bluuebunny/crossref_metadata_2025_split huggingface dataset:
Thrombin-Induced Connective Tissue Growth Factor Expression in Human Lung Fibroblasts Requires the ASK1/JNK/AP-1 Pathway
Authors: Yu Chung-Chi , Hsu Ming-Jen , Kuo Min-Liang , Chen Robert Fu-Chean , Chen Mei-Chieh , Bai Kua-Jen , Yu Ming-Chih , Chen Bing-Chang , Lin Chien-Huang 
Published: June 2009

Abstract
Thrombin plays an important role in lung inflammatory diseases. Thrombin can induce connective tissue growth factor (CTGF) expression in lung fibroblasts. However, little is known about the signaling pathway in thrombin-induced CTGF expression. In this study, we investigated the role of apoptosis signal-regulating kinase 1 (ASK1) in thrombin-induced CTGF expression in human lung fibroblasts. Thrombin caused a concentration- and time-dependent increase in CTGF expression in WI-38 cells and primary lung fibroblasts. Thrombin-induced CTGF expression and CTGF-luciferase activity were inhibited by a protease-activated receptor 1 antagonist (SCH79797), the dominant-negative mutants (DNs) of ASK1 and JNK1/2, and an AP-1 inhibitor (curcumin). Thrombin caused ASK1 Ser967 dephosphorylation, the dissociation of ASK1 and 14-3-3, and a subsequent increase in ASK1 activity. Thrombin induced increases in JNK phosphorylation and kinase activity, which were attenuated by ASK1DN. Furthermore, SCH79797 diminished the thrombin-induced ASK1 and JNK activities. Thrombin-induced CTGF-luciferase activity was predominately controlled by the sequence −747 to −184 bp upstream of the transcription start site of the human CTGF promoter and was attenuated by transfection with the deleted AP-1 binding site construct. Thrombin caused increases in c-Jun phosphorylation, the formation of an AP-1-specific DNA-protein complex, and the recruitment of c-Jun to the CTGF promoter. Furthermore, thrombin-mediated AP-1 activation was inhibited by ASK1DN, JNK1/2DN, and SP600125. These results suggest for the first time that thrombin, acting through protease-activated receptor 1, activates the ASK1/JNK signaling pathway, which in turn initiates c-Jun/AP-1 activation and recruitment of c-Jun to the CTGF promoter and ultimately induces CTGF expression in human lung fibroblasts.
Dok-4 Is a Novel Negative Regulator of T Cell Activation
Authors: Gérard Audrey , Ghiotto Marguerite , Fos Camille , Guittard Geoffrey , Compagno Daniel , Galy Anne , Lemay Serge , Olive Daniel , Nunès Jacques A. 
Published: June 2009

Abstract
Dok-4 (downstream of tyrosine kinase-4) is a recently identified member of the Dok family of adaptor proteins, which are characterized by an amino-terminal pleckstrin homology domain, a phosphotyrosine-binding domain, and a carboxyl-terminal region containing several tyrosines and poly-proline-rich motifs. Two members of the Dok family, Dok-1 and Dok-2, have already been described as negative regulators in T cells. However, the function of Dok-4, which is also expressed in T cells, remains unknown. In this study, we report that Dok-4 is phosphorylated after TCR engagement and shuttled within the cytoplasm of T cells before being recruited to the polarized microtubule organizing center after the formation of the immunological synapse. Loss-of-function experiments using RNA interference constructs show that Dok-4 is a negative regulator of ERK phosphorylation, IL-2 promoter activity, and T cell proliferation. Exogenous expression of wild-type Dok-4 induces a significant activation of Rap1, which is involved in the regulation of ERK. The pleckstrin homology domain of Dok-4 is required both for its cytoplasmic shuttling and relocalization as well as for its inhibitory properties on T cell activation. Thus, Dok-4 represents a novel negative regulator of T cells.
Pyrin Critical to Macrophage IL-1β Response to Francisella Challenge
Authors: Gavrilin Mikhail A. , Mitra Srabani , Seshadri Sudarshan , Nateri Jyotsna , Berhe Freweine , Hall Mark W. , Wewers Mark D. 
Published: June 2009

