Effects of butylated hydroxyanisole on the aryl hydrocarbon hydroxylase of rats and mice

The effects of butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) on the aryl hydrocarbon hydroxylase (AHH) activities in the liver, lung and skin of rats and mice have been studied to examine the possible mechanisms of the anticarcinogenic actions of these compounds. Both compounds inhibit the hydroxylase activities of hepatic microsomes and nuclei, with BHA a more potent inhibitor than BHT. The AHH of lung microsomes is inhibited to a lesser extent by BHA and BHT than that of the liver. The AHH activities of both liver and lung microsomes become less susceptible to the inhibition after pretreatment of the animals with 3-methylcholanthrene (MC) or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) but phenobarbital (PB) pretreatment does not produce such an effect. In skin homogenates, however, the AHH activities of control rats and mice are not inhibited by BHA and BHT. The only skin sample which is inhibited by BHA and BHT is that from TCDD-pretreated mice. It has been established that the extent of inhibition with different samples is related to the concentration of BHA in the incubation but not to the amounts or specific activities of microsomes used. Double reciprocal plots suggest that BHA exerts a mixed inhibition on the hydroxylase of liver microsomes with a K_i of 7.7 μM. Analysis of the metabolites of benzo[a]pyrene (BP) shows that BHA inhibits the formation of various metabolites uniformly without changing the regio-selectivity of the enzyme system. The mechanism of inhibition has also been studied with a reconstituted AHH system consisting of cytochrome P-450 (P-450), reductase and phospholipid. The system with P-450 isolated from PB-induced microsomes is inhibited to a much greater extent than that with MC-induced P-450. The results indicate that the inhibitory action of BHA is dependent on the species of the animal, tissue types and treatment with inducers.     

Effets du rubidium et du césium sur la croissance et la nutrition minerale des characees

Résumé Le rubidium et le césium introduits à l'état de chlorure dans le milieu de culture ont à faible dose un effet stimulant sur la croissance de Chara fragilis et de Chara vulgaris. La résistance de ces végétaux à l'action toxique des deux ions est accrue par l'addition de potassium au milieu.Les analyses chimiques confirment que le rubidium et le césium sont antagonistes vis-à-vis du potassium et du sodium alors qu'ils ne modifient pas de manière significative le taux de calcium.

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Chara fragilis and Chara vulgaris were cultivated in a natural medium containing rubidium and caesium as chloride.The growth of Characeae was increased after culture in the solutions containing Rb and Cs in small amount. The resistance to the toxic effects of these two ions is enhanced if potassium chloride is added to the medium.Quantitative analyses indicate that Rb and Cs decrease the rate of Na and K but have no significative influence on the rate of Ca.

Université de Dijon, Laboratoire de Nutrition minérale des Végétaux     

Inhibition of calcium and glucose uptake by murine leukemia L5178Y cells treated with antiserum

Studies were performed to determine whether decreases in transport of calcium and glucose might be among the earliest changes triggered by the antigen-antibody reactions occurring on the cell surface of murine leukemia L5178Y cells after treatment with rabbit antisera. After treatment with antisera, in the absence of complement, these cells exhibited a decreased uptake of ⁴⁵Ca, 2-deoxy[³H]glucose, and 3-0-methyl[³H]glucose. These changes occurred rapidly, within 2 minutes after the addition of antiserum, in contrast to the previously reported inhibitory effects of antiserum on DNA, RNA, and protein synthesis, which became demonstrable only after 4 to 8 hours. The kinetics of uptake of the radioactive substrates was biphasic, with a very rapid initial uptake followed by less rapid linear uptake. The precise mechanism of cell growth inhibition remains to be elucidated, but one of the initial effects of antiserum treatment may be a perturbation at the cell membrane such that transport of specific nutrients is decreased, resulting in the observed effects on macromolecular synthesis.     

