Dopamine D2-like receptors in the rat kidney: effect of denervation

A new method for estimation of agonist-affinity (KA) and relative efficacy was introduced. This method afforded a procedure by which relative efficacy may be estimated while the actual KA values of agonist-receptor complexes are unknown. The relative efficacy may be estimated by employing a newly defined drug parameter, namely the eES value. The eES value is related to drug efficacy and is defined in such a manner that an isolated eES is a meaningful quantity which may indicate whether or not spare receptors are present in an agonist-effector system. The estimation of eES was based on the fact that fixed agonist-competitive antagonist combinations mimic partial agonists and mediate submaximal concentration-effect curves. However, for the practical estimation of eES one may employ data acquired from agonistic concentration-effect curves determined in the absence and presence of increasing concentrations of a competitive antagonist. This procedure was illustrated by utilizing theoretical concentration-effect curves and applied practically by estimating eES and KA values acquired from sets of carbachol and salbutamol curves. The sets of carbachol and salbutamol concentration-effect curves were determined in the absence and presence of their respective competitive antagonists, namely tripitramine and pindolol.     

Intracellular Ca2+ elevation and contraction due to prostaglandin F2alpha in rat aorta

Prostaglandin F2alpha was tested to determine (a) whether its effect on intracellular Ca2+ levels ([Ca2+]i) and force in vascular smooth muscle was mediated through activation of the thromboxane A2 and/or prostaglandin receptor, and (b) the relative roles of Ca2+ influx via L-type and non-L-type Ca2+ channels in prostaglandin receptor-mediated contraction. [Ca2+]i and force were measured simultaneously in fura-2-loaded rat aortic strips. The thromboxane A2 receptor antagonist, SQ29548 ([1S]-1a,2b(5Z),3b,4a-7-(3-[2-[(phenylamino)carbonyl] hydrazinomethyl)-7-oxobicyclo-[2.2.1]hept-2-yl-5-heptenoic acid), prevented the prostaglandin F2alpha-induced plateau [Ca2+]i elevation and force by 80-90%, while abolishing these responses due to the thromboxane A2 receptor agonist, U46619 (9,11-dideoxy-9alpha,11alpha-methanoepoxy prostaglandin F2alpha). Prostaglandin F2alpha (+ SQ29548)-induced plateau [Ca2+]i elevation and force were not inhibited by verapamil. Ni2+, a non-selective cation channel blocker, in the presence of verapamil, abolished the prostaglandin F2alpha (+ SQ29548)-elevated [Ca2+]i, while the contraction was only partially inhibited. These results suggest that, in rat aorta, (1) elevated [Ca2+]i and force due to high prostaglandin F2alpha concentrations largely results from thromboxane A2 receptor activation, and (2) the prostaglandin component of the prostaglandin F2alpha-induced contraction is dependent on Ca2+ influx via non-L-type channels.     

Effects of chronic administration of captopril on vascular reactivity of the rat aorta]

The various methods used to treat cutaneous leishmaniasis (CL) have not given consistent results. The aim of the present study was to compare the efficacy of a solution of meglumine antimoniate (MA; 85 mg Sb/ml) given intralesionally (i.l.) with that of the same solution given intramuscularly (i.m.). Eighty CL patients, with a total of 147 lesions, were randomly allocated into the two treatment groups. Forty were injected i.m. with MA (15 mg Sb/kg.day) on 6 days/week until 12 injections had been given to each. The lesions of the other 40 patients were infiltrated with MA (0.2-0.8 ml/lesion) every other day for 30 days. After 15 days' therapy, none of the lesions on those treated i.m. had fully healed (although five lesions showed some improvement) whereas two lesions on those treated i.l. had fully healed and 10 showed good improvement. After 30 days, 46 (68%) of the 68 lesions on those treated i.m. had healed completely, 11 (16%) had improved, and five (8%) worsened. The corresponding values for the 66 lesions on those treated i.l. were 48 (73%), 10 (15%) and three (5%). There was no statistically significant difference between the two treatment groups, either in terms of satisfactory response (lesions fully healed or improved) or unsatisfactory response (lesions unchanged or advanced), when assessed on day 30 (P > 0.5). Intralesional antimony is a rapidly effective, safe and economical method of treating simple CL, particularly in patients with cardiac, liver or renal disease, for whom antimonials are contra-indicated.     

