Enantiomeric cannabinoids: stereospecificity of psychotropic activity

Summary The 1,1-dimethylheptyl homolog of (–)-(3R,4R)-7-hydroxy-delta-6-tetrahydrocannabinol (compoundII) is highly psychotropic in mice, rats and pigeons. The (+)-(3S,4S) enantiomer (III) was found to be psychotropically inactive at doses up to several thousand times those of the ED₅₀ of (II).We thank Dr A. Breuer and Mrs H. Amsalem for help with the syntheses. The research reported above was supported in Tucson by NIH grant NS 15441; in Uppsala by grants from the Swedish Medical Research Council (5757) and the Swedish Council for the Planning and Coordination of Research (84/2082); in Jerusalem by the Szold Foundation.Presented in part at a meeting at the US National Institute on Drug Abuse, Washington, D.C., October 1986, see NIDA Research Monographs79 (1987) 15.     

Influence of α-MSH and ACTH on cortical bone remodelling in hypophysectomized rats

Summary Hypophysectomy increases both periosteal resorption and endosteal apposition along the femur diaphysis in rat. Administration of -MSH decreased the periosteal resorption but had no effect on the endosteal apposition. ACTH had only minor effects on the endosteum. Thus, -MSH and ACTH, in the doses used, have different effects on cortical bone in rat. The effect of -MSH on cortical bone could be an effect of the hormone alone or by its stimulation of other factors.     

Effect of halogenmethylenebisphosphonates on bone cells in culture and on bone resorption in vivo

Summary Dihalogenmethylenebisphosphonates increase alkaline phosphatase activity and fatty acid oxidation in calvaria cells in culture (Cl₂MBP>Br₂MBPF₂MBP). The monohalogen C1MBP and the non-halogenated analogues are less active on phosphatase and inactive on or inhibitory towards fatty acid oxidation. The three dihalogenbisphosphonates and C1MBP inhibit bone resorption in vivo, Cl₂MBP most strongly.Acknowledgments. We thank Miss M.-L. Aebersold, Miss J. Portenier, Mrs I. Tschudi and Mrs Ch. Marti for their skilled technical assistance. This work has been supported by the Swiss National Science Foundation (grant 3.937.82), by the Procter & Gamble Company, Cincinnati, USA, by the Istituto Gentili S.p.A., Pisa, Italy, and by the Ausbildungs- und Förderungsfonds der Arbeitsgemeinschaft für Osteosynthese (AO), Chur, Switzerland.     

Beta1-adrenoceptor mediated salivary gland enlargement in the rat

Summary In the rat, prolonged activation of ₁-adrenoceptors causes a gain in salivary gland weight, whereas prolonged blockade of these receptors causes a reduction in weight.This work was supported by grants to J.E. from the Medical Faculty in Lund and the Swedish Medical Research Council (05927).     

Neuronal nicotinic receptors: insights gained from gene knockout an knocking mutant mice

Neuronal nicotinic acetylcholine receptors are ligand-gated ion channels that subserve a range of functions in the brain and peripheral nervous system. They are pentamers variously composed of (₂– ₁₀) and subunits ( ₂– ₄). Pharmacological and ligand-binding studies have shown that the different subunits vary in their distribution and channel properties, but precise delineation of the in vivo function of individual subunits has been hampered by lack of subunit-specific antagonists. The development of transgenic mice with targeted deletions of specific subunits (knockout mice) or mutations in critical receptor domains (knockin mice) has extended understanding of nicotinic receptors, revealing that some subunits are necessary for viability, whereas others mediate modulatory effects on learning and memory, locomotion, anxiety, nociception, dopaminergic neurotransmission, seizure threshold, development of the visual system and autonomic function. In some cases, studies of transgenic mice have confirmed expectations derived from pharmacological and expression studies, but in other cases, compensation by related subunits has revealed a degree of functional redundancy not predicted by previous approaches.Received 19 September 2002; received after revision 12 November 2002; accepted 11 December 2002     

Die Wirkung von Progesteron und Progesteronmetaboliten auf das Wachstum von embryonalem Knorpel in vitro

Summary Embryonic chick bones were cultured in vitro in the presence of progesterone and of a series of progesterone metabolites. It was found that only 5-pregnane-3,20-dione, 3-hydroxy-5-pregnane-20-one, and ⁴-pregnene-20-ol-3-one influence the growth of bones in vitro in a manner similar to progesterone. All other investigated progesterone metabolites exert either a different or no demonstrable action on bone growth.     

