coagulation and fibrinolysis. TAFI leads to potent inhibition of tissue plasminogen activator–induced fibrinolysis. The relevance of TAFI in thromboembolic disease is unclear. We have investigated the risk of ischemic stroke (IS) in relation to plasma levels of functional TAFI.

Methods—

In a case-control study, we enrolled 264 individuals; 114 had IS, and 150 were recruited as controls who were age and sex matched and had no history of arterial disease. The individuals supplied information on their personal and family histories of cardiovascular diseases and conventional cardiovascular risk factors. Functional TAFI assays were performed by use of a method based on the activation of TAFI with thrombin-thrombomodulin and the measure of the TAFI activity generated. Other hemostatic parameters assayed were factor VIIIc, anti-phospholipid antibodies,fibrinogen, factor V Leiden, and the prothrombin gene G20210A mutations (PT20210A).

Results—

Functional TAFI levels were significantly higher in patients with IS (113.7±25%; range, 57% to 209%) than in controls (102.6±19%). The odds ratio for IS in patients with functional TAFI levels >120% was 5.7 (95% confidence interval, 2.3 to 14.1).

Conclusions—

We found that functional TAFI levels in plasma (>120%) increased the risk of IS ≈6-fold. Further studies should elucidate the physiological role of TAFI in arterial disease and possibly provide clues to therapeutic approaches.
Risk Factors for Progression of Atherosclerosis Measured at Multiple Sites in the Arterial Tree
Authors: van der Meer Irene M. , Iglesias del Sol Antonio , Hak A. Elisabeth , Bots Michiel L. , Hofman Albert , Witteman Jacqueline C.M.
Published: September 2003

Background and Purpose—

Studies investigating determinants of atherosclerotic disease progression are relatively rare. Moreover, although atherosclerotic disease can be assessed noninvasively in different vascular beds, previous studies have not considered progression of atherosclerosis at >1 site. The present study was designed to identify risk factors for progression of atherosclerosis measured at multiple sites in the arterial tree.

Methods—

The Rotterdam Study is a population-based cohort study of 7983 men and women ≥55 years of age. Carotid plaques and intima-media thickness were assessed by ultrasound, aortic atherosclerosis by x-ray, and lower-extremity atherosclerosis by the ankle-arm index. Data on progression of atherosclerosis over an average period of 6.5 years were available for 3409 participants. Associations of established cardiovascular risk factors with mild, moderate, and severe progression of atherosclerosis were investigated through multinomial regression analysis.

Results—

Age, smoking, total cholesterol, and systolic blood pressure and/or hypertension were strong, independent predictors of moderate and severe progression of atherosclerosis at multiple sites. Diabetes mellitus predicted only severe progression of atherosclerosis. Associations of sex with progression of atherosclerosis were remarkably modest.

Conclusions—

Age, smoking, total cholesterol, and systolic blood pressure and/or hypertension strongly predict progression of extracoronary atherosclerosis in the elderly, but sex remarkably does not. These results emphasize the need for prevention of progression of extracoronary atherosclerotic disease in men and women alike.
Hypoxic Induction of Endoglin via Mitogen-Activated Protein Kinases in Mouse Brain Microvascular Endothelial Cells
Authors: Zhu Yonghua , Sun Yunjuan , Xie Lin , Jin Kunlin , Sheibani Nader , Greenberg David A.
Published: September 2003

Background and Purpose—

Endoglin (CD105) is a membrane glycoprotein that is mutated in hereditary hemorrhagic telangiectasia (Osler-Rendu-Weber disease) and shows increased expression in proliferating endothelial cells during angiogenesis.

Methods—

We investigated the effect of hypoxia on endoglin expression in murine cerebral microvascular endothelial (bEND.3) cells in vitro and the possible involvement of mitogen-activated protein kinase (MAPK) pathways.

Results—

Hypoxia increased endoglin mRNA and protein expression in bEND.3 cells, which was associated with phosphoactivation of extracellular signal–related kinase (ERK), p38 MAPK, and Jun amino-terminal kinase (JNK). Inhibitors of p38 decreased hypoxic induction of endoglin expression, as did dominant negative MAPK kinase 3 (MKK3), which activates p38. In contrast, constitutively active MKK3 or JNK1 potentiated the hypoxic induction of endoglin.

Conclusions—

These results indicate that hypoxia induces the expression of endoglin at both the mRNA and protein levels and that induction is regulated by the p38 and perhaps also JNK pathways.
Assessment of Cerebrovascular Autoregulation in Head-Injured Patients
Authors: Steiner Luzius A. , Coles Jonathan P. , Johnston Andrew J. , Chatfield Doris A. , Smielewski Peter , Fryer Tim D. , Aigbirhio Franklin I. , Clark John C. , Pickard John D. , Menon David K. , Czosnyka Marek
Published: September 2003

Background and Purpose—

Cerebrovascular autoregulation is frequently measured in head-injured patients. We attempted to validate 4 bedside methods used for assessment of autoregulation.

Methods—

PET was performed at a cerebral perfusion pressure (CPP) of 70 and 90 mm Hg in 20 patients. Cerebral blood flow (CBF) and cerebral metabolic rate for oxygen (CMR
o
2
) were determined at each CPP level. Patients were sedated with propofol and fentanyl. Norepinephrine was used to control CPP. During PET scanning, transcranial Doppler (TCD) flow velocity in the middle cerebral artery was monitored, and the arterio-jugular oxygen content difference (AJD
o
2
) was measured at each CPP. Autoregulation was determined as the static rate of autoregulation based on PET (SROR
PET
) and TCD (SROR
TCD
) data, based on changes in AJD
o
2
, and with 2 indexes based on the relationship between slow waves of CPP and flow velocity (mean velocity index, Mx) and between arterial blood pressure and intracranial pressure (pressure reactivity index, PRx)

Results—

We found significant correlations between SROR
PET
and SROR
TCD
(
r
2
=0.32;
P
<0.01) and between SROR
PET
and PRx (
r
2
=0.31;
P
<0.05). There were no significant associations between PET data and autoregulation as assessed by changes in AJD
o
2
. Global CMR
o
2
was significantly lower at the higher CPP (
P
<0.01).

