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‣ Arginine induced acute pancreatitis alters the actin cytoskeleton and increases heat shock protein expression in rat pancreatic acinar cells

Tashiro, M; Schafer, C; Yao, H; Ernst, S; Williams, J
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /08/2001 Português
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Arginine induced acute pancreatitis was evaluated as a novel and distinct form of experimental pancreatitis with particular attention to the actin cytoskeleton and expression of heat shock or stress proteins. Arginine induced a dose related necrotising pancreatitis in rats, as shown by histological evaluation, and an increase in serum amylase. Severe pancreatitis induced by 4.5 g/kg arginine was accompanied by dramatic changes in the actin cytoskeleton, as visualised with rhodamine phallodin. Intermediate filaments were also disrupted, as visualised by cytokeratin 8/18 immunocytochemistry. Arginine pancreatitis was accompanied by a stress response with a large increase in the small heat shock protein HSP27, as well as HSP70, peaking at 24 hours and localised to acinar cells. There was a lower increase in HSP60 and HSP90 and no effect on GRP78. HSP27 was also shifted to phosphorylated forms during pancreatitis. A lower dose of arginine (3.0 g/kg) induced less pancreatitis but a larger increase in HSP70 and HSP27 expression and phosphorylation of HSP27. Thus HSP expression can be overwhelmed by severe damage. The present work in conjunction with earlier work on caerulein induced pancreatitis indicates that changes in the actin cytoskeleton are an early component in experimental pancreatitis.


Keywords: experimental pancreatitis; pancreas; actin cytoskeleton; cytokeratins; heat shock proteins; HSP27; rat

‣ Effects of L-arginine on lower limb vasodilator reserve and exercise capacity in patients with chronic heart failure

Kanaya, Y; Nakamura, M; Kobayashi, N; Hiramori, K
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /05/1999 Português
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OBJECTIVE—To determine whether the reactive hyperaemic response of the lower limb increases with improved exercise capacity after acute supplementation with L-arginine, the precursor of nitric oxide, in patients with chronic heart failure.
METHODS—19 patients with chronic heart failure were enrolled in the study. Rest calf blood flow and femoral occlusion induced calf blood flow changes were measured by venous occlusion plethysmography before and after intravenous infusion of 10% L-arginine solution (5 ml/kg for 30 minutes) or placebo. Postexercise calf blood flow was also measured after the experimental infusion. During both postinfusion periods, several exercise capacity indices were determined by a symptom limited cardiopulmonary exercise test using a bicycle ergometer.
RESULTS—Baseline calf blood flow, systemic blood pressure, and heart rate showed no significant changes in either of the two experimental conditions. However, the occlusion induced blood flow response was significantly enhanced by L-arginine infusion (mean (SEM) peak flow, 19.6 (1.5) v 28.9 (3.1) ml/min/dl calf tissue; p < 0.01), but not by placebo (peak flow, 19.1 (1.4) v 20.9 (1.8) ml/min/dl calf tissue; NS). Calf blood flow response after exercise was also higher after L-arginine infusion than after placebo (peak flow...

‣ Effects of L- and D-arginine on the basal tone of human diseased coronary arteries and their responses to substance P

Tousoulis, D; Tentolouris, C; Crake, T; Katsimaglis, G; Stefanadis, C; Toutouzas, P; Davies, G
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /05/1999 Português
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OBJECTIVE—To assess the effects of substance P administration alone and in combination with L- and D-arginine in patients with normal angiograms and in patients with coronary artery disease.
DESIGN—Intracoronary infusions of (a) normal saline, (b) the receptor mediated nitric oxide stimulant substance P (5.6 and 27.8 pmol/min) before and after L- or D-arginine (50 and 150 µmol/min), and (c) glyceryl trinitrate (250 µg bolus) were given to 17 patients with coronary artery disease and stable angina, and to six patients with normal angiograms. The diameter of angiographically normal proximal and distal segments and coronary stenoses were measured by computerised quantitative angiography.
RESULTS—L-arginine administration was associated with significant dilatation of stenoses (p < 0.01) of proximal segments of both "normal" (p < 0.05) and diseased (p < 0.01) arteries, and of distal segments of diseased arteries (p < 0.01). No significant changes were associated with D-arginine administration. Dose dependent dilatation of all segments including stenoses, was observed with substance P both before and after L-arginine infusion (p < 0.01). The magnitude of dilatation of stenoses and all segments of both "normal" and diseased coronaries was greater after L-arginine (p < 0.05) but not D-arginine and substance P infusion...