Evidence for a role of IFN γ in control ofListeria monocytogenes in T cell deficient mice

Summary The role of interferon (IFN) in controlling chronic infections ofListeria monocytogenes (Listeria) was studied in athymic C57BL/6nu/nu mice, and by treating thymectomized C57BL/6 +/+ mice with monoclonal rat CD4 and CD8-specific monoclonal antibodies (Mab). Mice treated with a combination of the two T cell subset antibodies were similar to athymic, nude mice in being able to contol Listeria infection, keeping the titers below 3–5 log₁₀ bacteria per organ, but they could not eliminate them completely. Treatment with antibodies to IFN of nude or CD4⁺ + CD8⁺-T cell-depleted mice suffering from chronic Listeria infection caused a marked increase of Listeria titers, in liver and spleen. This result implies a role of IFN in maintaining anti-Listeria resistance in mice lacking mature T cells.     

Effects of selective dopamine D1 and D2 antagonists on male rat sexual behavior

Summary The effects of selective dopamine (DA) D₁ and D₂ antagonists on male rat sexual behavior were investigated. The D₁ antagonist (+)SCH-23390, 25–100 g kg^–1 s.c. –20 min, and the D₂ antagonist raclopride, 0.1–1.6 mg kg^–1 s.c., –20 min, decreased both the number of mounts and intromissions preceding ejaculation. No statistically significant effects in the time up to ejaculation or in the time up to the first intromission were noted, whereas both compounds produced a statistically significant increase in the post-ejaculatory interval. The effect can generally be characterized as psychomotor inhibition, and no evidence was obtained for a specific role of DA D₁ or D₂ receptors in the mediation of male rat sexual behavior.The expert technical assistance of Ms Elisabeth Wallin is gratefully acknowledged. The figures were skilfully prepared by Ms Madelene Kröning at the Department of Psychology. This study received support from the Bank of Sweden tercentenary Foundation, The Swedish MRC and Wilhelm and Martina Lundgren Foundation.     

Kinetic arguments for the existence of a single form of intestinal ornithine decarboxylase during the postnatal development of normal and sparse-fur mutant mice and after EGF treatment

Summary The K_m for ornithine is remarkably constant during the course of postnatal development in both normal and spf mutant mice even if a large but transient increase in ornithine decarboxylase (ODC) activity is noted. Four hours after EGF injection (4 g/g b.wt) to 17-day-old normal and spf mice, a marked stimulation of ODC activity is observed but K_m remains unaffected. These data argue against the existence of multiple forms of ODC in the intestinal mucosa of mice.Supported by grant MA-8923 from the Medical Research Council of Canada. The author is Chercheur-boursier du Fonds de la Recherche en Santé du Québec.     

In vivo effects of immunostimulating lipopeptides on mouse liver microsomal cytochromes P-450 and on paracetamol-induced toxicity

Summary Immunomodulating lipopeptides lauroyl-L-Ala--D-Glu-LL-A2pmNH2-Gly (RP 44.102) and lauroyl-L-Ala--D-Glu-LL-A2pmNH2 (RP 56.142) were found to protect mice against the hepatotoxicity of paracetamol, which is due to cytochrome P-450 dependent formation of toxic metabolites and radicals. In fact they decreased the amount of hepatic microsomal cytochrome P-450, and the level of CCl₄-induced lipid peroxidation. In contrast lauroyl-L-Ala--D-Glu-DD-A2pmNH2 (RP 53.204), which only differs by the configuration of the two chiral carbons of A2pm (diaminopimelic acid) and is not an immunomodulating agent, failed to protect against poisoning by paracetamol and had no effect on the level of hepatic cytochrome P-450 or the microsomal CCl₄-induced lipid peroxidation. This provides a clear connection between the immunostimulating properties of a compound and its effects on xenobiotic biotransformations.     

Relevance of specific activity in experimental erythrocytocide by55Fe

Summary Iron loads between 0.20 g and 26 g, added to 5Ci⁵⁹Fe, were followed for up to 150 days in mice. Relative organ uptake increased as a function of iron load in liver and kidneys while it decreased in bone marrow and blood. Several weeks after injection, all load-related differences disappeared.Research carried out under the auspices of the US Department of Energy and by NIH GRANT HL-15685-02.     

