Bacteriophages isolated from activated sludge processes and their polyvalency

In this study, bacteriophages were isolated from activated sludge and their host range was studied. Bacterial isolates were obtained from an activated sludge process treating urban sewage, and bacteriophages were obtained by plaque assay using the bacterial isolates obtained in this study as the host. Out of 15 bacteria isolated, 9 supported plaque formation. The host range test was conducted with a combination of 8 bacteriophage isolates and 9 bacterial isolates. All of the 8 bacteriophages tested were found to form plaques on more than 1 host, and 4 of them formed plaques on both gram-positive and gram-negative bacterial isolates. Three of the 8 bacteriophages failed to form plaques on their original bacterial host. The experimental result indicates that bacteriophages are an active part of the activated sludge microbial ecosystem, having a very close ecological relationship with their host bacteria.     

Evidence that activation of Src family kinase is not required for fertilization-associated [Ca2+]i oscillations in mouse eggs

Recent evidence in marine invertebrate, frog, and zebrafish eggs suggests the involvement of a Src family kinase (SFK) in fertilization-induced Ca2+ release. In the present study, we have investigated whether activation of an SFK is required for initiation of intracellular Ca2+ ([Ca2+]i) oscillations in mouse fertilization. We detected a Hck-like protein and tyrosine-phosphorylated proteins in soluble and insoluble sperm fractions, respectively. However, the presence of these proteins did not correspond to the active fractions of porcine sperm extracts (pSE). Moreover, [Ca2+]i oscillations induced by pSE in mouse eggs were unaltered by pre-incubation of pSE with specific SFK inhibitors such as 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazol[3,4-d]-pyrimidine (PP2) or lavendustin A, despite the fact that the inhibitors were shown to be active both in vivo and in vitro. Another SFK inhibitor, peptide A, blocked oscillations when incubated with pSE prior to injection into eggs, but this inhibition required more than ten times the concentration reportedly required to inhibit SFK activity. In addition, pre-injection or pre-incubation of eggs with these inhibitors did not affect the ability of pSE to trigger [Ca2+]i oscillations in mouse eggs. Microinjection of a recombinant c-Src protein or mRNAs encoding constitutively active Src proteins did not induce [Ca2+]i release. Finally, when sperm and eggs, both of which were pre-treated with PP2, were fertilized, [Ca2+]i oscillations occurred normally. We can therefore conclude that activation of an SFK is neither necessary nor sufficient for triggering fertilization-induced [Ca2+]i oscillations.     

Calbindin D28K-like immunoreactive nerve fibres in the predentine of rat molar teeth

Immunoelectron-microscopy was applied to reveal the existence of nerve fibres and terminals showing calbindin D28k (CB)-like immunoreactivity (IR) in the rat molar tooth pulp. In the root pulp, thick, smooth-surfaced CB-IR nerve fibres were in bundles accompanying the blood vessels. In the coronal pulp, the fibres arborized repeatedly and extensively. CB-IR nerve fibres had a predominantly thick, smooth-surfaced appearance, though parts appeared thin and beaded. Occasionally some thin, varicose CB-IR nerve fibres ran along the odontoblasts, penetrating into the predentine alongside the dentinal tubules. They could be traced for approx. 10-20 microns into the predentine from the pulp-predentine border. Immunoelectron-microscopy revealed that only some of the nerve terminals in the predentine showed CB-IR, and that predentinal CB-IR nerve terminals were located close to the odontoblast processes. No synaptic structures were observed between them. The presence of CB-IR nerve terminals in the predentine suggests that many, if not all, CB-IR nerve fibres could be nociceptors. The CB could be involved in Ca2+ homeostasis during the activation of nociceptors.     

Electron cyclotron resonance plasma production by using pulse mode microwaves and dependences of ion beam current and plasma parameters on the pulse condition

We measure the ion beam current and the plasma parameters by using the pulse mode microwave operation in the first stage of a tandem type ECRIS. The time averaged extracted ion beam current in the pulse mode operation is larger than that of the cw mode operation with the same averaged microwave power. The electron density n(e) in the pulse mode is higher and the electron temperature T(e) is lower than those of the cw mode operation. These plasma parameters are considered to cause in the increase of the ion beam current and are suitable to produce molecular or cluster ions.     

