Halocyamines: novel antimicrobial tetrapeptide-like substances isolated from the hemocytes of the solitary ascidian Halocynthia roretzi

Two novel antimicrobial tetrapeptide-like substances, halocyamine A and B, were isolated from the solitary ascidian Halocynthia roretzi by a procedure including extraction steps, chromatographies on coarse and fine HP-20 columns, and preparative reversed-phase high-performance liquid chromatography. The structures of halocyamine A and B were determined to be L-histidyl-L-6,7-dihydroxyphenylalanylglycyl-6-bromo-8,9-didehy drotryptamine and L-threonyl-L-6,7-dihydroxyphenylalanyl-L-histidyl-6-bromo-8,9- didehydrotryptamine, respectively, by spectral analyses and degradation studies. Besides antimicrobial activities against several kinds of bacteria and yeasts, both of them showed cytotoxic activities against neuronal cells cultured from rat fetal brain, mouse neuroblastoma N-18 cells, and human hepatoma Hep-G2 cells. They were only detected in the "morula"-like cells, which are of the most abundant cell type among the hemocytes of H. roretzi.     

Superoxide dismutases enhance the rate of autoxidation of 3-hydroxyanthranilic acid

The autoxidation of 3-hydroxyanthranilate to cinnabarinate at 37 degrees C and at pH 7.4 is hastened by superoxide dismutase (SOD). The Cu,Zn-containing enzyme from bovine erythrocytes and the Mn-containing enzyme from Escherichia coli were equally effective in this regard; whereas the H2O2-inactivated Cu,Zn enzyme was ineffective. Catalase appears to augment the effect of superoxide dismutase, because it prevents the bleaching of cinnabarinate by H2O2. It follows that O2-, which is a product of the autoxidation, slows the net autoxidation by engaging in back reactions and that SOD increases the rate of autoxidation by removal of O2- and thus by prevention of these back reactions.     

Hydrogen exchange kinetics of nucleic acids: Double and triple helices with Hoogsteen-type basepairs

The kinetics of hydrogen-tritium exchange reaction have been followed by a Sephadex technique of a double-helical poly(ribo-2-methylthio-adenylic acid)·poly(ribouridylic acid) complex with the Hoogsteen-type basepair. Only one hydrogen in every 2-methylthio-adenine·uracil basepair has been found to exchange at a measurably slow rate, 0.023 s^?1 (at 0°C), which is, however, much greater than that for a double-helix with the Watson-Crick type A·U pair. The kinetics of hydrogen-tritium exchange were also examined by triple-helical poly(rU)·poly(rA)·poly(rU) which involves both the Watson-Crick and Hoogsteen basepairings. Here, three hydrogens in every U·A·U base triplet have been found to exchange at a relatively slow rate, 0.0116 s^?1 (at 0°C). The kinetics of hydrogen-deuterium exchange reactions of these polynucleotide helices have also been followed by a stopped-flow ultraviolet absorption spectrophotometry at various temperatures. On the basis of these experimental results, the mechanism of the hydrogen exchange reactions in these helical polynucleotides was discussed. In the triple helix, the rate-determining process of the slow exchange of the three (one uracil-imide and two adenine-amino) hydrogens is considered to be the opening of the Watson-Crick part of the U·A·U triplet. This opening is considered to take place only after the opening of the Hoogsteen part of the triplet.     

Global stability of stationary solutions to a nonlinear diffusion equation in phytoplankton dynamics

We consider a nonlinear diffusion equation proposed by Shigesada and Okubo which describes phytoplankton growth dynamics with a selfs-hading effect.We show that the following alternative holds: Either (i) the trivial stationary solution which vanishes everywhere is a unique stationary solution and is globally stable, or (ii) the trivial solution is unstable and there exists a unique positive stationary solution which is globally stable. A criterion for the existence of positive stationary solutions is stated in terms of three parameters included in the equation.     

Schistosoma japonicum: Immediate hypersensitivity reactions to egg antigen in humans and rodents

Immediate hypersensitivity reactions in schistosoma japonicum infections were examined in both man and experimental animals. In man higher reaction to soluble egg antigen than to adult worm antigen was detected by the use of the radioallergosorbent test (RAST). Blood-collecting filter paper can be used in RAST for seroepidemiological study in place of a skin test. Reaginic antibody formation against egg antigen was detected at the approximate time of egg deposition in strains of mice, Mongolian gerbils, cotton rats, and laboratory rats by the use of passive cutaneous anaphylaxis or Prausnitz-Küstner-type skin tests. At the same time circumoval precipitin tests were positive. Results with athymic nude mice suggest that these reactions are T-cell dependent. No detectable reagin synthesis against adult worm antigen was found in the animals so far examined, confirming stronger allergenic reaction to egg antigen than to that of adult worms in S. japonicum infections in man and animals.     