Repair of rat gastric mucosa: effect of 16,16-dimethyl prostaglandin E2

Coadministration with the human immunodeficiency virus (HIV) protease inhibitor ritonavir was investigated as a method for enhancing the levels of other peptidomimetic HIV protease inhibitors in plasma. In rat and human liver microsomes, ritonavir potently inhibited the cytochrome P450 (CYP)-mediated metabolism of saquinavir, indinavir, nelfinavir, and VX-478. The structural features of ritonavir responsible for CYP binding and inhibition were examined. Coadministration of other protease inhibitors with ritonavir in rats and dogs produced elevated and sustained plasma drug levels 8 to 12 h after a single dose. Drug exposure in rats was elevated by 8- to 46-fold. A > 50-fold enhancement of the concentrations of saquinavir in plasma was observed in humans following a single codose of ritonavir (600 mg) and saquinavir (200 mg). These results indicate that ritonavir can favorably alter the pharmacokinetic profiles of other protease inhibitors. Combination regimens of ritonavir and other protease inhibitors may thus play a role in the treatment of HIV infection. Because of potentially substantial drug level increases, however, such combinations require further investigation to establish safe regimens for clinical use.     

Effect of beraprost sodium, a stable prostaglandin I2 analogue, on platelet aggregation in diabetes mellitus

Test results concerning platelet behavior in vitro, particularly aggregation, are frequently abnormal in diabetic patients. The concept has therefore arisen that platelet hyper-reactivity is one factor contributing to diabetic microangiopathy. We report here the antiplatelet effect of beraprost sodium, a chemically stable prostaglandin I2 analogue made in Japan, in 6 diabetic patients. Platelet aggregation induced by adenosine 5'-diphosphate (10 microM) after administration of beraprost sodium (40 micrograms every 8 h for 14 days) was significantly decreased as compared with levels before beraprost sodium administration. These results indicate the possibility that the occurrence of vascular complications in diabetes mellitus can be suppressed by long-term administration of beraprost sodium.     

Dexamethasone inhibits the pyrogenic activity of prostaglandin F2 alpha, but not prostaglandin E2

The effect of dexamethasone on prostaglandin (PG) E2- and PGF2 alpha-induced fever was studied in rats. Intracerebroventricular injection of PGE2 and PGF2 alpha (500 ng) induced increases in body temperature (maximal temperature rises of 0.97 +/- 0.13 degrees C and 0.78 +/- 0.18 degrees C, respectively, vs. vehicle 0.12 +/- 0.09 degrees C) of unrestrained rats maintained within the thermoneutral zone. PGE2-induced fever peaked earlier and the defervescence was faster when compared to the response induced by PGF2 alpha. Subcutaneous pre-administration of dexamethasone (0.5 mg/kg) did not affect PGE2-induced fever (maximal temperature rise of 1.00 +/- 0.08 degrees C), but completely prevented the pyrogenic activity of PGF2 alpha (maximal temperature rise of 0.16 +/- 0.16 degrees C). Neither PGE2- nor PGF2 alpha-induced fever was significantly altered (maximal temperature rises of 0.90 +/- 0.11 degrees C and 0.64 +/- 0.14 degrees C, respectively) by intraperitoneal administration of indomethacin (2 mg/kg). These results demonstrate for the first time that glucocorticoids, in addition to inhibiting endotoxin- and cytokine-induced fever, can also modulate the pyrogenic activity of some prostaglandins, possibly via suppression of the synthesis of corticotropin-releasing factor, indicating that multiple mechanisms may be involved in the antipyretic activity of these steroids.     

Protective effect of prostaglandin E2 on cerulein-induced rat pancreatitis]

In order to clarify the effect of prostaglandin E2 (PGE2) on cerulein-induced rat pancreatitis, we investigated the interaction of PGE2 with cerulein or secretin. Intravenous infusion of 10 micrograms/kg.h cerulein inhibited external secretion of the pancreas from one hour and caused macroscopic edema at 3 hours. Administration of PGE2 relieved the inhibitory effect of supramaximal dose of cerulein and decreased the pancreatic edema. The 100 micrograms/kg.hr PGE2 had no significant effect on the pancreatic juice volume and amylase secretion stimulated with 0.2 micrograms/kg.hr of cerulein. Intravenous injection of 100 micrograms/kg PGE2 inhibited both the volume and amylase secretion of pancreatic juice stimulated with 1 U/kg.h of secretin. The protective effect of PGE2 on cerulein-induced pancreatitis was not the stimulation on secretion but caused the cytoprotective effect of PG such as stabilization of cytoplasmic and lysosomal membrane.     

