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This study was designed: (1) to test the hypothesis that the pressor response to sodium chloride loading in patients with essential hypertension is associated with disorder of divalent cations (calcium and magnesium); and (2) to clarify the relationship between intracellular free calcium concentration and serum levels of calcium-regulating factors and intracellular magnesium concentration. With sodium chloride loading, mean blood pressure, urinary calcium and magnesium excretions and platelet calcium concentration were increased, and serum total and ionized calcium, magnesium concentrations and erythrocyte magnesium concentration were decreased. Change in mean blood pressure was correlated with changes in parathyroid hormone (r = 0.49, P less than 0.05), serum total and ionized calcium (r = -0.50, P less than 0.05) and erythrocyte magnesium (r = -0.56, P less than 0.05) and platelet calcium concentrations (r = 0.46, P less than 0.05). In addition, change in platelet calcium concentration was related to changes in parathyroid hormone (r = 0.44, P = 0.05), serum total and ionized calcium (r = -0.66, P less than 0.01) and erythrocyte magnesium concentration (r = -0.49, P less than 0.05). It is concluded that the pressor effect of excessive sodium chloride intake on blood pressure in patients with essential hypertension is associated with a disorder of divalent cations and that alteration of the intracellular free calcium concentration with sodium chloride loading may occur through handling of serum total and ionized calcium, parathyroid hormone and/or intracellular magnesium concentration.

We examined metoprolol oxidation capacity in 218 unrelated, healthy Korean subjects using the 8 h urinary metabolic ratio (MR) of metoprolol to alpha-hydroxymetoprolol after an oral dose of 100 mg metoprolol tartrate. The results were compared with those from 295 Japanese and 107 mainland Chinese whose metoprolol oxidation capacities were assessed in a similar manner. The frequency of occurrence of poor metabolisers (PMs) was 0.5% (1/218) in the Korean, 0.7% (2/295) in the Japanese and 0% in the Chinese population. However, the respective mean (+/- s.d.) MRs (0.84 +/- 1.14 and 0.87 +/- 0.90) in the Korean and Japanese extensive metabolisers (EMs) were significantly (P less than 0.001) less than that in the Chinese EMs (2.81 +/- 2.35), and the mode of the distribution histogram and the probit plot of data for Chinese EMs were shifted to the right compared with those for Korean and Japanese EMs. The results indicate that Koreans, like Japanese and mainland Chinese, have a much lower frequency of the PM phenotype of debrisoquine/sparteine-type oxidation compared with that reported for Caucasian populations. Chinese EMs appear to have a lower capacity to metabolise metoprolol to alpha-hydroxymetoprolol compared with Korean or Japanese EMs.

To examine predictors for the efficacy of antihypertensive agents, we investigated the effects of nifedipine and captopril on blood pressure (BP) and humoral factors in patients with essential hypertension. Eleven essential hypertensive patients (mean age: 54) were treated with long acting nifedipine at 20 to 40 mg/day for 8 weeks and 25 essential hypertensives (mean age: 51) were treated with captopril at 37.5 to 75 mg/day. Blood pressure was measured every 2 weeks. Plasma renin activity (PRA), and plasma concentrations of aldosterone, epinephrine and norepinephrine were determined before and at the end of treatment. Both nifedipine and captopril decreased BP (nifedipine: mean BP 119 +/- 3 to 101 +/- 2 mm Hg, captopril: 124 +/- 2 to 100 +/- 2, P less than .01 for each), whereas neither of them affected heart rate. The 8-week treatment of nifedipine showed no significant effect on humoral factors. Captopril increased PRA by 63% (P less than .05) and decreased plasma epinephrine by 42% (P less than .01) and norepinephrine by 35% (P less than .01). The change in mean BP was positively correlated with pretreatment PRA (r = 0.68, P less than .01) in nifedipine-treated patients and inversely with pretreatment norepinephrine (r = -0.53, P less than .01) in captopril treatment. The results suggest that both nifedipine and captopril were effective antihypertensive agents and that the long term treatment of nifedipine is more effective in essential hypertensives with lower PRA, while captopril is more effective in those with higher plasma norepinephrine concentration.

A case of hydrocephalus showing slit ventricle syndrome after multiple shunt revisions was treated with a programmable pressure valve, and intracranial pressure was monitored with a telemetric sensor. High pressure setting produced positive and normal values of intracranial pressure in this patient. We report our experiences.

