bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential expression of matrix metalloproteinases and their inhibitors in human and mouse lung development
Authors: Ryu Julie , Vicencio Alfin G. , Yeager Michael E. , Kashgarian Michael , Haddad Gabriel G. , Eickelberg Oliver
Published: June 2005

Lung development is a highly orchestrated process characterized by timed expression and activation of growth factor and protease/antiprotease systems. This interplay is essential in regulating vasculogenesis, alveolarization, and epithelial to mesenchymal transition during lung development. Alterations in the proteolytic/antiproteolytic balance of the lung have been associated with several respiratory diseases characterized by changes in the lung extracellular matrix (ECM). Here, we characterized the expression pattern of matrix metalloproteases (MMP) and their inhibitors, the tissue inhibitors of metalloproteases (TIMP), in human and mouse lung development. Using MMP/TIMP expression arrays, RT-PCR, Western Blotting, and ELISA analyses, we demonstrate that fetal human lung is characterized by a dominant proteolytic profile with high MMP-2 and little TIMP-3 expression. Adult human lung, in contrast, exhibits a more anti-proteolytic profile with decreased MMP-2 and increasedTIMP-3 expression. MMP-14, MMP-20,TIMP-1, and TIMP-2 were constitutively expressed, irrespective of the developmental stage. Similar results were obtained using mouse lungs of different developmental stages, with the addition that in mouse lung, TIMP-2 and TIMP-3 were upregulated as lung development progressed. Exposure of neonatal mice to chronic hypoxia (10% O2), a stimulus that leads to an arrest of lung development, resulted in upregulation of MMP-2 with a concomitant downregulation of TIMP-2.These results provide a comprehensive analysis of MMP and TIMP expression during human and mouse lung development. MMP-2, TIMP-2, and TIMP-3 may be key regulatory enzymes during lung development, possibly through their complex action on ECM components, membrane receptor ectodomain shedding, and growth factor bioactivity.
Modulation of tissue factor expression by rapamycin and FK-506 in lipopolysaccharide-stimulated human mononuclear cells and serum-stimulated aortic smooth muscle cells
Authors: Hammal Soumeya , Ameziane Nejma , Labro Marie-Thérèse , de Prost Dominique , Ollivier Véronique
Published: June 2005

Inflammation is a key pathogenic component of atherosclerosis; it also promotes thrombosis, a process underlying acute coronary events and stroke. Cells present in atherosclerotic plaque show abnormal tissue factor (TF) expression. Macrolides, in addition to their antimicrobial properties, have antiinflammatory effects that might help prevent atherothrombosis. The aim of this study was to determine the effect of an immunosuppressant macrolide, rapamycin (Sirolimus), on the expression of TF and its inhibitor (TFPI) by monocytic cells (human blood mononuclear and THP-1 cells) and human aortic smooth muscle cells, in comparison with FK-506 and azithromycin. In monocytic cells, rapamycin and FK-506 inhibited LPS-induced TF activity, antigen and mRNA expression through a transcriptional mechanism involving NF-κB. In smooth muscle cells, rapamycin and azithromycin had no effect on serum-induced TF expression, while FK-506 increased serum-induced TF protein and mRNA expression. TFPI levels in the culture supernatants of serum-stimulated smooth muscle cells were not modified by any of the three macrolides. Rapamycin slightly inhibits TFPI induction by LPS in monocytic cells. In addition to its recently established efficacy in the prevention of stent restenosis, the inhibitory effect of rapamycin on theTF pathway might have interesting therapeutic implications.
A major role for P2X1 receptors in the early collagen-evoked intracellular Ca2+ responses of human platelets
Authors: Fung C. Y. Eleanor , Brearley Charles A. , Farndale Richard W. , Mahaut-Smith Martyn P.
Published: June 2005

In the platelet, ATP-gated P2X1 receptors have been reported to amplify functional responses to collagen, however the relative importance of early Ca2+ mobilisation events is unknown. We now report that selective desensitisation of P2X1 receptor activity leads to a major reduction in the initial intracellular Ca2+ responses to a wide range of collagen concentrations (0.25–2 μg ml-1). Peak [Ca2+]i increases were reduced to 8.5 and 55% of control, and the maximum rate of rise was reduced to 12 and 33% of control, at low and high collagen concentrations, respectively. This P2X1-dependent acceleration and enhancement of collagen-stimulated Ca2+ responses was not observed in the absence of extracellular Ca2+. These results demonstrate a major role for ATP-gated Ca2+ influx in the early collagen-evoked Ca2+ signals and can at least partly explain the important contribution of P2X1 receptors to arterial thrombosis.
Diagnostic strategy using a modified clinical decision rule and D-dimer test to rule out pulmonary embolism in elderly in- and outpatients
Authors: Kamphuisen Pieter W. , van Mierlo Patricia J. W. B. , Büller Harry R. , Söhne Maaike
Published: June 2005

