Ultrastructure of muscle spindles in C57BL/6Jdy 2J/dy 2J dystrophic mice

Summary The sensory organs of skeletal muscles, the muscle spindles, were examined using electron microscopy indy ^2J/dy ^2J dystrophic mice. Despite widespread damage to the extrafusal (skeletomotor) fibres the intrafusal (spindle) fibres appeared normal and seemed resistant to the aetiological factors for murine dystrophy.This work was supported by grants from the Medical Research Council and the Science Research Council of Great Britain.     

Dystrophic mutation (dy2J) affecting regulation of lactate dehydrogenase (LDH) and pyruvate kinase (PK) in C57BL/6J mice

Summary The genotype difference (dystrophic vs nondystrophic) in the LDH isozymes is observed in kidney. These differences are evident only at birth and at early developmental stages (before the expression of dystrophic symptoms). The tissue specific genotype differences for PK are limited to the thigh muscle (M form) and heart (L form), after the onset of the condition. These differences may reflect the pleiotropic effect of the dy^2J locus during the temporal regulation of these and other enzymes implicated in muscular dystrophy (MD).This research was financed by a Natural Sciences and Engineering Research Council Canada grant to S.M.S.     

On the mechanism of inhibition of p27 degradation by 15-deoxy-Δ12,14-prostaglandin J 2 in lymphoblasts of Alzheimer’s disease patients

It has been proposed that neuroinflammation, among other factors, may trigger an aberrant neuronal cell cycle re-entry leading to neuronal death. Cell cycle disturbances are also detectable in peripheral cells from Alzheimer’s disease (AD) patients. We previously reported that the anti-inflammatory 15- deoxy-Δ^12,14-prostaglandin J ₂ (15d-PGJ ₂) increased the cellular content of the cyclin-dependent kinase inhibitor p27, in lymphoblasts from AD patients. This work aimed at elucidating the mechanisms of 15d-PGJ ₂-induced p27 accumulation. Phosphorylation, half-life, and the nucleo-cytoplasmic traffic of p27 protein were altered by 15d-PGJ2 by mechanisms dependent on PI3K/Akt activity. 15d-PGJ ₂ prevents the calmodulin-dependent Akt overactivation in AD lymphoblasts by blocking its binding to the 85-kDa regulatory subunit of PI3K. These effects of 15d-PGJ ₂ were not mimicked by 9,10-dihydro-15-deoxy-Δ^12,14- prostaglandin J ₂, suggesting that 15d-PGJ ₂ acts independently of peroxisome proliferator-activated receptor γ activation and that the α,β-unsaturated carbonyl group in the cyclopentenone ring of 15d-PGJ ₂ is a requisite for the observed effects. Received 14 July 2008; received after revision 2 September 2008; accepted 12 September 2008     

Ultrastructure of muscle spindles in C57BL/6J dy2J/dy2J dystrophic mice.

The sensory organs of skeletal muscles, the muscle spindles, were examined using electron microscopy in dy2J/dy2J dystrophic mice. Despite widespread damage to the extrafusal (skeletomotor) fibres the intrafusal (spindle) fibres appeared normal and seemed resistant to the aetiological factors for murine dystrophy.     

Dystrophin-dependent efficiency of metabolic pathways in mouse skeletal muscles

Muscles from themdx mouse (X-linked genetic disorder similar to Duchenne muscular dystrophy) lack dystrophin-associated transsarcolemmal proteins¹ and show reduced maintenance metabolic rates². Here, microcalorimetric comparisons of metabolic stimulation by exogenous substrates in isolated muscles revealed substrate-selective limitation of chemical reaction rates through both glycolytic and TCA-cycle pathways, identical in slow- and fast-twitchmdx muscles. This systemic approach, as opposed to comparisons of single-enzyme activities, sheds new light on the function of dystrophin and associated proteins. The in vivo efficiency of metabolic pathways may depend on stabilization of enzyme complexes by dystrophin-associated elements of the cytoskeleton.     

Glucagon treatment of muscular dystrophic mice: A lack of effect

Zusammenfassung Nachweis, dass Körpergewicht, Dystrophie und Sterblickeit bei männlichen und weiblichen muskeldystrophischen Mäusen (dy/dy) nach 100tägiger Behandlung mit Glukagon (2 oder 10 g pro Tag, s.c.) unverändert blieben.     

Inhibition of riboflavin-sensitized photoinactivation of taka-amylase a by some inhibitors

Zusammenfassung Durch Gegenwart von Co⁺⁺ oder J^– oder Tryptophan oderp-Phenylendiamin wird die durch Riboflavin sensibilisierte Photoinaktivierung der Taka-Amylase A stark gehemmt. Die möglichen Mechanismen der Hemmungswirkung dieser Inhibitoren werden diskutiert.     

