MAP kinase links the fertilization signal transduction pathway to the G1/S-phase transition in starfish eggs

The mechanism by which fertilization initiates S-phase in the zygote is examined by manipulating the activity of MAP kinase in mature starfish eggs. These unfertilized eggs, which are arrested at G1-phase after the completion of meiosis, have high MAP kinase activity but undetectable cdc2 kinase activity. Either fertilization or inhibition of protein synthesis causes a decrease in MAP kinase activity, which is followed by DNA synthesis. Inactivation of MAP kinase with its specific phosphatase, CL100, initiates DNA synthesis in the absence of fertilization, while constitutive activation of MAP kinase with MEK represses the initiation of DNA synthesis following fertilization. Thus, in unfertilized mature starfish eggs, a capacity for DNA replication is already acquired, but entry into S-phase is negatively regulated by MAP kinase activity that is supported by a continuously synthesized protein(s) but not by cdc2 kinase. Upon fertilization, downregulation of MAP kinase activity is necessary and sufficient for triggering the G1/S-phase transition.     

Visibility of Vortex Core in Fermionic Superfluid with Population Imbalance

In Fermionic superfluids with a vortex, at T=0, the depletion of the atomic density appears in the core region, which is strongly associated with the discreteness of the core-bound state, called the Caroli-de Gennes-Matricon state. In imbalanced superfluid, however, it is found by the microscopic study based on the Bogoliubov-de Gennes approximation that this quantum depletion is progressively filled out in majority spin species as population imbalance increases. In contrast, the minority species keeps the depletion, which enables the direct observation of “superfluidity”, because the quantized vortex is a hallmark of superfluidity.     

Josephson Vortex Flow and Plasma Amplification in High-T c Superconductors

We investigate Josephson vortex flow states in two device arrangements considered to amplify the excited Josephson plasma. First, we study an intrinsic Josephson junction embedded inside a wave guide. The simulation result in the system reveals that the amplitude of the excited electric field almost linearly grows along the junction plane at the resonant voltage with the Josephson plasma. Second, we simulate a system composed of two separated Josephson junctions embedded inside a long wave guide and confirm that the excited electric field is amplified by the synchronization between two junction sites.     

Farnesol-induced generation of reactive oxygen species via indirect inhibition of the mitochondrial electron transport chain in the yeast Saccharomyces cerevisiae

The mechanism of farnesol (FOH)-induced growth inhibition of Saccharomyces cerevisiae was studied in terms of its promotive effect on generation of reactive oxygen species (ROS). The level of ROS generation in FOH-treated cells increased five- to eightfold upon the initial 30-min incubation, while cells treated with other isoprenoid compounds, like geraniol, geranylgeraniol, and squalene, showed no ROS-generating response. The dependence of FOH-induced growth inhibition on such an oxidative stress was confirmed by the protection against such growth inhibition in the presence of an antioxidant such as alpha-tocopherol, probucol, or N-acetylcysteine. FOH could accelerate ROS generation only in cells of the wild-type grande strain, not in those of the respiration-deficient petite mutant ([rho0]), which illustrates the role of the mitochondrial electron transport chain as its origin. Among the respiratory chain inhibitors, ROS generation could be effectively eliminated with myxothiazol, which inhibits oxidation of ubiquinol to the ubisemiquinone radical by the Rieske iron-sulfur center of complex III, but not with antimycin A, an inhibitor of electron transport that is functional in further oxidation of the ubisemiquinone radical to ubiquinone in the Q cycle of complex III. Cellular oxygen consumption was inhibited immediately upon extracellular addition of FOH, whereas FOH and its possible metabolites failed to directly inhibit any oxidase activities detected with the isolated mitochondrial preparation. A protein kinase C (PKC)-dependent mechanism was suggested to exist in the inhibition of mitochondrial electron transport since FOH-induced ROS generation could be effectively eliminated with a membrane-permeable diacylglycerol analog which can activate PKC. The present study supports the idea that FOH inhibits the ability of the electron transport chain to accelerate ROS production via interference with a phosphatidylinositol type of signal.     

Depletion of glutathione as a cause of the promotive effects of polygodial, a sesquiterpene on the production of reactive oxygen species in Saccharomyces cerevisiae

The pungent sesquiterpenoid unsaturated dialdehyde, polygodial, exhibited a strong yeastcidal activity against the cells of Saccharomyces cerevisiae, in which production of reactive oxygen species (ROS) at a significant level could be detected with a fluorescent probe. The production of ROS in polygodial-treated cells was further confirmed by its elimination and the accompanying protection against yeastcidal effects in the presence of antioxidants such as L-ascorbate and alpha-tocopherol (alpha-TOH). Polygodial could accelerate ROS production only in cells of the wild-type grande strain but not in those of the respiratory-deficient petite mutant (rho0), indicating the role of the mitochondrial electron transport chain in the production of ROS. Unlike the case with antimycin A which accelerates ROS production by directly targeting the mitochondrial electron flow, polygodial caused depletion of cytoplasmic and mitochondrial glutathione which functions in estiminating ROS inevitably generated during aerobic growth. Polygodial-mediated depletion of intracellular glutathione was possibly dependent on a direct interaction between its enal moiety and the sulfhydryl group of the cysteine in glutathione by a Michael-type reaction.     