Excluding or confirming pulmonary embolism remains a diagnostic challenge. In elderly patients pulmonary embolism is associated with substantial co-morbidity and mortality, and many elderly patients with suspected pulmonary embolism are inpatients. The safety and efficacy of the combination of a clinical probability (CDR) and d-dimer test in excluding pulmonary embolism in this group is unclear. We retrospectively analysed data of two prospective studies of consecutive in-and outpatients with suspected pulmonary embolism. The patients were categorized into three age groups: <65 years, 65–75 years and >75 years. The sensitivity, negative predictive value and the proportion of patients with the combination of a non-high CDR score according to Wells (≤ 4) and a normal d-dimer result were calculated for each group. In 747 consecutive patients with suspected pulmonary embolism, sensitivity and negative predictive value of a non-high CDR and a normal d-dimer result in outpatients (n=538) of all age categories (<65 years, 65–75 years and >75 years) were both 100%.These tests were, however, less reliable for inpatients(n=209), irrespective of their age (sensitivity 91% [ CI: 79–98%], negative predictive value 88 % [CI: 74–96%].The proportion of both in-and outpatients >75 years with a non-high CDR and a normal d-dimer concentration was only 14%,whereas this was 22% in patients 65–75 years and 41% among in-and outpatients <65 years, respectively. In elderly outpatients the combination of a non-high CDR and a normal d-dimer result is a safe strategy to rule out pulmonary embolism. However, in inpatients this algorithm is not reliable to safely exclude pulmonary embolism. In addition, the proportion of patients >65 years in which this strategy excludes pulmonary embolism is markedly lower compared to younger patients.
Oral contraceptives, hormone replacement therapy, thrombophilias and risk of venous thromboembolism: a systematic review
Authors: Wu Olivia , Robertson Lindsay , Langhorne Peter , Twaddle Sara , Lowe Gordon D. O. , Clark Peter , Greaves Mike , Walker Isobel D. , Brenkel Ivan , Regan Lesley , Greer Ian A.
Published: June 2005

Combined oral contraceptives, oral hormone replacement therapy and thrombophilias are recognised risk factors for venous thromboembolism in women. The objective of this study was to assess the risk of thromboembolism among women with thrombophilia who are taking oral contraceptives or hormone replacement therapy, conducting a systematic review and metaanalysis. Of 201 studies identified, only nine met the inclusion criteria. Seven studies included pre-menopausal women on oral contraceptives and two studies included peri-menopausal women on hormone replacement therapy. For oral contraceptive use, significant associations of the risk of venous thromboembolism were found in women with factor V Leiden (OR 15.62; 95%CI 8.66 to 28.15); deficiencies of antithrombin (OR 12.60; 95%CI 1.37 to 115.79), protein C (OR 6.33; 95%CI 1.68 to 23.87), or protein S (OR 4.88; 95%CI 1.39 to 17.10), elevated levels of factor VIIIc (OR 8.80; 95%CI 4.13 to 18.75); and factor V Leiden and prothrombin G20210A (OR 7.85; 95%CI 1.65 to 37.41). For hormone replacement therapy, a significant association was found in women with factor V Leiden (OR 13.16; 95%CI 4.28 to 40.47).Although limited by the small number of studies, the findings of this study support the presence of interaction between thrombophilia and venous thromboembolism among women taking oral contraceptives. However, further studies are required to establish with greater confidence the associations of these, and other, thrombophilias with venous thromboembolism among hormone users.
Fibrillar type I collagens enhance platelet-dependent thrombin generation via glycoprotein VI with direct support of α2β1 but not αIIbβ3 integrin
Authors: Lecut Christelle , Jandrot-Perrus Martine , Feijge Marion A. H. , Cosemans Judith M. E. M. , Heemskerk Johan W. M.
Published: June 2005

The role of collagens and collagen receptors was investigated in stimulating platelet-dependent thrombin generation. Fibrillar type-I collagens, including collagen from human heart, were most potent in enhancing thrombin generation, in a way dependent on exposure of phosphatidylserine (PS) at the platelet surface. Soluble, non-fibrillar type-I collagen required pre-activation of integrin α2β1 with Mn2+ for enhancement of thrombin generation. With all preparations, blocking of glycoprotein VI (GPVI) with 9O12 antibody abrogated the collagen-enhanced thrombin generation, regardless of the α2β1 activation state. Blockade of α2β1 alone or antagonism of autocrine thromboxane A2 and ADP were less effective. Blockade of αIIbβ3 with abciximab suppressed thrombin generation in platelet-rich plasma, but this did not abolish the enhancing effect of collagens. The high activity of type-I fibrillar collagens in stimulating GPVI-dependent procoagulant activity was confirmed in whole-blood flow studies, showing that these collagens induced relatively high expression of PS. Together, these results indicate that: i) fibrillar type-I collagen greatly enhances thrombin generation, ii) GPVI-induced platelet activation is principally responsible for the procoagulant activity of fibrillar and non-fibrillar collagens, iii) α2β1 and signaling via autocrine mediators facilitate and amplify this GPVI activity, and iv) αIIbβ3 is not directly involved in the collagen effect.
Isolated integrin β3 subunit cytoplasmic domains require membrane anchorage and the NPXY motif to recruit to adhesion complexes but do not discriminate between β1– and β3-positive complexes
Authors: Ponsonnet Lionel , Foletti Alessandro , Alghisi Gian Carlo , Rüegg Curzio
Published: June 2005