Abstract
Relative to monocytes, human macrophages are deficient in their ability to process and release IL-1β. In an effort to explain this difference, we used a model of IL-1β processing and release that is dependent upon bacterial escape into the cytosol. Fresh human blood monocytes were compared with monocyte-derived macrophages (MDM) for their IL-1β release in response to challenge with Francisella novicida. Although both cell types produced similar levels of IL-1β mRNA and intracellular pro-IL-1β, only monocytes readily released processed mature IL-1β. Baseline mRNA expression profiling of candidate genes revealed a remarkable deficiency in the pyrin gene, MEFV, expression in MDM compared with monocytes. Immunoblots confirmed a corresponding deficit in MDM pyrin protein. To determine whether pyrin levels were responsible for the monocyte/MDM difference in mature IL-1β release, pyrin expression was knocked down by nucleofecting small interfering RNA against pyrin into monocytes or stably transducing small interfering RNA against pyrin into the monocyte cell line, THP-1. Pyrin knockdown was associated with a significant drop in IL-1β release in both cell types. Importantly, M-CSF treatment of MDM restored pyrin levels and IL-1β release. Similarly, the stable expression of pyrin in PMA-stimulated THP-1-derived macrophages induces caspase-1 activation, associated with increased IL-1β release after infection with F. novicida. In summary, intracellular pyrin levels positively regulate MDM IL-1β responsiveness to Francisella challenge.
Keratinocyte Growth Factor Improves Allogeneic Bone Marrow Engraftment through a CD4+Foxp3+ Regulatory T Cell-Dependent Mechanism
Authors: Bruinsma Marieke , van Soest Peter L. , Leenen Pieter J. M. , Löwenberg Bob , Cornelissen Jan J. , Braakman Eric 
Published: June 2009

Abstract
Keratinocyte growth factor (KGF) protects mice from acute graft-vs-host disease and graft rejection by cytoprotective and yet incompletely understood immunological mechanisms. Recently, we showed that administration of KGF induces selective peripheral expansion of CD4+Foxp3+ regulatory T cells (Treg). In this study, we set out to assess whether the peripheral expansion of Treg accounts for the immunomodulatory effects of KGF after bone marrow (BM) transplantation. To exclude potentially confounding cytoprotective and thymopoietic effects of KGF, we applied KGF to congenic wild-type mice that served as T cell provider mice for T and B cell-deficient RAG-1−/− mice that were subsequently transplanted with allogeneic BM. Treatment of congenic T cell provider mice with KGF significantly improved engraftment and reduced graft rejection in BMT recipients. CD4+Foxp3+ Treg remained increased for 4 wk, while expansion of congenic CD3+ T cells was inhibited. To assess a causal relationship between expansion of Treg and improved BM engraftment, congenic Scurfy mice, which lack Foxp3+ Treg, served as T cell provider mice and were treated with KGF. KGF-treatment of Scurfy mice did not affect engraftment nor did it inhibit the expansion of congenic T cells. These data demonstrate that administration of KGF to the T cell provider mice improves engraftment of allogeneic BM through a CD4+Foxp3+ Treg-dependent mechanism.
Defective Regulatory T Cells In Patients with Severe Drug Eruptions: Timing of the Dysfunction Is Associated with the Pathological Phenotype and Outcome
Authors: Takahashi Ryo , Kano Yoko , Yamazaki Yoshimi , Kimishima Momoko , Mizukawa Yoshiko , Shiohara Tetsuo 
Published: June 2009

