Triphasic vascular effects of thiol compounds and their oxidized forms on dog coronary arteries

The vascular effects of 2-mercaptoethanol, cysteamine, L-cysteine, glutathione (GSH), cystamine and oxidized GSH (GSSG) on the isometric tension of isolated dog coronary arterial strips were examined, and these effects were compared with the triphasic response induced by dithiothreitol (DTT); a rapid and weak contraction (phase A), an intervening slow relaxation (phase B) and a slowly-developing strong contraction (phase C) which we previously reported. The responses of the arteries induced by 2-mercaptoethanol, cysteamine and L-cysteine consisted of phases A, B and C. The order of contractile potency (ED₅₀ of phase C) was DTTL-cysteine>2-mercaptoethanolcysteamine, while the order of relaxant potency (ED₅₀ of phase B) was DTT>cysteamine2-mercaptoethanol. GSSG and cystamine mainly produced relaxation, which corresponded to phase B. The phase C contraction was specific to the reduced forms of thiols, except for GSH, which produced only relaxation. The participation of endothelial cells was not essential for the contracting or relaxing effects of the thiol compounds. The phase C contraction was depressed by W-7, a calmodulin antagonist, while phase A was not. Therefore calmodulin-dependent protein kinases may participate in phase C, not in phase A.     

Some citrus chitinases also possess chitosanase activities

Several acidic chitinase and chitosanase isoforms were found in 4-week-old nonembryogenic sweet orange (Valencia [Citrus sinensis (L.) Osbeck]) callus tissue. Two isoforms (designated A1-CF1 and A1-CF2) were purified to homogeneity using HPLC size exclusion, anion exchange, and chromatofocusing techniques. Both hydrolase isoforms exhibited activity with either colloidal chitin or solubilized shrimp shell chitosan. Specific activities for the purified isoforms could not be calculated because of the lack of protein and contamination of ampholytes. However, the specific activities for chitinase and chitosanase after anion exchange were respectively 404 nmol GlcNAc per min per mg protein and 2,475 nmol GlcN per min per mg protein. The M_r for both enzymes was 30,500. The homogeneous proteins cross-reacted in western blots with antiserum against a basic class I potato leaf chitinase.     

Transplantation of human cortex with Alzheimer's disease into rat occipital cortex; a model for the study of Alzheimer disease

Summary Senile dementia of the Alzheimer type (SDAT) is a major problem in the human senescent population. As this pathology cannot be reproduced in animals, research into its development is greatly impeded. The technique of implantation of the nervous tissue has been utilized in order to establish an animal model and to test the possible existence of a transmissible agent. When human temporal cortex with Alzheimer's disease is implanted in the occipital cortex of 7-week-old rats, human cerebral tissue containing abundant tangles induces in the receiver cortex a reactive fibrous gliosis. In the processes of the astrocytes, twisted filaments are evident among bundles of normal filaments. These alterations could be induced by the metabolising of abnormal filament subunits or by some infectious agent introduced by the implant.This study is supported by grant No. 2.4517.82 of Fonds de la Recherche Fondamentale Collective of Belgium.     

Evaluation of monoaminergic receptors in the genetically epilepsy prone rat

Summary The intensity of sound-induced convulsions in the genetically epilepsy-prone rat (GEPR) was reduced in a dose related fashion by intracerebroventricular administration of dobutamine, (₁ agonist), terbutaline (₂ agonist) or phenylephrine (₁ agonist). BHT-920 (₂ agonist) did not cause a dose-related decrease in sound-induced convulsion intensity. Binding studies showed that whole brain and receptor densities (B_max) were normal while the K_d was increased for the ligand in GEPR brain.Acknowledgment. We are most grateful to Boehringer Ingelheim for generously supplying BHT 920. We are also indebted to Ciba-Geigy Corporation for the gift of terbutaline hydrochloride and phentolamine hydrochloride. The work was supported in part by NIH grant NS 16829.     

Physiological state of submitochondrial particles and their susceptibility to Triton X-100

Summary The solubilizing effect of Triton X-100 on beef heart submitochondrial particles (ETP_H) has been studied under various physiological conditions. Coupled, uncoupled and azide-inhibited ETP_H particles have been studied. Quantitative and qualitative differences are found in the proteins solubilized by the detergent from ETP_H particles under the various conditions tested.Acknowledgments: This work was supported in part by a grant from a Spanish Comisión Asesora para la Investigación Cientifica y Técnica. M.C. Barbero was recipient of a scholarship from the Basque Government.     

Isolation and partial characterization of cuticular collagen from the parasitic nematodeGaigeria pachyscelis

Summary The cuticle from adultGaigeria pachyscelis was isolated by solubilizing the internal tissues with sodium dodecyl sulphate (SDS) at 37°C. Cuticular protein was extracted with guanidine-HCl and -mercaptoethanol and purified by ammonium sulphate fractionation and DEAE-cellulose chromatography. SDS-polyacrylamide gel electrophoresis of purified protein revealed 2 polypeptides with apparent mol. wts of 58,000 and 74,000. As judged from their hydroxyproline content both of them are collagenous in nature. Results of gel filtration indicate that cuticular collagen exists in two forms, a non-associated form at low concentration and an associated form at high concentration.Acknowledgments. We thank Drs L.N. Singh and H.C. Tewari for providing the necessary facilities. Laboratory assistance of Mr Ram Kishore is highly appreciated.     

Effect of hormones and cyclic AMP on γ-glutamyltranspeptidase activity of rat mammary gland explants

Summary -Glutamyl transpeptidase activity was assayed in midpregnant rat mammary gland explants at 14, 22 and 38 h, in the presence and absence of insulin, prolactin and corticosterone. With these 3 hormones the explants attained the characteristics of a secretory gland after 22 h of tissue culture, at which time the enzyme exhibited its maximal activity. The addition of dibutyryl cyclic AMP in the presence of the 3 hormones produced a significant increase in enzyme activity, which was maximal with a 1 mM concentration of the cyclic nucleotide. A similar effect was observed when theophylline or theophylline plus dibutyryl cyclic AMP were added to the culture medium.This work was supported by grant B1138-8333 from the Departamento de Desarrollo de la Investigación, Universidad de Chile.     

Oxygen consumption in rat skeletal muscle at various rates of oxygen delivery

Summary Resting oxygen consumption of the vascularly isolated gracilis anticus muscle of the rat perfused by natural circulation via the femoral artery with diluted or undiluted blood depends on oxygen delivery (the product of flow rate and arterial oxygen concentration-DO₂) only when DO₂ falls below 16 l/min/g.     

High-affinity binding ofβ-alanine to cerebral synaptosomes might involve glycine-receptors

Summary High-affinity, Na⁺-independent binding of -alanine to a synaptosomal fraction of rat brain was potently inhibited by glycine and by some other -amino acids, but not by taurine or GABA. This binding mechanism, which was also sensitive to both bicuculline and strychnine, might involve synaptic receptors for both -alanine and glycine.This study was supported in part by Centro Ramón y Cajal and Fundacion Juan March.