A new mutant protein of recombinant human granulocyte colony-stimulating factor (rhG-CSF) was produced for the studies on receptors for human G-CSF. The mutant protein [(Tyr1, Tyr3]rhG-CSF), the biological activity of which was almost equal to that of rhG-CSF, was prepared by the replacement of threonine-1 and leucine-3 of rhG-CSF with tyrosine. The radioiodinated preparation of the mutant protein showed high specific radioactivity and retained full biological activity for at least 3 weeks. The binding capacity of the radioiodinated ligand was compared with that of [35S]rhG-CSF. Both radiolabeled ligands showed specific binding to murine bone marrow cells. Unlabeled rhG-CSF and human G-CSF purified from the culture supernatant of the human bladder carcinoma cell line 5637 equally competed for the binding of labeled rhG-CSFs in a dose-dependent manner, demonstrating that the sugar moiety of human G-CSF made no contribution to the binding of human G-CSF to target cells. In contrast, all other colony-stimulating factors and lymphokines examined did not affect the binding. Scatchard analysis of the specific binding of both labeled ligands revealed a single class of binding site with an apparent dissociation constant (Kd) of 20-30 pM and 100-200 maximal binding sites per cell. These data indicate that the radioiodinated preparation of the mutant protein binds the same specific receptor with the same affinity as [35S]rhG-CSF. The labeled mutant protein also showed specific binding to human circulating neutrophils.(ABSTRACT TRUNCATED AT 250 WORDS)

The anti-angiogenic activity of AGM-1470, a new synthetic analog of fumagillin isolated from Aspergillus fumigatus, was extensively examined both in vitro and in vivo using four different types of assay and compared to that of the fumagillin parent. Locally administered AGM-1470 inhibited the angiogenesis in the chick embryo chorioallantoic membrane assay and the rat corneal assay. In the rat sponge implantation assay, systemically administered AGM-1470 inhibited angiogenesis induced by basic fibroblast growth factor. Furthermore, in the rat blood vessel organ culture assay, AGM-1470 (1-1,000 ng/ml) was found to selectively inhibit the capillary-like tube formation of endothelial cells with a minimal effect on the non-endothelial cell growth. AGM-1470 showed more potent anti-angiogenic activity and less toxicity than the fumagillin parent. Therefore, AGM-1470 is much better than the fumagillin parent as anti-angiogenic compound.

The number and the affinity of granulocyte colony-stimulating factor (G-CSF) receptors expressed by blast cells in acute myeloblastic leukaemia (AML) were determined using radiolabelled recombinant human G-CSF (rhG-CSF). Eighteen of 20 patients demonstrated specific binding, and Scatchard analysis revealed a single class of high affinity (Kd 15-130 pM) G-CSF receptors on the AML blasts. The number of G-CSF receptors varied from 55 to 1,200 per cell (mean 278). In the remaining two patients, specific binding was not observed. The number of G-CSF receptors did not differ significantly between various AML subtypes, but the mean receptor number was the highest on type M2 blasts. A chemical cross-linking study revealed that the G-CSF receptor has an approximate molecular weight of 140,000. Autoradiography showed heterogeneity of the distribution of G-CSF receptors on the AML blasts obtained from a single patient. The number of colonies stimulated by the addition of rhG-CSF varied from 0 to 566 per dish, and blast colony formation was observed in eight of 20 patients. The population mean of G-CSF receptor number expressed by blasts that formed colonies on stimulation with rhG-CSF was significantly higher than that on blasts which did not form colonies. These results suggest that a proliferative response of AML blasts to G-CSF may be predicted when the blasts express a large number of G-CSF receptors. Accordingly, it may be safer to restrict the clinical use of G-CSF to AML patients who have blasts with a low G-CSF receptor expression and no response to G-CSF in blast colony assay.

Although the existence of salt sensitivity in essential hypertensives has been well known, the precise mechanism(s) has not yet been elucidated. The aim of this study was to clarify the relation between the responses in blood pressure, extra- and intracellular cations and calcium-regulating hormones to oral NaCl loading in essential hypertensives. After oral NaCl loading, mean blood pressure, urinary excretions of calcium and magnesium, and PLT[Ca2+]i were significantly increased. [Ca2+]o and E[Mg]i were decreased. The changes (delta) in mean blood pressure by NaCl loading positively correlated with delta PLT[Ca2+]i and delta PTH, and negatively with delta[Ca2+]o and delta E[Mg]i. Delta PLT[Ca2+]i positively correlated with delta PTH and negatively with delta[Ca2+]o and delta E[Mg]i. From these results, the blood pressure response to oral NaCl loading is associated with the alternation of [Ca2+]i metabolism in which the changes in magnesium metabolism and calcium-regulating hormones may be involved.

