Phenytoin-induced DNA synthesis and inositol 1,4,5-triphosphate formation in L-929 fibroblasts

Summary Culture of L-929 fibroblasts in the presence of phenytoin (2.5–5.0 g/ml) increased DNA synthesis, as indicated by increased [³H]thymidine uptake, while a higher dose (20 g/ml) inhibited DNA synthesis. In like manner, a low dose of phenytoin (5.0 g/ml) was effective in increasing inositol 1,4,5-trisphosphate formation while a higher dose (10 g/ml) tended to inhibit this activity. These data suggest that the formation of inositol phosphate second messengers may play a role in phenytoin-induced fibroblast proliferation and connective tissue growth.     

Effect of aloe lectin on deoxyribonucleic acid synthesis in baby hamster kidney cells

A homogeneous glycoprotein (mol. wt 40,000) containing 34% carbohydrate was isolated from Aloe arborescens var. natalensis. At a concentration of 5 micrograms/ml, this glycoprotein was shown to stimulate deoxyribonucleic acid (DNA) synthesis in baby hamster kidney (BHK) cells and to have the properties of a lectin which reacts with sheep blood cells. The chemical and physical properties of the glycoprotein (aloe lectin) are also discussed.     

Influence of beta-aminoproprionitrile (BAPN) on cell growth and elastic fiber formation in cultures of auricular chondrocytes

Auricular chondrocytes isolated from 4-day-old rabbits and grown in vitro for 14 days, proliferated rapidly and produced a conspicuous network of elastic fibers. Beta-aminoproprionitrile (BAPN), which in vivo inhibits cross-linking of elastin, decreased the formation of elastic fibers at a concentration of 10-20 micrograms/ml and prevented formation at 40 micrograms/ml. At a concentration of 5 micrograms/ml only the so-called patches of elastin appeared to be absent. The inhibitory effect of BAPN on cell growth did not exceed 10%, which indicates that BAPN is only slightly harmful to auricular chondrocytes and can safely be used in studies on elastin deposition by these cells in vitro.     

Lack of pressor effect of dopamine in the pentobarbital-anesthetized rat

The blood pressure and heart rate responses to intravenous dopamine infusion at 2.5, 5.0 and 10.0 micrograms.min-1 X 100 g-1 were studied in conscious and pentobarbital-anesthetized Sprague-Dawley rats. In the conscious rats, dopamine caused a significant dose-related increase in the mean arterial blood pressure which was abolished in the anesthetized rats. The heart rate increased significantly only at the highest dose infused. The responses to equipressor doses of noradrenaline (40 ng.min-1 X 100 g-1) and phenylephrine (1.0 micrograms.min-1 X 100 g-1) were also suppressed in the anesthetized rats. The results suggest that pentobarbital anesthesia depresses the blood pressure response to dopamine infusion in the rat through a depression of activation of alpha-adrenoceptors.     

The effect of nicotine on ethanol-induced gastric ulcers in rats

Nicotine, in concentrations of 5 and 25 micrograms/ml drinking water, given ad libitum for 10 days, dose-dependently increased lesion formation and worsened ethanol-induced ulceration in rat stomachs. Daily fluid intake and b.wt gain were not adversely affected by nicotine pretreatment.     

Effect of actinomycin D on Trypanosoma cruzi

Viable metacyclic forms of T. cruzi, Y strain, treated with an adequate dose of actinomycin D (50 micrograms Act-D/ml/10(7) parasites/ml for 72 h at 28 degrees C) showed the following properties: 1) they lost their ability to replicate in culture medium, in blood and in tissues of normal mice and were no longer able to incorporate tritiated thymidine; 2) they could not penetrate into Vero cells and could not replicate inside normal macrophages; 3) they retained their immunogenicity and the ability to protect mice against a virulent infection; 4) they did not induce histological lesions as described in chronic experimental Chagas' disease.     

The dopamine autoreceptor agonist B-HT 920 markedly stimulates sexual behavior in male rats

B-HT 920, a selective agonist at dopamine (DA) autoreceptors, strongly increased the incidence of penile erections (PE) in male rats, an effect which was dose-related and antagonized by haloperidol. B-HT 920 at 100 and 200 micrograms/kg i.p. significantly altered the copulatory pattern of sexually active male rats, reducing the number of mounts and intromissions as well as the latency to the first ejaculation, a stimulant effect which was confirmed in sluggish males at a dose of 100 micrograms/kg.     

Influence of phenobarbital and TCDD on the hepatic metabolism of TCDD in the dog

The influence of phenobarbital and TCDD pretreatment on the formation and biliary excretion of TCDD-metabolites following single doses of 3H-TCDD was investigated. Without pretreatment, 24.5% of the absorbed 3H-TCDD dose was excreted in the bile within 110 h. Phenobarbital did not influence this rate, whereas a single dose of 10 micrograms of unlabeled TCDD/kg b.wt nine days earlier resulted in a doubling of the amount of radioactive material eliminated in the bile (47.4%).     