Integrin adhesion receptors consist of non-covalently linked α and β subunits each of which contains a large extracellular domain, a single transmembrane domain and a short cytoplasmic tail. Engaged integrins recruit to focal structures globally termed adhesion complexes. The cytoplasmic domain of the β subunit is essential for this clustering. β1 and β3 integrins can recruit at distinct cellular locations (i.e. fibrillar adhesions vs focal adhesions, respectively) but it is not clear whether individual β subunit cytoplasmic and transmembrane domains are by themselves sufficient to drive orthotopic targeting to the cognate adhesion complex. To address this question, we expressed fulllength β3 transmembrane anchored cytoplasmic domains and truncated β3 cytoplasmic domains as GFP-fusion constructs and monitored their localization in endothelial cells. Membrane-anchored full-length β3 cytoplasmic domain and a β3 mutant lacking the NXXY motif recruited to adhesion complexes, while β3 mutants lacking the NPXY and NXXY motifs or the transmembrane domain did not. Replacing the natural β subunit transmembrane domain with an unrelated (i.e. HLA-A2 α chain) transmembrane domain significantly reduced recruitment to adhesion complexes. Transmembrane anchored β3 and cytoplasmic domain constructs, however, recruited without discrimination to β1– and β3-rich adhesions complexes. These findings demonstrate that membrane anchorage and the NPXY (but not the NXXY) motif are necessary for β3 cytoplasmic domain recruitment to adhesion complexes and that the natural transmembrane domain actively contributes to this recruitment. The β3 transmembrane and cytoplasmic domains alone are insufficient for orthotopic recruitment to cognate adhesion complexes.
Ultrasound thrombolysis
Authors: Pfaffenberger Stefan , Devcic-Kuhar Branka , Kastl Stefan P. , Maurer Gerald , Wojta Johann , Gottsauner-Wolf Michael , Huber Kurt
Published: June 2005

Cardiovascular diseases are a major cause of mortality in the developed world. Efficacy of thrombolysis plays an important role in the management of acute myocardial infarction and cerebral insult both in the acute event and in the long-term outcome of these patients. New adjunctive strategies have been tested, therefore, to make thrombolytic therapies more effective and safer. Ultrasound Thrombolysis is a technique which showed promising results under in vitro conditions and in animal studies. Now clinical trials have to prove if it is also feasible for clinical application. This report gives an overview on different technical approaches and their current performances in the clinical setting. All original articles are chronologically ordered in tables providing detailed information on each study concerning experimental design, acoustical parameters and thrombolysis outcome.
Clinical features of heparin-induced thrombocytopenia including risk factors for thrombosis
Authors: Farner Beate , Kroll Hartmut , Kohlmann Thomas , Warkentin Theodore E. , Eichler Petra , Greinacher Andreas
Published: June 2005

Immune mediated heparin induced thrombocytopenia (HIT) is a prothrombotic adverse effect of heparin. However, only a subgroup of patients with HIT develops thromboembolic complications. We aimed to identify risk factors for developing HITassociated thrombosis. We analyzed a registry of patients with clinical suspicion of HIT who tested positive using a sensitive functional assay. Patient information was obtained by a standardized questionnaire. By multivariate analysis the association of age, gender, type of patient population, and magnitude of the platelet count decline with the frequency, type (venous or arterial), and temporal pattern of thrombotic events was assessed. In 408 HIT patients we observed predominance of venous thrombosis (2.4:1), with 40% of patients developing a pulmonary embolism. However, in the subgroup of post-cardiovascular surgery patients there was predominance of arterial thrombosis (1:8.5). The type of arterial thrombosis (limb artery thrombosis > thrombotic stroke > myocardial infarction) was the converse of that observed with typical atherothrombotic clots in non-HIT populations. In 59.8% of patients HIT-related thrombosis manifested either on the same day a platelet count decrease >50% was documented (26.3%) or before the decrease in platelet counts (33.5%).The most important risk factors for thrombosis were orthopedic/trauma surgery and the magnitude of platelet count decrease. HIT-associated thrombosis occurs in a considerable proportion of patients before platelet counts decrease by more than 50%.
Ruling out deep venous thrombosis in primary care
Authors: Moons Karel G. M. , Hoes Arno W. , Oudega Ruud
Published: June 2005

In primary care, the physician has to decide which patients have to be referred for further diagnostic work-up. At present, only in 20% to 30% of the referred patients the diagnosis DVT is confirmed. This puts a burden on both patients and health care budgets. The question arises whether the diagnostic work-up and referral of patients suspected of DVT in primary care could be more efficient. A simple diagnostic decision rule developed in primary care is required to safely exclude the presence of DVT in patients suspected of DVT, without the need for referral. In a cross-sectional study, we investigated the data of 1295 consecutive patients consulting their primary care physician with symptoms suggestive of DVT, to develop and validate a simple diagnostic decision rule to safely exclude the presence of DVT. Independent diagnostic indicators of the presence of DVT were male gender, oral contraceptive use, presence of malignancy, recent surgery, absence of leg trauma, vein distension, calf difference and D-dimer test result. Application of this rule could reduce the number of referrals by at least 23% while only 0·7% of the patients with a DVT would not be referred. We conclude that by using eight simple diagnostic indicators from patient history, physical examination and the result of D-dimer testing, it is possible to safely rule out DVT in a large number of patients in primary care, reducing unnecessary patient burden and health care costs.
A subpopulation of human B lymphocytes can express a functional Tissue Factor in response to phorbol myristate acetate
Authors: Mechiche Houria , Cornillet-Lefebvre Pascale , Nguyen Philippe
Published: June 2005

Tissue Factor (TF, CD142) is a transmembrane glycoprotein that belongs to the cytokine receptor superfamily. This multifunctional protein is constitutively present in most tissues. Among circulating blood cells, TF expression is known to be restricted to monocytes which do not constitutively produce TF but express TF in response to various stimuli. Here, we report that highly purified B lymphocytes can support a procoagulant activity (PCA) in response to phorbol myristate acetate (PMA).Using flow cytometry (FCM), we observed that a subpopulation of B lymphocytes (CD19+) can expressTF and anionic phospholipids in response to PMA. TF protein was identified and characterized by immunofluorescence, immunocytochemistry and electronic microscopy. Using RT-PCR, we identified the presence of TF mRNA in response to PMA. In conclusion, B lymphocytes are a potential source of functional TF in human.
Preoperative fibrin monomer measurement allows risk stratification for high intraoperative blood loss in elective surgery
Authors: Gabi Konrad , Rohner Mirjam , Gähler Anita , Szadkowski Christine , Schnider Thomas W. , Lange Jochen , Riesen Walter , Korte Wolfgang
Published: June 2005