Drusenoid Pigment Epithelial Detachment: Genetic and Clinical Characteristics

Few studies report drusenoid pigment epithelial detachment (DPED) in Asians. In this multicenter study, we report the clinical and genetic characteristics of 76 patients with DPED, and, for comparison, 861 patients with exudative age-related macular degeneration (AMD) were included. On the initial presentation, the mean best-corrected visual acuity was 0.087 ± 0.17 (logMAR unit), and mean DPED height and width were 210 ± 132 and 1633 ± 1114 µm, respectively. Fifty-one (67%) patients showed macular neovascularization in the contralateral eye. The risk allele frequency of both ARMS2 A69S and CFH I62V was significantly higher in DPED than in typical AMD and polypoidal choroidal vasculopathy (PCV) (ARMS2 A69S risk allele frequency: DPED 77% vs. typical AMD 66% vs. PCV 57%, CFH I62V risk allele frequency: DPED 87% vs. typical AMD 73% vs. PCV 73%), although the risk allele frequency of both genes was similar between the DPED group and retinal angiomatous proliferation (RAP) group (ARMS2 A69S: p = 0.32, CFH I62V, p = 0.11). The prevalence of reticular pseudodrusen (RPD) was highest in RAP (60%), followed by DPED (22%), typical AMD (20%), and PCV (2%). Although the prevalence of RPD differs between DPED and RAP, these entities share a similar genetic background in terms of ARMS2 and CFH genes.     

Position Dependent Response of Superconducting MgB2 Neutron Detectors Studied by Pulsed Laser Irradiation

The MgB₂ detector consists of 200?nm thick MgB₂ thin film meander structure of 3?μm line width protected by 300?nm thick SiO layer. The electrical connections were provided by 150?nm thick Nb. The devices were placed in a 4?K refrigerator to investigate the behavior at temperatures below T _c. The position-dependent response of the MgB₂ detector can be investigated by scanning a laser spot with the aid of the XYZ piezo-driven stage and an optical fiber with a focusing lens. We found that the inhomogeneity in a detector is very critical to specify the detection efficiency since the detector has the sensitivity only in a narrow temperature regime near T _c.     

PNA‐Encoded Synthesis (PES) of a 10 000‐Member Hetero‐Glycoconjugate Library and Microarray Analysis of Diverse Lectins

Identification of selective and synthetically tractable ligands to glycan‐binding proteins is important in glycoscience. Carbohydrate arrays have had a tremendous impact on profiling glycan‐binding proteins and as analytical tools. We report a highly miniaturized synthetic format to access nucleic‐acid‐encoded hetero‐glycoconjugate libraries with an unprecedented diversity in the combinations of glycans, linkers, and capping groups. Novel information about plant and bacterial lectin specificity was obtained by microarray profiling, and we show that a ligand identified on the array can be converted to a high‐affinity soluble ligand by straightforward chemistry.     

Biochemical Studies of Mitochondrial Malate: Quinone Oxidoreductase from Toxoplasma gondii

Toxoplasma gondii is a protozoan parasite that causes toxoplasmosis and infects almost one-third of the global human population. A lack of effective drugs and vaccines and the emergence of drug resistant parasites highlight the need for the development of new drugs. The mitochondrial electron transport chain (ETC) is an essential pathway for energy metabolism and the survival of T. gondii. In apicomplexan parasites, malate:quinone oxidoreductase (MQO) is a monotopic membrane protein belonging to the ETC and a key member of the tricarboxylic acid cycle, and has recently been suggested to play a role in the fumarate cycle, which is required for the cytosolic purine salvage pathway. In T. gondii, a putative MQO (TgMQO) is expressed in tachyzoite and bradyzoite stages and is considered to be a potential drug target since its orthologue is not conserved in mammalian hosts. As a first step towards the evaluation of TgMQO as a drug target candidate, in this study, we developed a new expression system for TgMQO in FN102(DE3)TAO, a strain deficient in respiratory cytochromes and dependent on an alternative oxidase. This system allowed, for the first time, the expression and purification of a mitochondrial MQO family enzyme, which was used for steady-state kinetics and substrate specificity analyses. Ferulenol, the only known MQO inhibitor, also inhibited TgMQO at IC₅₀ of 0.822 μM, and displayed different inhibition kinetics compared to Plasmodium falciparum MQO. Furthermore, our analysis indicated the presence of a third binding site for ferulenol that is distinct from the ubiquinone and malate sites.