bluuebunny/crossref_metadata_2025_split huggingface dataset:
Over‐expression of bHLH genes facilitate neural formation of mouse embryonic stem (ES) cells in vitro
Authors: Kanda Seiji , Tamada Yasuko , Yoshidome Atsuko , Hayashi Itoko , Nishiyama Toshimasa 
Published: March 2004

AbstractMouse embryonic stem (ES) cells are useful tools for investigating differentiation into neurons and glial cells in vitro. In order to induce ES cells to differentiate into neural cells, many researchers have investigated the efficiency of induction. Embryoid body (EB) formation and retinoic acid are potent differentiation inducers known to be a trigger at the early stage of development. Basic helix‐loop‐helix (bHLH) is one of the important transcription factors, which is essential for premature neural formation. In NeuroD2 and Mash1‐transfected cells, neural formation was observed at day 6 after the plating of embryoid bodies in culture. Nestin was detected in NeuroD2‐ and Mash1‐transfected cells at day 10, and strong signal was detected in Mash1 transfectants by RT‐PCR analysis. Map2 and Nurr1 were also detected strongly at the early stage in transfected cells compared with the wild type control, especially in the Mash1 transfectant. In immunocytochemical analysis, Tuj1‐positive neurons were detected at high frequency in Mash1 transfectants and some cells were stained by tyrosine hydrogenase (TH), a marker of dopaminergic neurons. These results demonstrate that bHLH has a potential activity at an early stage for ES cells and can induce effective and rapid neural differentiation in vitro.
Assessing joint maintenance shops in the Taiwanese Army using data envelopment analysis
Authors: Sun Shinn 
Published: March 2004

AbstractThis paper proposes an alternative data envelopment analysis (DEA) method for assessing the performance of joint maintenance shops (JMSs) in the Taiwanese Army over two 6‐month periods in 2000. It further examines the possibility of using DEA over time as a basis for continuous improvement. To help productivity improvements, the DEA results for the first time period were implemented at the five selected JMSs located in Taipei, Taoyuan, Taichung, Chaiyi, and Kaoshiung. The findings show that most of previously inefficient JMSs, on average, have become relative more efficient through DEA recommendation remedial actions. Three JMSs increased their technical and pure technical efficiencies; and one JMS increased its scale efficiency. Few JMSs were operating at increasing returns to scale, indicating a potential to expand. As such, it can be a valuable benchmarking tool for the JMS managers and assist in efficiently using scare resources to produce productivity. Finally, the Army has accepted DEA as a performance measurement tool for JMSs.
NASA Rover finds evidence of a once wet and habitable Mars
Authors: Showstack Randy 
Published: March 2004

NASA's Opportunity rover has found strong evidence that a rock outcrop in Mars' Meridiani Planum region once was a wet and habitable environment, rover science team members announced at a 2 March news briefing at NASA headquarters in Washington, D.C.“We have concluded that the rocks here were once soaked in liquid water”said Steven Squyres, Mars Exploration Rover principal investigator and astronomy professor at Cornell University. He said it was not yet possible to indicate how long ago or for how long the rocks were wet.
University of Connecticut Geology Department faces dissolution
Authors: Showstack Randy 
Published: March 2004

The University of Connecticut's board of trustees is expected to vote in favor of dissolving the school's department of geology and geophysics at its 23 March meeting.The board has been prompted to act for several reasons, including a perceived lack of cohesion and direction within the department. An Internet petition drive to save the geology department had garnered nearly 3,800 signatures by 6 March. However, some individuals, including department chair Raymond Joesten, view dissolution as a potentially positive measure that would allow geology and geophysics studies in the university to move forward.
Back to the future
Authors: Witton Leah Christen 
Published: March 2004

Past Global Changes (PAGES) is a worldwide program that seeks to facilitate international and interdisciplinary collaborations aimed at understanding the Earth's past environment to make sound predictions for the future. PAGES' scope of interest includes the physical climate system, biogeochemical cycles, ecosystem processes, biodiversity and human dimensions.PAGES is a core project of the International Geosphere‐Biosphere Programme (IGBP) and is funded by the U.S. National Science Foundation, the National Oceanic and Atmospheric Administration, and the Swiss National Science Foundation; and is overseen by a Scientific Steering Committee comprised of members chosen to provide a range of scientific expertise and a balance of nationalities. It is run by a small International Project Office that serves as a critical interface between scientists and science initiatives. The Office manages projects; organizes and supports meetings and courses; aids in synthesizing research results in collaboration with scientists at large; publishes a free, international science newsletter; and provides and maintains a Web site that offers a variety of additional services. PAGES has over 3000 member scientists in more than 100 countries around the world. Membership is free and open to anyone. The PAGES Web site, www.pages‐igbp.org, receives over 1500 hits a day.
General anesthetic octanol and related compounds activate wild‐type and delF508 cystic fibrosis chloride channels
Authors: Marcet Brice , Becq Frédéric , Norez Caroline , Delmas Patrick , Verrier Bernard 
Published: March 2004