Conclusions—

Despite some variability, SROR
TCD
and PRx may provide useful approximations of autoregulation in head-injured patients. At least with our methods, CMR
o
2
changes with the increase in CPP; hence, flow-metabolism coupling may affect the results of autoregulation testing.
Long-Term Mortality and Recurrent Stroke Risk Among Chinese Stroke Patients With Predominant Intracranial Atherosclerosis
Authors: Wong Ka Sing , Li Huan
Published: September 2003

Background and Purpose—

The goal of this study was to document the long-term outcome of ischemic stroke patients in a population with predominant intracranial atherosclerosis and risk factors for a recurrent event.

Methods—

Intracranial and extracranial arteries of consecutive patients with acute ischemic stroke were studied prospectively with transcranial Doppler and duplex ultrasound. All patients were followed up regularly for the development of recurrent stroke, cardiac event, or death.

Results—

We included 705 patients with acute ischemic stroke, of whom 345 were documented ultrasonographically as having large-artery lesions. The follow-up period was up to 42 months (mean, 28±5 months). One hundred seventeen patients (17%) died of any cause, and 199 (28%) suffered further cerebrovascular cardiac events. The 3.5-year cumulative mortality rate was 20.8%; for cerebrovascular event, it was 29.5%. The annual recurrent stroke rates during the first year were 10.9% for patients without vascular lesion, 17.1% for intracranial atherosclerosis only, and 24.3% for both intracranial and extracranial atherosclerosis; for the second year, the rates were 7.5%, 8.6%, and 7.7%, respectively. More occurrence of death (log rank, 5.19;
P
=0.02) or cerebrovascular event (log rank, 9.68;
P
=0.002) was found among patients with than those without vascular lesions. Patients with both intracranial and extracranial arterial lesions were at highest risk of death (log rank, 9.64;
P
=0.008) and cerebrovascular event (log rank, 11.56;
P
=0.003). When death and further vascular event were combined as poor outcomes in a Cox proportional-hazards regression model, number of abnormal arteries, advanced age, diabetes, atrial fibrillation, and previous stroke were significant predictors.

Conclusions—

Patients with intracranial atherosclerosis, especially coexisting extracranial carotid disease, are at higher risk of suffering death or further vascular event. Our findings provide important data for planning future randomized clinical trials for this high-risk group of stroke patients.
Association Between Interleukin-1A Polymorphism and Cerebral Amyloid Angiopathy–Related Hemorrhage
Authors: McCarron M.O. , Stewart J. , McCarron P. , Love S. , Vinters H.V. , Ironside J.W. , Mann D.M.A. , Graham D.I. , Nicoll J.A.R.
Published: September 2003

Background and Purpose—

It has been suggested that the interleukin-1A (
IL-1A
) allele 2 is a risk factor for Alzheimer’s disease (AD). Because cerebral amyloid angiopathy–related hemorrhage (CAAH) often coexists with AD, we examined the
IL-1A
polymorphism in CAAH.

Methods—

In a case-control study, patients with pathologically verified CAAH, AD patients without intracerebral hemorrhage, and neuropathologically normal control subjects were studied. DNA was extracted from brain tissue, and
IL-1A
was genotyped. Logistic regression was used to examine the
IL-1A
polymorphism in CAAH patients with and without AD compared with AD and non-AD control subjects.

Results—

There were 42 patients with CAAH, 232 AD patients, and 167 non-AD control subjects. In age-adjusted analyses, there was no association between possession of
IL-1A
allele 2 and risk of CAAH compared with AD control subjects (odds ratio [OR], 0.94; 95% confidence interval [CI], 0.45 to 1.97;
P
=0.87) or non-AD control subjects (OR, 0.94; 95% CI, 0.47 to 1.87;
P
=0.86). Stratifying for the presence of apolipoprotein E ε2 or ε4 demonstrated the known increased risk of CAAH from these lipoprotein E alleles. Subgroup analyses demonstrated a nonsignificant excess of the
IL-1A
2,2 genotype in patients with CAAH and AD compared with those CAAH patients who did not have histological evidence indicating AD (OR, 2.17; 95% CI, 0.15 to 122.3;
P
=0.64). Comparisons between CAAH patients with AD and AD control subjects and between CAAH patients without AD and non-AD control subjects did not demonstrate an association between CAAH and possession of either the
IL-1A
allele 2 or the 2,2 genotype.

