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

Dihalogenmethylenebisphosphonates increase alkaline phosphatase activity and fatty acid oxidation in calvaria cells in culture (Cl2MBP greater than Br2MBP approximately equal to F2MBP). The monohalogen ClMBP and the non-halogenated analogues are less active on phosphatase and inactive on or inhibitory towards fatty acid oxidation. The three dihalogenbisphosphonates and ClMBP inhibit bone resorption in vivo, Cl2MBP most strongly.     

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.     

Immunological blocking of exogenous and endogenous secretin in the dog

Summary In mongrel dogs with chronic gastric and duodenal fistula the biological activity of secretin on exocrine pancreatic secretion could be blocked by preincubating the secretin injected with rabbit antisecretin antibody. In addition, the activity of endogenous secretin released by acid was markedly reduced by application of antibody. Since no such effect was observed after testmeal stimulation, secretin is most probably not released in a significant amount by the liquid meal used in this experiment.This work was supported by grant No. 3.816-0.76 of the Swiss National Foundation for Scientific Research.We express our thanks to Miss G. van Hees, Mrs L. Jecker and Dr F. Meyer for their excellent technical assistance, Dr W. W. Rittmann for the surgical support, Dr G.A. Stalder for general advice, Dres R. Studer and D. Gillessen for providing secretin Roche, and Mrs C. Frei for secretarial work.     

Ultrastructural studies of Morris hepatoma cells reversely transformed by ginsenosides

Summary Ginsenosides, which were extracted from Panax ginseng, C. A. Meyer, induced well the development of subcellular organelles in cultured Morris hepatoma cells (MH₁C₁).Acknowledgments. We wish to thank Mr H. Konishi, Mrs H. Makino, Mrs Y. Nakayabu and Miss N. Honjo for their excellent technical assistance.     

Reciprocal changes in serum free thyroxine fraction and thyrotropin level after administration of thiocyanate to rats

Summary The administration of thiocyanate to rats caused a significant increase of serum free thyroxine fraction, which coincided with the significant decrease of TSH level. The other components (AFT₄, T₄, T₃) in serum at this time were decreased or unchanged. The finding suggests the role of free thyroxine fraction in feed-back regulation of TSH secretion.Acknowledgment. We thank Dr A. Parlow and the NIAMDD, Rat Pituitary Hormone Distribution Program, for material for rat TSH immunoassay; Dr J. Nauman (Inst. Postgrad. Med., Warsaw, Poland) for T₃ antibody, and to Ing. J. Sadlo, Mrs M. t'astná and Miss R Fajkoová for technical assistance.     

Behaviour of sodium-potassium-activated adenosine triphosphatase in rat kidney tissue by folic acid

Zusammenfassung Männliche Wistarratten zeigten nach i.v. Gabe von Folsäure einen Aktivitätsabfall der Enzyme, die in den aktiven Ionentransport eingreifen: (Na⁺K⁺) ATPase (60% gegenüber Kontrollen), ICDH (56%) und MDH (48%) und a.Ptase (66%).

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With technical assistance of Mrs. I.Bieder and Miss M.Kirstein.

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With the support of the Schweiz. Nationalfonds zur Förderung der wissenschaftlichen Forschung No. 4809.     

Fatty acid metabolism in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

Peroxisomes are essential subcellular organelles involved in a variety of metabolic processes. Their importance is underlined by the identification of a large group of inherited diseases in humans in which one or more of the peroxisomal functions are impaired. The yeast Saccharomyces cerevisiae has been used as a model organism to study the functions of peroxisomes. Efficient oxidation of fatty acids does not only require the participation of peroxisomal enzymes but also the active involvement of other gene products. One group of important gene products in this respect includes peroxisomal membrane proteins involved in metabolite transport. This overview discusses the various aspects of fatty acid -oxidation in S. cerevisiae. Addressed are the various enzymes and their particular functions as well as the various transport mechanisms to take up fatty acids into peroxisomes or to export the -oxidation products out of the peroxisome to mitochondria for full oxidation to CO₂ and H₂O.Received 19 February 2003; received after revision 27 March 2003; accepted 27 March 2003     