The effects of recombinant human erythropoietin (r-HuEPO) on haematological parameters were studied in rats in which uraemia and anaemia had been induced by gentamicin, an aminoglycoside antibiotic and a nephrotoxic agent. After the occurrence of slight polycythaemia, the red blood cell count, haematocrit and haemoglobin concentration decreased by 20-30% compared with those of the control (saline-injected) rats. At the end of gentamicin treatment, the endogenous serum EPO level had decreased to about 40% compared with that of control rats. Gentamicin-treated rats showed marked elevation of blood urea nitrogen, extensive tubular necrosis in the kidney and haemosiderin deposition in the spleen. In the osmotic fragility test, the fragility of erythrocytes significantly increased compared with that of control rats. These findings indicate that the anaemia induced by gentamicin is due not only to a deficiency of EPO but also to an enhancement of fragility of erythrocytes in an azotaemic environment. The administration of r-HuEPO during anaemia markedly increased red blood cell count, haematocrit and haemoglobin concentration. It is suggested that a gentamicin-treated rat is a useful and convenient anaemic model and r-HuEPO is useful for treatment of anaemia in acute renal failure.

We measured the effects of recombinant human granulocyte-colony stimulating factor (rhG-CSF) on the adherence of human neutrophils by using a dacron fiber system to assay the adhesive ability of neutrophils. rhG-CSF enhanced neutrophil adherence to dacron fibers. N-formyl-methionyl-leucyl-phenylalanine (fMLP) induced neutrophil-neutrophil interaction (neutrophil aggregation) in addition to neutrophil-dacron interaction, whereas rhG-CSF did not cause neutrophil aggregation. These results indicated that rhG-CSF increases the adhesive ability of neutrophils without neutrophil-neutrophil interaction, and the action of rhG-CSF in neutrophil activation is different from the neutrophil activation caused by fMLP.

Single injection of recombinant human granulocyte colony-stimulating factor (rhG-CSF) immediately induced a decrease in the number of circulating neutrophils in rats. This neutropenia occurred 10 minutes after the injection but disappeared 40 minutes after injection. This transient neutropenia was dose-dependently induced by rhG-CSF and also induced by repeated injections. We studied the kinetics of circulating neutrophils in transient neutropenia. rhG-CSF markedly decreased the number of 3H-diisopropylfluorophosphate (3H-DFP) labeled neutrophils in the circulation 10 minutes after injection but the labeled neutrophils recovered to near the control level 40 minutes after the injection. These results indicate that the neutrophil margination accounts for the neutropenia and the marginated neutrophils return to the circulation.

Prostaglandin (PG) E2, known as a bone-resorption factor, was released as a predominant arachidonate metabolite in the culture medium of an osteoblastic cell line cloned from mouse calvaria (MC3T3-E1). Epidermal growth factor (EGF) (10 ng/ml) prominently enhanced endogenous PGE2 synthesis, requiring the simultaneous presence of unidentified factor(s) contained in bovine serum. PGE2 synthesis increased after a lag phase for 1-2 h and reached a maximum level at about 3 h after EGF addition. EGF-stimulated PGE2 synthesis was almost completely blocked by 10 microM cycloheximide or 1 microM actinomycin D. Furthermore, when the cells were pretreated with EGF, the microsomes exhibited an increased activity of fatty acid cyclooxygenase (arachidonic acid----PGH2), whereas the activity of PGE synthase (PGH2----PGE2) remained unchanged. These results suggested an EGF-mediated induction of cyclooxygenase. Following increased PGE2 synthesis, DNA synthesis increased and alkaline phosphatase activity decreased in a slower response to EGF. PGE2 (above 0.1 microM) added to the cells could replace EGF. However, such effects of EGF on the osteoblasts could not be attributed totally to an autocrine function of PGE2 produced by stimulation with EGF because these effects of EGF were not abolished by indomethacin, which blocked the PGE2 synthesis.