Intimate coupling of creatine phosphokinase and myofibrillar adenosinetriphosphatase

ATPase and creatine phosphokinase (CPK) activities of isolated cardiac myofibrils were determined with ³²P γ-labeled ATP alone and with the addition of phosphorylcreatine (PC). With ATP and PC as substrates the label in the inorganic phosphate formed is greatly diluted indicating that the ATP formed by PC through CPK can reach the ATPase active site more readily than labeled ATP from the medium. The tight coupling of the ATPase and CPK activities further strengthens our view that PC serves an important role as high energy carrier between the energy producing sites (mitochondria) and the energy utilizing sites (myofibrils).     

A direct fluorometric assay of benzo[a]pyrene hydroxylase.

A technique to measure the activity of pyruvate carboxylase spectrophotometrically in crude liver homogenates is described. The assay is based on the transformation of oxaloacetate, which is formed during the carboxylation reaction, into citrate in the presence of excess acetyl CoA and citrate synthase. After removal of pyruvate with KBH₄ and of protein with HClO₄, citrate is cleaved with citrate lyase into oxaloacetate and acetate, and oxaloacetate then is measured spectrophotometrically. Optimal concentrations of pyruvate, Mg²⁺, ATP, and KHCO₃ for the carboxylation reaction and the V_max were in good correlation with the data found by others using [¹⁴C]pyruvate.     

Decrease of estrogen receptors induced by 17 beta-estradiol and progesterone in cultured rabbit endometrial cells

A whole cell technique to measure estradiol receptors in cultured rabbit endometrial cells is described. The estradiol receptors measured appear to be composed of at least two components: one component has high affinity but low capacity, while the other component has low affinity but high capacity. Using the assay, the effects of estradiol and progesterone pretreatment were examined on the estradiol receptor levels. It was found that both of the hormones decreased the number of estrogen receptors in the cultured cells. The finding that estradiol decreased its own receptors was unexpected and its possible relevance is discussed.     

Immunohistochemical localization of a gastrin-like peptide in the brain of an amphibian,Xenopus laevis Daud.

Summary The indirect immunofluorescence technique was used to demonstrate a substance reacting with gastrin antisera in the brain of Xenopus laevis.Immunoreactive material was found in two sites: (1) In the caudal hypothalamus more precisely in the nucleus infundibularis ventralis, (NIV) of the pars ventralis of the tuber cinereum, (PVTC). The fluorescent axons of the reactive parikarya of the NIV give rise to two symmetrical tracts which run rostro-ventrally and join, in the infundibular floor, the preoptico-hypophysial tract, where they form an uneven median tract coursing caudally and running along the medio-tuberal area before entering the external zone of the median eminence. (2) In the anterior preoptic area (APOA), where numerous nerve fibers and endings form a dense network near the preoptic recess. The exact origin of these terminals has not yet been determined.Control of immunohistochemical specificity shows that the labeling by gastrin antisera is suppressed by gastrin (2–17), but also by cholecystokinin (CCK) and pentagastrin (Peptavlon). These results indicate that the immunoreactive substance revealed belongs to the gastrin group and has an antigenic determinant composed of the amino acid sequence or a portion thereof common to gastrin, CCK and Peptavlon (Trp-Met-Asp-Phe).It should be emphasized that, in the brain of Xenopus laevis, both gastrinimmunoreactive sites correspond to the sites of uptake of steroid hormones (Kelley et al., 1975; Morrell et al., 1975).Supported by the D.G.R.S.T., Contrat n 77.7.0648     

Ultrastructural development of the neurohemal organ joined to the ecdysial gland after imaginal moulting in the male isopod Sphaeroma serratum fab. (Crustacea Flabellifera)