Abstract
Toxic epidermal necrolysis (TEN) and drug-induced hypersensitivity syndrome (DIHS) represent two ends of a spectrum of severe drug eruptions: DIHS is unique in that severe epidermal damage seen in TEN is absent, sequential reactivations of herpesviruses occur, and autoimmunity often ensues. To investigate whether changes in regulatory T (Treg) cell function would contribute to variability in the clinical manifestations, we examined the frequency, phenotype, and function of Treg cells both during the acute stage and again long after clinical resolution of both diseases. Dramatic expansions of functional Treg cells were found in the acute stage of DIHS. In contrast, Treg function was profoundly impaired in TEN, although present in normal frequency. Skin homing addressins were more preferentially expressed on Treg cells in DIHS than in TEN. Indeed, Treg cells were more abundantly present in the skin lesions of DIHS. Surprisingly, Treg cells contracted upon resolution of DIHS became functionally deficient, whereas their functional defects in TEN were restored upon recovery. These findings indicate that a transitory impairment in their function during the acute stage of TEN may be related to severe epidermal damage, while a gradual loss of their function after resolution of DIHS may increase the risk of subsequently developing autoimmune disease.
Regulation of Th17 Differentiation by Epidermal Fatty Acid-Binding Protein
Authors: Li Bing , Reynolds Joseph M. , Stout Robert D. , Bernlohr David A. , Suttles Jill 
Published: June 2009

Abstract
Epidermal fatty acid-binding protein, E-FABP, a lipid chaperone, has been shown to regulate the inflammatory function of macrophages and dendritic cells. Herein, we demonstrate that T cell expression of E-FABP promotes Th17 differentiation, while counterregulating development of FoxP3+ regulatory T cells (Tregs). In response to immunization with myelin oligodendrocyte glycoprotein peptide (MOG35–55), E-FABP-deficient mice generated reduced levels of Th17 cells and elevated levels of Tregs, as compared with wild-type mice. Likewise, naive CD4+ T cells isolated from E-FABP-deficient mice showed reduced expression of IL-17 and enhanced expression of FoxP3, in vitro, when subjected to Th17 or Treg polarizing conditions, respectively. It has been demonstrated previously that IL-21, induced by IL-6, stimulates the expression of the nuclear receptors retinoic acid-related orphan receptor (ROR)γt and RORα, which in turn induce expression of IL-17. We found that the impaired Th17 differentiation by E-FABP-deficient CD4+ T cells was associated with lower levels of IL-21 expression in response to IL-6, as well as reduced expression of RORγt and RORα. However, E-FABP-deficient CD4+ T cells expressed significantly higher levels of the nuclear receptor peroxisome proliferator-activating receptor (PPAR)γ than did wild-type CD4+ T cells, and treatment with the PPARγ antagonist GW9662 restored expression of IL-21, RORγt, RORα, and IL-17 by E-FABP-deficient T cells to wild-type levels. The negative influence of E-FABP deficiency on IL-17 expression was attributed to PPARγ-mediated suppression of IL-6-induced STAT3 activity. Thus, taken together, our data indicate that expression of E-FABP by CD4+ T cells contributes to the control of IL-6 stimulation of the IL-21/ROR/IL-17 pathway and to the Th17/Treg counterbalance.
Hypoxia Prolongs Monocyte/Macrophage Survival and Enhanced Glycolysis Is Associated with Their Maturation under Aerobic Conditions
Authors: Roiniotis John , Dinh Hang , Masendycz Paul , Turner Amanda , Elsegood Caryn L. , Scholz Glen M. , Hamilton John A. 
Published: June 2009