We recently demonstrated that patients with increased blood loss due to intraoperative coagulopathy show a persistent pre-, intra- and postoperative increase in fibrin monomer concentration. We thus tested the hypothesis that preoperative fibrin monomer concentrations can be used as a risk indicator for intraoperative blood loss in a study designed for diagnostic test evaluation in 168 patients admitted to the surgical service of our hospital. Intraoperative blood loss increased with preoperative fibrin monomer concentration (median blood loss of 50, 100, 200 and 400 ml in preoperative fibrin monomer quartile groups 1 to 4, p<0.001,ANOVA on ranks; interquartile comparisons p < 0.05 (4/6), MannWhitney Rank Sum test). In contrast, intraoperative blood loss was unrelated to preoperative values of prothrombin time, activated partial thromboplastin time and platelet count. By multivariate (logistic regression) analysis, only fibrin monomer remained a significant predictor of intraoperative blood loss > 500 ml when age, gender, BMI, fibrin monomer and the different types of surgical procedures (tumor surgery, vascular surgery, cholecystectomy, gastric banding, varicous vein surgery and hernia repair) were included as independent variables. Most importantly, accuracy evaluation showed that preoperative fibrin monomer concentration < 3 μg/l excluded intraoperative blood loss > 500 ml with 92% sensitivity and 95% negative predictive value. These results support our hypothesis that preoperative fibrin monomer concentrations are related to intraoperative blood loss in elective surgery. Fibrin monomer should be further investigated for it’s potential to serve as a routine tool for preoperative risk stratification of intraoperative bleeding.
Bridging anticoagulation with low-molecular-weight heparin after interruption of warfarin therapy is associated with a residual anticoagulant effect prior to surgery
Authors: Woods Karen , Foster Gary A. , Crowther Mark A. , Douketis James D.
Published: June 2005

Bridging anticoagulation with low-molecular-weight heparin (LMWH) is common in patients who require temporary interruption of warfarin before surgery or a procedure, but whether such patients have a residual anticoagulant effect just before a procedure is not known. Consecutive patients who received bridging anticoagulation with LMWH had anti-Xa levels measured just before a procedure. The proportion of patients with a residual anticoagulant effect, defined as an anti-Xa level ≥0.10 IU/ml, was determined. Multivariable regression analysis identified predictors of a residual anticoagulant effect, expressed as an odds ratio (OR) and corresponding 95% confidence interval (CI). A pre-procedure residual anticoagulant effect was detected in 12 of 73 (16%) patients overall, in 11 of 37 (30%) patients who received therapeutic-dose LMWH, and in 1 of 36 patients (3%) who received low-dose LMWH. Receiving therapeutic- dose LMWH (OR = 118.8; 95% CI: 5.8, 999.9), and increasing age (OR = 4.0; 95% CI: 1.3, 12.5) were predictors of a residual pre-procedure anticoagulant effect. In patients who require bridging anticoagulation with LMWH, a residual anticoagulant effect from LMWH is detected in 1 of 6 patients, and receiving therapeutic-dose LMWH is the strongest predictor of such an effect.
Variability of anti-β2 glycoprotein I antibodies measurement by commercial assays
Authors: Tincani Angela , Sanmarco Marielle , de Moerloose Philippe , Boffa Marie-Claire , Reber Guido
Published: June 2005

The aim of this study was to evaluate the agreement in assay results between commercial kits for the measurement of anti- β2glycoprotein I antibodies. Ten manufacturers provided one IgG and one IgM kit to three testing centres. Samples from patients with primary (n=13) or secondary (n=3) antiphospholipid syndrome (APS), from lupus patients without APS features (n=6) and from normal individuals (n=2) were tested in the three centres according to manufacturers’ instructions. Dilutions in normal serum of a pool made from positive patients’ samples (Forum Calibrators) and dilutions of humanized monoclonal antibodies (MoAbs) were used as additional calibrators. The calibration curves obtained with each calibrator differed widely between kits. The rate of positivity of patients’ samples varied from 7 to 16 for IgG and from 2 to 17 for IgM, depending on the kit. Perfect agreement occurred in 12/22 samples for IgG and 5/22 samples for IgM. Samples from normals were found negative by all kits. Between kits, cutoff values varied up to five fold when expressed in Forum Calibrators arbitrary units and up to three fold when expressed in MoAbs equivalents. Examination of discrepant samples indicated that about half of the discrepancies, scoring 8:2 and 9:1, involved the same few kits. In highly discrepant samples, some kits appeared as high responders as compared to others. In conclusion, with the exception of a few kits, agreement in assay results was acceptable. In conclusion, additional efforts are however necessary, especially concerning the way to assess the cutoff point and the adoption of a reference calibrator, in order to improve standardization of the assays.
An association between oxygen desaturation and D-dimer in patients with obstructive sleep apnea syndrome
Authors: Shitrit David , Peled Nir , Shitrit Ariella Bar-Gil , Meidan Silvia , Bendayan Daniel , Sahar Gidon , Kramer Mordechai R.
Published: June 2005

