Mutagenicity of anthraquinones in the Salmonella preincubation test

The mutagenicities of 15 naturally occurring anthraquinones were examined in Salmonella typhimurium strains TA98, TA100 and TA2637 by the preincubation method. 7 of the 15 compounds tested, i.e., chrysazin, emodin, islandicin, alizarin, chrysophanol, 2-hydroxyanthraquinone and emodic acid, were strong mutagens in strain TA2637 with metabolic activation. All of these compounds contain 1-3 hydroxyl groups, and some also have methyl groups. Cynodontin, an anthraquinone with 4 hydroxyl groups and 1 methyl group, was only slightly mutagenic in strain TA2637. 2-Hydroxyanthraquinone, alizarin, emodin, islandicin and chrysazin were also mutagenic in strain TA100 with S9 mix. All the bisanthraquinones tested, i.e., skyrin, (+)rugulosin, (-)luteoskyrin, (-)rubroskyrin and sennoside A, were non-mutagenic in this test system with or without metabolic activation. Unsubstituted anthraquinone and anthrone were also non-mutagenic. These results show that hydroxyl substituents are necessary for the mutagenicity of anthraquinones, the optimal substitutions being 1-3 hydroxyl groups per molecule. The 4th hydroxyl group, in the compound cynodontin reduces the mutagenicity considerably.     

High-Molecular-Weight Hyaluronic Acids Inhibit Chemotaxis and Phagocytosis but Not Lysosomal Enzyme Release Induced by Receptor-Mediated Stimulations in Guinea Pig Phagocytes

The effects of high-molecular-weight (HMW) hyaluronic acids (HAs) of 1.9 × 10⁶ Da, 8 × 10⁵ Da and 3 × 10⁵ Da on the receptor-mediated functions of guinea pig peritoneal phagocytes were studied. HMW-HAs of 1.9 × 10⁶ Da (HA190) and 8 × 10⁵ Da (HA80) effectively inhibited the chemotactic activity of polymorphonuclear leukocytes (PMNs) for formyl-Met-Leu-Phe (fMLP). The degree of inhibition was dose-dependent and the concentrations of HA190 and HA80 required for 50% inhibition were 0.5–1.5 mg/ml and 1.5–2.5 mg/ml, respectively. HMW-HA of 3 × 10⁵ Da (HA30) hardly affected the chemotaxis within a concentration range of 0.5–5.0 mg/ml. The phagocytic activities of PMNs and macrophages (Møs) for serum-opsonized zymosan (SOZ) and polystyrene latex particles were also inhibited by these HAs in a dose- and molecular-weight-dependent manner and HA190 was again the most inhibitory. By contrast, the release of lysosomal enzyme from Møs stimulated with SOZ was not significantly affected by HMW-HAs at any concentration used. Furthermore, the binding of [³H]fMLP with PMNs and the rosette formation of Møs with SOZ were not influenced by the presence of HMW-HAs. These findings suggested that the binding of HMW-HAs to the HA receptors on PMNs and Møs might produce certain intracellular signals which would be responsible for the suppression of the chemotaxis and the phagocytosis but not for the release of lysosomal enzyme. For the generation of such signals, higher-molecular-weight HMW-HAs would be more effective than lower one.     

Detection of a serum class I molecule in rat with anti-rat liver beta-2-microglobulin

Rat serum was fractionated on a column of Sephacryl-300 and tested with a rabbit anti-rat beta-2-microglobulin (B2m) antiserum. This antiserum was directed against B2m purified from rat liver, and its specificity was confirmed by immunoprecipitation procedures. The antiserum recognized three peaks in the fractionated rat serum: a 200- to 300-kd (kilodalton) fraction, a 40- to 70-kd component, and the free 12-kd B2m. Indirect immunoprecipitation from the 200- to 300-kd fraction led to the identification of a 43-kd polypeptide associated with B2m. A xenoantiserum against RT1 class I antigen also precipitated a similar polypeptide from the same fraction, but this molecule differed in size and antigenic specificity from the one precipitated by anti-rat B2m.This work was supported in part by Grant 59010101 from the Ministry of Education, Science, and Culture, Japan.     

