Effects of the umuC36 mutation on ultraviolet-radiation-induced base-change and frameshift mutations in Escherichia coli

The effects of the umuC36 mutation on the induction of base-change and frameshift mutations were studied. An active umuC gene was necessary in either the uvr⁺ or uvr^? strains of Escherichia coli K12 for UV- and X-ray-induced mutations to His⁺, ColE^R and Spc^R, which are presumably base-change mutations, but it was not essential for ethyl methanesulphonate or N-methyl-N′-nitro-N-nitrosoguanidine-induced His⁺ mutations. In contrast, only 1 out of 13 trp^? frameshift mutations examined was UV reversible, and the process of mutagenesis was umuC⁺-dependent, whereas a potent frameshift mutagen, ICR191, effectively induced Trp⁺ mutations in most of the strains regardless of the umu⁺ or umuC genetic background. These results suggest that base substitutions are a major mutational type derived from the umuC⁺-dependent pathway of error-prone repair.     

A rapid and simple method for the determination of base substitution and frameshift specificity of mutagens

A rapid method for the determination of mutagenic specificity has been developed which makes use of the ochre mutation (TAA) in the his-4 gene of Escherichiacoli. Reversion to His⁺ may occur by suppressor mutation (Type I) or by mutation within the his-4 gene (Type II). The Type I mutations may be further subdivided with respect to the type of suppressor mutation by their ability to suppress nonsense mutants of bacteriophage T4, thus allowing the identification of the responsible base substitution (Kato et al., 1980). The system has the ability to identify mutagens which produce A:T → G:C transitions since only Type II mutants can arise through this base substitution; and in fact, the system confirms the A:T → G:C specificity of the mutagen, N⁴-hydroxycytidine (Janion and Glickman, 1980) since only Type II mutants were induced by treatment with this base analogue.When this system was further tested with several additional mutagens, the results indicate that ethyl methanesulphonate, methyl nitrosourea and ethyl nitrosourea produce primarily Type I revertants which were primarily G:C → A:T transitions. UV-light, γ-rays, 4NQO and methyl methanesulphonate produced all types of base substitutions. The tester strain was further improved by introducing a series of sequenced trp^? frameshift mutations, thus allowing the simultaneous monitoring of frameshift and base-substitution mutations.     

Comparative in vitro studies of Tiazofurin and a selenazole analog

2-beta-D-Ribofuranosylselenazole-4-carboxamide, a selenazole analog of the antitumor agent Tiazofurin, is severalfold more cytotoxic to murine tumor cells in culture than Tiazofurin. Like Tiazofurin, the cytotoxicity of the selenazole analog is reversed by exogenous guanosine, and both nucleosides specifically inhibit IMP dehydrogenase activity in cultured P388 cells. The dose-dependency for this inhibition correlates with the relative cytotoxicities of both drugs, indicating that a more potent inhibition of IMP dehydrogenase by the selenazole analog is primarily responsible for its increased cytotoxicity. The specific inhibition of the isolated enzyme by potential metabolites of the selenazole analog is discussed.     

A partial characterization of suppressor cells in rat fetal liver cells

Rat fetal liver cells (FLC) obtained at 18–20 days gestation suppressed mixed lymphocyte reactions(MLR) of adult lymph node cells. The suppression was not strain specific: both syngeneic and allogeneic FLC were capable of suppressing the MLR. The same suppressor activity was observed with fetal spleen cells but not with fetal thymus cells. Removal of phagocytic cells from FLC failed to inhibit the suppressor activity. The suppressor cells were separated into two different types by BSA density gradient: one is radiosensitive, the other radioresistant. A stronger suppressor activity was observed in radiosensitive cells. The suppressor cells belonged to the fraction agglutinated with peanut agglutinin. The data suggest that the suppressor cells in rat FLC may be a proliferating blastoid-type cell rather than mature lymphocytes or mature macrophages.     