Ginkgo biloba extract inhibits oxygen species production generated by phorbol myristate acetate stimulated human leukocytes

Summary A Ginkgo biloba extract (Gbe) containing flavonoids, among other compounds, was tested for the release of activated oxygen species ( , H₂O₂, OH^.) during the stimulation of human neutrophils (PMNs) by a soluble agonist. The extract slows down O₂-consumption (respiratory burst) of stimulated cells by its inhibitory action on NADPH-oxidase, the enzyme responsible for the reduction of O₂ to . Consequently, superoxide anion ( ) and hydrogen peroxide (H₂O₂) production is significantly decreased when the PMNs stimulation is done in the presence of the extract at concentrations of 500, 250 and 125 g/ml. Moreover, the hydroxyl radical generation (OH^.) is very much decreased at concentrations as low as 15.6 g Gbe/ml, which indicates that the extract also has free radical scavenging activity. Gbe is able at least to reduce very severel the activity of myeloperoxidase contained in neutrophils. This enzyme, secreted into the intra and extracellular medium, catalyzes the oxidation of chloride (Cl^–) by H₂O₂ to yield strong oxidants (HOCl, chloramines) which are implicated in inflammatory processes     

Effect of cadmium on polyribosome structure and function in mouse liver

Summary Cadmium chloride injected to mice (20 moles/kg) provokes in the livers a degradation of polyribosomes and diminishes their protein synthetic ability measured in vitro. CdCl₂ added in a final concentrations between 30 and 100 M to the protein synthetic cell-free system derived from livers of control mice inhibits its activity.Acknowledgments. We are grateful to Prof. P. Sokoli for helpful discussion. The work was supported by Wellcome Trust grant and by a gift of chemicals from Kemika, Zagreb. We are grateful to both these organizations.     

The p38α MAPK positively regulates osteoblast function and postnatal bone acquisition

Bone continuously remodels throughout life by coordinated actions of osteoclasts and osteoblasts. Abnormalities in either osteoclast or osteoblast functions lead to bone disorders. The p38 MAPK pathway has been shown to be essential in controlling osteoblast differentiation and skeletogenesis. Although p38α is the most abundant p38 member in osteoblasts, its specific individual contribution in regulating postnatal osteoblast activity and bone metabolism is unknown. To elucidate the specific role of p38α in regulating osteoblast function and bone homeostasis, we generated mice lacking p38α in differentiated osteoblasts. Osteoblast-specific p38a knockout mice were of normal weight and size. Despite non-significant bone alterations until 5?weeks of age, mutant mice demonstrated significant and progressive decrease in bone mineral density from that age. Adult mice deficient in p38a in osteoblasts displayed a striking reduction in cancellous bone volume at both axial and appendicular skeletal sites. At 6?months of age, trabecular bone volume was reduced by 62?% in those mice. Mutant mice also exhibited progressive decrease in cortical thickness of long bones. These abnormalities correlated with decreased endocortical and trabecular bone formation rate and reduced expressions of type 1 collagen, alkaline phosphatase, osteopontin and osteocalcin whereas bone resorption and osteoclasts remained unaffected. Finally, osteoblasts lacking p38α showed impaired marker gene expressions and defective mineralization in vitro. These findings indicate that p38α is an essential positive regulator of osteoblast function and postnatal bone formation in vivo.     

Net transport of cholesterol from cells of the human EA.hy 926 endothelial cell line to high density lipoproteins

EA.hy 926 cells, a human endothelial cell line, show characteristics of differentiated endothelial cells. The cells express saturable binding of apo E-free¹²⁵I-high density lipoprotein₃ (HDL₃). B_max increased from 71 to 226 ng HDL₃ bound/mg cell protein after cholesterol loading of the confluent endothelial cells with cationized low density lipoprotein (LDL). The affinity did not change after cholesterol enrichment (K_d was 37 g HDL₃ protein/ml for control cells and 31 g/ml, for loaded cells). Incubation of cholesterol-loaded EA.hy 926 cells with native HDL and LDL had different effects on cellular cholesterol levels. Incubation with HDL decreased both esterified and unesterified cellular cholesteryl, but LDL did not change total cellular cholesterol However, LDL tended to increase cellular cholesteryl esters, with a concomitant decrease of unesterified, cellular cholesterol. Incubation of endothelial cells with both HDL and LDL also resulted in decreased total cellular cholesterol levels. These data show that cationized LDL-loaded human endothelial EA.hy 926 cells can be used to study the net transport of cellular cholesterol to HDL, the first step in reverse cholesterol transport.     