Cl−/HCO 3 − exchanger is operative in isolated enterocytes from rat jejunum

Enterocytes isolated from rat jejunum were tested for the existence of a Cl^–/HCO ₃ ^– exchange, previously evidenced in basolateral membrane vesicles but not in brush border. Cells were found to retain functional integrity and transport capabilities long enough to allow Cl^– fluxes to be measured. Both efflux and uptake experiments indicate that a Cl^–/HCO ₃ ^– antiport, inhibited by 4,4-diisothiocyanostilbene-2-2-disulfonic acid (DIDS), is functional under resting conditions.     

Inhibition of testosterone biosynthesis by medrogestone

Résumé In vitro, la medrogestone bloque la synthèse des hormones dans les gonades par inhibition de la 3-hydroxystéroide ^4,5-isomérase. In vivo, la medrogestone réduit le niveau de testostérone dans le testicules de rat.

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We wish to thank Mrs.S. gosselin, Miss N. Jagarinec andMrs. F. Fitzsimons for their technical assistance andDr. G. Schilling for analyses by gas-liquid chromatography.     

Synthesis of maleimide derivative of cortisol for enzyme coupling in cortisol enzyme immunoassay

Summary A new cortisol derivative, cortisol-21-m-maleimidobenzoate (CMB), was synthesized and conjugated with sulfhydryl groups of -galactosidase (BG). Both CMB-BG and CHS-BG conjugates have a high immunoreactivity to cortisol antibody, and although CHS-BG does not displace well with the added cortisol, CMB-BG does.Acknowledgments. This work was supported in part by Grant 884M from the Council for Tobacco Research. We wish to thank Miss Laura Morbach and Mrs Tatsue Monji for their assistance on this project.Address reprint requests to A. Castro.     

Identification of swelling-activated Cl<Superscript>-</Superscript> current in rabbit cardiac Purkinje cells

The presence and functional role of the swelling-activated Cl^- current (I_Cl(swell)) in rabbit cardiac Purkinje cells was examined using patch-clamp methodology. Extracellular hypotonicity (210 or 135 mOsm) activated an outwardly rectifying, time-independent current with a reversal potential close to the calculated Cl^- equilibrium potential (E_Cl). The magnitude of this current was related to tonicity of the superfusate. The current was blocked by 0.5 mM 4,4-diisothiocyanostilbene-2,2-disulfonic acid (DIDS). These features are comparable to those of I_Cl(swell) found in sinoatrial nodal, atrial, and ventricular myocytes. I_Cl(swell) activation at 210 and 135 mOsm depolarized the resting membrane potential with 6 and 10 mV and shortened the action potential by 18 and 33%, respectively. DIDS partially reversed I_Cl(swell)-induced action potential changes. We conclude that I_Cl(swell) is present in Purkinje cells and its activation leads to action potential shortening and resting membrane potential depolarization, both of which can promote the development of reentrant arrhythmias.Received 20 January 2004; received after revision 17 February 2004; accepted 25 February 2004     

Lag period of action of 25-hydroxycholecalciferol on bone collagen metabolism in vitamin D deficient rats

Zusammenfassung Nachweis, dass die Freisetzung von Hydroxyprolin bei Knochenfragmenten bereits 6 h nach i.v. Verabreichung von 2,5 g 25-Hydroxycalciferol signifikant erhöht war, während eine entsprechend zugeführte Dosis von Vitamin D₃ erst 13 h später deutlich wirkte.

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We thank Dr.E. Kodicek and Dr.E. M. Cruickshank of Dunn Nutritional Laboratory for help and criticism, and Mrs.J. Jobbágy for technical assistance.     

Effects of anaesthesia and chronic catheterization on circulating levels of prostaglandins (PGE2 and PGF2a in dogs

Summary Chronic catheterization of aorta and inferior vena cava in dogs did not significantly affect circulating levels of prostaglandins (PGE₂ and PGF_2a ). Pentobarbital (30 mg/kg i.v.) anaesthesia produced a significant decrease in PGF_2a .This work was supported by INSERM (grant No. 76.5.183.5).We wish to thank Miss Laure Eloy and Mrs Annick Soubrier for their technical assistance.     