Binding of simian virus 40 large tumor antigen to phospholipid vesicles

SV40 T antigen is the initiator protein of SV40 DNA replication. We examined the interaction of purified SV40 T antigen with phospholipids by (i) centrifugation analysis with phospholipid vesicles, (ii) filter binding assay and footprint analysis of T antigen binding to the replication origin of SV40 DNA and (iii) analysis of the initiation of SV40 DNA replication in vitro. In all cases, cardiolipin showed affinity for T antigen and inhibited its DNA binding capacity. Phosphatidylglycerol with unsaturated fatty acids also inhibited the binding of T antigen to the replication origin of SV40 DNA, whereas phosphatidylglycerol with saturated fatty acids did not. This finding suggested the importance of unsaturated fatty acids for the interaction of T antigen with phospholipids. Other phospholipids including phosphatidylserine, phosphatidylinositol and phosphatidylethanolamine showed little or no affinity for T antigen.     

An autoradiographic,biochemical, and morphological study of the harderian gland of the mouse

Biochemical and morphological properties of the Harderian gland of the mouse were examined by combining autoradiographic, biochemical, and electron microscopic techniques. Autoradiographs show that the radioactive carbon from [U-¹⁴C]glucose injected into the abdominal cavity is completely incorporated into the acid-insoluble substances within 30 minutes. The results of chemical analysis show that the main components of this gland are glyceryl ether diesters and phospholipids. Scanning electron microscopy shows numerous lipid droplets in the secretory cells and alveolar lumina. Myoepithelial cells lie between the secretory cell base and the basement membrane and have a basket-like distribution of processes as confirmed by hydrochloric acid and collagenase digestions. Myofilaments are demonstrated in the cytoplasm. Two types of secretory cells (A and B) comprise the alveolar epithelium and can be differentiated under the electron microscope. The cytoplasm of both contains numerous vacuoles. The vacuoles are almost empty in A cells, which are a more numerous constituent of the alveolar epithelium than B cells. However, the vacuoles of the B cells contain densely osmiophilic material. In both, cell types show a merocrine mode of secretion. Unmyelinated nerve cell endings occur in the interstices of the connective tissue, and contain clear or cored vesicles.     

Ontogeny of DNA synthetic rhythms in chick (Gallus domesticus) liver.

1. The diurnal pattern of DNA synthesis and mitotic activity in neonatal (1-4-day-old) chick liver were investigated under various feeding and lighting regimens. 2. In the meal-fed chicks under the condition of light-dark cycle, DNA synthesis exhibited a 12 hr cycle; the peaks occurring at 9:00 and 21:00. 3. Fasting caused a gradual decrease in the 21:00 peaks. 4. The changes in the lighting regimen to 24 hr continuous lighting also caused a profound change in the DNA-synthetic pattern, suggesting a complex interplay of feeding and lighting regimens in the manifestation of the DNA-synthetic rhythm in neonatal chick liver.     

LHRH increases plasma 7B2 concentration in normal human subjects

We studied the response of plasma 7B2 to LHRH and ovine corticotropin releasing hormone (o-CRH) in healthy young subjects. The plasma 7B2 concentration significantly increased from 78.3 +/- 7.5 (mean +/- SEM) to 102.0 +/- 6.0 ng/L (142.7 +/- 12.7% of the basal value; P less than 0.01) following iv administration of LHRH in seven young subjects. On the other hand, no increase in plasma 7B2 was found after iv administration of o-CRH in six young subjects. These results, together with our previous report of no increase in plasma 7B2 after administration of TRH and GHRH in young subjects, suggest that pituitary 7B2 may be present in gonadotrophs and be released only by LHRH in physiological conditions.