The bone marrow of a patient with IgA monoclonal gammopathy associated with gastric carcinoma was massively infiltrated with extracellular globules of varying sizes from 2 to 110 micron. These exposed globules were not found in the metastatic lesions heavily infiltrated with plasma cells, but were exclusively confined to the bone marrow. They were indistinguishable from Russell bodies and composed of monoclonal immunoglobulin (IgA, lambda). Of interest is the fact that, despite a small number of plasma cells, there was an intramedullary extensive infiltration of exposed globules. The following conditions may be related to these changes. (1) The globules were not removed rapidly by effectual phagocytosis. (2) The environment of the bone marrow may be favorable for their remaining. In our case, however, the possibility that the plasma cells around the tumor were not of the same clonal origin as the marrow plasma cells cannot be ruled out. (3) The marrow plasma cells may produce immunoglobulin more vigorously, whereas their survival time be shorter than usual. As a result, these exposed globules released on the death of the cells accumulated massively in the bone marrow.

Turnover rates of blood glucose and lactate, along with the rates of their interconversion, were calculated from changes of their specific activities in rats infused with [U-14C] glucose and [U-14C] lactate based on a simple flux model of the Cori cycle. In the fasted state, most (75%) of the glucose metabolized was converted to lactate, whereas only a small fraction (35%) of the lactate metabolized was converted to glucose. Of the glucose metabolized, 27% returned from the lactate pool to the glucose pool. Feeding stimulated the outflow of glucose carbon from the Cori cycle via the action of secreted insulin. Insulin-induced stimulation of blood glucose removal was associated with increased glucose production, whereas anti-insulin serum-induced stimulation of glucose production was associated with increased glucose removal. It is argued that a "compensatory" mechanism independent of insulin and epinephrine might be in operation to minimize the glycemic changes that would otherwise occur in response to drastic metabolic changes.

We explored the effects of 12-hour infusion of atrial natriuretic peptide (alpha-rANP:rat, 1-28) on arterial acid-base balance, using 5/6 nephrectomized rats with chronic renal failure. Before the infusion, nephrectomized rats had a higher mean arterial blood pressure, greater urine volume, and lower creatinine clearance than the normal controls, but they did not show a significant difference in arterial hydrogen ion concentration (pH), plasma bicarbonate concentration (HCO3-), partial pressure of carbon dioxide (PCO2), plasma base excess (BE), or plasma ANP concentration. alpha-rANP infusion produced a continuous blood pressure reduction in both nephrectomized and control rats. Urine volume and urinary sodium and potassium excretion tended to increase at 2-hour infusion, but not at 12-hour infusion. In the controls alpha-rANP significantly increased pH from 7.47 to 7.50, and decreased PCO2 by 14%. In contrast, in nephrectomized rats alpha-rANP significantly decreased pH from 7.48 to 7.44, HCO3- by 13%, and BE from -0.07 to -3.22 meq/l. Rats with chronic renal failure had greater reduction in HCO3- than the controls (p less than 0.05). There was no difference in plasma ANP level between the two groups. Thus, it is indicated that the long-term infusion of alpha-rANP reduces pH in rats with chronic renal failure, thereby adversely affecting the acid-base balance.

The present study was undertaken to evaluate the effects of single and repeated doses of triazolam (1 mg/kg IP) and desipramine (DMI) (10 mg/kg IP) alone and in combination on certain pharmacologic responses and brain catecholamine levels in rats. The significant findings were: (1) triazolam and DMI given alone and in combination were synergistic with pentobarbital (PB) hypnosis; (2) repeated treatment with DMI alone repressed body weight gain; triazolam potentiated this anorectic effect; (3) triazolam induced muscle relaxation acutely, but was not affected by concomitant DMI treatment; and (4) triazolam and DMI given in combination produced hypothermia although the drugs given separately did not. Development of tolerance to hypothermia and to anorectic effects produced by triazolam and DMI significantly decreased norepinephrine (NE) levels in the hypothalamus as compared to the repeated treatment with triazolam, and significantly increased NE and dopamine (DA) levels in the striatum and DA levels in the cortex, as compared to the single treatments.