The effect of sodium nitroprusside on the membrane potential and mechanical tension of the smooth-muscle cells in the rat aorta

The rat aorta smooth muscles were contracted and depolarised with high potassium or phenylephrin solution which was followed by a relaxation and repolarisation after sodium nitroprusside administration. The latter effect was decreased az a combined action of phenylephrin and high potassium solution. Nitroprusside seems to act through a cGMP-dependent potassium transient activation of the smooth muscle cell membrane.     

Differential effect of tetrandrine on aortic relaxation and chronotropic activity in rat isolated aorta and atria

Investigation of the pathogenesis of arteriosclerosis and/or atherosclerosis has been progressed using molecular biology. New concepts have been developed and, receptors and substances have been found clinically and experimentally, which have led us to create new methods of evaluating or diagnosing the grade of atherosclerosis lesion. Dealing with the new concepts or knowledge in this symposium, this introductory paper describes an overview of pathogenesis of atherosclerosis, from which the new methods of evaluating or diagnosing lesion has been exploited. The injury to the endothelium leads to endothelial cell dysfunction, which initiates the acceleration of LDL oxidation and increases adherence of monocytes, macrophages and T lymphocytes, migrating subendothelially and causing large foam cells to develop because of lipid accumulation. Macrophages and platelets release many growth factors, which accelerate the growth of vascular smooth muscle cells, forming fibrous plaque. In these pathogenic processes of atherosclerosis, angiotensin II participates in releasing growth factor for cell proliferation and hepatocyte growth factor (HGF) participates in revascularization of the sclerotic lesion, suggesting a candidate marker for atherosclerosis. Hyperlipidemia and hypercoagulation are the major factors in advanced atherosclerosis. Using new methods to evaluate or diagnose lesions, further therapy and prevention for atherosclerosis will progress in future.     

Protective effect of a prostaglandin I2 analog, TEI-7165, on ischemic neuronal damage in gerbils

In this study we have analyzed short- and long-term changes in extracellular signal-regulated kinase (ERK) 1 and 2 activity during 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced differentiation of human promyelocytic leukemia cells. Immunoprecipitation of HL-60 cellular extracts with an ERK antibody followed by in vitro myelin basic protein phosphorylation demonstrated a rapid reduction in total ERK activity by 70%. Mitogen-activated protein kinase substrate peptide phosphorylation also demonstrated that this reduction was sustained during differentiation. Immunoblot analysis revealed that ERK1 and ERK2 are the predominant ERK isoforms present in HL-60 cells and that over a 96-h period ERK1 protein was gradually reduced by 60% while ERK2 protein showed only a small, insignificant reduction. Therefore, the large, rapid decrease in total ERK activity could not be attributed to the gradual reductions in ERK1 or ERK2 amounts. Immunoblot analysis with two different phosphotyrosine antibodies revealed a rapid decrease in ERK1 phosphotyrosine and a concurrent transient increase in ERK2 phosphotyrosine. These contrasting changes in phosphorylated ERKs were paralleled by respective shifts in mobility during SDS-PAGE analysis. Together these results indicate that the rapid reduction in total ERK activity is due to rapid tyrosine and possible threonine dephosphorylation of ERK1 but not of ERK2. These results also indicate that ERK1 and ERK2 are regulated by distinct mechanisms during TPA-induced HL-60 differentiation, suggesting that their biological roles are nonredundant.     

Effects of prostaglandin E2 and oxytocin on the electrical activity of hormone-treated and pregnant rat myometria