AbstractIn chronic inflammatory lesions macrophages are abundant and adapt to the low oxygen concentrations often present there. In low oxygen some cell types die by apoptosis, as reported for macrophage cell lines, while others survive better as they shift their metabolism to anaerobic glycolysis. It was found here that hypoxia prolongs the survival of murine bone marrow-derived macrophages, either in the absence or presence of low CSF-1 (M-CSF) concentrations. Although Akt activity increased in bone marrow-derived macrophages in the low oxygen conditions, the levels of both anti- and proapoptotic Bcl-2 family members decreased. Glycolysis was enhanced as judged by increased glucose uptake, glucose transporter expression, lactate dehydrogenase mRNA expression, and lactate secretion. Human monocytes responded similarly to low oxygen, and a number of genes associated with glycolysis were shown by microarray analysis and quantitative PCR to be up-regulated. Interestingly, human monocyte-derived macrophages showed evidence of enhanced glycolysis even under aerobic conditions. It is proposed that certain monocyte/macrophage populations survive better under conditions of low oxygen, thereby contributing to their increased numbers at sites of chronic inflammation and tumors; it is also proposed that as macrophages differentiate from monocytes they begin to adopt a glycolytic metabolism allowing them to adapt readily when exposed to low oxygen conditions.
IL-10 Deficiency Unleashes an Influenza-Specific Th17 Response and Enhances Survival against High-Dose Challenge
Authors: McKinstry K. Kai , Strutt Tara M. , Buck Amanda , Curtis Jonathan D. , Dibble John P. , Huston Gail , Tighe Michael , Hamada Hiromasa , Sell Stewart , Dutton Richard W. , Swain Susan L. 
Published: June 2009

Abstract
We examined the expression and influence of IL-10 during influenza infection. We found that IL-10 does not impact sublethal infection, heterosubtypic immunity, or the maintenance of long-lived influenza Ag depots. However, IL-10-deficient mice display dramatically increased survival compared with wild-type mice when challenged with lethal doses of virus, correlating with increased expression of several Th17-associated cytokines in the lungs of IL-10-deficient mice during the peak of infection, but not with unchecked inflammation or with increased cellular responses. Foxp3− CD4 T cell effectors at the site of infection represent the most abundant source of IL-10 in wild-type mice during high-dose influenza infection, and the majority of these cells coproduce IFN-γ. Finally, compared with predominant Th1 responses in wild-type mice, virus-specific T cell responses in the absence of IL-10 display a strong Th17 component in addition to a strong Th1 response and we show that Th17-polarized CD4 T cell effectors can protect naive mice against an otherwise lethal influenza challenge and utilize unique mechanisms to do so. Our results show that IL-10 expression inhibits development of Th17 responses during influenza infection and that this is correlated with compromised protection during high-dose primary, but not secondary, challenge.
Des codes pour rire
Authors: Béru Laurent 
Published: June 2009

Résumé Type d’humour originaire des États-Unis, le stand-up , qui s’apparente à de « l’improvisation préparée », s’est invité sur une chaîne audiovisuelle française (Canal Plus), au cours des étés 2006 et 2007, sous la forme d’un programme satirique : le Jamel Comedy Club . Présentée et produite par l’acteur comique Jamel Debbouze, cette émission hebdomadaire diffusée en prime time a permis l’exposition médiatique d’une toute nouvelle génération d’humoristes français, dont la majorité provient des quartiers populaires et/ou appartient aux minorités ethnoraciales. Derrière le divertissement apparent, une certaine critique de la société française apparaît. L’un des objectifs des auteurs de ce programme était en effet le suivant : faire rire les téléspectateurs autour de problèmes sociaux difficiles qui touchent certains lieux et certaines populations en France, notamment la discrimination ethnoraciale et la ségrégation sociospatiale.
Discours journalistique et parole ordinaire : analyse d’un rendez-vous manqué
Authors: Croissant Valérie , Touboul Annelise 
Published: June 2009

Résumé Participative, critique ou productive, la qualification de la parole des usagers dans l’espace médiatique varie d’autant plus que ces phénomènes semblent prendre de l’ampleur ces dernières années. Cet article se propose d’analyser deux types d’espaces médiatiques dans lesquels la parole des usagers est diffusée : les forums des sites Internet de presse et les chroniques du médiateur de Radio France. Il s’agit ici d’interroger les modalités de mise en scène de la parole ordinaire dans les médias. Dans leur manière de rendre compte de cette parole d’usagers, ces espaces participent en fait de la délimitation du champs journalistique, et donc de l’accès à la sphère publique par une différenciation des discours. Le cadre événementiel de cette analyse est le référendum français sur la constitution européenne de 2005.
Vaches et blés sur le papier
Authors: Joly Nathalie 
Published: June 2009