Absorption tests on protein-deficient rats. Absorption of S35-tethionine and I131-mriolein

Zusammenfassung Die Wirkung der Eiweissmangeldi?t auf die Resorption von S³⁵-Methionin und J¹³¹-Triolein wurde untersucht. Die Resorption von Methionin wurde nicht wesentlich beeinflusst, w?hrend die Fettresorption nach 24t?gigem Eiweissmangel signifikant vermindert ist.      

An increase in thyroid radioiodine uptake following the administration of cyproterone acetate

Zusammenfassung Die Wirkung des Cyproteron acetates auf die J¹³²-Aufnahme der Rattenschilddrüse wurde untersucht. Die Radiojodaufnahme der Schilddrüse wurde durch diese Substanz erhöht, wobei gleichzeitig eine mässige Zunahme des Schilddrüsengewichtes festgestellt werden konnte. Gleichzeitige Verabreichung von Oestradiol hob diesen Effekt auf.     

CD20-related signaling pathway is differently activated in normal and dystrophic circulating CD133+ stem cells

Among the heterogeneous population of circulating hematopoietic and endothelial progenitors, we identified a subpopulation of CD133⁺ cells displaying myogenic properties. Unexpectedly, we observed the expression of the B-cell marker CD20 in blood-derived CD133⁺ stem cells. The CD20 antigen plays a role in the modulation of intracellular calcium homeostasis through signaling pathways activation. Several observations suggest that an increase in intracellular calcium concentration ([Ca²⁺]_i) could be involved in the etiology of the Duchenne muscular dystrophy (DMD). Here, we show that a CD20-related signaling pathway able to induce an increase in [Ca²⁺]_i is differently activated after brain derived neurotrophic factor (BDNF) stimulation of normal and dystrophic blood-derived CD133⁺ stem cells, supporting the assumption of a “CD20-related calcium impairment-affecting dystrophic cells. Presented findings represent the starting point toward the expansion of knowledge on pathways involved in the pathology of DMD and in the behavior of dystrophic blood-derived CD133⁺ stem cells. Received 15 October 2008; received after revision 27 November 2008; accepted 05 December 2008     

The effect of various anions on the contractility of the guinea-pig papillary muscle

Zusammenfassung Ersatz der Chlorionen des Suspensionsmediums durch NO ₃ ^– , Br^– oder J^–verstärkt die Kontraktionskraft des Herzmuskels im Gegensatz zu der des Skelettmuskels durch Vergrösserung der Anstiegssteilheit der Kontraktion. Dagegen wirkt F^– (5 mM) am Herzen positiv inotrop durch Verlängerung der Anstiegszeit der Kontraktion.     

Molecular mechanisms of BK channel activation

Large conductance, Ca²⁺-activated potassium (BK) channels are widely expressed throughout the animal kingdom and play important roles in many physiological processes, such as muscle contraction, neural transmission and hearing. These physiological roles derive from the ability of BK channels to be synergistically activated by membrane voltage, intracellular Ca²⁺ and other ligands. Similar to voltage-gated K⁺ channels, BK channels possess a pore-gate domain (S5–S6 transmembrane segments) and a voltage-sensor domain (S1–S4). In addition, BK channels contain a large cytoplasmic C-terminal domain that serves as the primary ligand sensor. The voltage sensor and the ligand sensor allosterically control K⁺ flux through the pore-gate domain in response to various stimuli, thereby linking cellular metabolism and membrane excitability. This review summarizes the current understanding of these structural domains and their mutual interactions in voltage-, Ca²⁺ - and Mg²⁺ -dependent activation of the channel. Received 25 September 2008; received after revision 23 October 2008; accepted 24 October 2008     

The metabolic fate of [35S]-diallyl disulphide in mice

Summary Diallyl disulphide (DADS) is a major constituent of garlic oil. Uptake of [³⁵S]-labelled diallyl disulphide by mouse liver is highest at 90 min after treatment. 2h after treatment with [³⁵S]-DADS, 70% of the radioactivity is present in the liver cytosol of which 80% is metabolized to sulphate.     

Ketoconazole,an inhibitor of calcium transport in skeletal muscle sarcoplasmic reticulum

Summary Ketoconazole, an antimycotic agent, inhibits calcium binding and accumulation, and induces calcium release in sarcoplasmic reticulum. The Mg²⁺-ATPase and the (Ca²⁺+Mg²⁺)-ATPase activities are stimulated at low but inhibited at high concentrations of ketoconazole.The author wishes to thank Dr K. S. Cheah for discussion and Mr C. C. Ketteridge for preparing the sarcoplasmic reticulum and carrying out the ATPase assays.     

On the use of a peptidase inhibitor (trasylol) for storage of I125-labelled peptide hormones (HGH,TSH, ACTH,insulin, glucagon,PTH)

Zusammenfassung Von den untersuchten J¹²⁵-markierten Peptidhormonen sind HGH und TSH auch ohne Trasylol gut haltbar. Die mässige Degradation von Insulin und die rasch zunehmende Degradation von PTH und Glukagon kann durch Trasylol (500 E/ml) verhindert werden. Für ACTH ist eine Kombination von Trasylol mit 0.5% Mercaptoethanol erforderlich.     