1. The membrane properties of the rat myometrium, during gestation and following ovarian hormone treatment, have been investigated with the micro-electrode technique. 2. Spontaneously generated bursts of electrical activity alternating with silent periods were recorded from non-pregnant, pregnant and post-partum myometria. The membrane potential was highest during the middle stage of gestation, but the spike amplitude within a burst was not uniform. In the final stage of gestation and during parturition, the membrane potential was low and the spikes within a burst were of low frequency and uniform amplitude. 3. During parturition and post-partum, a gradual depolarization of the membrane, accompanied by an increase in membrane resistance, occurred before the generation of a burst. 4. Excitability of the membrane fluctuated from a peak just before the generation of a burst to a low after the cessation of a burst. 5. Displacement of the membrane potential by electrical current or by lowering the temperature modified the slope spontaneous depolarization, but the fluctuations of excitability persisted. The Q10 value for the frequency of spontaneous bursts, measured between 36 and 30 degrees C, was 3-8. 6. Hyperpolarization of the membrane increased the maximum rate of rise of the spike, but beyond -70 mV, the rate of rise was reduced. Half-inactivation of spike generation of spike generation occurred at a membrane potential less negative than the interburst potential, indicating that the current carrying system was not fully activated during parturition. 7. In both normal and spayed rats, oestradiol hyperpolarized the membrane and the burst of spikes was generated hyperpolarized the membrane and the burst of spikes was generated on a sustained depolarization. Progesterone slightly hyperpolarized the membrane and burst discharges occurred without a sustained depolarization. Simultaneous treatment with progesterone and oestradiol produced a plateau potential of long duration during burst discharges. 8. The thickness of the muscle layer, length constant of the tissue and time constant of the membrane were measured during gestation and from spayed rats under various hormonal conditions. The length constant of the tissue was increased by oestradiol and was further increased by simultaneous treatment withoestradiol and progesterone. The increase in tissue thickness appeared to have the most marked influence on the length constant. 9. The resting and active membrane properties of the progresterone treated myometrium were similar to those observed during the middle stages of gestation. The oestradiol-treated myometrium did not resemble that during the last stages of gestation and parturition, which was simulated by combination of the two hormones, oestradiol preceding progesterone.     

Effect of inhibiting renal kallikrein on prostaglandin E2, water, and sodium excretion

To test the hypothesis that renal kinins act as natriuretic and diuretic hormones, we examined the effect of inhibiting glandular kallikrein on renal function in normotensive unanesthetized rats during normal sodium intake. To inhibit kallikrein at both the luminal and basolateral sides of the distal nephron, we used Fab fragments of monoclonal antibodies to rat urinary kallikrein (Fab-kallikrein). Fab fragments have advantages over intact IgG: they are filtered through the glomerulus and reach the lumen of the distal nephron, where kallikrein is localized and urinary kinins are released. Furthermore, the Fab fragment-antigen complex does not activate the complement system, avoiding the side effects associated with intact antibodies. Fab-kallikrein effectively blocked generation of kinins in the nephron lumen, decreasing urinary kininogenase activity (kallikrein) by 74% to 85% and kinin excretion by 76% to 79%. Fab-kallikrein induced a 30% decrease in urine volume and a 20% to 40% decrease in urinary sodium excretion but did not alter blood pressure, glomerular filtration rate, or renal blood flow. Although urinary prostaglandin E2 excretion also tended to decrease, this change was slower and of lesser magnitude than those of kinin and kininogenase excretion and did not attain statistical significance after Bonferroni's correction. In controls injected with either vehicle or Fab fragments of monoclonal antibodies to ricin (a vegetable protein not present in mammals), none of these parameters decreased significantly. We conclude that renal kinins participate in the short-term regulation of water and sodium excretion in normotensive unanesthetized rats, acting as diuretic and natriuretic hormones.(ABSTRACT TRUNCATED AT 250 WORDS)     

Zinc (Zn2+) and fish immune response effect on carp IL2-like production and activity

Supernatants obtained from PHA-activated pronephric carp lymphocytes were able to provoke the growth of carp blasts but not from fresh lymphocytes without mitogens. These supernatants contain certainly soluble growth factor(s) such as IL2. The addition of Zn2+ (1 mM) to lymphocyte cultures could not enhance supernatants' ability to promote blast proliferation. When Zn2+ and PHA (50 micrograms/ml) were added, respectively, supernatants obtained could enhance significant tritiated thymidine incorporation in blasts. Zn2+ by itself is not able to provoke the growth of blasts in vitro. In contrast, the addition of Zn2+ and PHA-stimulated lymphocyte supernatants in vitro to blasts or nonstimulated lymphocyte cultures enhances their mitotic capacity.     

Vasorelaxing activity of resveratrol and quercetin in isolated rat aorta

PURPOSE: To assess the severity of adverse reactions to contrast media in outpatient computed tomographic (CT) examinations in a conventional clinical setting. MATERIALS AND METHODS: In 4,936 patients, CT was performed with four protocols: ionic contrast medium with sodium meglumine as the cation (in one protocol, contrast material was warmed to 35 degrees C before injection; in another protocol, it was administered at ambient temperature); warmed, ionic contrast medium with nonsodium pure meglumine as the cation; and warmed, nonionic iopamidol. RESULTS: Adverse reactions to ionic contrast material statistically significantly decreased (P<.05) when it was warmed before administration. Reactions to ionic contrast media without a sodium cation were statistically significantly fewer (P<.001) than reactions to those with a sodium cation. In all protocols, pediatric patients had fewer reactions than adult patients. CONCLUSION: In outpatient CT examinations, nonionic, warmed contrast medium was the best option because no severe reactions resulted from its use. Prevalence of adverse reactions was comparable to that in controlled randomized studies.     

