bluuebunny/crossref_metadata_2025_split huggingface dataset:
Supernatants of Pseudomonas aeruginosa induce the Pseudomonas‐specific antibiotic elafin in human keratinocytes
Authors: Meyer‐Hoffert Ulf , Wichmann Nils , Schwichtenberg Lars , White Paul C. , Wiedow Oliver 
Published: July 2003

Abstract: Elafin is a skin‐derived serine‐protease inhibitor. It is thought to be important to prevent human leukocyte elastase‐mediated tissue damage and might play an important role in maintaining the integrity of the human epidermis. Recent studies have provided evidence for an antimicrobial activity of elafin against P. aeruginosa. As gram‐negative infections typically occur in barrier‐disrupted skin we were interested to determine whether supernatants of the gram‐negative bacteria P. aeruginosa and Escherichia coli were capable of inducing elafin expression.Supernatants of various P. aeruginosa strains stimulated elafin mRNA‐expression and protein release, whereas supernatants of E. coli did not induce elafin expression. In non‐differentiated cells the relative increase of elafin mRNA was much higher (100‐fold) than in differentiated cells (sixfold), although the latter exhibited higher constitutive mRNA‐expression (150‐fold). However, concentrations of secreted elafin were similar in differentiated and non‐differentiated cells after stimulation. We could not confirm a bactericidal effect against P. aeruginosa as described previously but observed that its growth was inhibited as demonstrated for different strains in liquid cultures. Growth of E. coli was not affected by elafin.In conclusion, the data presented in this paper suggest that elafin represents an innate immune response factor induced by secreted products of P. aeruginosa. Besides its elastase inhibitory potency elafin is an antimicrobial agent against P. aeruginosa.
Adalimumab, a fully human anti‐TNF‐α monoclonal antibody, treatment does not influence experimental UV response in the skin of rheumatoid arthritis patients
Authors: Tjioe M. , Gerritsen M. J. P. , Den Broeder A. A. , Van Hooijdonk C. A. E. M. , Kroot E. J. A. , Van Riel P. L. C. M. , Barrera P. , Van De Kerkhof P. C. M. 
Published: July 2003

Abstract: TNF‐α is known to play an important role in UV‐induced immunomodulation and photodamage. It plays a role in UVB‐mediated induction of apoptosis and is a strong inducer of the c‐Jun N‐terminal kinase (JNK) pathway, which eventually leads to the loss of dermal collagen and elastin content. Recently chimeric anti‐TNF‐α has been introduced as a therapy for rheumatoid arthritis.The aim of the present study was to investigate the effect of anti‐TNF‐α treatment on UV‐induced DNA damage, apoptosis, and induction of matrix metallo proteinases.Twelve patients with rheumatoid arthritis were included and irradiated with 2 MED broadband UVB before and after administration of 0.5 mg/kg anti‐TNF‐α monoclonal antibody. Twenty‐four hours after irradiation biopsies were taken. Frozen and paraffin sections were stained for p53, c‐Jun, phosphorylated c‐Jun, sunburn cells and MMP‐1.No significant changes were observed in the expression of p53 and sunburn cells and MMP‐1 content after treatment with anti‐TNF‐alpha, whereas a slight but significant decrease in c‐Jun and phosphorylated c‐Jun expression was noted (P = 0.0250 and P = 0.0431, respectively).Our results showed no influence of anti‐TNF‐α on UV response at therapeutic doses in patients with rheumatoid arthritis.
Functional testing of keratin 14 mutant proteins associated with the three major subtypes of epidermolysis bullosa simplex
Authors: Sørensen Charlotte B. , Andresen Brage S. , Jensen Uffe B. , Jensen Thomas G. , Jensen Peter K. A. , Gregersen Niels , Bolund Lars 
Published: July 2003