Cystic fibrosis transmembrane conductance regulator (CFTR) Cl− channel is defective during cystic fibrosis (CF). Activators of the CFTR Cl− channel may be useful for therapy of CF. Here, we demonstrate that a range of general anesthetics like normal‐alkanols (n‐alkanols) and related compounds can stimulate the Cl− channel activity of wild‐type CFTR and delF508‐CFTR mutant.
The effects of n‐alkanols like octanol on CFTR activity were measured by iodide (125I) efflux and patch‐clamp techniques on three distinct cellular models: (1) CFTR‐expressing Chinese hamster ovary cells, (2) human airway Calu‐3 epithelial cells and (3) human airway JME/CF15 epithelial cells which express the delF508‐CFTR mutant.
Our data show for the first time that n‐alkanols activate both wild‐type CFTR and delF508‐CFTR mutant. Octanol stimulated 125I efflux in a dose‐dependent manner in CFTR‐expressing cells (wild‐type and delF508) but not in cell lines lacking CFTR. 125I efflux and Cl− currents induced by octanol were blocked by glibenclamide but insensitive to 4,4′‐diisothiocyanatostilbene‐2,2′‐disulfonic acid, as expected for a CFTR Cl− current.
CFTR activation by octanol was neither due to cell‐to‐cell uncoupling properties of octanol nor to an intracellular cAMP increase. CFTR activation by octanol requires phosphorylation by protein kinase‐A (PKA) since it was prevented by H‐89, a PKA inhibitor.
n‐Alkanols chain length was an important determinant for channel activation, with rank order of potencies: 1‐heptanol<1‐octanol<2‐octanol<1‐decanol. Our findings may be of valuable interest for developing novel therapeutic strategies for CF.

British Journal of Pharmacology (2004) 141, 905–914. doi:10.1038/sj.bjp.0705684
α1‐Adrenoceptor subtypes involved in vasoconstrictor responses to exogenous and neurally released noradrenaline in rat femoral resistance arteries
Authors: Zacharia Joseph , Hillier Chris , MacDonald Allan 
Published: March 2004

The α1‐adrenoceptor subtypes involved in responses to exogenous and neurally released noradrenaline in rat femoral resistance arteries were characterised using a small vessel myograph, with antagonists prazosin (nonsubtype selective), 5‐methyl‐urapidil (α1A‐selective), BMY 7378 (α1D‐selective) and the alkylating agent chloroethylclonidine (preferential for α1B‐).
Prazosin and 5‐methyl‐urapidil produced rightward shifts of the exogenous noradrenaline concentration – response curve (CRC) with pA2 values of 9.2 and 9.1 respectively, in agreement with the presence of α1A‐adrenoceptors. BMY 7378 (1 μM) shifted the noradrenaline CRC with an apparent pKB of 6.7, in agreement with the presence of α1A‐, but not α1D‐, adrenoceptors. Chloroethylclonidine at 1 μM had no effect and at 10 μM produced only a small reduction (c. 20%) in the maximum response to noradrenaline, indicating little, if any, contribution from α1B‐adrenoceptors.
Responses of the rat femoral resistance arteries to electrical field stimulation (EFS) at 5–30 Hz for 10 s and 0.05 ms pulse width were principally due to α1‐adrenoceptor stimulation. Prazosin and 5‐methyl‐urapidil inhibited EFS‐mediated responses with pIC50s of 9.3 and 8.2, respectively, consistent with the α1A‐adrenoceptor being the predominant subtype. Responses to EFS at 10–30 Hz were relatively insensitive to BMY 7378 (pIC50, 6.5–6.7), while responses to 5 Hz were inhibited with a significantly higher pIC50 of 8.02, suggesting the contribution of α1D‐adrenoceptors. Chloroethylclonidine had no effect on responses to EFS, ruling out the contribution of an α1B‐subtype.
In the presence of cocaine, the predominant subtype involved in responses to EFS was the α1A‐adrenoceptor, with a contribution from α1D‐adrenoceptors at low frequency, as seen in the absence of cocaine. However, there was also a significant increase in the sensitivity to BMY 7378 at higher frequencies, suggesting that a further small α1D‐adrenoceptor component may be uncovered in the presence of cocaine.
The present study has shown a predominant role of the α1A‐adrenoceptor in contractions due to exogenous noradrenaline and to neurally released noradrenaline in rat femoral resistance arteries. α1D‐Adrenoceptors are not involved in responses to exogenous noradrenaline but appear to be activated by neurally released noradrenaline at a low frequency of stimulation and at higher frequencies in the presence of neuronal‐uptake blockade.