Effect of epidermal growth factor on growth and maturation of fetal and neonatal rat small intestine in organ culture

Small intestinal explants from pre- and post-natal rats were incubated in an organ culture system in the absence and presence of epidermal growth factor (EGF). The rate of synthesis of small intestinal DNA and protein as well as the activity of lactase and alkaline phosphatase increased rapidly between 17 and 20-day gestational age, whereafter they declined. The maximal incorporation of 3H-thymidine and 14C-alanine into DNA and protein, respectively, was significantly stimulated by EGF (100 ng/ml). EGF had no effect on the activity of either lactase or alkaline phosphatase in the small intestinal explants.     

Isometachromin, a new cytotoxic sesquiterpenoid from a deep water sponge of the family Spongiidae.

Isometachromin (1), a new sesquiterpene-quinone that is related structurally to metachromin C (2), and the known compounds ilimaquinone (3) and 5-epi-ilimaquinone (4), were isolated from a deep water sponge in the family Spongiidae; the structure of isometachromin was elucidated by spectral methods. Isometachromin exhibits in vitro cytotoxicity against the human lung cancer cell line A549 (IC50 = 2.6 micrograms/ml), but not against P388 murine leukemia (IC 50 > or equal to 10 micrograms/ml) and also exhibits antimicrobial activity.     

Effects of 18-hydroxydeoxycorticosterone on central nervous system excitability

The effects of 18-hydroxydeoxycorticosterone (18-OH-DOC) on central nervous system excitability were studied in adrenalectomized rats. Sixty-four evoked potentials (EP) recorded from the pontine reticular formation were averaged before and after the injection of vehicle and hormone. 750 micrograms of 18-OH-DOC dissolved in 0.5 ml of a 4:1 saline Cremophor-EL solution were injected i.v. A decrease of 55.7 +/- 6.1% in the amplitude of the EPs was observed with the hormone 16.3 min +/- 2.7 (SE) after injection. Amplitude values returned to baseline levels 38 min +/- 6.8 (SE) after injection. The secretion of 18-OH-DOC is greatly increased by ACTH and might modulate central nervous system function.     

Exogenous fibronectin modifies the aggregation of collagen-stimulated human platelets

Fibronectins (FN) are adhesive glycoproteins whose role in platelet aggregation is unclear. Addition of 3, 6 and 12 micrograms/ml of human plasma FN in vitro to isolated human platelets, which had been freed from plasma FN by gel filtration and subsequently stimulated with collagen, inhibited the last stage of platelet aggregation. With 3 and 6 micrograms/ml of FN a shortening of the lag-time was also observed. These data showed that FN may play a role in platelet-collagen interaction as well as in platelet-platelet interaction.     

Antidiuretic effects of oxytocin in the Brattleboro rat

The antidiuretic activity of oxytocin (OT) was measured in Brattleboro rats with congenital diabetes insipidus. A dose dependent antidiuretic response was found in animals receiving chronic infusions of 0.1 micrograms/h, 1.0 micrograms/h, and 5 micrograms/h of OT. OT infused at the rate of 5 micrograms/h over a 7-day period completely reversed the symptoms of diabetes insipidus. The results support the concept that OT serves as a weak agonist of vasopressin at the level of the kidney and at pharmacological levels exhibits antidiuretic activity.     

The use of 5-fluorocytosine and ketoconazole in the culture of the erythrocytic stages of Plasmodium falciparum and some tumor cell lines

In vitro culture systems are often contaminated by bacteria and fungi. It is therefore often necessary to supplement culture media with agents such as penicillin/streptomycin, gentamycin or amphotericin B. The latter cannot be used in the in vitro culture of erythrocytic stages of P. falciparum, and thus anti-fungal agents have not been regularly used in this system. We describe the prophylactic use of 5-fluorocytosine (5-FC) and ketoconazole (KTZ) in tissue cultures at concentrations up to 300 and 10 micrograms/ml respectively which have no effect on the growth of P. falciparum (FCR-3 strain). A melanoma cell line (C32) and a line of uterine carcinoma (C41) were also unaffected by similar concentrations of 5-FC and KTZ. When dissolved in complete culture medium (RPMI 1640) with 10% human plasma, the minimum inhibitory concentration of 5-FC for a susceptible strain of Candida remained below 2 micrograms/ml. These experiments suggest that 5-FC (at 50 micrograms/ml) alone or in combination with KTZ (at 1 microgram/ml) is a useful addition to the armamentarium of antimicrobials available to the tissue culture biologist for a variety of cell culture systems.     