Abstract: Epidermolysis bullosa simplex (EBS) is a group of autosomal dominantly inherited skin disorders characterized by the development of intra‐epidermal skin blisters on mild mechanical trauma. The three major clinical subtypes (Weber‐Cockayne, Koebner and Dowling‐Meara) are all caused by mutations in either the keratin 5 (KRT5) or keratin 14 (KRT14) gene.Previously, we identified three novel KRT14 missense mutations in Danish EBS patients associated with the three different forms of EBS (1). The identified KRT14 mutations represent the full spectrum of the classical EBS subtypes. In the present study we investigated these mutations in a cellular expression system in order to analyse their effects on the keratin cytoskeleton. KRT14 expression vectors were constructed by fusing the nucleotide sequence encoding the FLAG reporter peptide to the 3′ end of the KRT14 cDNA sequences. The expression vectors were transiently transfected into normal human primary keratinocytes (NHK), HaCaT or HeLa cells in order to analyze the ability of the mutant K14 proteins to integrate into the existing endogenous keratin filament network (KFN).No effect on the keratin cytoskeleton was observed upon transfection of NHK with the various K14 constructs neither with nor without a subsequently induced heat‐stress. In contrast, all constructs, including wild‐type K14, caused collapse of the endogenous KFN in a small fraction of the transfected HeLa and HaCaT cells. However, overexpression of the mutation associated with the most severe form of the disease, EBS Dowling‐Meara, resulted in a higher number of transfected HaCaT cells with KFN collapse (P < 0.001). Thus, although a background KFN perturbance was observed upon transfection with the wild‐type K14 construct, the mutant protein associated with the most severe form of EBS worsened the KFN perturbation significantly compared with the mutant proteins associated with the milder forms of the disease and the normal K14 protein. This shows that the clinical severity of disease‐associated mutations identified in patients can be tested using this expression system, although it can not at present be used to discriminate between the milder forms.Assessment of the endogenous K14 protein expression in NHK and HaCaT cells indicated that the higher level of endogenous keratin expression in NHK might make these cells more resistant to perturbation of the keratin cytoskeleton by overexpressed K14 protein than HaCaT cells.
Effects of cyclooxygenase inhibitor pretreatment on nitric oxide production, nNOS and iNOS expression in rat cerebellum
Authors: DiGirolamo G , Farina M , Riberio M L , Ogando D , Aisemberg J , de Los Santos A R , Martí M L , Franchi A M 
Published: July 2003

The therapeutic effect of nonsteroidal anti‐inflammatory drugs (NSAIDs) is thought to be due mainly to its inhibition of cyclooxygenase (COX) enzymes, but there is a growing body of research that now demonstrates a variety of NSAIDs effects on cellular signal transduction pathways other than those involving prostaglandins.
Nitric oxide (NO) as a free radical and an agent that gives rise to highly toxic oxidants (peroxynitrile, nitric dioxide, nitron ion), becomes a cause of neuronal damage and death in some brain lesions such as Parkinson and Alzheimer disease, and Huntington's chorea.
In the present study, the in vivo effect of three NSAIDs (lysine clonixinate (LC), indomethacine (INDO) and meloxicam (MELO)) on NO production and nitric oxide synthase expression in rat cerebellar slices was analysed. Rats were treated with (a) saline, (b) lipopolysaccharide (LPS) (5 mg kg−1, i.p.), (c) saline in combination with different doses of NSAIDs and (d) LPS in combination with different doses of NSAIDs and then killed 6 h after treatment.
NO synthesis, evaluated by Bred and Snyder technique, was increased by LPS. This augmentation was inhibited by coadministration of the three NSAIDs assayed. None of the NSAIDs tested was able to modify control NO synthesis.
Expression of iNOS and neural NOS (nNOS) was detected by Western blotting in control and LPS‐treated rats. LC and INDO, but not MELO, were able to inhibit the expression of these enzymes.
Therefore, reduction of iNOS and nNOS levels in cerebellum may explain, in part, the anti‐inflammatory effect of these NSAIDs and may also have importance in the prevention of NO‐mediated neuronal injury.

British Journal of Pharmacology (2003) 139, 1164–1170. doi:10.1038/sj.bjp.0705315
Modulation of neuronal nicotinic receptor function by the neuropeptides CGRP and substance P on autonomic nerve cells
Authors: Di Angelantonio Silvia , Giniatullin Rashid , Costa Valeria , Sokolova Elena , Nistri Andrea 
Published: July 2003

One classical example of how neuropeptides can affect the function of ligand‐gated receptors is the modulation of neuronal nicotinic receptors (nAChRs) by substance P. The present review updates current understanding of this action by substance P and compares it with other neuropeptides more recently found to modulate nAChRs in the autonomic nervous system.
Calcitonin gene‐related peptide (CGRP) and its N‐terminal fragments have been shown to exert complex inhibitory as well facilitatory actions on nAChRs. Fragments such as CGRP1–4, CGRP1–5 and CGRP1–6 rapidly and reversibly enhance agonist sensitivity of nAChRs without directly activating those receptors. Longer fragments or the full‐length peptide potently inhibit responses mediated by nAChRs via an apparently competitive‐type antagonism. This phenomenon differs from the substance P‐induced block, which is agonist use‐dependent and preferential towards large nicotinic responses.
It is argued that the full‐length peptides CGRP and substance P might play distinct roles in the activity‐dependent modulation of cholinergic neurotransmission, by inhibiting background noise in the case of CGRP or by reducing excessive excitation in the case of substance P. Hence, multiple neuropeptide mechanisms may represent a wide array of fine‐tuning processes to regulate nicotinic synaptic transmission.
The availability of novel CGRP derivatives with a strong enhancing action on nAChRs may offer new leads for the drug design targeted for potentiation of nAChRs in the autonomic nervous system as well as in the brain, a subject of interest to counteract the deficit of the nAChR function associated with neurodegenerative diseases like Alzheimer's and Parkinson's diseases.