Translocation of colicin E1 through cytoplasmic membrane of Escherichia coli

The product of the malE—lacZ gene fusion was reported to compete with some proteins including outer membrane lipoprotein in the protein translocation across the Echerichia coli membrane. The fusion product also inhibited colicin E1 export. Furthermore, globomycin, which accumulated prolipoprotein in the membrane, inhibited the translocation of colicin E1 in the wild-type cells, but not in lipoprotein-negative mutant cells. Since colicin E1 contains the internal signal-like sequence [Proc. Natl. Acad. Sci. USA (1982) 79, 2827–2831], these results suggest that colicin E1 is exported by the aid of this sequence at a common site for maltose-binding protein and lipoprotein translocation.     

Versatile Method for Evaluating a Cell Culture by Various Morphological Techniques

Cellulose acetate is a versatile material for evaluating cells grown under identical conditions by various morphological techniques. This inexpensive material is transparent, easily cut to size and shape, nontoxic to cell cultures, and resistant to most chemicals used in histochemistry and in scanning and transmission electron microscopy. Samples may be obtained during and after the culture process. Cellulose acetate slides can be mounted directly over glass slides for direct observation and are easily peeled off plastic blocks for electron microscopy, leaving the cells behind. Relative disadvantages include its autofluorescence and a tendency to soften in strong acids or pure solutions of organic solvents such as xylene and propylene oxide.     

Migration inhibitory action of bacterial lipopolysaccharide on progenitor cells of monocyte/macrophage lineage growing in culture in the presence of colony-stimulating factor (CSF-1)

Addition of lipopolysaccharide (LPS) to the culture of mouse myeloid stem cells (CFU_c) increased the incidence of compact colonies and decreased that of dispersed ones in the presence of colony-stimulating factor (CSF-1) which had not such an effect by itself even in high concentrations. Although colony morphology was thus changed, nearly all colonies were composed of monocytes. The incidence of compact colonies increased with the increase of LPS concentration but plateaued at about 50%. Bone marrow cells of LPS-tolerant mice responded to LPS in vitro to a slightly decreased extent. The activity of LPS was decreased by alkaline or acid hydrolysis of the LPS molecule and inhibited by polymixin B, but not by indomethacin, α-L-fucose, nor by α-methyl-D-mannoside. Other immunopotentiating substances, such as OK-432, Lentinan, and Levamisole, had no effect on the colony morphology. Both muramyl dipeptide and poly(I)poly(C) were also ineffective. Furthermore, the action of LPS was not abolished by the use of heat-inactivated serum in the culture. LPS was no longer stimulative for the induction of lysosomal enzymes in the CSF-stimulated culture, although it greatly enhanced the enzyme induction in the unstimulated culture. These results indicate that the cells of monocyte/macrophage lineage develop the capacity for migration before they become responsive to LPS, and that the LPS-responding monocytic cells can proliferate even in a state of confluence induced by LPS.     

Effects of inhibitors of DNA synthesis on spontaneous and ultraviolet light-induced sister-chromatid exchanges in Chinese hamster cells

Effects of inhibitors of DNA synthesis on spontaneous and ultraviolet light (UV)-induced sister-chromatid exchanges (SCE) were examined in a Chinese hamster cell line, V79 B-1. The inhibitors used were hydroxyurea (HU), 1-beta-D-arabinofuranosylcytosine (ara-C), aphidicolin (APC), 2',3'-dideoxythymidine triphosphate (ddTTP), neocarzinostatin (NCS), novobiocin (NB) and cycloheximide (CHX). HU, ara-C, and APC increased spontaneous SCE frequency, and had a synergistic effect on UV-induced SCE frequency. DdTTP, NCS and NB failed to show any statistically significant effect on either spontaneous or UV-induced SCE frequencies, though NCS and NB did slightly increase both spontaneous and UV-induced SCE frequencies. On the contrary, CHX decreased spontaneous SCE frequency, and more drastically, also UV-induced SCE frequency. These results are interpreted with respect to the replicating fork of DNA, a structure postulated to be involved in the formation of spontaneous and UV-induced SCE. A new model for SCE formation is proposed.     

Isolation, homogeneity, and properties of core particle from pyocin R1.

By a mild alkaline treatment of pyocin R1, the core particle was released from the contracted sheath. After sucrose density-gradient centrifugation, core-rich fractions were treated with anti-sheath serum and by a second density-gradient centrifugation, purified core particles were isolated. Homogeneity of the preparation was confirmed by observation under the electron microscope, immuno-precipitation reaction, and sucrose density-gradient centrifugation. The core particle exhibited a sedimentation coefficient of 37S. The quaternary structure of the core consists of a single kind of subunit protein with a molecular weight of 18,000. No contamination by other proteins was detected by SDS-disc electrophoreses. Amino acid analysis revealed that the core is rich in glycine, alanine, valine, leucine, aspartic acid (or asparagine), glutamic acid (or glutamine), and serine. This amino acid composition bears some resemblance to that of T-even bacteriophage tail-core.