Conclusions—

The
IL-1A
allele 2 or 2,2 genotype does not appear to be a major risk factor for CAAH.
A retrospective 11-year analysis of obstetric patients with idiopathic thrombocytopenic purpura
Authors: Webert Kathryn E. , Mittal Richa , Sigouin Christopher , Heddle Nancy M. , Kelton John G.
Published: September 2003

Numerous studies have examined the outcomes of infants born to mothers with idiopathic thrombocytopenic purpura (ITP). Fewer studies have discussed the morbidity of obstetric patients with ITP. We describe a retrospective study of 92 women with ITP during 119 pregnancies over an 11-year period. Most women had thrombocytopenia during pregnancy. At delivery, women in 98 pregnancies (89%) had platelet counts lower than 150 × 109/L; most had mild to moderate thrombocytopenia. For many, the pregnancy was uneventful; however, women had moderate to severe bleeding in 25 pregnancies (21.5%). Women in 37 pregnancies (31.1%) required treatment to increase platelet counts. During delivery, 44 women (37.3%) received epidural analgesia without complications, with most having a platelet count between 50 and 149 × 109/L. Most deliveries (82.4%) were vaginal. Bleeding was uncommon at delivery. Infant platelet counts at birth ranged from 12 to 436 × 109/L; 25.2% of infants had platelet counts lower than 150 × 109/L, and 9% had platelet counts lower than 50 × 109/L. Eighteen infants (14.6%) required treatment for hemostatic impairment. Two fetal deaths occurred. One was caused by hemorrhage. ITP in pregnancy carries a low risk, but mothers and infants may require therapy to raise their platelet counts. (Blood. 2003;102:4306-4311)
Farnesyltransferase inhibitor R115777 in myelodysplastic syndrome: clinical and biologic activities in the phase 1 setting
Authors: Kurzrock Razelle , Kantarjian Hagop M. , Cortes Jorge E. , Singhania Neil , Thomas Deborah A. , Wilson Edward F. , Wright John J. , Freireich Emil J. , Talpaz Moshe , Sebti Saïd M.
Published: September 2003

R115777 is a potent farnesyltransferase (FTase) inhibitor with substantial antitumor activity in preclinical models. We conducted a phase 1 study (3 + 3 design) of R115777 in patients with myelodysplastic syndrome (MDS). R115777 was administered twice daily (3-weeks-on/1-week-off schedule for 8 weeks) (starting dosage, 300 mg by mouth twice daily; total, 600 mg). Maintenance therapy at the dose/schedule tolerated during induction could be continued until toxicity or lack of benefit. Twenty-one patients with MDS were treated (median age, 66 years). Four (19%) patients had ras mutations (n-ras,3; k-ras, 1). Objective responses (hematologic improvement, 3; partial remission, 2; or complete remission, 1) were seen in 6 of 20 (30%) evaluable patients, only 2 of whom had ras mutations. Response sequences were unusual in some patients who had increases in platelet counts without intervening aplasia. Other responders demonstrated an initial, albeit modest, myelosuppressive effect. The maximum tolerated dose was 400 mg by mouth twice a day. The most frequent side effect was myelosuppression. Dose-limiting toxicities (fatigue and confusion) occurred at 900 mg by mouth total daily dose. R115777 inhibited HDJ-2 prenylation and suppressed the activity of FTase, but not of the related geranylgeranyltransferase I enzyme, in peripheral blood mononuclear cells. Modulation of Akt, Erk, and signal transducer and activator of transcription 3 (STAT3) phosphorylation was variable, and responses occurred even without their down-regulation. Reductions in serum tumor necrosis factor-α (TNF-α) levels by day 7 showed a trend toward correlation with response (P = .09). We conclude that, at doses that are well tolerated, R115777 markedly inhibits the FTase target and has antitumor activity in MDS. (Blood. 2003;102:4527-4534)
Autoantibodies frequently detected in patients with aplastic anemia
Authors: Hirano Naoto , Butler Marcus O. , von Bergwelt-Baildon Michael S. , Maecker Britta , Schultze Joachim L. , O'Connor Kevin C. , Schur Peter H. , Kojima Seiji , Guinan Eva C. , Nadler Lee M.
Published: September 2003

Although accumulating evidence strongly suggests that aplastic anemia (AA) is a T cell-mediated autoimmune disease, no target antigens have yet been described for AA. In autoimmune diseases, target autoantigens frequently induce not only cellular T-cell responses but also humoral B-cell responses. We hypothesized that the presence of antigen-specific autoantibodies could be used as a “surrogate marker” for the identification of target T-cell autoantigens in AA patients. We screened a human fetal liver library for serologic reactivity against hematopoietic stem/progenitor cell antigens and isolated 32 genes. In 7 of 18 AA patients, an immunoglobulin G (IgG) antibody response was detected to one of the genes, kinectin, which is expressed in all hematopoietic cell lineages tested including CD34+ cells. No response to kinectin was detected in healthy volunteers, multiply transfused non-AA patients, or patients with other autoimmune diseases. Epitope mapping of IgG autoantibodies against kinectin revealed that the responses to several of the epitopes were shared by different AA patients. Moreover, CD8+ cytotoxic T cells raised against kinectin-derived peptides suppressed the colony formation of granulocyte macrophage colony-forming units (CFU-GMs) in an HLA class I-restricted fashion. These results suggest that kinectin may be a candidate autoantigen that is involved in the pathophysiology of AA. (Blood. 2003;102:4567-4575)
Treosulfan and fludarabine: a new toxicity-reduced conditioning regimen for allogeneic hematopoietic stem cell transplantation
Authors: Casper Jochen , Knauf Wolfgang , Kiefer Thomas , Wolff Daniel , Steiner Beate , Hammer Ulrich , Wegener Rudolf , Kleine Hans-Dieter , Wilhelm Stefan , Knopp Agnes , Hartung Gernot , Dölken Gottfried , Freund Mathias
Published: September 2003