‣ Vasomotor effects of L- and D-arginine in stenotic atheromatous coronary plaque

Tousoulis, D; Davies, G; Tentolouris, C; Goumas, G; Stefanadis, C; Toutouzas, P
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /09/2001 Português
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OBJECTIVE—To examine the effects of exogenous L- and D-arginine on coronary stenosis vasomotion in relation to stenosis morphology.
DESIGN—Intracoronary infusions of normal saline, L- and D-arginine (50 and 150 µmol/min), and glyceryl trinitrate (250 µg bolus) were given in 24 patients with coronary artery disease and stable angina. Coronary stenoses were classified as smooth or complex (irregular borders). The diameter of the coronary stenoses and their adjacent reference segments was measured by computed quantitative angiography.
RESULTS—During L-arginine infusion a larger proportion of complex stenoses than smooth stenoses dilated by ⩾ 10% (p < 0.01), and the magnitude of dilatation was greater at the site of complex stenoses (p < 0.05). Irrespective of the type of morphology there was a positive correlation (p < 0.01) between the severity of stenoses and the magnitude of vasodilatation to L-arginine. The response to glyceryl trinitrate was similar in the two groups. No significant change was found in either group in response to D-arginine.
CONCLUSIONS—In patients with coronary artery disease, coronary stenoses—particularly those of complex morphology—dilate in response to the administration of L-arginine but not D-arginine. This finding is consistent with partial deficiency of the substrate for nitric oxide synthesis...

‣ Expression of nitric oxide synthases and effects of L-arginine and L-NMMA on nitric oxide production and fluid transport in collagenous colitis

Perner, A; Andresen, L; Normark, M; Fischer-Hansen, B; Sorensen, S; Eugen-Olsen, J; Rask-Madsen, J
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /09/2001 Português
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BACKGROUND AND AIMS—Luminal nitric oxide (NO) is greatly increased in the colon of patients with collagenous and ulcerative colitis. To define the source and consequence of enhanced NO production we have studied expression of NO synthase (NOS) isoforms and nitrotyrosine in mucosal biopsies from these patients. In addition, effects on colonic fluid transfer caused by manipulating the substrate of NOS were studied in patients with collagenous colitis.
PATIENTS—Eight patients with collagenous colitis, nine with active ulcerative colitis, and 10 with uninflamed bowel were included.
METHODS—Expression of NOS isoforms was quantified by western blotting. Inducible NOS (iNOS) and nitrotyrosine were localised by immunohistochemistry. Modulation of NOS activity by topical NG-monomethyl-L-arginine (L-NMMA) or L-arginine was assessed during perfusion of whole colon. Plasma and perfusate nitrite/nitrate (NOx) was measured by Griess' reaction.
RESULTS—Both in collagenous and ulcerative colitis, expression of iNOS was 102-103 higher (p<0.001) than in uninflamed bowel and localised primarily to the epithelium. Endothelial NOS was evenly expressed in all groups while neuronal NOS was undetectable. Nitrotyrosine was markedly expressed in active ulcerative colitis but rarely detected in collagenous colitis and never in uninflamed bowel. In collagenous colitis...

