Photopharmacological Manipulation of Mammalian CRY1 for Regulation of the Circadian Clock

CRY1 and CRY2 proteins are highly conserved components of the circadian clock that controls daily physiological rhythms. Disruption of CRY functions are related to many diseases, including circadian sleep phase disorder. Development of isoform-selective and spatiotemporally controllable tools will facilitate the understanding of shared and distinct functions of CRY1 and CRY2. Here, we developed CRY1-selective compounds that enable light-dependent manipulation of the circadian clock. From phenotypic chemical screening in human cells, we identified benzophenone derivatives that lengthened the circadian period. These compounds selectively interacted with the CRY1 photolyase homology region, resulting in activation of CRY1 but not CRY2. The benzophenone moiety rearranged a CRY1 region called the "lid loop"located outside of the compound-binding pocket and formed a unique interaction with Phe409 in the lid loop. Manipulation of this key interaction was achieved by rationally designed replacement of the benzophenone with a switchable azobenzene moiety whose cis-trans isomerization can be controlled by light. The metastable cis form exhibited sufficiently high half-life in aqueous solutions and structurally mimicked the benzophenone unit, enabling reversible period regulation over days by cellular irradiation with visible light. This study revealed an unprecedented role of the lid loop in CRY-compound interaction and paves the way for spatiotemporal regulation of CRY1 activity by photopharmacology for molecular understanding of CRY1-dependent functions in health and disease

Preliminary results of the TH99 geological and geophysical survey in the Cooperation Sea and Prydz Bay area

Geophysical and geological surveys were carried out in the 1999-2000 austral summer season for the TH99 cruise aboard the R/V Hakurei-maru. The survey includes the abyssal basin and the continental rise area of the Cooperation Sea (CS) and Prydz Bay (PB), offshore of Mac Robertson Land and Princess Elizabeth Land, Antarctica. In the northern abyssal plain of the CS, a mantle-like sequence (G) appears at a depth of 10 s two-way travel time (TWT), or approximately 13 km. The sequence disappears south of 64°S, in the southern part of the CS. These observations suggest that the southern part of the CS and PB is underlain by intermediate type crust that is characteristic of oceanic and continental material. The deep structure of most of the PB continental shelf is not clear due to strong seafloor multiples. More than 7 km of thick sedimentary sequences in the northwest area of PB are confirmed by the MCS survey. Very thick sedimentary sequences, interpreted as prerift, rift and drift sediment can be observed under thick pelagic sediment. Geological samples, which were recovered in the CS and PB, offer important evidence on the recent history of drift sediments caused by iceberg floes. Two cores reached 0.78 Ma (Bruhnes-Matsuyama (B-M) boundary) in magneto-stratigraphic measurements, and they gave distinct age determinations for recent sediments

The Role of Heparan Sulfate Proteoglycans as an Attachment Factor for Rabies Virus Entry and Infection

Rabies virus (RABV) is the causative agent of fatal neurological disease. Cellular attachment is the initial and essential step for viral infections. Although extensive studies have demonstrated that RABV uses various target cell molecules to mediate infection, no specific molecule has been identified as an attachment factor for RABV infection. Here we demonstrate that cellular heparan sulfate (HS) supports RABV adhesion and subsequent entry into target cells. Enzymatic removal of HS reduced cellular susceptibility to RABV infection, and heparin, a highly sulfated form of HS, blocked viral adhesion and infection. The direct binding between RABV glycoprotein and heparin was demonstrated, and this interaction was shown to require HS N- and 6-O-sulfation. We also revealed that basic amino acids in the ectodomain of RABV glycoprotein serve as major determinants for the RABV-HS interaction. Collectively, our study highlights a previously undescribed role of HS as an attachment factor for RABV infection

Performance of a Biodegradable Composite with Hydroxyapatite as a Scaffold in Pulp Tissue Repair

Vital pulp therapy is an important endodontic treatment. Strategies using growth factors and biological molecules are effective in developing pulp capping materials based on wound healing by the dentin-pulp complex. Our group developed biodegradable viscoelastic polymer materials for tissue-engineered medical devices. The polymer contents help overcome the poor fracture toughness of hydroxyapatite (HAp)-facilitated osteogenic differentiation of pulp cells. However, the composition of this novel polymer remained unclear. This study evaluated a novel polymer composite, P(CL-co-DLLA) and HAp, as a direct pulp capping carrier for biological molecules. The biocompatibility of the novel polymer composite was evaluated by determining the cytotoxicity and proliferation of human dental stem cells in vitro. The novel polymer composite with BMP-2, which reportedly induced tertiary dentin, was tested as a direct pulp capping material in a rat model. Cytotoxicity and proliferation assays revealed that the biocompatibility of the novel polymer composite was similar to that of the control. The novel polymer composite with BMP-2-induced tertiary dentin, similar to hydraulic calcium-silicate cement, in the direct pulp capping model. The BMP-2 composite upregulated wound healing-related gene expression compared to the novel polymer composite alone. Therefore, we suggest that novel polymer composites could be effective carriers for pulp capping