New conditioning regimens are being explored to reduce toxicity and enable allogeneic bone marrow transplantation in patients not eligible for conventional transplantation. We have investigated treosulfan, an alkylating agent, with the aim of developing an efficient and reliable but less-toxic conditioning regimen. A series of 30 patients who were not eligible for standard conditioning therapy received transplants from HLA-matched related (n = 14) or unrelated (n = 16) donors after administration of treosulfan 10 g/m2 intravenously daily for 3 days and fludarabine 30 mg/m2 intravenously daily for 5 days. Patients receiving grafts from unrelated donors also were given rabbit antithymocyte globulin 10 mg/kg intravenously daily for 3 days. All patients achieved prompt neutrophil and platelet recovery. Extramedullary toxicity was generally mild with Common Toxicity Criteria (CTC) grade 3 or 4 attributable to the conditioning seen only with transaminases. Complete donor chimerism was achieved by 90% of the patients. Acute graft-versus-host disease (GVHD) grade III or IV developed in 14% of the patients and chronic GVHD in 39%. An estimated overall survival rate of 73% and an event-free survival rate of 49% have been reached after a median of 22 months (range, 7.4-33.4 months). In summary, the combination of treosulfan and fludarabine is a safe and efficient conditioning regimen.
Differential requirement for A2a and A3 adenosine receptors for the protective effect of inosine in vivo
Authors: Gomez Gregorio , Sitkovsky Michail V.
Published: September 2003

Inosine is an endogenous nucleoside with immunosuppressive properties that is known to inhibit the accumulation of proinflammatory cytokines and protect mice from endotoxin-induced inflammation and lung tissue damage. There are no known receptors specific for inosine, but A3 adenosine receptors (A3Rs) have been shown to bind inosine, resulting in mast cell degranulation and increased vascular permeability. The present study specifically addresses the requirement for A2aR and/or A3R for the protective effect of inosine in 2 experimental in vivo models of inflammatory disease. The data show that A3R is essential for protection against ConA-induced fulminant hepatitis since only A3R-expressing mice were protected by inosine whereas wild-type and A2aR-deficient mice exhibited severe liver damage even after administration of inosine. In addition, we show in a model of LPS-induced endotoxemia that inosine protected both A2aR-/- and A3R-/- mice from inflammation, but not A2aA3R double-null mice, indicating that in this model both A2aR and A3R were used by inosine. Thus, we demonstrate that A2a and A3 adenosine receptors are differentially utilized by inosine for the down-regulation of tissue damage under different inflammatory conditions in vivo. (Blood. 2003;102:4472-4478)
Insights into the multistep transformation of MGUS to myeloma using microarray expression analysis
Authors: Davies Faith E. , Dring Ann M. , Li Cheng , Rawstron Andrew C. , Shammas Masood A. , O'Connor Sheila M. , Fenton James A.L. , Hideshima Teru , Chauhan Dharminder , Tai Isabella T. , Robinson Elizabeth , Auclair Daniel , Rees Karen , Gonzalez David , Ashcroft A. John , Dasgupta Ranjit , Mitsiades Constantine , Mitsiades Nicholas , Chen Lan B. , Wong Wing H. , Munshi Nikhil C. , Morgan Gareth J. , Anderson Kenneth C.
Published: September 2003

To define specific pathways important in the multistep transformation process of normal plasma cells (PCs) to monoclonal gammopathy of uncertain significance (MGUS) and multiple myeloma (MM), we have applied microarray analysis to PCs from 5 healthy donors (N), 7 patients with MGUS, and 24 patients with newly diagnosed MM. Unsupervised hierarchical clustering using 125 genes with a large variation across all samples defined 2 groups: N and MGUS/MM. Supervised analysis identified 263 genes differentially expressed between N and MGUS and 380 genes differentially expressed between N and MM, 197 of which were also differentially regulated between N and MGUS. Only 74 genes were differentially expressed between MGUS and MM samples, indicating that the differences between MGUS and MM are smaller than those between N and MM or N and MGUS. Differentially expressed genes included oncogenes/tumor-suppressor genes (LAF4, RB1, and disabled homolog 2), cell-signaling genes (RAS family members, B-cell signaling and NF-κB genes), DNA-binding and transcription-factor genes (XBP1, zinc finger proteins, forkhead box, and ring finger proteins), and developmental genes (WNT and SHH pathways). Understanding the molecular pathogenesis of MM by gene expression profiling has demonstrated sequential genetic changes from N to malignant PCs and highlighted important pathways involved in the transformation of MGUS to MM. (Blood. 2003;102:4504-4511)
C/EBPϵ interacts with retinoblastoma and E2F1 during granulopoiesis
Authors: Gery Sigal , Gombart Adrian F. , Fung Yuen K. , Koeffler H. Phillip
Published: September 2003

CCAAT enhancer binding protein epsilon (C/EBPϵ) is a myeloid specific transcription factor that is essential for terminal granulocytic differentiation. Retinoblastoma (Rb) and E2F1 are critical cell cycle regulators that also have been implicated in several differentiation systems. Here, we demonstrate that C/EBPϵ interacts with Rb and E2F1 during granulocytic differentiation in NB4 and U937 human myeloid cells and in 32Dcl3 murine myeloid precursor cells. The interaction between C/EBPϵ and Rb enhances C/EBPϵ-mediated transcription of myeloid specific genes both in reporter assays and endogenously. The C/EBPϵ-E2F1 interaction results in repression of E2F1-mediated transcriptional activity. Finally, overexpression of C/EBPϵ in human myeloid cells leads to down-regulation of c-Myc. We propose that the interactions between C/EBPϵ, a tissue-specific transcription factor, and the broad-spectrum proteins, Rb and E2F1, are important in C/EBPϵ-induced terminal granulocytic differentiation.
Increased erythrocyte adhesion in mice and humans with hereditary spherocytosis and hereditary elliptocytosis
Authors: Wandersee Nancy J. , Olson Scott C. , Holzhauer Sandra L. , Hoffmann Raymond G. , Barker Jane E. , Hillery Cheryl A.
Published: September 2003