‣ Expression of kinin B1 and B2 receptors in immature, monocyte-derived dendritic cells and bradykinin-mediated increase in intracellular Ca2+ and cell migration

Bertram, Cornelia M.; Baltic, Svetlana; Misso, Neil L A; Bhoola, Kanti D.; Foster, Paul S; Thompson, Philip J; Fogel-Petrovic, Mirjana
Fonte: Federation of American Societies for Experimental Biology Publicador: Federation of American Societies for Experimental Biology
Tipo: Artigo de Revista Científica
Português
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The kinins, bradykinin (BK) and Lysdes[ Arg9]-BK, are important inflammatory mediators that act via two specific G protein-coupled kinins, B1 and B2 receptors (B2R). Kinins influence the activity of immune cells by stimulating the synthesis of cytokines,

‣ L-arginine and vitamin D: novel immunotherapies in tuberculosis

Ralph, Anna; Kelly, Paul; Anstey, Nicholas
Fonte: Elsevier Publicador: Elsevier
Tipo: Artigo de Revista Científica
Português
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Worsening drug resistance and the need for prolonged treatment in tuberculosis (TB) require innovative solutions including investigation of inexpensive, safe adjunctive immunotherapies. L-arginine, the precursor of nitric oxide, and vitamin D recently hav

‣ Site-specific labelling of proteins with a rigid lanthanide-binding tag

Su, Xun-Cheng; Huber, Thomas; Dixon, Nicholas; Otting, Gottfried
Fonte: Wiley-VCH Verlag GMBH Publicador: Wiley-VCH Verlag GMBH
Tipo: Artigo de Revista Científica
Português
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This paper describes a generic method for the site-specific attachment of lanthanide complexes to proteins through a disulfide bond. The method is demonstrated by the attachment of a lanthanide-binding peptide tag to the single cysteine residue present in

‣ E Protein Domain III Determinants of Yellow Fever Virus 17DVaccine Strain Enhance Binding to Glycosaminoglycans,Impede Virus Spread, and Attenuate Virulence

Lee, Eva; Lobigs, Mario
Fonte: American Society for Microbiology Publicador: American Society for Microbiology
Tipo: Artigo de Revista Científica
Português
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The yellow fever virus (YFV) 17D strain is one of the most effective live vaccines for human use, but the in vivo mechanisms for virulence attenuation of the vaccine and the corresponding molecular determinants remain elusive. The vaccine differs phenotyp

‣ Understanding the role of inflammatory cytokines in malaria and related diseases

Clark, Ian A; Alleva, Lisa; Budd, Alison; Cowden, William
Fonte: Elsevier Publicador: Elsevier
Tipo: Artigo de Revista Científica
Português
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It is now broadly accepted for infectious disease in general that it is not the invading organism, but the body's unbridled response to it-the "cytokine storm"-that causes illness and pathology. Nevertheless, many researchers still regard the harmful effe

‣ Species Variability in Cardiovascular Research: The Example of Adrenocorticotrophin-Induced Hypertension

Whitworth, Judith; Zhang, Yi; Mangos, George; Kelly, John J
Fonte: Blackwell Science Asia Publicador: Blackwell Science Asia
Tipo: Artigo de Revista Científica
Português
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1. Lawrie Beilin has contributed greatly to international hypertension research through both animal and human studies. 2. Animals are used in biomedical research to gain insights that can be extrapolated ultimately to humans. 3. A simple experimental mani

‣ Apocynin but Not-L-Arginine Prevents and Reverses Dexamethasone-Induced Hypertension in the Rat

Hu, Lexian; Zhang, Yi; Lim, Pek (Chloe); Miao, Yuchun; Tan, Chrismin; McKenzie, Katja; Schyvens, Chris; Whitworth, Judith
Fonte: Elsevier Publicador: Elsevier
Tipo: Artigo de Revista Científica
Português
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Background: Dexamethasone (Dex)-hypertension in rats is associated with increased oxidative stress. We investigated effects of the NAD(P)H oxidase inhibitor apocynin and the nitric oxide (NO) precursor l-arginine on Dex-hypertension to determine the relat

‣ Inhibition of Nuclear Import Mediated by the Rev-Arginine Rich Motif by RNA Molecules