D-dimer, a degradation product of fibrin, is being increasingly used as a marker and prognostic factor in various thrombotic diseases. Previous reports have shown that obstructive sleep apnea is associated with platelet activation and hypercoagulability. The aim of the study was to assess the potential role of the plasma D-dimer test in patients with obstructive sleep apnea. We designed a prospective group comparison study in a tertiary- care, university-affiliated medical center. One hundred and three patients of mean age 57 years (range 50–76 years) with symptoms suggestive of obstructive sleep apnea were included. Polysomnography was performed in all cases, and blood was collected for plasma D-dimer measurement by MiniQuant turbidmetric assay. The demographic and polysomnograph data were compared between patients with normal and high (>250 ng/ml) D-dimer levels. The group with higher D-dimer values had lower mean minimal oxygen saturation (72.1±16.4 vs. 81.7±11.6%, p=0.008) and a longer mean period of oxygen saturation below 90% (84.1±86.2 vs. 38.5±70.8 minutes, p=0.032). There was no correlation of respiratory disturbance index and sleep architecture with D-dimer values. We concluded that sleep apnea syndrome is associated with fibrinolytic activity. Oxygen desaturation seems to be one of the mediatory factors in the putative connection between obstructive sleep apnea and hypercoagulability state.
Thrombocytopenia and platelet hypoaggregation induced by Bothrops asper snake venom
Authors: Soto Mónica , Escalante Teresa , Loría Gilbert D. , Arni Raghuvir , Gutiérrez José María , Rucavado Alexandra
Published: June 2005

Thrombocytopenia and platelet dysfunction occur in patients bitten by Bothrops sp snakes in Latin America. An experimental model was developed in mice to study the effects of B. asper venom in platelet numbers and function. Intravenous administration of this venom induces rapid and prominent thrombocytopenia and ex vivo platelet hypoaggregation. The drop in platelet numbers was primarily due to aspercetin, a protein of the C-type lectin family which induces von Willebrand factor-mediated platelet aggregation/agglutination. In addition, the effect of class P-III hemorrhagic metalloproteinases on the microvessel wall also contributes to thrombocytopenia since jararhagin, a P-III metalloproteinase, reduced platelet counts. Hypoaggregation was associated with the action of procoagulant and defibrin(ogen)ating proteinases jararacussin-I (a thrombin-like serine proteinase) and basparin A (a prothrombin activating metalloproteinase). At the doses which induced hypoaggregation, these enzymes caused defibrin(ogen)ation, increments in fibrin(ogen) degradation products and D-dimer and prolongation of the bleeding time. Incubation of B. asper venom with batimastat and α2-macroglobulin abrogated the hypoaggregating activity, confirming the role of venom proteinases in this effect. Neither aspercetin nor the defibrin(ogen)ating and hypoaggregating components induced hemorrhage upon intravenous injection. However, aspercetin, but not the thrombin-like or the prothrombin-activating proteinases, potentiated the hemorrhagic activity of two hemorrhagic metalloproteinases in the lungs.
In vitro expression demonstrates impaired secretion of the γAsn319, Asp320 deletion variant fibrinogen
Authors: Kani Satomi , Terasawa Fumiko , Lord Susan T. , Tozuka Minoru , Ota Hiroyoshi , Katsuyama Tsutomu , Okumura Nobuo
Published: June 2005

The hypodysfibrinogenemia Otsu is caused by the two-residue deletion,γAsn319 and γAsp320. Analysis of plasma or purified fibrinogen from the heterozygous propositus revealed that the amount of variant γ-chain was lower than that of normal γ-chain. In order to examine the basis for this difference, we transfected Chinese hamster ovary cells and established stable cell lines that expressed both chains, γΔ/γN, only the normal chain, γN, and only the variant chain, γΔ. We measured fibrinogen concentration of confluent cultures by ELISA. We found the ratios of the concentrations in the media to the concentrations in the cell lysates of γΔ, γΔ/γN, and γN-cells were 0.42, 0.60, and 1.00, respectively. We measured the concentrations of the γΔ and γN chains by densitometric analysis of samples following separation by SDS-PAGE and found the fraction of γΔ-chains in cell lysates was always greater than the fraction in the respective culture media. We examined the kinetics of fibrinogen synthesis, assembly and secretion in pulse-chase experiments, and found that the γΔ-chain was assembled into intact fibrinogen at a rate similar to assembly of the γN-chain into normal fibrinogen, but was secreted into the medium at a slightly slower rate than normal fibrinogen. Considered together, these experiments indicate secretion of the variant fibrinogen was slightly impaired. These results suggest that the reduced level of γΔ319,320 fibrinogen in the plasma of the Otsu patient arises from modestly impaired secretion of this variant fibrinogen.
Participating or Just Talking? Sustainable Development Councils and the Implementation of Agenda 21
Authors: Rosenberg Jonathan , Thomas Linus Spencer
Published: June 2005

Sustainable Development Councils were among the few specific recommendations for institution building to come out of Rio in 1992. At their best the councils manifest Agenda 21's call for new participatory arrangements. At their worst they represent the frustrations and unmet challenges of the thirteen years since Rio. The article compares attempts to establish councils in three Caribbean states: Grenada, Dominica, and St. Lucia. The cases offer lessons in the survivability of deliberative bodies concerned with sustainable development policy and raise questions about their efficacy. We conclude that such bodies survive when members derive significant if intangible benefits; and that by surviving, they help optimize limited human resources for the implementation of international environmental conventions and provide needed venues for deliberation and accountability. But the relationship between efficacy and survivability is not linear and councils may have to avoid direct challenges to government decision-makers and established relationships between the state and private sector.
Servants at the Gate: Eunuchs at the Court of al-Muqtadir
Authors: El-Cheikh Nadia Maria
Published: June 2005