British Journal of Pharmacology (2003) 139, 1061–1073. doi:10.1038/sj.bjp.0705337
Muscarinic regulation of cardiac ion channels
Authors: Harvey Robert D , Belevych Andriy E 
Published: July 2003

The parasympathetic component of the autonomic nervous system plays an important role in the physiological regulation of cardiac function by exerting significant influence over the initiation as well as propagation of electrical impulses, in addition to being able to regulate contractile force. These effects are mediated in whole or in part through changes in ion channel activity that occur in response to activation of M2 muscarinic cholinergic receptors following release of the neurotransmitter acetylcholine. The coupling of M2 receptor activation to most changes in cardiac ion channel function can be explained by one of two general paradigms. The first involves direct G protein‐dependent regulation of ion channel activity. The second involves indirect regulation of ion channel activity through modulation of cAMP‐dependent responses. This review focuses on recent advances in our understanding of the mechanisms by which M2 muscarinic receptor activation both inhibits and facilitates cAMP‐dependent ion channel responses in the heart.British Journal of Pharmacology (2003) 139, 1074–1084. doi:10.1038/sj.bjp.0705338
An angiotensin II type 1 receptor antagonist, olmesartan medoxomil, improves experimental liver fibrosis by suppression of proliferation and collagen synthesis in activated hepatic stellate cells
Authors: Kurikawa Nobuya , Suga Miwa , Kuroda Shoko , Yamada Keisuke , Ishikawa Hirokazu 
Published: July 2003

We studied the effect of a new angiotensin II type 1 (AT1) receptor antagonist, olmesartan medoxomil (olmesartan), on the fibrogenic responses in rat hepatic stellate cells (HSCs) and liver fibrogenesis.
Olmesartan (1 mg kg−1 per day) was orally administered to fibrotic rats, induced by bile duct ligation. Liver hydroxyproline content, the mRNA expression of collagen α1(I) and α‐smooth muscle actin (α‐SMA), and plasma levels of transforming growth factor‐β1 (TGF‐β1) were significantly reduced by olmesartan treatment, suggesting that olmesartan improved liver fibrosis. Interestingly, AT1 receptors were found to be expressed in α‐SMA‐positive cells in the fibrotic area of livers in bile duct‐ligated rats by immunohistochemical analysis. Olmesartan treatment reduced the number of these cells.
In vitro experiments showed that angiotensin II (Ang II) treatment induced proliferation and collagen synthesis, and upregulated the profibrogenic cytokines, TGF‐β1 and connective tissue growth factor (CTGF), in rat primary HSCs. Olmesartan blocked all these effects of Ang II.
Based on these results, since activated HSCs were found to express AT1 receptors and Ang II is thought to play an important role in the pathogenesis of liver fibrosis by binding to these receptors, olmesartan may act as a potent antifibrotic drug to suppress the proliferation, collagen synthesis and the expression of profibrogenic cytokines in activated HSCs by blocking these receptors.

British Journal of Pharmacology (2003) 139, 1085–1094. doi:10.1038/sj.bjp.0705339
Quantitative autoradiography of adenosine receptors in brains of chronic naltrexone‐treated mice
Authors: Bailey Alexis , Hawkins Rachel M , Hourani Susanna M O , Kitchen Ian 
Published: July 2003

Manipulation of μ opioid receptor expression either by chronic morphine treatment or by deletion of the gene encoding μ opioid receptors leads to changes in adenosine receptor expression. Chronic administration of the opioid receptor antagonist naltrexone leads to upregulation of μ receptor binding in the brain.
To investigate if there are any compensatory alterations in adenosine systems in the brains of chronic naltrexone‐treated mice, we carried out quantitative autoradiographic mapping of A1 and A2A adenosine receptors in the brains of mice treated for 1 week with naltrexone (8 mg−1 kg−1 day−1), administered subcutaneously via osmotic minipump.
Adjacent coronal brain sections were cut from chronic saline‐ and naltrexone‐treated mice for the determination of binding of [3H] D‐Ala2‐MePhe4‐Gly‐ol5 enkephalin ([3H] DAMGO), [3H]1,3‐dipropyl‐8‐cyclopentylxanthine ([3H] DPCPX) or [3H] 2‐[p‐(2‐carbonylethyl)phenylethylamino]‐5′‐N‐ethylcarboxamidoadenosine ([3H] CGS21680) to μ, A1 and A2A receptors, respectively.
A significant increase in μ and A1 receptor binding was detected in chronic naltrexone‐treated brains. The changes in μ receptors were significant in several regions, but changes in A1 were relatively smaller but showed significant upregulation collectively. No significant change in A2A receptor binding was detected in chronic naltrexone‐treated brains.
The results show that blockade of opioid receptors causes upregulation of A1 receptors, but not A2A receptors, by as yet undefined mechanisms.