British Journal of Pharmacology (2004) 141, 915–924. doi:10.1038/sj.bjp.0705690
Differential regulation by AT1 and AT2 receptors of angiotensin II‐stimulated cyclic GMP production in rat uterine artery and aorta
Authors: Hannan Ruth E , Gaspari Tracey A , Davis Elizabeth A , Widdop Robert E 
Published: March 2004

In the present study we determined whether angiotensin II (Ang II) could increase cyclic GMP levels in two blood vessels that exhibit markedly different angiotensin II receptor subtype expression: rat uterine artery (UA; AT2 receptor‐predominant) and aorta (AT1 receptor‐predominant), and investigated the receptor subtype(s) and intracellular pathways involved.
UA and aorta were treated with Ang II in the absence and presence of losartan (AT1 antagonist; 0.1 μM), PD 123319 (AT2 antagonist; 1 μM), NOLA (NOS inhibitor; 30 μM), and HOE 140 (B2 antagonist; 0.1 μM), or in combination.
Ang II (10 nM) induced a 60% increase in UA cyclic GMP content; an effect that was augmented with PD 123319 and HOE 140 pretreatment, and abolished by cotreatment with losartan, as well as by NOLA.
In aorta, Ang II produced concentration‐dependent increases in cyclic GMP levels. Unlike effects in UA, these responses were abolished by PD 123319 and by NOLA, whereas losartan and HOE 140 caused partial inhibition.
Thus, in rat UA, Ang II stimulates cyclic GMP production through AT1 and, to a less extent, AT2 receptors. In rat aorta, the Ang II‐mediated increase in cyclic GMP production is predominantly AT2 receptor‐mediated. In both preparations, NO plays a critical role in mediating the effect of Ang II, whereas bradykinin has differential roles in the two vessels. In UA, B2 receptor blockade may result in a compensatory increase in cyclic GMP production, whilst in aorta, bradykinin accounts for approximately half of the cyclic GMP produced in response to Ang II.

British Journal of Pharmacology (2004) 141, 1024–1031. doi:10.1038/sj.bjp.0705694
Agonist‐dependence of recovery from desensitization of P2X3 receptors provides a novel and sensitive approach for their rapid up or downregulation
Authors: Sokolova Elena , Skorinkin Andrei , Fabbretti Elsa , Masten Lara , Nistri Andrea , Giniatullin Rashid 
Published: March 2004

Fast‐desensitizing P2X3 receptors of nociceptive dorsol root ganglion (DRG) neurons are thought to mediate pain sensation. Since P2X3 receptor efficiency is powerfully modulated by desensitization, its underlying properties were studied with patch‐clamp recording.
On rat cultured DRG neurons, 2 s application of ATP (EC50=1.52 μM), ADP (EC50=1.1 μM) or α,β‐meATP (EC50=1.78 μM) produced similar inward currents that fully desensitized, at the same rate, back to baseline. Recovery from desensitization was much slower after ATP and ADP than after α,β‐meATP and, in all cases, it had sigmoidal time course.
By alternating the application of ATP and α,β‐meATP, we observed complete cross‐desensitization indicating that these agonists activated the same receptors. This notion was confirmed by the similar antagonism induced by 2′, 3′‐O‐(2,4,6,trinitrophenyl)‐adenosine triphosphate (TNP‐ATP).
Recovery from desensitization elicited by ATP was unexpectedly shaped by transient application of α,β‐methylene‐adenosine triphosphate (α,β‐meATP), and vice versa. Thus, short‐lasting, full desensitization produced by α,β‐meATP protected receptors from long‐lasting desensitization induced by subsequent ATP applications. ATP and ADP had similar properties of recovery from desensitization.
Low nM concentrations of α,β‐meATP (unable to evoke membrane currents) could speed up recovery from ATP‐induced desensitization, while low nM concentrations of ATP enhanced it. Ambient ATP levels were found to be in the pM range (52±3 pM).
The phenomenon of cross‐desensitization and protection was reproduced by rP2X3 receptors expressed by rat osteoblastic cell 17/2.8 or human embryonic kidney cell 293 cells, indicating P2X3 receptor specificity.
It is suggested that transient application of an agonist that generates rapid recovery from desensitization, is a novel, powerful tool to modulate P2X3 receptor responsiveness to the natural agonist ATP.