Fineberg, Konstantin; Fineberg, Tali; Graessmann, Adolf; Luedtke, Nathan; Tor, Yitzhak; Rui, Lixin; Jans, David A; Loyter, Abraham
Fonte: American Chemical Society Publicador: American Chemical Society
Tipo: Artigo de Revista Científica
Português
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The HIV-1 Rev protein plays a pivotal role in viral replication, and therefore, inhibition of its function should block the progression of the virus-induced immune deficiency syndrome (AIDS). Here, RNA molecules have been shown to inhibit import of the HIV-1 Rev protein into nuclei of permeabilized cells. Nuclear uptake of biotinylated recombinant His-tagged Rev-GFP was assessed in nuclear extracts from digitonin-permeabilized cells by binding to either importin β-receptors or nickel molecules immobilized on a microtiter plate. Using this method together with fluorescence microscopy, we determined that nuclear import of Rev is inhibited by the addition of a reticulocyte lysate which routinely is used as a source of nuclear import receptors. This inhibition was released by treatment with the RNase enzyme. Also t-RNA molecules and the oligoribonucleotide RRE IIB, namely, the second stem structure of the Rev responsive element (RRE) of the viral RNA, inhibit Rev nuclear import. Similar results were obtained when BSA molecules with covalently attached Rev-arginine rich motif (ARM) peptides were used as a nuclear transport substrate, indicating that the nuclear import inhibition of the Rev protein is due to the presence of the ARM domain. Binding experiments revealed that the RNA molecules inhibit the interaction between the ARM region and importin β...

‣ L-Arginine Transport in Humans with Cortisol-Induced Hypertension

Chin-Dusting, Jaye PF; Ahlers, Belinda A; Kaye, David M; Kelly, John; Whitworth, Judith
Fonte: Lippincott Williams & Wilkins Publicador: Lippincott Williams & Wilkins
Tipo: Artigo de Revista Científica
Português
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A deficient L-arginine-nitric oxide system is implicated in cortisol-induced hypertension. We investigate whether abnormalities in L-arginine uptake contribute to this deficiency. Eight healthy men were recruited. Hydrocortisone acetate (50 mg) was given

‣ Effects of oral L-arginine on plasma nitrate and blood pressure in cortisol-treated humans

Kelly, John; Williamson, Paula M; Martin, Allison; Whitworth, Judith
Fonte: Lippincott Williams & Wilkins Publicador: Lippincott Williams & Wilkins
Tipo: Artigo de Revista Científica
Português
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Objective: The aim of this study was to determine whether cortisol-induced hypertension can be reversed by coadministration of oral L-arginine. Study design: Three studies were undertaken in healthy male human subjects. The first study addressed the effect of oral L-arginine loading on plasma arginine concentration. Study 2 addressed the effect of coadministration of cortisol with L-arginine on plasma Larginine concentrations. Study 3 was a randomized placebo crossover control comparing the effects of cortisol 80 mg/day co-administered with a placebo to cortisol 80 mg/day co-administered with L-arginine 21 g/day. Methods: Blood pressure was measured by a random Hawksley sphygmomanometer. Plasma nitrate/nitrite concentrations were measured by a modified Greiss reaction. Plasma arginine and citrulline concentrations were measured by an automated amino acid analyser. Results: Plasma arginine concentrations were doubled by oral doses of 15 g/day and 21 g/day of L-arginine (study 1). Co-administration of cortisol did not alter plasma arginine concentrations in subjects taking 21 g of Larginine per day (study 2). Co-administration of L-arginine 21 g/day with cortisol 80 mg/day did not prevent cortisolinduced increases in blood pressure or cortisol-induced falls in plasma nitrate/nitrite concentrations. Conclusion: Cortisol-induced hypertension is accompanied by a fall in plasma nitrate/nitrite concentrations. Oral L-arginine administration does not prevent cortisol-induced falls in plasma nitrate/nitrite concentrations or increases in blood pressure. We propose that cortisol-induced reductions in nitrate/nitrite production occur at a point distal to L-arginine availability in the nitric oxide synthase pathway.