Biosynthesis of 1-aminocyclopropanecarboxylic acid: Steric course of the reaction at the C-4 position

Summary Under the action of the appropriate synthase from ripe tomatoes a 11 mixture of (3S, 4R)-[3,4-²H₂] and (3R, 4S)-[3,4-²H₂]-(2S)-adenosylmethionine is transformed into a 11 mixture of the two meso forms of [²H₂]-1-aminocyclopropanecarboxylic acid, a result which proves the operation of an inversion mechanism and which is consistent with direct nucleophilic displacement of the leaving group in the substrate.     

Serum-induced stimulation of snRNA synthesis in mouse 3T3 fibroblasts

Summary Small nuclear RNAs (snRNAs) from quiescent and serum-stimulated 3T3 cultures, labeled with [³H]uridine ([³H]U), were electrophoresed in polyacrylamide-urea slab gels and revealed by staining with ethidium bromide and by fluorography, Judged by labeling with [³H]U, synthesis of 7S and U1-U6 RNAs was very low or absent in quiescent cultures. The serum-induced transition of 3T3 cells from a resting to a growing state was accompanied by an early, apparently sequential stimulation of snRNA synthesis; stimulated synthesis of 7S, U1, U2, U3, U4 and U6 RNAs coincided in time with serum-induced stimulation of 45S pre-ribosomal RNA (pre-rRNA) and heterogeneous nuclear RNA (hnRNA) synthesis.     

Membrane transport of Na and K and cell dehydration in sickle erythrocytes

The cellular concentration of Hb S plays a central role in the kinetic of Hb S polymerization and cell sickling. Blood of patients with homozygous sickle cell (SS) anemia contains a variable fraction of cells which are markedly dehydrated and have increased Hb S concentration. Since a decrease in cellular Hb S concentration reduces Hb S polymerization and sickling, the study of the processes leading to sickle cell dehydration has important pathophysiological and therapeutic implications. Sickle cell dehydration is due to cellular loss of K and Cl. K loss in sickle cells can take place via either the Ca²⁺-activated K⁺ channel, or the K?Cl cotransport, or the combined effect of oxidative damage and deformation of the red cell membrane. Inhibitors of K transport through these pathways could be used to prevent dehydration of sickle cells in vivo, provided that they can be administered safely.     

The role of glycine in the biosynthesis of steroids

Zusammenfassung Während der Biosynthese von Cholesterol mit homogenisierter Rattenleber wird [2-¹⁴C] des Glycins viel besser eingebaut als [1-¹⁴C].Saccharomyces cerevisiae produziert radioaktives Squelen (ausser Ergosterol mit Radioaktivität des Ringsystems) mit [2-¹⁴C] Glycin und mit [3-¹⁴C] Serin, aber nicht mit [1-¹⁴C] Glycin.

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Studies on Biosynthesis. Part VI. For Part V, seeA. K. Bose, K. S. Khanchandani andB. L. Hungund, Experientia,27, 1403 (1971). b) Presented at the 164th National Meeting of the American Chemical Society, New York, August, 1972.

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The support of this research by Stevens Institute of Technology and Sandoz Foundation is gratefully acknowledged. We wish to thank Drs.P. T. Funke, M. S. Manhas, P. K. Bhattacharyya, M. Anchel andH. Levey for valuable discussions and help with some of the experiments.     

Glucocorticoids suppress cystathionine gamma-lyase expression and H2S production in lipopolysaccharide-treated macrophages

Hydrogen sulfide (H₂S) plays an important role in inflammation. We showed that macrophages expressed the H₂S-forming enzyme cystathionine gamma-lyase (CSE) and produced H₂S. Lipopolysaccharide (LPS) stimulated the CSE expression and H₂S production rate. l-cysteine reduced LPS-induced nitric oxide (NO) production. CSE inhibitor blocked the inhibitory effect of l-cysteine. CSE knockdown increased, whereas CSE overexpression decreased LPS-induced NO production. Dexamethasone suppressed LPS-induced CSE expression and the H₂S production rate as well as NO production. l-arginine increased, whereas N^G-nitro-l-arginine methyl ester (l-NAME) decreased LPS-induced CSE expression and H₂S production. Dexamethasone plus l-NAME significantly decreased LPS-induced CSE expression and H₂S production compared to l-NAME. Our results suggest that macrophages are one of the H₂S producing sources. H₂S might exert anti-inflammatory effects by inhibiting NO production. Dexamethasone may directly inhibit CSE expression and H₂S production, besides the NO-dependent way. Inhibition of H₂S and NO production may be a mechanism by which glucocorticoids coordinate the balance between pro- and anti-inflammatory mediators during inflammation.