British Journal of Pharmacology (2004) 141, 1048–1058. doi:10.1038/sj.bjp.0705701
Effects of chloride substitution on electromechanical responses in the pulmonary artery of Dahl normotensive and hypertensive rats
Authors: Bieger Detlef , Duggan Jennifer A , Tabrizchi Reza 
Published: March 2004

We have investigated the in vitro interaction between chloride ions and endothelium as revealed by alterations in vascular contractility and smooth muscle cell membrane potential in isolated pulmonary arteries from Dahl salt‐resistant normotensive and salt‐sensitive hypertensive rats.
Exposure to nitro‐L‐arginine methyl ester (L‐NAME) of tissues from normotensive but not hypertensive rats augmented contractions to cirazoline. While chloride removal did not alter cirazoline‐induced contractions, it completely abolished the augmentation by L‐NAME in normotensive rats. However, in hypertensive rats, removal of chloride ions significantly attenuated contractions elicited by cirazoline, and L‐NAME effectively reversed this inhibition.
Methacholine‐induced endothelium‐dependent relaxations of the same magnitude were evident in both normotensive and hypertensive rats. However, basal cyclic GMP levels were found to be significantly higher (7.8‐fold) in blood vessels of normotensive rats compared to hypertensive rats.
The resting membrane potential in pulmonary arteries of hypertensive rats (−52.1±1.04 mV) revealed a significant hyperpolarisation when compared with that of normotensive rats (−46.4±1.58 mV). Cirazoline did not produce a significant depolarisation in blood vessels of either normotensive or hypertensive rats. Perfusion with chloride‐free solution resulted in a modest but significant hyperpolarisation (−8.0 mV) in the blood vessels of hypertensive but not in normotensive rats.
We conclude that salt‐dependent hypertension in Dahl rats is accompanied by functional and biochemical changes in low‐pressure blood vessels. These changes can, in part, be attributed to impairment in the basal, but not methacholine‐stimulated, release of nitric oxide, and to altered chloride ion handling.

British Journal of Pharmacology (2004) 141, 1068–1076. doi:10.1038/sj.bjp.0705703
Nociceptin inhibits airway microvascular leakage induced by HCl intra‐oesophageal instillation
Authors: Rouget Céline , Cui Yong Yao , D'Agostino Bruno , Faisy Christophe , Naline Emmanuel , Bardou Marc , Advenier Charles 
Published: March 2004

Gastro‐oesophageal acid reflux may cause airway responses such as cough, bronchoconstriction and inflammation in asthmatic patients. Our previous results suggest that microvascular leakage induced, in the guinea‐pig airways, by intra‐oesophageal hydrochloric acid (HCl) infusion was mainly dependent on the release of tachykinins. Nociceptin, an endogenous ligand of the opioid receptor NOP, has been shown to inhibit bronchoconstriction and cough in guinea‐pig or cat by inhibiting tachykinin release.
The purpose of this study was to investigate the effects of nociceptin on the intra‐oesophageal HCl‐induced airway microvascular leakage evaluated by Evans blue dye extravasation measurement in anaesthetised guinea‐pigs pretreated with propranolol, atropine and phosphoramidon.
Infusion of intra‐oesophageal HCl led to a significant increase in plasma extravasation in the main bronchi and trachea. This increase was abolished when animals underwent a bilateral vagotomy.
Airway microvascular leakage was inhibited by nociceptin (3–30 μg kg−1 i.v.) in a dose‐dependent manner (maximal inhibition at the dose of 30 μg kg−1: 19.76±1.13 vs 90.92±14.00 ng mg−1 tissue for nociceptin and HCl infusion, respectively, in the main bronchi, P<0.01). The NOP receptor agonist [Arg14,Lys15]N/OFQ mimicked the inhibitory effect of nociceptin, but at a 10‐fold lower dose (3 μg kg−1 i.v). The NOP receptor antagonist J‐113397 had no effect on plasma protein extravasation by itself, but was able to block the inhibitory effect of nociceptin.
Morphine (1 mg kg−1) had a similar inhibitory effect as that of nociceptin. Naloxone pretreatment abolished the effect of morphine, but was enable to block the inhibitory effect of nociceptin.
Under similar conditions, nociceptin, in the previous range of concentration, was unable to counteract the airway microvascular leakage induced by substance P (SP).
These results suggest that airway plasma extravasation induced by intra‐oesophageal HCl instillation might be inhibited by specific stimulation of the NOP receptor with nociceptin. Nociceptin is likely to act at a pre‐junctional level, by inhibiting tachykinin release, since it was unable to prevent SP‐induced airway plasma extravasation.