Asymmetric binding of cytochrome b5 to the membrane of human erythrocyte ghosts

The intact, amphipatic form of cytochrome $\text{b}{}_5$ could bind to unsealed ghosts, but not to resealed ghosts, suggesting that the cytochrome could bind only to the inner (cytoplasmic) surface of the ghost membrane. This was further confirmed by the finding that the cytochrome could bind to closed, inside-out vesicles prepared from the ghosts. This asymmetric binding was not due to the exclusive localization of sialic acid and sugar chains on the outer surface of the ghosts membrane, because the cytochrome could not bind to ghosts even after enzymatic removal of these components. Although liposomes consisting of phosphatidylcholine or both phosphatidylcholine and sphingomyelin could effectively bind the cytochrome, this binding capacity was progressively decreased as increasing amount of cholesterol was included in the composition of phosphatidylcholine liposomes. Removal of cholesterol from resealed ghosts by incubation with egg phosphatidylcholine liposomes resulted in the binding of cytochrome $\text{b}{}_5$ to the outer surface of the treated ghosts. The possibility is discussed that the asymmetric binding is due to preferential localization of cholesterol in the outer leaflet of the lipid bilayer that constitutes the ghost membrane.     

Paleopathology of Toyotamaphimeia machikanensis (Diapsida,Crocodylia) from the Middle Pleistocene of Central Japan

The type specimen of Toyotamaphimeia machikanensis (Diapsida, Crocodylia) from the Osaka Group (Plio-Pleistocene) of Osaka Prefecture, Central Japan, shows multiple abnormalities. The anterior third of the mandible was amputated and shows signs of healing. The right tibia and fibula were fractured, and each of them was later fused in dislocated position with callus formation. Additionally, there are two puncture marks on a dermal scute. Based on the morphologies of these pathologies, the fauna of the community in which this animal lived, and the behavior of extant crocodilians, it is suggested that this individual survived its injuries sustained during multiple intraspecific fights for territoriality or mating.     

Presence of mast cell precursors in the yolk sac of mice

Concentration of mast-cell precursors in hematopoietic tissues of mouse embryos was evaluated by a limiting dilution method. Cells from yolk sacs, livers, and bodies of (WB x C57BL/6)F1 (hereafter called WBB6F1)- +/+ embryos were injected directly into the skin of adult WBB6F1-W/Wv mice which were genetically depleted of tissue mast cells. Concentration of mast-cell precursors was calculated from the proportion of injection sites at which mast cells did not appear. Since the concentration of mast-cell precursors in the yolk sac was about 30 times as great as that of embryonic body at Day 9.5 of the pregnancy, the mast-cell precursors seemed to be generated within the yolk sac. The concentration in the yolk sac reached the maximum level at Day 11, and then dropped markedly at Day 13. In contrast, mast-cell precursors increased from Day 11 to Day 15 in the fetal liver. As a result, the concentration of 11-day yolk sacs was comparable to that of 15-day fetal liver. Although intravenous injection of 15-day fetal liver cells (2 x 10(6)) rescued the general mast-cell depletion of WBB6F1-W/Wv mice, the intravenous injection of the same number of 11-day yolk sac cells did not rescue it. In contrast with fetal livers, yolk sacs scarcely contained hematopoietic stem cells which were measured by spleen colony formation. Therefore, the mast-cell precursors of the yolk sac may not originate from such stem cells.     

Replacement of large copepods by small ones with eutrophication of embayments: cause and consequence

Species and size compositions of the planktonic copepod community were compared between two eutrophic embayments, Tokyo Bay and Osaka Bay. Within these bays, the median body weight of the community was lowest at the innermost station and increased gradually offshore. In Tokyo Bay, which is more eutrophic than Osaka Bay, the dominant species was Oithona davisae and the median carbon weight of the community was ca 0.1 µg. In Osaka Bay, Paracalanus sp., Calanus sinicus and Corycaeus spp. were dominant and the median weight was 1–2 order of magnitude higher (2–78 µg) than in Tokyo Bay. Some 40 years ago, when eutrophication was less prominent, Acartia omorii, Paracalanus sp. and Microsetella norvegica comprised a considerable portion of the community in Tokyo Bay. The proliferation of O. davisae might have been caused with the recent change in food particle composition and/or dissociated life cycle of this species from the benthic anoxic layer. Decrease in the size composition of the copepod community may make the embayment ecosystem unfavorable for planktivorous fish, but not for jellyfish, e.g. Aurelia and Bolinopsis. This may also result in an acceleration of the flux of biogenic materials from the embayment to the outer ocean.     