Mice with disruptions of the red blood cell (RBC) cytoskeleton provide severe hemolytic anemia models in which to study multiorgan thrombosis and infarction. The incidence of cerebral infarction ranges from 70% to 100% in mice with α-spectrin deficiency. To determine whether mutant RBCs abnormally bind adhesive vascular components, we measured adhesion of mouse and human RBCs to immobilized human thrombospondin (TSP) and laminin (LM) under controlled flow conditions. Mutant RBCs had at least 10-fold higher adhesion to TSP compared with normal RBCs (P < .006). Mutant relative to unaffected RBC adhesion to LM was significantly (P < .01) increased as well. Treatment of RBCs with the anionic polysaccharide dextran sulfate inhibited mutant RBC adhesion to TSP (P < .001). Treatment of RBCs with antibodies to CD47 or the CD47-binding TSP peptide 4N1K did not inhibit TSP adhesion of RBCs. Previously, we have shown that infarcts in α-spectrin–deficient sph/sph mice become histologically evident beginning at 6 weeks of age. TSP adhesion of RBCs from 3- to 4- and 6- to 8-week-old sph/sph mice was significantly higher than RBCs from adult mice (> 12 weeks old; P < .005). While the mechanism of infarction in these mice is unknown, we speculate that changes in RBC adhesive characteristics contribute to this pathology.
Identification of novel CTL epitopes of CMV-pp65 presented by a variety of HLA alleles
Authors: Kondo Eisei , Akatsuka Yoshiki , Kuzushima Kiyotaka , Tsujimura Kunio , Asakura Shoji , Tajima Kohei , Kagami Yoshitoyo , Kodera Yoshihisa , Tanimoto Mitsune , Morishima Yasuo , Takahashi Toshitada
Published: September 2003

Cytomegalovirus (CMV)–specific T-cell immunity plays an important role in protection from CMV disease in immunocompromised patients. Identification of cytotoxic T-lymphocyte (CTL) epitopes is essential for monitoring T-cell immunity and also for immunotherapy. In this and previous studies, CMV-pp65–specific CTL lines were successfully generated from all of 11 CMV-seropositive healthy donors, using pp65-transduced CD40-activated B (CD40-B) cells as antigen-presenting cells. By use of enzyme-linked immunospot (ELISPOT) assays, individual CTL epitopes could be mapped with truncated forms of the pp65 gene. For human leukocyte antigen (HLA) alleles with a known binding motif, CTL epitopes within the defined regions were predicted by computer algorithm. For HLA alleles without a known binding motif (HLA-Cw*0801, -Cw*1202, and -Cw*1502), the epitopes were alternatively identified by step-by-step truncations of the pp65 gene. Through this study, a total of 14 novel CTL epitopes of CMV-pp65 were identified. Interestingly, 3 peptides were found to be presented by 2 different HLA class I alleles or subtypes. Moreover, use of CD40-B cells pulsed with a mixture of synthetic peptides led to generation of pp65-specific CTL lines from some of seronegative donors. The study thus demonstrated an efficient strategy for identifying CTL epitopes presented by a variety of HLA alleles.
Thrombospondin 1 as a scavenger for matrix-associated fibroblast growth factor 2
Authors: Margosio Barbara , Marchetti Daniela , Vergani Veronica , Giavazzi Raffaella , Rusnati Marco , Presta Marco , Taraboletti Giulia
Published: September 2003

The antiangiogenic factor thrombospondin 1 (TSP-1) binds with high affinity to several heparin-binding angiogenic factors, including fibroblast growth factor 2 (FGF-2), vascular endothelial growth factor (VEGF), and hepatocyte growth factor/scatter factor (HGF/SF). The aim of this study was to investigate whether TSP-1 affects FGF-2 association with the extracellular matrix (ECM) and its bioavailability. TSP-1 prevented the binding of free FGF-2 to endothelial cell ECM. It also promoted the mobilization of matrix-bound FGF-2, generating a TSP-1/FGF-2 complex. The region of TSP-1 responsible for these activities was located within the 140-kDa antiangiogenic and FGF-2 binding fragment, whereas the 25-kDa heparin-binding fragment was inactive. Matrix-released FGF-2/TSP-1 complex had a reduced ability to bind to and induce proliferation of endothelial cells. TSP-1 depleted the ECM laid by FGF-2-overproducing tumor cells of its FGF-2-dependent mitogenic activity for endothelial cells. Besides FGF-2, TSP-1 also inhibited VEGF and HGF/SF binding to the ECM and mobilized them from the ECM. Our study shows that TSP-1 acts as a scavenger for matrix-associated angiogenic factors, affecting their location, bioavailability, and function. (Blood. 2003; 102:4399-4406)
Naturally occurring anti–IFN-γ autoantibody and severe infections with Mycobacterium cheloneae and Burkholderia cocovenenans
Authors: Höflich Conny , Sabat Robert , Rosseau Simone , Temmesfeld Bettina , Slevogt Hortense , Döcke Wolf-Dietrich , Grütz Gerald , Meisel Christian , Halle Elke , Göbel Ulf B. , Volk Hans-Dieter , Suttorp Norbert
Published: September 2003