This paper investigates the eunuch's institution in the court of the Abbasid caliph al-Muqtadir and seeks, first, to delineate the variety of functions that the eunuchs held in the early fourth/tenth century Abbasid court both in the harem and in ceremonial; second, it investigates the careers of the eunuchs Sāfī al-Huramī and Muflih al-Khādim al-Aswad, exploring their sources of authority and their various networks which allowed them to exercise a high degree of political influence. Le but de ce travail est d'étudier l'institution "Eunuque" au coeur de la cour Abbaside du caliphe al-Muqtadir. En premier lieu nous soulignons les fonctions multiples que les eunuques performaient dans la cour abbaside (début 4/10ème siècle) tant au sein du harem pendant le ceremonial royal. En second lieu nous explorons plus en détail les carrières des eunuques Sāfī al-Huramī et Muflih al-Khādim al-Aswad, identifiant les sources de leur pouvoir et les réseaux differents qui leur permirent d'éxercer une veritable influence politique.
Patronage in the Ancient Near East
Authors: Westbrook Raymond
Published: June 2005

Patronage is generally assumed by scholars to have been a universal feature of ancient Near Eastern societies, but has been neglected as a topic of serious investigation. The purpose of this study is to offer, without prior assumptions, textual evidence that establishes the existence of the concept of patronage. The approach is to present case studies from various parts of the region which are best explained by the presence of patronage. For these purposes patronage is narrowly de fined on the basis of ancient Roman and contemporary anthropological models. Les historiens du Proche-Orient ancien supposent que le patronage était un phénomène universel dans la région, sans que ce sujet n'ait fait l'objet d'une étude approfondie. Dans cet article je propose de présenter sans présomptions préalables des preuves textuelles que le concept de patronage existait. L'approche est de présenter des cas concrets provenants de plusieurs parties de la région qui s'expliquent au mieux par la présence du patronage. À ces fins, j'adopte une définition étroite du patronage, à la base de modèles romains anciens et antropologiques modernes.
Market Welfare in the Early-Modern Ottoman Economy—A Historiographic Overview with Many Questions
Authors: Shechter Relli
Published: June 2005

In early-modern Ottoman economy the notion of market welfare proposed here meant a system that partially sti fled competition and efficiency for the sake of economic stability and equity for those established within its boundaries. Such a system worked even in the face of political decentralization when economic regulation from "above" (Istanbul and "the state") was seemingly on the wane. Discussing available research and raising questions for future study, the article examines forms of regulation from the "middle" by local officials/notables, courts, and economic institutions in cities throughout the Empire and the role of consumers in economic regulation. The article further suggests why economic opening in a later era of integration into the world economy gradually put an end to an inward-looking, early-modern economic life. Dans l'économie ottomane du début de l'ère moderne (17e–18e siècles), la notion, proposée ici, de bien-être par le marché signifiait un système qui décourageait voire étouffait partiellement compétition et efficacité dans le but d'assurer une stabilité et une équité économique à ceux compris dans son périmètre. Un tel système fonctionnait toujours même dans un contexte de décentralisation politique, alors que la régulation économique venue « d'en haut » (d'Istanbul et de « l'Etat » ) semblait être en train de s'affaiblir. Considérant la recherche effectuée à ce sujet et soulevant des nouvelles questions pour une future enquête, le présent article étudie des formes de régulation émanant du « milieu » et établies localement par des fonctionnaires/dignitaires, des tribunaux, et des institutions économiques dans des villes à travers l'Empire, ainsi que le rôle joué par les consommateurs dans ce processus. L'article tente aussi d'expliquer pourquoi l'ouverture économique durant la période postérieure, celle de l'intégration dans l'économie mondiale, a mis fin, progressivement, à une vie économique tournée vers l'intérieur qui a caractérisé le début de l'ère moderne.
Banding Selection Scores in Human Resource Management Decisions: Current Inaccuracies and the Effect of Conditional Standard Errors
Authors: Bobko Philip , Roth Philip L. , Nicewander Alan
Published: June 2005

The personnel selection literature has recently included discussion of statistically based banding as a way to handle some differences in test scores when assessing job applicants. Banding uses classical test theory and an estimated standard error of measurement to create bands of individual scores, and these bands are treated as equivalent with respect to top-down selection. However, such banding operationally assumes that standard errors of measurement are homogeneous, whereas a focus on the top score logically and statistically implies the use of a conditional standard error. Other methods, such as item response theory and binomial error models, are therefore more appropriate for computing bands. Via example and analysis, the authors demonstrate that more accurately computed bands are substantially narrower under a variety of circumstances than currently computed bands. Bands as currently constructed will label an inaccurate excess of individuals as equivalent, particularly if the test is relatively easy.
Potential Problems in the Statistical Control of Variables in Organizational Research: A Qualitative Analysis With Recommendations
Authors: Becker Thomas E.
Published: June 2005

The author examines statistical control in a random sample of 60 articles published in four top journals during 2000 to 2002. Authors’bases for including control variables, clarity regarding measures and methods, and reporting of results were recorded. Potential problems included a lack of explanations for inclusion, unclear descriptions of measures and methods, incomplete reporting, and other flaws. Implications for interpreting results, replication, future reviews, and effect sizes are discussed. Twelve recommendations for addressing these issues are offered.
Introducing Cognizer™: A Comprehensive Computer Package for the Elicitation and Analysis of Cause Maps
Authors: Clarkson Gail P. , Hodgkinson Gerard P.
Published: June 2005

