Induction and inhibition of oocyte maturation by EDCs in zebrafish

BACKGROUND: Oocyte maturation in lower vertebrates is triggered by maturation-inducing hormone (MIH), which acts on unidentified receptors on the oocyte surface and induces the activation of maturation-promoting factor (MPF) in the oocyte cytoplasm. We previously described the induction of oocyte maturation in fish by an endocrine-disrupting chemical (EDC), diethylstilbestrol (DES), a nonsteroidal estrogen. METHODS: In this study, stimulatory and inhibitory effects of EDCs and natural steroids on oocyte maturation were examined in zebrafish. For effective agents, some details about the mechanism in induction or inhibition of maturation were examined. Possible groups of DES interacting with the MIH receptor are discussed based on relative potency of steroids to induce maturation. RESULTS: Among agents tested, tamoxifen (TAM) and its metabolite 4-hydroxytamoxifen (4-OHT) showed stimulatory activity similar to DES. The time courses of the change in germinal vesicle breakdown and an intracellular molecular event (the synthesis of cyclin B) induced by TAM were indistinguishable from those induced by MIH. In contrast, pentachlorophenol (PCP) had a potent inhibitory effect on MIH-induced oocyte maturation. PCP inhibited not only MIH-induced maturation but also DES- and TAM-induced maturation. Methoxychlor also inhibited maturation when oocytes were pre-treated with this agent. CONCLUSION: These results suggest that EDCs act as agonists or antagonists in the induction of oocyte maturation in fish

Identification of α-type subunits of the Xenopus 20S proteasome and analysis of their changes during the meiotic cell cycle

BACKGROUND: The 26S proteasome is the proteolytic machinery of the ubiquitin-dependent proteolytic system responsible for most of the regulated intracellular protein degradation in eukaryotic cells. Previously, we demonstrated meiotic cell cycle dependent phosphorylation of α4 subunit of the 26S proteasome. In this study, we analyzed the changes in the spotting pattern separated by 2-D gel electrophoresis of α subunits during Xenopus oocyte maturation. RESULTS: We identified cDNA for three α-type subunits (α1, α5 and α6) of Xenopus, then prepared antibodies specific for five subunits (α1, α3, α5, α6, and α7). With these antibodies and previously described monoclonal antibodies for subunits α2 and α4, modifications to all α-type subunits of the 26S proteasome during Xenopus meiotic maturation were examined by 2D-PAGE. More than one spot for all subunits except α7 was identified. Immunoblot analysis of 26S proteasomes purified from immature and mature oocytes showed a difference in the blots of α2 and α4, with an additional spot detected in the 26S proteasome from immature oocytes (in G2-phase). CONCLUSIONS: Six of α-type subunits of the Xenopus 26S proteasome are modified in Xenopus immature oocytes and two subunits (α2 and α4) are modified meiotic cell cycle-dependently

Dynamic Finite Element Analysis of Impulsive Stress Waves Propagating from the Greater Trochanter of the Femur by a Sideways Fall

Fall accidents are a common cause of femoral fracture in the elderly. The greater trochanter of the femur is often subjected to impact loading by a sideways fall, and thus it is worth studying the impulsive stress waves propagating in the femur. In this study, the impulsive stress was analyzed by the dynamic finite element method using a 3-dimensional model of the femur, and the influence of the fall configuration on the stress was discussed. The stress was concentrated around the femoral neck during the propagation of the stress wave, and the tensile maximum principal stress changed into compressive minimum principle stress on the anterior and medial sides of the neck. On the other hand, the compressive minimum principal stress changed into tensile maximum principle stress on the lateral side of the neck. The largest maximum principal stress during the impact loading was always larger in the neck than in the impact region. The largest absolute value of the minimum principal stress was found in the neck or the impact region depending on the fall configuration. The largest absolute values of the maximum and minimum principal stress were nearly equal, indicating that the bone fracture due to the tensile stress may occur around the femoral neck

Photo-response of the conductivity in functionalized pentacene compounds

We report the first investigation of the photo-response of the conductivity of a new class of organic semiconductors based on functionalized pentacene. These materials form high quality single crystals that exhibit a thermally activated resistivity. Unlike pure pentacene, the functionalized derivatives are readily soluble in acetone, and can be evaporated or spin-cast as thin films for potential device applications. The electrical conductivity of the single crystal materials is noticeably sensitive to ambient light changes. The purpose, therefore, of the present study, is to determine the nature of the photo-response in terms of carrier activation vs. heating effects, and also to measure the dependence of the photo-response on photon energy. We describe a new method, involving the temperature dependent photo-response, which allows an unambiguous identification of the signature of heating effects in materials with a thermally activated conductivity. We find strong evidence that the photo-response in the materials investigated is predominantly a highly localized heating mechanism. Wavelength dependent studies of the photo-response reveal resonant features and cut-offs that indicate the photon energy absorption is related to the electronic structure of the material.Comment: Preprint: 18 pages total,7 figure