Résumé En agriculture comme ailleurs, l’activité des professionnels apparaît de plus en plus équipée par toute sorte d’instruments destinés à noter, à enregistrer ou à mesurer. Pour cerner les enjeux et la nature exacte des changements signalés par ce constat, l’article propose de s’intéresser aux pratiques ordinaires de l’écrit dans le métier d’agriculteur et de les resituer dans une perspective historique. En décrivant plus particulièrement une pratique extrêmement répandue dans le monde agricole, la tenue d’agendas qui recueillent une chronique quotidienne des occupations sur la ferme, il éclaire par ce biais la diversité des modelages dont les écrits du travail ont pu faire l’objet de la part des institutions d’enseignement et de vulgarisation tout au long des xix e et xx e siècles et dont le sens général est celui d’une adaptation à un ordre économique et technique et d’une rationalisation des méthodes de gestion. Il met également en évidence les ressources cognitives associées à cette mise en mémoire du travail agricole, les réglages de l’activité qu’elle autorise, les formes de jugement et de réflexivité qu’elle soutient. C’est donc à une réflexion sur les modes d’appropriation des pratiques de l’écrit à laquelle l’auteur invite, nourrie d’exemples de détournements et de reformulations que les agriculteurs ont régulièrement opérés face aux cadrages normatifs qui leur étaient proposés. Une telle perspective s’avère particulièrement utile à la saisie des transformations en cours concernant les écrits du travail. Confrontés depuis quelques années à des exigences croissantes de traçabilité de leurs pratiques, les agriculteurs sont davantage sommés que par le passé de recourir à des canevas d’écriture standardisés. Se pose alors une série de questions sur les modalités de l’articulation entre des écrits confectionnés « pour soi » et des écrits destinés à « faire preuve » que l’auteur esquisse en conclusion, pour préciser des orientations de recherche à venir.
P1 (Adenosine) Purinoceptor Assays
Authors: Mustafa S. Jamal , Ansari Habib R. , Abebe Worku 
Published: June 2009

AbstractP1 purinoceptors, or adenosine (ADO) receptors, mediate the biological effects of the endogenous nucleoside, ADO and its analogs. ADO works through four receptor subtypes: A1, A2A, A2B, and A3. Isolated tissue assays used for the pharmacological characterization of ADO receptors based on functional responses are described in this unit. The guinea pig atrium, pig coronary artery, guinea pig aorta ,and mouse aorta have been used for the characterization of ADO receptor subtypes. Curr. Protoc. Pharmacol. 45:4.7.1‐4.7.13. © 2009 by John Wiley & Sons, Inc.
Models of Anxiety: Stress‐Induced Hyperthermia (SIH) in Singly Housed Mice
Authors: Groenink Lucianne , Vinkers Christiaan , van Oorschot Ruud , Olivier Berend 
Published: June 2009

AbstractDescribed in this unit is the stress‐induced hyperthermia (SIH) test in mice in a single‐housed format. This protocol has proven reliable in detecting the anxiolytic properties of test compounds. In this test, SIH is quantified in singly housed mice using a rectal temperature measurement as the stressor. Rectal temperature is measured twice at a 10‐min interval. Due to the stress experienced during the first temperature measurement, the temperature of the second measurement (T2) is ∼0.8° to 1.5°C higher than that of the first (T1). This difference in temperature (ΔT = T2 − T1) is defined as the SIH response. The SIH response is reduced by different classes of anxiolytics. The SIH test is simple and robust, it does not require training of animals, and test compound effects on motor behavior, feeding, and nociception do not affect test outcome. Furthermore, it is one of few anxiety tests that focuses on the physiological component of anxiety. Curr. Protoc. Pharmacol. 45:5.16.1‐5.16.12. © 2009 by John Wiley & Sons, Inc.
Acute Seizure Tests in Epilepsy Research: Electroshock‐ and Chemical‐Induced Convulsions in the Mouse
Authors: Giardina William J. , Gasior Maciej 
Published: June 2009