Impact of eutrophication on shallow marine benthic foraminifers over the last 150 years in Osaka Bay, Japan

High-resolution foraminiferal analysis was conducted on a short sediment core from the inner part of Osaka Bay, Japan. Changes in foraminiferal assemblages were associated with eutrophication, bottom water hypoxia, and changes in red tide-causing algae. Before the 1920s, the calcareous species Ammonia beccarii, and the agglutinated species Eggerella advena and Trochammina hadai were rare, but calcareous foraminifers in general were abundant. Between the 1920s and 1940s, calcareous foraminifers decreased abruptly in abundance, while A. beccarii, E. advena and T. hadai increased in abundance. This faunal change corresponded in time to an increase in nutrients flowing in through the Yodo River, and bottom water hypoxia related to eutrophication. In the 1960s and 1970s, A. beccarii, E. advena and T. hadai further increased in abundance to become dominant, and many calcareous foraminifers nearly disappeared, corresponding to increasing bottom water hypoxia related to the rapid increase in discharged nutrients during the high economic growth period from 1953 to 1971. After the 1990s, A. beccarii decreased rapidly in abundance and E. advena and Uvigerinella glabra increased in abundance. The main components of red tide-causing algae changed from dinoflagellates to diatoms in the 1980s through 1990s, thus there was a change in the food supply to the benthos, which may have caused the increase in abundance of E. advena and U. glabra.     

Variation in metal concentrations in the brown alga Undaria pinnatifida in Osaka Bay, Japan

In order to evaluate the usefulness of a biomonitoring system using seaweeds for assessing the geographic distribution of metal ions in coastal seawaters, the metal concentrations in the sporophytes of an annual kelp, Undaria pinnatifida (Harvey) Suringar, were collected at 15 localities in Osaka Bay, Japan and compared. About 160 cm² of the blade was cut out from the central part of clean sporophytes, rinsed in filtered seawater using an ultrasonic cleaning bath, and freeze‐dried. After digestion with 12% HNO₃ in a microwave apparatus, metal concentrations in the samples were determined by inductively coupled plasma mass spectrometry. The concentrations (dry weight basis) of most examined elements were in the parts per million range; Cr, 0.48–3.18; Ni, 0.77–5.94; Cu, 3.20–43.8; Zn, 11.3–86.8; Pb, 0.14–3.53. Comparisons of metal compositions of the U. pinnatifida samples from the northeastern area of the bay, which has a large urban population and highly developed industries inland, showed high concentrations of Cd, Pb and Cu compared with the samples from the southwestern area of the bay where the population and industries are much smaller. This suggests that U. pinnatifida metal loads can be used as a marker to track the geographic distributions of the metal concentrations in coastal seawaters, reflecting inland human activities such as shipbuilding and repairing in port areas, and can be used as a useful biomonitoring system of coastal environments for long‐term trend.     

Induction of the differentiation of memory T killer cells with factors released from macrophage-like cell lines

Macrophage-like cell lines J774.1 and WEHI-3 as well as peritoneal exudate macrophages have been demonstrated to produce factors which induce the differentiation of memory cells into specific T killer cells in the absence of an added antigen. LPS stimulation was required for J774.1 cells and peritoneal macrophages to produce the factors but not for WEHI-3 cells. Interferon seemed to be one of the responsible factors. However, macrophages seem to produce other active factors; one has a molecular weight (MW) of more than 80,000 and lacks thymocyte mitogenic activity; another, with a weak thymocyte mitogenic activity, has a MW of 38,000 to 44,000. The low MW thymocyte mitogenic factor (interleukin-1) showed weak T killer cell differentiating activity.     