Summary The present ultrastructural study deals with the lateral cephalic nerve plexus of Sphaeroma serratum, a neurohemal organ joined to the Y organ (ecdysial gland). This plexus acts as a storage centre for neurosecretory products from two sources: the two autochtonous cells (plexus cells) within the plexus itself, and the neurosecretory cells in various parts of the central nervous system, particulary the mandibular ganglion (A-cells).In prepuberal animals, plexus cells and subesophageal A-cells produce neurosecretory granules of two types measuring 1550±50Å and 1570±40Å respectively. Five categories of axon terminals were distinguished in the plexus. The granules found in two of these terminal types are believed to come from the plexus cells and from the mandibular ganglion A-cells.Cessation of production of neurosecretory granules in these A cells and plexus cells was observed in puberal animals, in the plexus with concomitant depletion and disappearance of different granule categories. The first axon terminals affected by this process are the two categories containing granules originating in the plexus and mandibular ganglion A-cells. Degeneration of the ecdysial gland in male Sphaeroma serratum might be connected with the cessation of granule formation in these two types of cell.

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Résumé Chez Sphaeroma serratum, la mue de puberté est suivie d'une dégénérescence de l'organe Y (glands de mue). Le plexus nerveux céphalique latéral, organe neurohémal accolé à cette glande a été l'objet de la présente étude ultrastructurale. Cet organe représente un centre de stockage de neurosécrétions qui proviennent d'une part, de deux cellules autochtones (cellules plexales) situées au sein même de ce plexus, d'autre part, de cellules neurosécrétrices situées dans le ganglion mandibulaire (cellules de type A).Chez les individus pubères, les cellules plexales et les cellules A du ganglion sous-oesophagien synthétisent des granules de neurosécrétion dont la taille est respectivement 1550±50Å et 1570±40Å. Il a été reconnu au sein du plexus 5 catégories de terminaisons dont les granules proviendraient pour deux d'entre elles des cellules plexales et des cellules A du ganglion mandibulaire. Chez les animaux pubères on observe un arrêt de la synthèse des granules de neurosécrétion au sein des cellules plexales et des cellules A du ganglion mandibulaire. Simultanément on enregistre dans le plexus la raréfaction puis la disparition des divers types de granules. Ce processus atteint en premier les terminaisons correspondant aux cellules plexales et aux cellules A du ganglion mandibulaire. La dégénérescence de la glande de mue chez les mâles pourrait être en relation avec l'arrêt de synthèse de ces cellules.

     

A spectrofluorimetric investigation of calf thymus DNA modified by BP diolepoxide and 1-pyrenyloxirane

7β,8α-Dihydroxy-9α,10α-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BP diolepoxide, $\text{1}$) and 1-pyrenyloxirane ($\text{2}$) bind chemically to calf thymus DNA. The fluorescence efficiency of pyrenyl groups in mutagen modified DNA varies appreciably with its conformation and decreases in the order: pyrenees, modified denatured DNA and modified native DNA. A particularly interesting observation is that the fluorescence efficiency of mutagen modified DNA intensifies substantially upon denaturation. Our results suggest that the pyrenyl groups in mutagen modified DNA are intercalated between the base pairs of DNA. Since both $\text{1}$ and $\text{2}$ are powerful frame-shifting mutagens for S. typhimurium TA-98, the intercalative covalent binding of these compounds to DNA may provide a molecular basis for their mutagenic activity.     

Studies on the red oxidase (cytochrome o) of Azotobacter vinelandii

In an attempt to isolate and to study the electron transport system of $\text{Azotobacter vinelandii}$, we have isolated and purified a membrane-bound cytochrome $\text{o}$. The cytochrome $\text{o}$, purified as a detergent (Triton X-100) and hemoprotein complex, contained 1.6 nmoles heme per mg of protein. Cold-temperature spectrum showed that no other cytochrome was associated with the purified preparation, and electrophoresis revealed that only one type of hemoprotein was obtained. The purified cytochrome $\text{o}$ reacted with both carbon monoxide and cyanide readily. Only in the reduced form did it combine with carbon monoxide, whereas the oxidized form reacted with cyanide. An “oxygenated” form of the cytochrome $\text{o}$ was demonstrated to be spectrally distinguishable from both the oxidized and the reduced forms.