α1-Adrenergic stimulation of ketogenesis and fatty acid oxidation is associated with inhibition of lipogenesis in rat hepatocytes

Summary The effect of norepinephrine on fatty acid synthesis (³H₂O incorporation into fatty acids), on fatty acid oxidation to CO₂ and on ketogenesis was studied in isolated hepatocytes of fed rats. After incubation with norepinephrine (50 M), lipogenesis was lower (5.7±1.1 nmoles³H₂O incorporated into fatty acids/mg dry weight/30 min) than in controls (7.5±1.7; n=6, p<0.02). In contrast, (1-¹⁴C) palmitate conversion into total ketone bodies was increased to 10.9±1.8 nmoles/mg/30 min with norepinephrine, vs 8.5±1.6 in controls (p<0.05), and more (1-¹⁴C) palmitate was converted to¹⁴CO₂ with norepinephrine than in controls (1.48±0.10 nmoles/mg/30 min vs 1.06±0.11, p<0.05). The inhibitory effect of norepinephrine on lipogenesis was abolished by addition of the ₁-receptor blocker prazosin, but not by ₂ or -blockers. The results demonstrate that the ketogenic effect of norepinephrine is coupled with an inhibitory effect on lipogenesis which may be explained by diminished activity of acetyl-CoA carboxylase, diminished formation of malonyl-CoA and decreased activity of carnitine palmitoyl transferase I.     

Preferential excretion of glycated albumin in C57BL-Ks-J mice: Effects of diabetes

Urinary excretion of glycated albumin was quantitated in genetically hyperglycemic mice (C57BL-Ks-J, db/db mice), a model for non-insulin-dependent diabetes mellitus, and compared with their non-diabetic littermates. The data indicated a preferential excretion of glycated albumin in non-diabetic mice. This phenomenon of editing of glycated albumin is decreased significantly in diabetic mice. Quantitative measurements of overall excretion of glycated albumin suggested that the loss of editing in diabetic mice is due to the dilution of glycated albumin by the unmodified albumin which is excreted in large amounts in diabetic mice. Therefore, the loss of editing observed in this model resembled the one we characterized in insulin-dependent diabetic humans and a streptozotocin-diabetic rat model³.     

Beitrag zur Frage der Eisenresorption

Summary In rabbits whose intestinal tract has been sterilised by means of penicilline and terramycine, there is no detectable iron resorption, even when the animals are made anaemic.Iron resorption can be restored by means of addition of terramycine-resistentE. coli or Enterococci to the diet.     

Influence of cyclic nucleotides on protein synthesis in vascular smooth muscle

Summary The incorporation of leucice-¹⁴C into protein in bovine mesenteric arteries was augmented by cyclic GMP (10^–3 M) and decreased by cyclic AMP (10^–3 M). There was no effect of 5 AMP (10^–3 M). The phosphodiesterase inhibiting drugs theophylline (10^–3 M) and papaverine (5×10^–5 g/ml) both decreased the leucine-¹⁴C incorporation.We are indebted to Mrs.Lena Burlin for hear assistance. Finacial support has been provided by the Swedish State Medical Research Council (No. 04X-101X-4498).     

Genotype-specific reduction in methyl nitrosourea (MNU) induced sister chromatid exchanges (SCE) in vivo during aging

Summary We studied mice from five strains (BALB/c, C3H/HeSnJ, C57BL/6J, Cs^b and 129/ReJ) at two ages (young, 10±1 weeks; and old, 67±3 weeks) for the induction of sister chromatid exchanges (SCEs) in vivo by methyl nitrosourea (MNU). The SCE frequency is genotype-specific. The F₁ phenotype resembles the low responding parent. SCE induction is significantly lower in the older animals of each strain than their younger counterparts, and the reduction of SCE/cell with old age is strain-specific. A general explanation for these results must include strain differences in relative mutagenic sensitivity, genotype-specific pattern of reduction in DNA repair and other such factors affecting SCE formation, with old age.