Occurrence and diversity of barnacles on international ships visiting Osaka Bay,Japan, and the risk of their introduction

Abstract

The occurrence and diversity of barnacles attached to the hulls of two intercontinental bulk carriers were studied at the port in Osaka Bay, Japan, to assess the potential risk of introduction of exotic species to the inner part of the bay. Barnacles were sampled from the bulbous bows, near the midship draft marks, and around the propeller posts and rudders. Twenty-two species were found, with 14 of these not previously recorded in Osaka Bay, including four species not previously recorded elsewhere in Japan. From an assessment of environmental similarity between Osaka Bay and native ranges, of the 14 species, Elminius modestus and Amphibalanus variegatus were considered to have the highest risk of introduction to the inner part of the bay.     

A preliminary simulation model of movement of larvae ofCulex pipiens molestus (Diptera: Culicidae) (Experimental studies on the dispersal of insects,II)

Experiments for analyzing the movement and dispersal of larvae of C. pipiens molestus were made and preliminaly simulation models were constructed. The models were based on two distributions, the distance and direction of movements between 1 minute intervals. The distribution of distance is represented by a unimodal curve, and that of angle by a bimodal one. The results of simulation reasonably depicted the actual process of movements. Nevertheless, the simulated time-dispersal curves overestimated the number of dispersed individuals. This might be due to mutual attraction among the larvae.     

Structural change of myofibrils during mitosis of newt embryonic myocardial cells in culture

Newt embryonic myocardial cells can undergo mitosis in culture. The successive changes in the striation pattern of sarcomeres of myofibrils during mitosis were studied by polarization microscopy without fixing or killing the cells. Birefringence of well-organized striation patterns, i.e., bright A-bands and dark I-bands, was clearly visible in interphase cells and did not show any detectable changes during incubation for 3 h or more. Electron microscopy showed the presence of well-organized myofibrils with Z-bands in these interphase cells. When myocardial cells entered the mitotic stage, the birefringence of striation pattern of their myofibrils gradually changed with the pattern in small parts of the myofibrils gradually becoming indistinct (called 'indistinct striation' in this paper). These indistinct regions increased in size during the mitotic stage. In addition, in some regions of the indistinct striation, the birefringence of sarcomeres gradually decreased and finally disappeared (called 'disappearance of sarcomeres' in this paper). No myocardial cells underwent mitosis without these disruptive changes of the myofibril striation patterns. In the post-mitotic stage, the well-organized striation of the myofibrils reappeared. Electron microscopy showed disorganized sarcomeres without Z-bands in the regions of indistinct striation, and no well-organized myofibrils in the regions where the sarcomeres had disappeared. Thus the well-organized myofibrils with Z-bands became transiently disorganized at least in some parts, during mitosis. They were then reorganized into daughter myocardial cells.     

Method for enzymatic determination of imidazole acetic acid

A method for enzymatic assay of imidazole acetic acid (ImAA) was developed, based on the strict substrate specificity of imidazole acetate monooxygenase from Pseudomonas sp. [Maki et al. (1969) J. Biol. Chem., 244., 2942–2950], which catalyzes concomitant conversion of NADH to NAD⁺. Thus, ImAA was determined by measuring decrease in absorbancy at 340 nm. Tissue extracts were partially purified and/or concentrated by column chromatography on Bio-Rad AG-1 before enzymatic assay. The lowest measurable level of ImAA by this method was 2 nmol.     