‣ The heterodimeric amino acid transporter 4F2hc/y+LAT2 mediates arginine efflux in exchange with glutamine

Broer, Angelika; Wagner, Carsten; Lang, Florian; Broer, Stefan
Fonte: Portland Press Publicador: Portland Press
Tipo: Artigo de Revista Científica
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The cationic amino acid arginine, due to its positive charge, is usually accumulated in the cytosol. Nevertheless, arginine has to be released by a number of cell types, e.g. kidney cells, which supply other organs with this amino acid, or the endothelial cells of the blood-brain barrier which release arginine into the brain. Arginine release in mammalian cells can be mediated by two different transporters, y+LAT1 and y+LAT2. For insertion into the plasma membrane, these transporters have to be associated with the type-II membrane glycoprotein 4F2hc. The present study elucidates the function and distribution of y+LAT2. In contrast to y+LAT1, which is expressed mainly in kidney epithelial cells, lung and leucocytes, y+LAT2 has a wider tissue distribution, including brain, heart, testis, kidney, small intestine and parotis. When co-expressed with 4F2hc in Xenopus laevis oocytes, y+LAT2 mediated uptake of arginine, leucine and glutamine. Arginine uptake was inhibited strongly by lysine, glutamate, leucine, glutamine, methionine and histidine. Mutual inhibition was observed when leucine or glutamine was used as substrate. Inhibition of arginine uptake by neutral amino acids depended on the presence of Na+, which is a hallmark of y+LAT-type transporters. Although arginine transport was inhibited strongly by glutamate...

‣ Site-specific Labelling with a Metal Chelator for Protein-structure Refinement

Pintacuda, Guido; Moshref, Ahmad; Leonchiks, Ainars; Sharipo, Anatoly; Otting, Gottfried
Fonte: Kluwer Academic Publishers Publicador: Kluwer Academic Publishers
Tipo: Artigo de Revista Científica
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A single free Cys sidechain in the N-terminal domain of the E. coli arginine repressor was covalently derivatized with S-cysteaminyl-EDTA for site-specific attachment of paramagnetic metal ions. The effects of chelated metal ions were monitored with 15N-HSQC spectra. Complexation of Co2+, which has a fast relaxing electron spin, resulted in significant pseudocontact shifts, but also in peak doubling which was attributed to the possibility of forming two different stereoisomers of the EDTA-Co2+ complex. In contrast, complexation of Cu2+ or Mn2+, which have slowly relaxing electron spins, did not produce chemical shift changes and yielded self-consistent sets of paramagnetic relaxation enhancements of the amide protons. T1 relaxation enhancements with Cu2+ combined with T2 relaxation enhancements with Mn2+ are shown to provide accurate distance restraints ranging from 9 to 25 Å. These long-range distance restraints can be used for structural studies inaccessible to NOEs. As an example, the structure of a solvent-exposed loop in the N-terminal domain of the E. coli arginine repressor was refined by paramagnetic restraints. Electronic correlation times of Cu2+ and Mn2+ were determined from a comparison of T1 and T2 relaxation enhancements.

‣ The role of corticosterone in corticotrophin (ACTH)-induced hypertension in the rat

Mangos, George; Turner, Steven; Fraser, Tafline; Whitworth, Judith
Fonte: Lippincott Williams & Wilkins Publicador: Lippincott Williams & Wilkins
Tipo: Artigo de Revista Científica
Português
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Objective: Corticotrophin (ACTH)-induced hypertension in the rat is prevented by L- but not D-arginine. We examined the effects of exogenous corticosterone in the male Sprague Dawley (SD) rat to determine whether ACTH-induced hypertension is mediated by corticosterone. Methods: Exogenous corticosterone (10, 20 or 40 mg/kg per day) or sham (polyethylene glycol (PEG) 1 ml/kg per day) was injected subcutaneously in divided doses (s/c b.d.) over 15 treatment days to 40 SD rats (n = 10 each group). Subsequently, the effects of L-arginine, D-arginine or L-arginine + N-nitro-L-arginine (NOLA) on corticosterone-induced hypertension (corticosterone 20 mg/kg per day) were examined. Systolic blood pressure (SBP) and metabolic parameters were measured every two days. Results: Twenty and 40 mg/kg per day of corticosterone increased SBP compared with sham (P < 0.01, P < 0.05 respectively, sham versus respective group). Forty mg/kg per day of corticosterone raised serum corticosterone concentration compared with sham (502 ± 20 versus 364 ± 25 ng/ml, P < 0.001). L-arginine prevented the rise in SBP produced by corticosterone (131 ± 3 to 131 ± 2 mmHg, control versus day 10) but D-arginine did not (129 ± 3 to 142 ± 4 mmHg on day 8, P < 0.01). NOLA blocked the effect of L-arginine and amplified the rise in blood pressure produced by corticosterone (130 ± 3 to 171 ± 6 mmHg on day 10...