British Journal of Pharmacology (2003) 139, 1187–1197. doi:10.1038/sj.bjp.0705340
Hepatic insulin sensitizing substance: a novel ‘sensocrine’ mechanism to increase insulin sensitivity in anaesthetized rats
Authors: Porszasz Robert , Legvari Gyorgyi , Pataki Tunde , Szilvassy Judith , Nemeth Jozsef , Kovacs Peter , Paragh Gyorgy , Szolcsanyi Janos , Szilvassy Zoltan 
Published: July 2003

We recently described the sensory nitrergic nature of the hepatic insulin sensitizing substance (HISS) mechanism linked to postprandial activation of anterior hepatic plexus fibres in rabbits. This study is designed to assess the involvement of the sensory pathways in this mechanism.
Selective sensory denervation of the anterior hepatic plexus (AHP) was achieved by a 3‐day perineurial treatment with 2% capsaicin solution in Wistar rats (230–250 g). After 1 week, hyperinsulinaemic (100 μU kg−1) euglycaemic (5.5 mmol kg−1) glucose clamp studies were performed to estimate insulin sensitivity.
The rats with regional AHP sensory denervation exhibited a significantly decreased insulin sensitivity, that is, 9.1±1.0 mg kg−1 min−1 glucose reinstalled euglycaemia vs 13.3±1.9 mg kg−1 min−1 glucose (P<0.01) in control rats.
Acute partial hepatic denervation by AHP cut was without effect on insulin sensitivity, whereas chronic hepatic denervation induced insulin resistance was similar to that achieved by regional AHP capsaicin treatment.
Intraportal administration of L‐NAME (10 mg kg−1) decreased, whereas capsaicin (0.3 mg kg−1 min−1) increased insulin sensitivity. Neither atropine (1 mg kg−1) nor acetylcholine (1–10 μg mg min−1) produced any significant effect. In animals with preceding regional capsaicin desensitization, none of the pharmacological manoeuvres modified the resulting insulin‐resistant state.
Cysteamine (200 mg kg−1 s.c.) is known to cause functional somatostatin depletion‐induced insulin resistance similar to that produced by either chronic partial hepatic denervation or perineurial AHP capsaicin desensitization. Intraportal capsaicin (0.3 mg kg−1 min−1) was unable to modify insulin resistance achieved by cysteamine.
We conclude that capsaicin‐sensitive sensory fibres play a crucial role in neurogenic insulin sensitization known as the HISS mechanism without involvement of anatomical reflex‐mediated circuits. The results also suggest that HISS is identical to somatostatin of AHP sensory neural origin.

British Journal of Pharmacology (2003) 139, 1171–1179. doi:10.1038/sj.bjp.0705342
Tityustoxin‐K(alpha) blockade of the voltage‐gated potassium channel Kv1.3
Authors: Rodrigues Aldo Rogelis A , Arantes Eliane C , Monje Francisco , Stuhmer Walter , Varanda Wamberto Antonio 
Published: July 2003

We investigated the action of TsTX‐Kα on cloned Kv1.3 channels of the Shaker subfamily of voltage‐gated potassium channels, using the voltage–clamp technique. Highly purified TsTX‐Kα was obtained from the venom of the Brazilian scorpion Tityus serrulatus using a new purification protocol. Our results show that TsTX‐Kα blocks Kv1.3 with high affinity in two expression systems.
TsTX‐Kα blockade of Kv1.3 channels expressed in Xenopus oocytes was found to be completely reversible and to exhibit a pH dependence. The KD was 3.9 nM at pH 7.5, 9.5 nM at pH 7.0 and 94.5 nM at pH 6.5.
The blocking properties of TsTX‐Kα in a mammalian cell line (L929), stably transfected to express Kv1.3, were studied using the patch–clamp technique. In this preparation, the toxin had a KD of 19.8 nM at pH 7.4.
TsTX‐Kα was found to affect neither the voltage‐dependence of activation, nor the activation and deactivation time constants. The block appeared to be independent of the transmembrane voltage and the toxin did not interfere with the C‐type inactivation process.
Taken as a whole, our findings indicate that TsTX‐Kα acts as a simple blocker of Kv1.3 channels. It is concluded that this toxin is a useful tool for probing not only the physiological roles of Kv1.2, but also those mediated by Kv1.3 channels.