Regulated interaction between polypeptide chain elongation factor-1 complex with the 26S proteasome during Xenopus oocyte maturation

BACKGROUND: During Xenopus oocyte maturation, the amount of a 48 kDa protein detected in the 26S proteasome fraction (p48) decreased markedly during oocyte maturation to the low levels seen in unfertilized eggs. The results indicate that the interaction of at least one protein with the 26S proteasome changes during oocyte maturation and early development. An alteration in proteasome function may be important for the regulation of developmental events, such as the rapid cell cycle, in the early embryo. In this study, we identified p48. RESULTS: p48 was purified by conventional column chromatography. The resulting purified fraction contained two other proteins with molecular masses of 30 (p30) and 37 (p37) kDa. cDNAs encode elongation factor-1γ and δ were obtained by an immuno-screening method using polyclonal antibodies against purified p48 complex, which recognized p48 and p37. N-terminal amino acid sequence analysis of p30 revealed that it was identical to EF-1β. To identify the p48 complex bound to the 26S proteasome as EF-1βγδ, antibodies were raised against the components of purified p48 complex. Recombinant EF-1 β,γ and δ were expressed in Escherichia coli, and an antibody was raised against purified recombinant EF-1γ. Cross-reactivity of the antibodies toward the p48 complex and recombinant proteins showed it to be specific for each component. These results indicate that the p48 complex bound to the 26S proteasome is the EF-1 complex. MPF phosphorylated EF-1γ was shown to bind to the 26S proteasome. When EF-1γ is phosphorylated by MPF, the association is stabilized. CONCLUSION: p48 bound to the 26S proteasome is identified as the EF-1γ. EF-1 complex is associated with the 26S proteasome in Xenopus oocytes and the interaction is stabilized by MPF-mediated phosphorylation

Debye relaxation in high magnetic fields

Dielectric relaxation is universal in characterizing polar liquids and solids, insulators, and semiconductors, and the theoretical models are well developed. However, in high magnetic fields, previously unknown aspects of dielectric relaxation can be revealed and exploited. Here, we report low temperature dielectric relaxation measurements in lightly doped silicon in high dc magnetic fields B both parallel and perpendicular to the applied ac electric field E. For B//E, we observe a temperature and magnetic field dependent dielectric dispersion e(w)characteristic of conventional Debye relaxation where the free carrier concentration is dependent on thermal dopant ionization, magnetic freeze-out, and/or magnetic localization effects. However, for BperpE, anomalous dispersion emerges in e(w) with increasing magnetic field. It is shown that the Debye formalism can be simply extended by adding the Lorentz force to describe the general response of a dielectric in crossed magnetic and electric fields. Moreover, we predict and observe a new transverse dielectric response EH perp B perp E not previously described in magneto-dielectric measurements. The new formalism allows the determination of the mobility and the ability to discriminate between magnetic localization/freeze out and Lorentz force effects in the magneto-dielectric response.Comment: 19 pages, 6 figure

Preventive Effects of Health-Food Drinks on the Obesity and DNA Damage of Mice Fed a High-Fat Diet under a Mild Stress

The body weight gain of mice fed an obesity-inducing diet is suppressed by a mild restraint stress, but at the same time, as previously reported, the stress induces DNA damage in the cells of multiple organs. In the present study, we attempted to prevent not only the obesity but also the DNA damage of mice fed an obesity-inducing diet under a mild restraint with commercially available health-food drinks such as fruit and vegetable juices, soymilks, vinegars and lactic acid bacteria drinks, which are rich in antioxidants. The body weight gain of young female mice fed a high-fat diet containing 20% fat for 4 weeks was considerably inhibited by restraint for 15 min per day in weeks 2 to 4 of 4-week period. The inhibition was further promoted with the concomitant administration of health-food drinks noted above, and was accompanied by a decrease of periovular fat, a major abdominal fat in the female mice. The definite loss of energy intake in the mice given health-food drinks was approximately compensated by the energy of the drink administered. On the other hand, the increase of DNA damage generated by restraint in the cells of five organs – the liver, pancreas, spleen, heart and bone marrow – was markedly suppressed with the administration of these drinks. The results suggest that a combination of mild stress and intake of suitable health-food containing some antioxidants may inhibit lifestyle-related diseases including hyperlipidemia and obesity, which may contribute to the inhibition of metabolic syndrome and childhood obesity