Managerial and organizational cognition (MOC) researchers have issued a number of calls over the years for large-scale studies involving the mass application of causal mapping techniques. A number of important advances in the development of procedures for the systematic elicitation and comparison of cause maps mean that such work has been technically feasible for some time. However, due to a dearth of suitable supporting computer software, very few researchers to date have responded to this key challenge, vital to the longer-term viability of the MOC field. Building on innovations by Langfield-Smith and Wirth and Markóczy and Goldberg, the authors report on the development of Cognizer™, a comprehensive computer package designed to meet the requirements of researchers lookingto elicit and compare large numbers of maps on a longitudinal or cross-sectional basis. The principal features of Cognizer are illustrated using a cross-sectional data set comprising 200 cause maps from five organizations.
Crossover Linear Modeling: Combining Multilevel Heterogeneities in Crossover Relationships
Authors: Han Jian
Published: June 2005

In organizational research, many multilevel structures are not fully nested but have a crossover structure. For example, in early recruitment study, the relationship between job applicants and recruiting companies has the crossover structure because each applicant is interested in applying multiple companies whereas each company is of interest to multiple applicants, and applicants and companies are not fully nested within each other. This article introduces the crossover linear modeling (CLM) method and argues that CLM is useful when dealing with multilevel crossover structures by combining the heterogeneities of all levels into one model. The article uses an example in early recruitment research to illustrate the use of CLM in modeling direct effects, cross-level effects, and interaction effects in a crossover data structure. The statistical analysis on an actual data set shows that CLM is able to identify both company-level and applicant-level heterogeneities. The article presents further examples to show that CLM can be applied to many organizational research settings when multilevel crossover structures are present.
Impaired Ig class switch in mice deficient for the X-linked lymphoproliferative disease gene Sap
Authors: Al-Alem Umaima , Li Cuiling , Forey Nathalie , Relouzat Francis , Fondanèche Marie-Claude , Tavtigian Sean V. , Wang Zhao-Qi , Latour Sylvain , Yin Luo
Published: June 2005

X-linked lymphoproliferative disease (XLP) is characterized by abnormal immune responses to Epstein-Barr virus attributed to inactivating mutations of the SAP gene. Previous studies showed immunoglobulin E (IgE) deficiency and low serum IgG levels in Sap-deficient mice before and after viral infections, which are associated with impaired CD4+ T-helper function. In the present work, we find that signaling lymphocytic activation molecule (SLAM)-associated protein (SAP) is expressed in B cells and this expression is down-regulated after stimulation with lipopolysaccharide (LPS) and interleukin 4 (IL-4). We demonstrate that B cells from Sap-deficient mice exhibit reduced IgG and IgA production in vitro. This impairment correlates with decreased circular transcript levels of Iα, Iγ2a, Iγ2b, and Iγ3 after stimulation, which indicate a defective Ig switch recombination in Sap-deficient B cells. While XLP is believed to cause defects in T, natural killer T (NKT), and natural killer (NK) cells, our results indicate that B cells are also affected. (Blood. 2005;106:2069-2075)
Paradoxic inhibition of human natural interferon-producing cells by the activating receptor NKp44
Authors: Fuchs Anja , Cella Marina , Kondo Takayuki , Colonna Marco
Published: June 2005

Natural killer (NK) cell-mediated cytotoxicity is triggered by multiple activating receptors associated with the signaling adaptor protein DNAX activation protein 12/killer cell-activating receptor-associated protein (DAP12/KARAP). Here, we show that one of these receptors, NKp44, is present on a subset of natural interferon-producing cells (IPCs) in tonsils. NKp44 expression can also be induced on blood IPCs after in vitro culture with interleukin 3 (IL-3). Crosslinking of NKp44 does not trigger IPC-mediated cytotoxicity but, paradoxically, inhibits interferon α (IFN-α) production by IPCs in response to cytosine-phosphate-guanosine (CpG) oligonucleotides. We find that IPCs in tonsils are in close contact with CD8+ T cells and demonstrate that a subset of memory CD8+ T cells produces IL-3. Therefore, IL-3-mediated induction of NKp44 on IPCs may be an important component of the ongoing crosstalk between the innate and adaptive immune response that allows memory CD8+ T cells to control the IPC response to virus. (Blood. 2005;106: 2076-2082)
Inhibition of mitochondrial respiration elevates oxygen concentration but leaves regulation of hypoxia-inducible factor (HIF) intact
Authors: Doege Kathrin , Heine Sandra , Jensen Inga , Jelkmann Wolfgang , Metzen Eric
Published: June 2005