Environmental risk incurred with the use of synthetic surfactants is dealt with in this paper. The background and necessity of risk management related to surfactant usage in the Lake Biwa basin are introduced, as well as a research scheme that acknowledges risks in three sub-processes--consumption and discharge, fate in aquatic environment, and ecotoxicological response of linear alkylbenzene sulfonate (LAS). The ayu (Plecoglossus altivelis) has been selected as the significant biological target in the basin. Results of a field survey of the behavior of LAS along streams flowing down to the lake are presented. Included are the estimation and verification of an original unit of surfactant consumption per capita per day based on LAS and MBAS concentrations observed in diurnal monitoring. A simulation model representing the flowdown process of LAS dynamically is formulated, with which longitudinal dispersion, settling, and modified biodegradation of LAS are evaluated in the field survey. On the basis of the research scheme described above, ecotoxicological laboratory tests on ayu have been carried out. The special significance of acute and subchronic effects on ayu in various life stages exposed to low concentrations of LAS can be recognized. It has been concluded that the LC50 of young ayu is not greater than 0.1 ppm under the disadvantageous condition of high temperature or extreme hardness even in normal ranges of environmental indicators. An advanced plan of risk management for surfactant usage is proposed based on methods of elevated risk, comparative risk, risk--benefit, and balanced risk. The occurrence and magnitude of risk phenomena in each subdivided basin adjacent to the lake are identified, taking into consideration features such as (1) the spawning place of ayu and aquafarms, (2) COD and MBAS concentrations and their tolerable levels in current regulation of stream pollution, (3) socioeconomic perspectives including recreational activities and voluntary actions to improve the neighborhood environment, and (4) future preparation of measures for resolving technological risk including sewerage construction. Because LAS concentrations in several rivers exceed the tolerable level of the most delicate life stage of ayu, latent environmental effects can be suggested. It is concluded therefore that it is necessary to establish a water quality goal related to surfactants that takes patterns of water use and conservation of the ecosystem in the local environment into consideration. These judgments are followed by formulation of the "requirement" for environmental management practice for the Lake Biwa administration.

Although the existence of so-called streptozocin hypertension seems well established, some reports have indicated that no rise in blood pressure (BP) occurred after streptozocin treatments. To ascertain the streptozocin-induced BP response, normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were treated with streptozocin, 40 to 45 and 35 mg/kg i.v., respectively, and BP was determined directly and indirectly every week for 3 to 4 weeks. Direct mean BP was determined without anesthesia or restraint through a cannula inserted into the rat's abdominal aorta. Indirect BP was determined at the tail without anesthesia after prewarming the rat in a holder. Compared with control values, indirect BP increased significantly in diabetic WKY 2 weeks after streptozocin treatment. In contrast, direct BP of these rats decreased, compared with control values. Indirect BP of diabetic SHR was as high as that of the controls, whereas direct BP of diabetic SHR decreased significantly 1 week after the treatment and thereafter, compared with control values. These discrepancies between the direct and indirect BP values may be caused by severe emaciation of diabetic rats. Extra pressure in the cuff may be necessary to occlude the bloodstream. These results indicate that under these conditions the value of BP obtained by the direct measurement is more reliable than that by the indirect one; therefore, we concluded that so-called streptozocin hypertension does not exist.

The effect of recombinant interleukin 1 Beta (IL-1(beta)) was investigated on osteoblastic cell line MC3T3-E1 cloned from mouse calvaria. IL-1(beta) stimulated cell proliferation which increased cell number and caused dose-related stimulation of DNA synthesis, with a maximal effect at a concentration of 12.5 U/ml; suppressed alkaline phosphatase activity and collagen synthesis maximally at 0.5 and 62.5 U/ml, respectively; and increased the amount of free [3H] hydroxyproline in the cultures, but the amount was quite low. Prostaglandin E2 synthesis was also stimulated dose dependently by the presence of IL-1(beta), with a maximal increase at 2.5 U/ml, at which concentration the prostaglandin E2 level in the medium was 1.61 +/- 0.10 ng/ml. The increased prostaglandin E2 synthesis did not affect either the IL-1(beta)-mediated change in DNA or collagen synthesis or alkaline phosphatase activity. These results extend the possibility that IL-1(beta) is to act as a regulator of bone formation.

We examined the effects of antihypertensive therapy with captopril, an angiotensin converting enzyme inhibitor, on exercise tolerance and humoral factors in 19 elderly patients (greater than 60 years old) with essential hypertension. Captopril (37.5-75 mg/day) was administered for 8 weeks. Fourteen of the 19 patients in whom captopril was effective took a treadmill exercise test according to Kattus' protocol. Exercise tolerance was increased in all patients (from 13.1 +/- 1.3 to 16.5 +/- 1.0 min, P less than 0.01). Captopril attenuated the rise in blood pressure during the exercise test but did not affect the heart rate. Resting values of plasma adrenaline decreased by 47% and noradrenaline by 17%, with no significant changes in plasma renin activity (PRA) or aldosterone. The change in mean blood pressure showed an inverse relationship to pretreatment plasma noradrenaline (r = -0.73, P less than 0.01). The results show that captopril is effective in the treatment of hypertensive elderly patients, and suggest that the sympathetic nervous system is involved in the mechanism of the antihypertensive response to captopril therapy.