British Journal of Pharmacology (2003) 139, 1180–1186. doi:10.1038/sj.bjp.0705343
Substance P increases neutrophil adhesion to human umbilical vein endothelial cells
Authors: Dianzani Chiara , Collino Massimo , Lombardi Grazia , Garbarino Giovanni , Fantozzi Roberto 
Published: July 2003

Adhesion of neutrophils (PMNs) to vascular endothelial cells (EC) is a critical step in recruitment and infiltration of leukocytes into tissues during inflammation. Substance P (SP), a neuropeptide released from sensory nerves, evoked PMN adhesion to EC. The NK receptor subtype(s) and the cell type(s) involved were investigated.
SP was coincubated with human PMNs and EC from the human umbilical vein (HUVEC); adhesion was quantitated by computerised microimaging fluorescence analysis.
The proadhesive effects of SP (range 10−18–10−6 M) were illustrated in a biphasic dose–response curve, with a maximum at 10−15 M (276±16% adhesion vs control; P<0.01) and another one at 10−10 M (200±18% adhesion vs control; P<0.01). Neurokinin A was less active and neurokinin B was inactive. The adhesion molecules LFA‐1 and OKM‐1, but not selectins, were involved according to results with selective mAbs.
The NK1 agonist [Sar9,Met(O2)11]SP reproduced the effects of SP, whereas the NK2 agonist [βAla8]‐neurokininA (4–10) acted at 10−13–10−8 M only. The NK3 agonist, senktide, was ineffective.
The NK1 antagonists, CP 96,345 and L 703,606 (both 10−6 M), abolished the effect of 10−15 M SP and inhibited that of 10−10 M SP by 56±5% (P<0.01). By comparison, the NK2 antagonist, SR 48,968 (10−7 M), partially antagonised the adhesion evoked by 10−10 M SP (% inhibition: 61±6; P<0.05).
Since preincubation of PMNs and HUVEC with SP gave the same results it is clear that both cell types contributed to its proadhesive effects.
These results indicate that SP induced a proadhesive effect during inflammatory processes, which was mediated by NK1 and NK2 receptors.

British Journal of Pharmacology (2003) 139, 1103–1110. doi:10.1038/sj.bjp.0705344
Role of c‐jun N‐terminal kinase in the induced release of GM‐CSF, RANTES and IL‐8 from human airway smooth muscle cells
Authors: Oltmanns Ute , Issa Razao , Sukkar Maria B , John Matthias , Chung K Fan 
Published: July 2003

Human airway smooth muscle cells (HASMC) contribute to airway inflammation in asthma by virtue of their capacity to produce several inflammatory mediators including IL‐8, GM‐CSF and RANTES. The intracellular signal pathway underlying the production of these cytokines in HASMC is not entirely elucidated.
We examined the role of the mitogen‐activated protein kinase (MAPK) c‐jun N‐terminal kinase (JNK) in TNFα‐ and IL‐1β‐induced GM‐CSF, RANTES and IL‐8 production in HASMC by using a novel specific inhibitor for JNK (SP600125).
Confluent HASMC were treated with TNFα or IL‐1β (10 ng ml−1) for 24 h in the presence or absence of SP600125 (1–100 μM). JNK activity was determined by a kinase assay. Phosphorylation of JNK, p38 MAPK and ERK was examined by Western blotting. Culture supernatants were assayed for GM‐CSF, RANTES and IL‐8 content by ELISA.
Maximum TNFα‐ or IL‐1β‐induced phosphorylation of JNK in HASMC occurred after 15 min and returned to baseline levels after 4 h. SP600125 inhibited TNFα‐ and IL‐1β‐induced JNK activity in HASMC as shown by the reduced phosphorylation of its substrate c‐jun. Furthermore, GM‐CSF, RANTES and to a lesser extent IL‐8 release from HASMC treated with TNFα and IL‐1β was inhibited dosedependently by SP600125.
JNK activation is involved in TNFα‐ and IL‐1β‐induced GM‐CSF, RANTES and IL‐8 production from HASMC. JNK may therefore represent a critical pathway for cytokine production in HASMC.

British Journal of Pharmacology (2003) 139, 1228–1234. doi:10.1038/sj.bjp.0705345