The transcription factor hypoxia-inducible factor-1 (HIF-1) is critical for erythropoietin and other factors involved in the adaptation of the organism to hypoxic stress. Conflicting results have been published regarding the role of the mitochondrial electron transport chain (ETC) in the regulation of HIF-1α. We assessed cellular hypoxia by pimonidazole staining and blotting of the O2-labile HIF-1 α-subunit in human osteosarcoma cell cultures (U2OS and 143B). In conventional, gas-impermeable cell culture dishes, ETC inhibitors had no effect on pimonidazole staining or HIF-1α abundance in a 20% O2 atmosphere; both parameters were undetectable. Pimonidazole staining and HIF activity were substantial in 0.1% O2 irrespective of ETC inhibition. At an intermediate oxygen concentration (3% O2) pimonidazole staining and HIF-α expression were detectable but strongly reduced after ETC inhibition in conventional cell cultures. All effects of ETC inhibition on HIF-1α regulation were eliminated in gas-permeable dishes. As shown in a 143B subclone deficient in mitochondrial DNA (206ρ0), genetic inactivation of the ETC led to similar responses with respect to HIF-1α regulation as ETC inhibitors. Our data demonstrate that reduction of oxygen consumption reduces the O2 gradient in conventional cell cultures, causing elevation of the cellular O2 concentration, which leads to degradation of HIF-α.
Respiratory rehabilitation after acute exacerbation of COPD may reduce risk for readmission and mortality – a systematic review
Authors: Puhan Milo A , Scharplatz Madlaina , Troosters Thierry , Steurer Johann
Published: June 2005

Background

Acute exacerbations of chronic obstructive pulmonary disease (COPD) represent a major burden for patients and health care systems. Respiratory rehabilitation may improve prognosis in these patients by addressing relevant risk factors for exacerbations such as low exercise capacity. To study whether respiratory rehabilitation after acute exacerbation improves prognosis and health status compared to usual care, we quantified its effects using meta-analyses.

Methods

Systematic review of randomized controlled trials identified by searches in six electronic databases, contacts with experts, hand-searches of bibliographies of included studies and conference proceedings. We included randomized trials comparing the effect of respiratory rehabilitation and usual care on hospital admissions, health-related quality of life (HRQL), exercise capacity and mortality in COPD patients after acute exacerbation. Two reviewers independently selected relevant studies, extracted the data and evaluated the study quality. We pooled the results using fixed effects models where statistically significant heterogeneity (p ≤ 0.1) was absent.

Results

We identified six trials including 230 patients. Respiratory rehabilitation reduced the risk for hospital admissions (pooled relative risk 0.26 [0.12–0.54]) and mortality (0.45 [0.22–0.91]). Weighted mean differences on the Chronic Respiratory Questionnaire were 1.37 (95% CI 1.13–1.61) for the fatigue domain, 1.36 (0.94–1.77) for emotional function and 1.88 (1.67–2.09) for mastery. Weighted mean differences for the St. Georges Respiratory Questionnaire total score, impacts and activities domains were -11.1 (95% CI -17.1 to -5.2), -17.1 (95% CI -23.6 to -10.7) and -9.9 (95% CI -18.0 to -1.7). In all trials, rehabilitation improved exercise capacity (64–215 meters in six-minute walk tests and weighted mean difference for shuttle walk test 81 meter, 95% CI 48–115).

Conclusion

Evidence from six trials suggests that respiratory rehabilitation is effective in COPD patients after acute exacerbation. Larger trials, however, are needed to further investigate the role of respiratory rehabilitation after acute exacerbation and its potential to reduce costs caused by COPD.
TGF-β1 induces human alveolar epithelial to mesenchymal cell transition (EMT)
Authors: Kasai Hidenori , Allen Jeremy T , Mason Roger M , Kamimura Takashi , Zhang Zhi
Published: June 2005

Background

Fibroblastic foci are characteristic features in lung parenchyma of patients with idiopathic pulmonary fibrosis (IPF). They comprise aggregates of mesenchymal cells which underlie sites of unresolved epithelial injury and are associated with progression of fibrosis. However, the cellular origins of these mesenchymal phenotypes remain unclear. We examined whether the potent fibrogenic cytokine TGF-β1 could induce epithelial mesenchymal transition (EMT) in the human alveolar epithelial cell line, A549, and investigated the signaling pathway of TGF-β1-mediated EMT.

Methods

A549 cells were examined for evidence of EMT after treatment with TGF-β1. EMT was assessed by: morphology under phase-contrast microscopy; Western analysis of cell lysates for expression of mesenchymal phenotypic markers including fibronectin EDA (Fn-EDA), and expression of epithelial phenotypic markers including E-cadherin (E-cad). Markers of fibrogenesis, including collagens and connective tissue growth factor (CTGF) were also evaluated by measuring mRNA level using RT-PCR, and protein by immunofluorescence or Western blotting. Signaling pathways for EMT were characterized by Western analysis of cell lysates using monoclonal antibodies to detect phosphorylated Erk1/2 and Smad2 after TGF-β1 treatment in the presence or absence of MEK inhibitors. The role of Smad2 in TGF-β1-mediated EMT was investigated using siRNA.

Results

The data showed that TGF-β1, but not TNF-α or IL-1β, induced A549 cells with an alveolar epithelial type II cell phenotype to undergo EMT in a time-and concentration-dependent manner. The process of EMT was accompanied by morphological alteration and expression of the fibroblast phenotypic markers Fn-EDA and vimentin, concomitant with a downregulation of the epithelial phenotype marker E-cad. Furthermore, cells that had undergone EMT showed enhanced expression of markers of fibrogenesis including collagens type I and III and CTGF. MMP-2 expression was also evidenced. TGF-β1-induced EMT occurred through phosphorylation of Smad2 and was inhibited by Smad2 gene silencing; MEK inhibitors failed to attenuate either EMT-associated Smad2 phosphorylation or the observed phenotypic changes.

Conclusion

Our study shows that TGF-β1 induces A549 alveolar epithelial cells to undergo EMT via Smad2 activation. Our data support the concept of EMT in lung epithelial cells, and suggest the need for further studies to investigate the phenomenon.