AbstractEpilepsy is a common (50 million patients worldwide) neurological disorder characterized by seizures that are caused by episodic abnormal electrical activity in the brain. Animal models play an essential role in epilepsy research including the discovery and development of new antiepileptic drugs. Described in this unit are protocols for traditional acute tests in which seizures are induced by either an electrical stimulation or a convulsant agent in non‐epileptic mice. Specifically, protocols for the following acute seizure tests are provided: the maximal electroshock induced test (MES), the maximal electroshock seizure threshold (MEST) test, the 6‐Hz seizure test, the subcutaneous pentylenetetrazol (s.c. PTZ) seizure test, and the intravenous pentylenetetrazol (i.v. PTZ) seizure test. These tests can be used to characterize anticonvulsant and/or proconvulsant properties of compounds in mice. The MES, s.c. PTZ, and 6‐Hz seizure tests represent the three most widely used animal tests in drug‐screening programs. Although the parameters of these tests are optimized for mice, the same tests (except for the 6‐Hz seizure test), with some modifications, can be used with rats.Curr. Protoc. Pharmacol. 45:5.22.1‐5.22.37. © 2009 by John Wiley & Sons, Inc.
Models of Aspects of Schizophrenia: Behavioral Sensitization Induced by Subchronic Phencyclidine Administration
Authors: Kalinichev Mikhail , Bate Simon T. , Jones Declan N.C. 
Published: June 2009

AbstractPresented in this unit is a protocol using subchronically administered phencyclidine (PCP) for establishing a behavioral sensitization model of aspects of schizophrenia. This model is validated using haloperidol and risperidone. The end‐point of the assay is locomotor hyperactivity, which is induced by PCP challenge following subchronic treatment with this NMDA receptor antagonist. The antipsychotics haloperidol, risperidone, and quetiapine all reduce hyperactivity in a dose‐dependent and selective manner. While the effects of other antipsychotics such as clozapine, olanzapine, and ziprasidone are similar to haloperidol, the interpretation of responses to them is often confounded by nonspecific effects during habituation.Curr. Protoc. Pharmacol. 45:5.54.1‐5.54.13. © 2009 by John Wiley & Sons, Inc.
Unnatural Nucleosides with Unusual Base Pairing Properties
Authors: Bergstrom Donald E. 
Published: June 2009

AbstractSynthetic modified nucleosides designed to pair in unusual ways with natural nucleobases have many potential applications in biology and biotechnology. This overview lays the foundation for future protocol units on synthesis and application of unnatural bases, with particular emphasis on unnatural base analogs that mimic natural bases in size, shape, and biochemical processing. Topics covered include base pairs with alternative H‐bonding schemes, dimensionally expanded base pairs, hydrophobic base pairs, metal‐ligated bases, degenerate bases, universal nucleosides, and triplex constituents. Curr. Protoc. Nucleic Acid Chem. 37:1.4.1‐1.4.32. © 2009 by John Wiley & Sons, Inc.
UV Melting of G‐Quadruplexes
Authors: Mergny Jean‐Louis , Lacroix Laurent 
Published: June 2009

AbstractAbsorbance versus temperature curves can provide information on the thermal stability of DNA or RNA quadruplexes. Quadruplex denaturation or renaturation can be followed by recording absorbance at 295 nm rather than 260 nm, the wavelength used to monitor duplex denaturation. This unit describes the use of absorbance versus temperature curves to determine melting temperatures (Tm values) and model‐dependent thermodynamic parameters. Curr. Protoc. Nucleic Acid Chem. 37:17.1.1‐17.1.15. © 2009 by John Wiley & Sons, Inc.