Development of modern benthic ecosystems in eutrophic coastal oceans: The foraminiferal record over the last 200 years,Osaka Bay,Japan

The ecosystem dynamics of a modern benthic community in Osaka Bay was studied by analyzing sediment cores and fossil foraminifera deposited during the past 200 years. The results suggest that the high-density/low-diversity assemblage has appeared in the early 1900s, coinciding with the eutrophication of the bay resulting from the Japanese industrial revolution. This assemblage proliferated during the period 1960 to 1970 when the eutrophication and bottom-water hypoxia were most pronounced. The development of the assemblage has been characterized by an increase in the relative and absolute abundance of eutrophication-tolerant species (Ammonia beccarii, Eggerella advena, and Trochammina hadai) and a decrease in many other foraminiferal species, such as Ammonia tepida, Elphidium, Miliolinella subrotunda, and Valvulineria hamanakoensis, that are unable to tolerate low-oxygen conditions. Approximately thirty years after the imposition of discharge restrictions in the 1970s, this assemblage continues to predominate in the inner part of the bay, and E. advena is currently found across the entire bay. These records make a significant contribution to understanding the long-term relationship between anthropogenic impact and ecosystem change.     

beta-Galactosidase deficient-type mucolipidosis: a complementation study of neuraminidase in somatic cell hybrids

By cell fusion with polyethylene glycol (PEG) a remarkable increase of neuraminidase activity was observed in the fused cells between ML-II and other two neuraminidase deficient disorders, ML-Gal (mucolipidosis, galactosidase deficient type) and ML-I (mucolipidosis I). No complementation was found in the combination of ML-I and ML-Gal. This result suggests that ML-I and ML-Gal may be allelic mutations and belong to the same genetic disorder as a primary neuraminidase deficiency.     

Monitoring of the toxic dinoflagellate Alexandrium catenella in Osaka Bay,Japan using a massively parallel sequencing (MPS)-based technique

Since 2002, blooms of Alexandrium catenella sensu Fraga et al. (2015) and paralytic shellfish toxicity events have occurred almost yearly in Osaka Bay, Japan. To better understand the triggers for reoccurring A. catenella blooms in Osaka Bay, phytoplankton community was monitored during the spring seasons of 2012–2015. Monitoring was performed using massively parallel sequencing (MPS)-based technique on amplicon sequences of the 18S rRNA gene. Dense blooms of A. catenella occurred every year except in 2012, however, there was no significant correlation with the environmental parameters investigated. Plankton community diversity decreased before and middle of the A. catenella blooms, suggesting that the decline in diversity could be an indicator for the bloom occurrence. The yearly abundance pattern of A. catenella cells obtained by morphology-based counting coincided with the relative sequence abundances, which supports the effectiveness of MPS-based phytoplankton monitoring.     

First record of an Early Barremian caprinid rudist from Japan – implications for the palaeobiogeography of the Caprinidae (Bivalvia)

Abstract: Pachytraga Paquier, 1900, the stratigraphically oldest genus of caprinine caprinid rudist, was previously known from only two chronospecies from a single lineage, that is the Hauterivian P. tubiconcha Astre, 1961 and the early Aptian P. paradoxa (Pictet and Campiche, 1869). Here, a new species, Pachytraga? tanakahitoshii, is erected on the basis of isolated left valves recovered from the Osaka and Sanchu areas, south‐west Japan. This species has a moderate shell size (antero‐posterior commissural diameter c. 30 mm), and its left valve is characterized by at least one possibly autapomorphic character (narrow anterior myophore, inclined inwards), as well as a mosaic of primitive (single longitudinal carina developed on the anterior side) and derived (simple marginal canals in the antero‐dorsal valve margin) characters of Mediterranean and Middle East Tethyan Pachytraga. The Japanese Pachytraga? represents the first probable record of this genus outside the Mediterranean/Middle Eastern Tethyan province, and its early Barremian age partly fills the ‘gap’ in its previously known stratigraphical record, although the evolutionary relationship of the Japanese form with Mediterranean and Middle East Tethyan Pachytraga remains unsolved. However, the discovery of early Barremian Pachytraga? in Japan indicates that the evolutionary history of the genus is more complex than previously thought and should thus be discussed in a broader palaeogeographical context that must now include the Pacific.