Recently various genetic defects in immunity mediated by interferon γ (IFN-γ) have been described, including mutations in the IFN-γ receptor 1 (IFN-γR1) and receptor 2 (IFN-γR2), signal transducer and activator of transcription 1 (STAT 1), and interleukin 12 receptor β 1 (IL-12Rβ1), and IL-12 p40 genes. These mutations are associated with the occurrence of severe infections with intracellular pathogens especially nontuberculous mycobacteria and vaccine-associated bacilli Calmette-Guérin (BCG). Here we report data on a previously healthy adult patient primarily presenting with severe infections with Burkholderia cocovenenans and subsequently Mycobacterium cheloneae. We found a strong inhibitory anti–IFN-γ activity in the patient's plasma and identified a highaffinity neutralizing anti–IFN-γ autoantibody. Unfortunately, the patient died due to severe sepsis before we knew the nature of the inhibitory activity. The application of alternative therapeutic approaches such as intravenous immunoglobulin or immunoadsorption may have been beneficial in this case. Screening for neutralizing anti–IFN-γ autoantibodies should supplement testing for IFN-γ and IL-12 pathway defects in patients with recurrent infections with intracellular pathogens, especially with nontuberculous mycobacteria.
Identification of a murine CD28 dileucine motif that suppresses single-chain chimeric T-cell receptor expression and function
Authors: Nguyen Phuong , Moisini Ioana , Geiger Terrence L.
Published: September 2003

Recent preclinical and clinical trials have demonstrated the therapeutic potential of T lymphocytes redirected with genetically engineered T-cell receptor (TCR) surrogates against infected, cancerous, or autoreactive cells. These surrogate TCRs link a ligand-recognition domain to signaling regions from the TCR. We previously compared the function of surrogate TCRs that include TCR or TCR and CD28 signaling regions. We found that primary murine T cells modified to specifically target Kb-restricted CD8+ T cells using either Kb-ζ or Kb-CD28-ζ receptors had similar functional activities, although the CD28-ζ receptor showed a 2-fold to 4-fold decreased expression. We have now identified a previously unrecognized dileucine motif in the murine CD28 signaling domain that accounts for this reduced expression. Inactivation of this motif increased chimeric receptor surface expression 2- to 5-fold. T cells expressing the dileucine-mutated CD28-ζ chimeric receptor demonstrated enhanced proliferation, cytokine production, and cytolytic activities. Further, cells expressing this dileucine-mutated receptor were highly effective in eliminating antigen-specific CD8+ T lymphocytes in vivo. These results therefore identify a critical motif limiting the function of receptor-modified T lymphocytes, demonstrate that inactivation of this motif enhances chimeric receptor function, and illustrate a potential novel application of receptor-modified T lymphocytes in the induction of immune tolerance. (Blood. 2003;102:4320-4325)
Dynamic tracking of human hematopoietic stem cell engraftment using in vivo bioluminescence imaging
Authors: Wang Xiuli , Rosol Michael , Ge Shundi , Peterson Denise , McNamara George , Pollack Harvey , Kohn Donald B. , Nelson Marvin D. , Crooks Gay M.
Published: September 2003

The standard approach to assess hematopoietic stem cell (HSC) engraftment in experimental bone marrow transplantation models relies on detection of donor hematopoietic cells in host bone marrow following death; this approach provides data from only a single time point after transplantation for each animal. In vivo bioluminescence imaging was therefore explored as a method to gain a dynamic, longitudinal profile of human HSC engraftment in a living xenogeneic model. Luciferase expression using a lentiviral vector allowed detection of distinctly different patterns of engraftment kinetics from human CD34+ and CD34+CD38- populations in the marrow NOD/SCID/β2mnull mice. Imaging showed an early peak (day 13) of engraftment from CD34+ cells followed by a rapid decline in signal. Engraftment from the more primitive CD34+CD38- population was relatively delayed but by day 36 increased to significantly higher levels than those from CD34+ cells (P < .05). Signal intensity from CD34+CD38--engrafted mice continued to increase during more than 100 days of analysis. Flow cytometry analysis of bone marrow from mice after death demonstrated that levels of 1% donor cell engraftment could be readily detected by bioluminescence imaging; higher engraftment levels corresponded to higher image signal intensity. In vivo bioluminescence imaging provides a novel method to track the dynamics of engraftment of human HSC and progenitors in vivo. (Blood. 2003;102: 3478-3482)
Sequence analysis of clonal immunoglobulin and T-cell receptor gene rearrangements in children with acute lymphoblastic leukemia at diagnosis and at relapse: implications for pathogenesis and for the clinical utility of PCR-based methods of minimal residual disease detection
Authors: Li Aihong , Zhou Jianbiao , Zuckerman David , Rue Montse , Dalton Virginia , Lyons Cheryl , Silverman Lewis B. , Sallan Stephen E. , Gribben John G.
Published: September 2003

Immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements provide clonal markers useful for diagnosis and measurement of minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL). We analyzed the sequences of Ig and TCR gene rearrangements obtained at presentation and relapse in 41 children with ALL to study clonal stability, which has important implications for monitoring MRD, during the course of the disease. In 42%, all original Ig and/or TCR sequences were conserved. In 24%, one original sequence was preserved but the other lost, and in 14% the original sequences were conserved with new sequences identified at relapse. In 20% only new sequences were found at relapse. Using primers designed from the novel relapse sequences, the relapse clone could be identified as subdominant clones in the diagnostic sample in 8 of 14 patients. Alteration of these clonal gene rearrangements is a common feature in childhood ALL. MRD detection should include multiple gene targets to minimize false-negative samples or include also multicolor flow cytometry. In some cases the leukemic progenitor cell might arise earlier in lineage before DHJH recombination but retain the capacity to further differentiate into cells capable of altering the pattern of Ig and/or TCR rearrangements. (Blood. 2003;102:4520-4526)
Reduced expression of NFAT-associated genes in UCB versus adult CD4+ T lymphocytes during primary stimulation
Authors: Kaminski Beth A. , Kadereit Suzanne , Miller Robin E. , Leahy Patrick , Stein Kevin R. , Topa David A. , Radivoyevitch Tomas , Veigl Martina L. , Laughlin Mary J.
Published: September 2003