A simple, sensitive, and highly reliable liquid chromatographic method using fluorescence detection is described for the simultaneous determination of metoprolol and alpha-hydroxymetoprolol in plasma and urine. This method involves a single extraction of the compounds with the internal standard pindolol from alkalinized plasma or urine into dichloromethane. A reconstituted aliquot with a mobile phase is injected onto a reversed-phase, Zorbax ODS column, and the detection is achieved by the excitation and emission wavelengths at 230 and 300 nm, respectively. The assay is reproducible and precise for metoprolol and alpha-hydroxymetoprolol in both plasma and urine samples, as judged by a coefficient of variation of less than 9.2% at all concentrations examined. The standard curves for metoprolol and alpha-hydroxymetoprolol are linear over 10-200 ng/ml in plasma and over 0.5-10 micrograms/ml in urine. The lower detection limit is 2 ng/ml for each of the compounds in plasma using a 0.5-ml sample. Preliminary data on the oxidation polymorphism of metoprolol in Japanese subjects are reported using the current assay method. In 183 Japanese subjects no poor metabolizer of metoprolol has been identified so far.

The effect of epinephrine on Cori cycle activity has been studied by a tracer kinetic technique. The subcutaneous injection of epinephrine into fasted adrenodemedullated rats activated gluconeogenesis and increased glucose utilization. Most of the glucose utilized in epinephrine-treated rats was converted to lactate, and the conversion to the metabolites other than lactate was markedly suppressed by epinephrine. As a result, not only hyperglycemia and hyperlactacidemia developed, but also the contribution of the Cori cycle to glucose metabolism increased after epinephrine. The glucose-alanine cycle, which is roughly one-third as active as the Cori cycle, was only slightly activated by epinephrine. No evidence was obtained, in vivo and in vitro, for the increased formation of lactate from muscle glycogen which decomposed in response to epinephrine. It is suggested that, in epinephrine-treated rats, blood glucose was converted to blood lactate predominantly while muscle glycogen selectively yielded the metabolites other than blood lactate. This suggests compartmentalization of glycolytic pathway in the peripheral tissues of epinephrine-treated rats.

A whole organ or a homogenate of rat pineal gland was incubated with arachidonic Acid. Two predominant metabolites were identified by mass spectrometry to be 12-hydroxy-5,8,10,14-eicosatetraenoic acid and 10-hydroxy-11,12-epoxy-5,8,14-eicosatrienoic acid. 15-Hydroxy-5,8,11,13-eicosatetraenoic acid was also formed in a smaller amount. In addition, peroxy acids appeared rapidly only at the initial stage of reaction. In various parts of rat brain the 12-lipoxygenase activity was by far the highest in pineal gland, and less than 5% of the activity was found in pituitary gland and hypothalamus.

Prostaglandin E2 (PGE2), a bone-resorption factor, was essentially the sole arachidonate metabolite in an osteoblastic cell line cloned from mouse calvaria (MC3T3-E1). When the cells were cultured in the presence of 2% newborn bovine serum, 1 microM epinephrine markedly stimulated PGE2 synthesis from endogenous arachidonic acid. The PGE2 synthesis commenced after a lag phase of 1-2 h, and reached a maximum at about 3 h after the addition of epinephrine. The effect of epinephrine was inhibited by propranolol, and epinephrine could be replaced by isoproterenol, suggesting beta-adrenergic stimulation of PGE2 production. A rapid increase in intracellular cAMP was observed upon the addition of epinephrine. When the intracellular cAMP level was raised using cholera toxin or forskolin, the PGE2 synthesis was also stimulated. The enhanced PGE2 synthesis was attributed to an increased level of cyclooxygenase, which was shown by immunoprecipitation of the enzyme using anti-cyclooxygenase antibody. Inhibitors of transcription and translation suppressed the epinephrine-dependent increase in cyclooxygenase activity. These findings suggest induction of cyclooxygenase involving cAMP via an as yet unclarified mechanism.

An improved purification procedure is described for the simultaneous assay of endogenous choline and acetylcholine by pyrolysis gas chromatography, particularly for providing a simple and effective method for propionylation of choline in the presence of acetylcholine. The reaction was carried out in acetonitrile solution prepared by dissolving the evaporated residue of the supernatant of brain homogenate. Thus samples for propionylation were prepared without the use of ion-exchange chromatography.