Estimation of source contribution to nitrate loading in road runoff using stable isotope analysis

The nitrogen load was determined in road runoff during rainfall events. Moreover, nitrate isotopes analysis was conducted to determine the contribution of nitrates from atmospheric deposition and leaching from road dust. The concentrations of NO₃-N in road runoff were higher than those in atmospheric deposits for each rainfall event, except one event with a long antecedent dry weather period. The δ¹⁸O-NO₃ in road runoff was lower than in atmospheric deposits and higher than in leachate from road dust; however, no difference in δ¹⁵N-NO₃ was observed. By using δ¹⁸O-NO₃ as an indicator for evaluating NO₃-N sources in road runoff, contribution ratios of NO₃-N from road dust were estimated to be 14–22%, 23–25%, and 22–34% for Event 1 to Event 3, respectively. These results indicated that the NO₃-N from the atmosphere accounts for more than half of the NO₃-N in road runoff.     

Hepatocyte growth factor is involved in the morphogenesis of tooth germ in murine molars

The patterns of gene expression for hepatocyte growth factor (HGF) and its receptor, c-Met, were revealed in the tooth germ of rat mandibular molars using RT-PCR. In situ hybridization demonstrated that the HGF gene was expressed only in the cells of the dental papilla of the tooth germ in vivo. The characteristic temporospatial distribution of HGF and c-Met during germ development was revealed using immunohistochemical studies in vivo. In order to demonstrate the functional role played by HGF in tooth development, HGF translation arrest by antisense phosphorothioate oligodeoxynucleotide (ODN) was carried out in vitro. In the control experiment, explants of tooth germs from embryonic 14 day mice were cultured in a modification of Trowell's system under serum-free and chemically defined conditions for two weeks. Other explants were cultured with 15mer antisense or sense ODN targeted to the HGF mRNA. Both the control and the sense-treated explants showed normal histological structure, as observed in vivo. On the other hand, antisense-treated explants exhibited an abnormal structure in which the enamel organs were surrounded by a thin layer of dentin and dental papilla, appearing 'inside-out' compared to the control and sense-treated explants, although the cytodifferentiation of ameloblasts and odontoblasts was not inhibited. The explants treated with recombinant human HGF combined with antisense ODN showed normal development, indicating that exogenous HGF rescued the explants from the abnormal structure caused by antisense ODN. The findings of a BrdU incorporation experiment suggested that the imbalance between the proliferation activity of the inner enamel epithelium and that of the dental papilla caused by HGF translation arrest results in the abnormal structure of the tooth germ. These results indicate that HGF is involved in the morphogenesis of the murine molar.     

Relationship between the energy distribution of electrostatic field change and lightning activity

A lightning strength parameter α (denoted as LSP or LAIDO), which is used for characterizing the phenomena of lightning activity, has been proposed, where α is defined by the characteristic energy obtained from the energy distribution of a radiated field component E in the electrostatic field changes produced by lightning discharge. Here, it is observed that the energy distribution of E_γ components is Maxwellian type. To prove more clearly the forementioned assumption from another viewpoint, changes of the amplitudes of the electrostatic field component E_S were measured and their distribution functions constructed. In the cloud-to-ground lightning discharges, the E_S components have a positive sign in the changes. However, those changes in the intercloud discharges are observed with either a negative or positive change depending on the distance between the striking point and the observation site. Therefore it is necessary to sum the numbers of occurrences of both positive and negative components for constructing an energy distribution of the E_S component. A fieldmill and a pair of doughnut-shaped electrostatic antennas are used in measuring the changes of the E_S component. As a result it is found that the changes of the amplitudes of the E_S components have a Maxwelliantype energy distribution. A characteristic energy β is defined which is obtained from the component of the electrostatic fields as a measure of LSP. Based on comparison of α and β it is clarified that β has the same characteristic nature with α. The lightning activity could be estimated from β.