The cellular and molecular mechanisms underlying the blunted allo-responsiveness of umbilical cord blood (UCB) T cells have not been fully elucidated. Protein expression of NFATc2 (nuclear factor of activated T cells c2), a critical transcription factor necessary for up-regulation of multiple cytokines known to amplify T-cell allogeneic responses, is reduced in UCB T cells. Affymetrix oligonucleotide microarrays were used to compare gene expression of primary purified CD4+ UCB T cells to adult peripheral blood CD4+ T cells (AB) at baseline, 6, and 16 hours of primary stimulation. NFAT-regulated genes exhibited lower expression in UCB CD4+ T cells including the following: granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin 3 (IL-3), IL-4, IL-5, IL-13, IL-2 receptor α (IL-2Rα; CD25), CD40L, and macrophage inflammatory protein 1 α (MIP-1α). Transcription factors involved in the NFAT pathway including C/EBPβ, JunB, and Fosl1 (Fra-1), as well as Th1- and Th2-related transcription factors STAT4 (signal transducers and activators of transcription 4), T-bet, and c-maf showed reduced expression in UCB compared with AB during primary stimulation. Reduced cytokine, chemokine, and receptor expression was also found in UCB. Gene array data were confirmed using RNase protection assays, flow cytometry, and quantitative multiplexed cytokine measurements. Reduced global expression of NFAT-associated genes, as well as cytokines and chemokines, in UCB CD4+ T cells may contribute to the decreased graft-versus-host disease (GVHD) observed after UCB transplantation. (Blood. 2003;102:4608-4617)
Maintaining the self-renewal and differentiation potential of human CD34+ hematopoietic cells using a single genetic element
Authors: Mulloy James C. , Cammenga Jorg , Berguido Francisco J. , Wu Kaida , Zhou Ping , Comenzo Raymond L. , Jhanwar Suresh , Moore Malcolm A. S. , Nimer Stephen D.
Published: September 2003

Hematopoiesis is a complex process involving hematopoietic stem cell (HSC) self-renewal and lineage commitment decisions that must continue throughout life. Establishing a reproducible technique that allows for the long-term ex vivo expansion of human HSCs and maintains self-renewal and multipotential differentiation will allow us to better understand these processes, and we report the ability of the leukemia-associated AML1-ETO fusion protein to establish such a system. AML1-ETO-transduced human CD34+ hematopoietic cells routinely proliferate in liquid culture for more than 7 months, remain cytokine dependent for survival and proliferation, and demonstrate self-renewal of immature cells that retain both lymphoid and myeloid potential in vitro. These cells continue to express the CD34 cell surface marker and have ongoing telomerase activity with maintenance of telomere ends, however they do not cause leukemia in nonobese diabetic-severe combined immunodeficiency (NOD/SCID) mice. Identification of the signaling pathways that are modulated by AML1-ETO and lead to the self-renewal of immature human progenitor cells may assist in identifying compounds that can efficiently expand human stem and progenitor cells ex vivo. (Blood. 2003; 102:4369-4376)
Up-regulation of SLAP in FLI-1-transformed erythroblasts interferes with EpoR signaling
Authors: Lebigot Ingrid , Gardellin Paola , Lefebvre Laurent , Beug Hartmut , Ghysdael Jacques , Quang Christine Tran
Published: September 2003

Rearrangement of the FLI-1 locus and ensuing overexpression of FLI-1 protein is an early event in Friend murine leukemia virus (F-MuLV)-induced erythroleukemia. When overexpressed in primary erythroblasts, FLI-1 converts erythropoietin (Epo)-induced terminal differentiation into a proliferative response. We found that SLAP, a gene encoding a recently described negative regulator of T-cell antigen receptor function during thymocyte development, is up-regulated both at the RNA and protein levels in FLI-1-transformed erythroblasts. Src-like adaptor protein (SLAP) was found in a specific complex with erythropoietin receptor (EpoR), a cytokine receptor essential to erythroid differentiation. Constitutive expression of SLAP severely impairs hemoglobinization and late survival during Epo-induced terminal differentiation of erythroblasts. This impairment is associated with the specific inhibition of several critical Epo-dependent signaling events, including signal transducer and activator of transcription 5 (STAT5) activation and up-regulation of the expression of the antiapoptotic BCL-X gene. Our data support a model by which FLI-1 inhibits normal erythroid differentiation through the deregulation of genes encoding adaptors/effectors that modify the signaling output of cytokine receptors normally required for terminal differentiation. (Blood. 2003; 102:4555-4562)
Acute graft-versus-host disease in patients with Fanconi anemia or acquired aplastic anemia undergoing bone marrow transplantation from HLA-identical sibling donors: risk factors and influence on outcome
Authors: Guardiola Philippe , Socié Gérard , Li Xiaxin , Ribaud Patricia , Devergie Agnès , Espérou Hélène , Richard Patrice , Traineau Richard , Janin Anne , Gluckman Eliane
Published: September 2003