Action of prostaglandin, PGF2alpha, on the uterus of the pregnant rat

In a cytogenetic study on the spermatogonia of Chinese hamster, cyclohexylamine (neutral sulphate) was evaluated for mutagenic effects in comparison with an untreated control group and a group treated with the mutagenic compound cyclophosphamide, by assessing spermatogonial metaphases of treated Chinese hamster for chromosomal structural changes. Each test group comprised 8 male hamsters selected at random. Approximately 100 metaphases from each animal were assessed. The doses were 5 X 150 mg cyclohexylamine sulphate (approx. 5 X 102 mg base) per kg body-weight orally, and 5 X 100 mg cyclophosphamide per kg body-weight orally. The individual doses were administered at intervals of 24 h. Preparations were made 24 h after the final treatment, essentially by the method of Hoo and Bowles [10]. Gaps, breaks, fragments, deletions and translocations were assessed as structural changes; frequencies were determined of the metaphases (a) with aberration(s) including gaps, (b)with aberration(s) less gaps and (c)with translocation(s). Aberrations occurred in the untreated negative control group (1.24% incl. gaps, 0.25% without gaps). Translocations were not seen in the untreated group. In the cyclochexylamine group, the frequencies of the aberrant metaphases were sometimes less than in the control group (0.87% including gaps, 0.37% without gaps). Statistically, the results were not significantly different from the control data. No translocations were seen after administration of cyclohexylamine. The positive cyclophosphamide control group clearly differed from the untreated control and from the cyclohexylamine group in the parameters (a) to (c); mainly, the results were highly significantly different from those obtained in the untreated control group. The frequencies of the aberrant metaphases were 3.41% including gaps and 1.99% without gaps. The frequency of the translocations was 0.71% (5 out of 704). Cyclohexylamine sulphate, administered 5 times at 150 mg/kg body-weight orally, had no mutagenic effect, whereas cyclophosphamide, adminstered 5 times at 100 mg/kg body-weight orally, had a chromosome-damaging effect on Chinese hamster spermatogonia.     

Phospholipase A2 activity and calpactin I levels in rat lymphokine-activated killer cells: correlation with the cytotoxic activity

The interaction of carnitine with human placental brush-border membrane vesicles was investigated. Carnitine was found to associate with the membrane vesicles in a Na(+)-dependent manner. The time course of this association did not exhibit an overshoot, which is typical of a Na+ gradient-driven transport process. The absolute requirement for Na+ was noticeable whether the association of carnitine with the vesicles was measured with a short time incubation or under equilibrium conditions, indicating Na(+)-dependent binding of carnitine to the human placental brush-border membranes. The binding was saturable and was of a high-affinity type with a dissociation constant of 1.37 +/- 0.03 microM. Anions had little or no influence on the binding process. The binding process was specific for carnitine and its acyl derivatives. Betaine also competed for the binding process, but other structurally related compounds did not. Kinetic analyses revealed that Na+ increased the affinity of the binding process for carnitine and the Na+/carnitine coupling ratio for the binding process was 1. The dissociation constant for the interaction of Na+ with the binding of carnitine was 24 +/- 4 mM. This constitutes the first report on the identification of Na(+)-dependent high-affinity carnitine binding in the plasma membrane of a mammalian cell. Studies with purified rat renal brush-border membrane vesicles demonstrated the presence of Na+ gradient-driven carnitine transport but no Na(+)-dependent carnitine binding in these membrane vesicles. In contrast, purified intestinal brush-border membrane vesicles posses neither Na+ gradient-driven carnitine transport nor Na(+)-dependent carnitine binding.     

Attenuation of ischemic liver injury by prostaglandin E1 analogue, misoprostol, and prostaglandin I2 analogue, OP-41483

Workplace AIDS training is a recent addition to many corporations' occupational health agenda. However, little is known about the objectives, content, and practices of AIDS training programs. A survey of 126 workplace AIDS trainers was conducted to determine the impact of the trainer's organizational affiliation (in-house, consultant, union, etc.) and personal motives on training program objectives, content, and practices. Results indicate that the organizational affiliation of trainers is significantly related to training objectives, topics, and practices, whereas strong personal motives for becoming an AIDS trainer is significantly associated with an emphasis on more controversial content areas and training practices. Findings are discussed in terms of applicability to other values-oriented training topics, applications to practice, and future research needs.