‣ ROLE OF NITRIC OXIDE IN ADRENOCORTICOTROPHIN-INDUCED HYPERTENSION: L-ARGININE EFFECTS REVERSED BY N-NITRO-L-ARGININE

Wen, Cheng; Li, Ming; Whitworth, Judith
Fonte: Blackwell Science Asia Publicador: Blackwell Science Asia
Tipo: Artigo de Revista Científica
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1. L-Arginine prevents adrenocorticotrophin (ACTH)-induced hypertension in the rat. To confirm that this effect is mediated through the nitric oxide (NO) system, we examined whether N-nitro-L-arginine (NOLA) could reverse the L-arginine-induced blockade of ACTH-induced hypertension. 2. Blood pressure and metabolic parameters were examined in sham-, ACTH., L-arginine + sham-, NOLA + sham-, ACTH + L-arginine- and ACTH + L-arginine + NOLA-treated Sprague-Dawley rats (n = 40). 3. Adrenocorticotrophin treatment increased systolic blood pressure (SBP), water intake and urine output and decreased bodyweight. N-Nitro-L-arginine alone increased SBP without affecting metabolic variables. L-Arginine alone did not affect blood pressure. The SBP was lower in L-arginine + ACTH- than ACTH-treated rats (P < 0.001), but was higher following ACTH + L-arginine + NOLA than ACTH + L-arginine (P < 0.05). 4. N-Nitro-L-arginine reversed the blood pressure-lowering effect of L-arginine in ACTH-induced hypertension in the rat, supporting the notion that NO plays a role in the hypertension.

‣ VASOPRESSIN V1A RECEPTOR ANTAGONISM DOES NOT REVERSE ADRENOCORTICOTROPHIN-INDUCED HYPERTENSION IN THE RAT

Fraser, Tafline; Turner, Steven; Wen, Cheng; Li, Ming; Burrell, L; Whitworth, Judith
Fonte: Blackwell Science Asia Publicador: Blackwell Science Asia
Tipo: Artigo de Revista Científica
Português
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1. The role of arginine vasopressin (AVP) was examined in adrenocorticotrophin (ACTH)-induced hypertension in Sprague-Dawley rats using the non-peptide AVP V(1a) receptor antagonist OPC 21268. 2. In an acute study, six rats were pretreated with ACTH for 11 days and direct arterial blood pressure (4 h), plasma osmolality and electrolyte concentrations were measured after OPC 21268 gavage. In a chronic study, 40 rats were randomly divided into four groups: (i) sham injection + sham gavage; (ii) ACTH + sham gavage; (iii) sham injection + OPC 21268; or (iv) ACTH + OPC-21268 for 16 days. Systolic blood pressure (SBP), water intake, urine volume (UV), urine osmolality and electrolytes, food intake, bodyweight and plasma osmolality and electrolyte concentrations were measured. 3. In the acute study, direct mean arterial blood pressure did not change with OPC 21268 (122±2 and 120±3 mmHg at 0 and 240 min, respectively). 4. In the chronic study, OPC 21268 did not affect ACTH-induced rises in blood pressure (from 125±2 (control) to 145±5 mmHg (group 4) compared with 122±3 (control) to 149±5 mmHg (group2)). Water intake and UV increased (from 29±2 to 83±6 mL/day; and from 5±1 to 36±5 mL/day, respectively) and the change in bodyweight decreased from 0±2 to -107±7 g. 5. These results suggest that AVP (at the V(1a) receptor) does not play a significant role in the maintenance of ACTH-induced hypertension.