To assess whether Fanconi anemia (FA) patients might be at risk for acute graft-versus-host disease (AGvHD) despite using low-intensity conditionings, we retrospectively analyzed the incidence of AGvHD and its impact on outcome in 37 FA patients and 73 patients with acquired aplastic anemia (AAA) that received transplants at Saint Louis Hospital from HLA-genotypic identical siblings with similar conditionings (thoraco-abdominal irradiation plus cyclophosphamide 20 [FA] or 150 mg/kg [AAA]). Despite being younger, FA patients had an increased risk of grades II to IV AGvHD (relative risk [RR], 2.00; P = .021), especially in younger patients (RR, 7.93; P = .014). The risks of requiring systemic corticosteroids to treat AGvHD and experiencing cortico-resistant AGvHD were significantly increased in FA patients. Although non-FA and FA patients had similar 10-year outcomes, acute and chronic GvHD had a biphasic effect on FA patient outcome with an additional cluster of lethal events starting by 5 years after transplantation. This late survival fall, restricted to FA patients, was closely related to head and neck carcinomas (15-year incidence: 53%). FA patients represent a group at risk regarding AGvHD when using irradiation-based conditionings. The impact of AGvHD on survival may not be limited to the early posttransplantation period and may be a major risk factor for head and neck carcinomas and late mortality in FA patients.
IL-10 regulates plasmacytoid dendritic cell response to CpG-containing immunostimulatory sequences
Authors: Duramad Omar , Fearon Karen L. , Chan Jean H. , Kanzler Holger , Marshall Jason D. , Coffman Robert L. , Barrat Franck J.
Published: September 2003

Immunostimulatory sequences (ISS) are short oligonucleotides containing unmethylated cytosine-phosphate-guanine (CpG) dinucleotides that stimulate innate immune responses through Toll-like receptor-9 on B cells and plasmacytoid dendritic cell (PDC) precursors. The anti-inflammatory cytokine interleukin (IL)-10 is predicted to be a potent inhibitor of many of the activities described for ISS, and this may impact the use of ISS in disease states characterized by elevated IL-10. As the activities of ISS on PDCs are central to many clinical applications of ISS, we have studied the effects of IL-10 on PDC stimulation by 3 distinct classes of ISS. IL-10 inhibited cytokine production and survival of ISS-activated PDCs; however, IL-12 induction was much more sensitive to inhibition than interferon (IFN)-α induction. Within the PDC population are cells that respond to ISS by producing either IL-12 or IFN-α but not both cytokines. IL-12-producing PDCs require costimulation through CD40 and appear more mature than IFN-α-producing PDCs. The 3 distinct classes of ISS differed with respect to induction of PDC maturation and T-cell priming capacity. IL-10 regulated PDC activation but did not inhibit the subsequent T-cell-priming ability of PDCs already activated by ISS. (Blood. 2003;102:4487-4492)
Does the tube-compensation function of two modern mechanical ventilators provide effective work of breathing relief?
Authors: Maeda Yoshiko , Fujino Yuji , Uchiyama Akinori , Taenaka Nobuyuki , Mashimo Takashi , Nishimura Masaji
Published: August 2003

Objective

An endotracheal tube (ETT) imposes work of breathing on mechanically ventilated patients. Using a bellows-in-a-box model lung, we compared the tube compensation (TC) performances of the Nellcor Puritan-Bennett 840 ventilator and of the Dräger Evita 4 ventilator.

Measurements and results

Each ventilator was connected to the model lung. The respiratory rate of the model lung was set at 10 breaths/min with 1 s inspiratory time. Inspiratory flows were 30 or 60 l/min. A full-length 8 mm bore ETT was inserted between the ventilator circuit and the model lung. The TC was set at 0%, 10%, 50%, and 100% for both ventilators. Pressure was monitored at the airway, the trachea, and the pleura, and the data were recorded on a computer for later analysis of the delay time, of the inspiratory trigger pressure, and of the pressure–time product (PTP). The delay time was calculated as the time between the start of inspiration and minimum airway pressure, and the inspiratory trigger pressure was defined as the most negative pressure level. The same measurements were performed under pressure support ventilation of 4 and 8 cmH2O.

The PTP increased according to the magnitude of inspiratory flow. Even with 100% TC, neither ventilator could completely compensate for the PTP imposed by the ETT. At 0% TC the PTP tended to be less with the Nellcor Puritan-Bennett 840 ventilator, while at 100% TC the PTP tended to be less with the Dräger Evita 4 ventilator. A small amount of pressure support can be equally effective to reduce the inspiratory effort compared with the TC.

Conclusion

Although both ventilators provided effective TC, even when set to 100% TC they could not entirely compensate for a ventilator and ETT-imposed work of breathing. The effect of TC is less than that of pressure support ventilation. Physicians should be aware of this when using TC in weaning trials.
Comparison of sufentanil with sufentanil plus magnesium sulphate for sedation in the intensive care unit using bispectral index
Authors: Memiş Dilek , Turan Alparslan , Karamanlıoğlu Beyhan , Oğuzhan Nihal , Pamukçu Zafer
Published: August 2003

Introduction

In intensive care unit patients we assessed, using bispectral index (BIS) monitoring, whether the addition of magnesium sulphate infusion could decrease the sufentanil infusion required to maintain sedation.

Patients and methods

A total of 30 adult patients who were expected to require machanical ventilation for 6 hours in the intensive care unit were randomly assigned to receive either sufentanil infusion or sufentanil plus magnesium infusion. We monitored BIS levels continously. BIS levels in the range 61–88 are required to maintain a state of sedation, and in both groups BIS levels were kept within this range by increasing or decreasing the sufentanil infusion. Hourly consumption of sufentanil was monitored. Cardiovascular, respiratory and biochemical data were recorded.

Results

There was no significant difference between the groups with respect to cardiovascular, respiratory and biochemical parameters. Magnesium infusion, when added to sufentanil infusion, decreased the consumption of sufentanil at all times accept during the first hour (P < 0.001). There was no significant difference in BIS values between the groups (P > 0.05).

Conclusion

This is the first clinical study to demonstrate that magnesium sulphate infusion decreases sufentanil requirements. Because of the limited number of patients included and the short period of observation, our findings must be confirmed by larger clinical trials of magnesium infusion titrated to achieve prespecified levels of sedation. Furthermore, randomized clinical studies are needed to determine the effects of magnesium infusion on opioids.