Failure of somatostatin to influence experimental tumor cell growth in vivo and in vitro

The influence of somatostatin on tumor cell growth was studied in vivo in mice (sarcoma 180 ascites tumor and Lewis lung tumor) and in vitro on nontransformed and polyoma-transformed cell lines. 4 or 20 micrograms/100 g of cyclic somatostatin and 4 micrograms/100 g of linear protamin Zn-bound somatostatin were injected s.c. twice daily in the in vivo study. Cyclic somatostatin (1, 4 or 10 micrograms/ml) was added twice daily to the cell cultures. Somatostatin administration influenced neither the survival of animals nor the growth rate of cultured cell lines.     

Nitroglycerin-induced desensitization of vascular smooth muscle may be mediated through cyclic GMP-disinhibition of phosphatidylinositol hydrolysis

The purpose of this study was to investigate the hypothesis that nitroglycerin-induced desensitization of vascular smooth muscle is mediated through cyclic GMP-disinhibition of phosphatidylinositol hydrolysis. Norepinephrine-induced contraction and increased levels of inositol monophosphate, a measure of phosphatidylinositol hydrolysis, in rat aorta. Prior treatment with nitroglycerin inhibited both the norepinephrine-induced contraction and the elevated levels of inositol monophosphate to the same relative magnitude. The nitroglycerin-induced inhibition of contraction and inositol monophosphate formation were prevented in tissues desensitized with nitroglycerin. These results suggest that: nitroglycerin may inhibit vascular smooth muscle contraction through cyclic GMP-inhibition of phosphatidylinositol hydrolysis and desensitization to the relaxant effects of nitroglycerin may be due to disinhibition of the hydrolysis.     

Serotonin-stimulated protein phosphorylation in aortic smooth muscle cells

The effects of serotonin on the formation of inositol phosphates and protein phosphorylation were examined in cultured smooth muscle cells. Serotonin stimulated the formation of [3H]inositol monophosphate, [3H]inositol bisphosphate and [3H]inositol trisphosphate. This effect was prevented by 5-HT2 specific antagonist, 6-methyl-1-(1-methylethyl)ergoline-8-carboxylic acid, 2-hydroxy-1-methylpropyl ester [Z]-2-butenedioate (LY53857). Serotonin stimulated the phosphorylation of many polypeptides, among which a 20 kDa polypeptide was the most prominent. The phosphorylation was also inhibited by LY53857. LY53857 alone produced no effects on protein phosphorylation. The 20 kDa polypeptides were also phosphorylated by the addition of 12-O-tetradecanoylphorbol-13-acetate. These results suggest that serotonin stimulates protein phosphorylation through 5-HT2 receptors and possibly activates protein kinase C in intact vascular smooth muscle cells.     

Interaction of cortisol and epinephrine in the regulation of leucine kinetics in man

To assess the interaction of the two major stress hormones epinephrine and cortisol in the regulation of leucine kinetics in man, epinephrine (50 ng/kg/min) was infused either alone or in combination with cortisol (2 micrograms/kg/min) into two groups of 6 postabsorptive normal male subjects during 180 min. Plasma leucine concentrations decreased by 28% (p less than 0.05) from baseline during epinephrine treatment (plasma levels 515 pg/ml); this was due to a decrease of leucine appearance (determined by 1-13C-leucine infusions) by 23% (p less than 0.025); leucine oxidation decreased by 29% (p less than 0.05). However, when plasma cortisol concentrations were elevated to supraphysiological levels (16.3 mumol/l) during epinephrine administration, the decreases of leucine plasma concentrations, appearance and oxidation were abolished. Plasma glucose and FFA concentrations were similarly elevated during both kinds of treatment. Since leucine appearance represents a measurement of total body protein breakdown and leucine disappearance into non-oxidative pathways reflects protein synthesis, the data indicate that plasma epinephrine concentrations during severe stress exert a protein anabolic effect in man which may counteract catabolic properties of elevated plasma cortisol.     

Effect of aloe lectin on deoxyribonucleic acid synthesis in baby hamster kidney cells

Summary A homogeneous glycoprotein (mol.wt 40,000) containing 34% carbohydrate was isolated fromAloe arborescens var.natalensis. At a concentration of 5 g/ml, this glycoprotein was shown to stimulate deoxyribonucleic acid (DNA) synthesis in baby hamster kidney (BHK) cells and to have the properties of a lectin which reacts with sheep blood cells. The chemical and physical properties of the glycoprotein (aloe lectin) are also discussed.     

Bombesin promotes pancreatic growth in suckling rats

The pancreatic growth promoting effect of long term administration of bombesin was investigated in suckling rats. The authors showed that bombesin given in 10 micrograms/kg b.wt doses s.c. every 8 h for 10 days from the day of parturition stimulated pancreatic growth: it increased pancreatic weight, protein and DNA content, trypsin and amylase activity and trypsin/DNA ratio. Conclusion: Bombesin is an effective stimulator of pancreatic growth in suckling rats.