British Journal of Pharmacology (2004) 141, 1077–1083. doi:10.1038/sj.bjp.0705704
Muscarinic M3 receptor‐dependent regulation of airway smooth muscle contractile phenotype
Authors: Gosens Reinoud , Bromhaar Mechteld M Grootte , Tonkes Annet , Schaafsma Dedmer , Zaagsma Johan , Nelemans S Adriaan , Meurs Herman 
Published: March 2004

Airway smooth muscle (ASM) cells are known to switch from a contractile to a proliferative and synthetic phenotype in culture in response to serum and growth factors. Phenotype switching in response to contractile agonists, however, is poorly characterised, despite the possible relationship between ASM phenotype and airway remodelling in asthma.
To investigate the effects of muscarinic receptor stimulation on ASM phenotype, we used organ‐cultured bovine tracheal smooth muscle (BTSM) strips, in which contractile responsiveness, contractile protein expression and proliferation were measured after pretreatment with methacholine.
Long‐term methacholine pretreatment (8 days) decreased maximal contraction and sensitivity to methacholine as well as to histamine and KCl. This decrease was dose‐dependent (pEC50=5.2±0.1). Pretreatment with the highest concentration of methacholine applied (100 μM) could suppress maximal histamine‐induced contraction to 8±1% of control. In addition, contractile protein expression (myosin, actin) was downregulated two‐fold. No concomitant increase in proliferative capacity was observed.
The M3/M2 muscarinic receptor antagonist DAU 5884 (0.1 μM) completely inhibited the observed decrease in contractility. In contrast, the M2/M3 muscarinic receptor antagonist gallamine (10 μM) was ineffective, demonstrating that M2 receptors were not involved.
Pretreatment (8 days) with 60 mM KCl could mimick the strong decreases in contractility. This was completely prevented by pretreatment with verapamil (1 μM).
Regulation of contractility was not affected by protein kinase C inhibition, whereas inhibitors of phosphatidyl inositol 3‐kinase and p42/p44 mitogen activated protein kinase were partially effective.
These results show that long‐term methacholine pretreatment (8 days) induces an M3 receptor‐dependent decrease in BTSM contractility without increased proliferative capacity.

British Journal of Pharmacology (2004) 141, 943–950. doi:10.1038/sj.bjp.0705709
Anti‐inflammatory effect of diosmectite in hapten‐induced colitis in the rat
Authors: González Raquel , Sánchez de Medina Fermin , Martínez‐Augustin Olga , Nieto Ana , Gálvez Julio , Risco Severiano , Zarzuelo Antonio 
Published: March 2004

Diosmectite is a natural silicate effectively used in the treatment of infectious diarrhoea. Its antidiarrhoeal properties involve adsorption of toxins and bacteria and modifications of the rheological characteristics of gastrointestinal mucus. Hence, the aim of this study was to test the intestinal anti‐inflammatory activity of diosmectite.
Diosmectite (500 mg kg−1 day−1, p.o.) was administered as a post‐treatment to rats with chronic trinitrobenzene sulphonic acid colitis. Colonic status was checked 1 and 2 weeks after colitis induction by macroscopic, histological and biochemical examination.
Diosmectite post‐treatment resulted in amelioration of the morphological signs (intestinal weight, macroscopic damage, necrosed area, histology) and biochemical markers (myeloperoxidase activity, glutathione levels, MUC2 expression, inducible nitric oxide synthase and interleukin‐1β (IL‐1β) and leukotriene B4 synthesis), as well as in the reduction of the severity of diarrhoea. The effect of the clay was comparable to that of sulphasalazine (50 mg kg−1 day−1).
Diosmectite exhibited a dose‐dependent capacity to adsorb proteins in vitro as well as a dose‐dependent inhibitory effect on the basolateral secretion of IL‐8 by lipopolysaccharide (LPS)‐stimulated HT29 cells.
Diosmectite had a dose‐dependent inhibitory effect on IL‐1β production by LPS‐stimulated THP‐1 cells.
The effect of diosmectite on MUC2 was post‐transcriptional, since mRNA levels were unaffected. However, diosmectite is able to upregulate MUC2 mRNA levels in HT29‐MTX cells.
Diosmectite has anti‐inflammatory activity administered as a post‐treatment. Possible mechanisms include adsorption of luminal antigens, increase of colonic mucin levels and possibly a direct modulatory action of cytokine production by mucosal cells.

British Journal of Pharmacology (2004) 141, 951–960. doi:10.1038/sj.bjp.0705710