Human myeloperoxidase activity is inhibited in vitro by quercetin. Comparison with three related compounds

Summary Quercetin is an effective inhibitor of human myeloperoxidase (MPO) activity, both with purified enzyme (IC₅₀=3.5 M) and in a system using stimulated human neutrophils. Quercetin is significantly more potent than three other related compounds (rutin, rutin sulfate and troxerutin) and than methimazole, a previously-known myeloperoxidase inhibitor. The inhibitory activity of quercetin is of the competitive type. Moreover, quercetin is directly able to scavenge hypochlorous acid (HOCl), a chlorinated species generated by the, MPO/H₂O₂/Cl^– system.     

Level of marker enzymes in spermatogenesis on administration of PGF2a in rats

Summary Intrapertioneal administration of PGF_2a in rats significantly increased testicular acid phosphatase (p<0.05), decreased hyaluronidase (p<0.05), whereas the activities of 5-nucleotidase, N-acetyl-B-glucosaminidase, B-galactosidase and uridine diphosphatase remained unaffected.Acknowledgment. We are grateful to Dr D. Sundaresan, for his interest in the study and to Mr B. M. Sharma for technical assistance.     

Induced regeneration of plants in tissue cultures ofBrassica oleracea

Résumé Dans les cultures de tissus repiqués de Chou moellier une néoformation de plantes entières se produit en conséquance de l'exclusion du 2,4-D du milien de culture.

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We thank Miss A.temberová and Mrs. M.Paldusová for valuable technical assistance.     

Effect of diphosphonates on ATP and Pi content,Pi uptake and energy charge of cultured calvaria cells

Summary In previous studies we have shown that glycolysis is strongly inhibited in cultured calvaria cells treated with ethane-1-hydroxy-1, 1-diphosphonate (EHDP) or dichloromethanediphosphonate (Cl₂MDP). This study shows that the energy charge of the adenylate pool, and the ATP and Pi content, were not changed by treatment with diphosphonates except for a slight decrease of ATP and Pi at 0.25 mM Cl₂ MDP. The uptake of Pi was diminished by 50% and 20% in cells treated for 6 days with Cl₂MDP or EHDP, respectively, but not when diphosphonates were present only during the uptake studies.Acknowledgments. We thank Miss B. Wyss for her skilled technical assistance. This work has been supported by the Swiss National Science Foundation (grant Nos 3.725.76 and 3.824.79), by the Procter & Gamble Company, USA, and by the Ausbildungs- und Förderungsfonds der Arbeitsgemeinschaft für Osteosynthese (AO), Chur, Switzerland.     

The ‘Spitzenkörper’, centre of the reducing power in the growing hyphal apices of two septomycetous fungi

Summary Redox dyes, such as methylene blue and neutral red, are stably reduced to their leucobase in the Spritzenkörper of the growing hyphal tips ofNeurospora crassa andMonilia fructigena.We acknowledge the support of the Fonds National Suisse de la Recherche Scientifique and thank both Mrs Ton-That for her electron micrograph and Miss E. Herz for her technical help.     

Effect of air ionization on blood serotonin in vitro

Summary The effect of negative and positive air ionisation on siliconized blood serotonin was studied in vitro. The experiments showed that within 10 min positive ionisation increased serotonin levels in total blood (+40%), plasma (+90%), erythrocytes (+50%) and thrombocytes (+200%). On the other hand, negative ionization (10 min) lowered the serotonin content of total blood (–30%), plasma (–42.5%), erythrocytes (–41.7%) and thrombocytes (–72.3%), thus confirming the Krueger Effect in vitro.This research was generously supported by a grant from the Joint Research Fund of the Hebrew University and Hadassah, and Dr. and Mrs.Leonard Appleby of Palmbeach, Florida.Acknowledgment: Our thanks are due to Mr.B. Shalita, Mrs.Zippora Adar and Mrs.Olga Fradkin for devoted technical assistance.