Development of Selective TGR5 Ligands Based on the 5,6,7,8-Tetrahydro-5,5,8,8-tetramethylnaphthalene Skeleton

TGR5, a G-protein-coupled receptor (GPCR), plays an important role in several physiological functions. TGR5 activation through bile acids induces an increase in energy expenditure. Therefore, synthetic TGR5 ligands could be useful for the treatment of obesity or dyslipidemia. In this study, we designed and synthesized a set of TGR5 ligands with a 5,6,7,8-tetrahydro-5,5,8,8-tetramethylnaphthalene (TMN) skeleton, and evaluated their TGR5 agonistic activity. We also investigated the selectivity of the synthesized compounds for TGR5 relative to the farnesoid X receptor (FXR) and retinoic acid receptor (RAR). Our results show that compound 4 b [N-(2-chlorophenyl)-5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenecarboxamide] exhibited potent TGR5 agonist activity with an IC₅₀ value of 8.4 nM without significant cytotoxicity. In addition, compound 4 b showed only slight agonistic activity toward FXR and RAR at 1 μM treatment. These data indicate that compound 4 b is a selective TGR5 agonist, and could be a promising therapeutic agent for dyslipidemia.     

Restoration of solid oxide fuel cell stacks after failure of partial cells

We investigated a solid oxide fuel cell stack that employs anode-supported planar cells in which two intermediate plates are installed every 10 cells to determine the influence of the separation and reconnection of the intermediate plates after high temperature operation. We showed that this separation and reconnection caused no significant degradation in stack performance. A 30-cell stack, which was constructed by removing two 10-cell sub-stacks from a 50-cell stack that had operated stably 1200 h, functioned well. The difference between the average voltages of the cells in the 50- and 30-cell stacks was less than 3% when the current density, fuel utilization, and oxygen utilization were 0.30 A cm⁻², 60%, and 15%, respectively. The 30-cell stack operated stably for about 1200 h with almost no degradation. These findings indicate that our stack can be restored after cells in the stack have broken down simply by removing the 10-cell sub-stacks that contain the broken cells and replacing them with undamaged 10-cell sub-stacks.     

Irradiation properties of T0 chopper components

We investigated the irradiation properties of the components of a T0 chopper. The organic materials in the rotor bearing grease, the magnetic fluids in seals, and the rubber in the timing belt, as well as the semiconductor materials in the rotation sensor and motor encoder, were all irradiated with high-energy γ-rays up to 100 kGy. No significant damage that shortens the lifetime of a T0 chopper was observed for the mechanical components. However, the semiconductor components were damaged by the irradiation. For the rotation sensor system detecting the rotor phase, the signal from a marker on the rotor shaft was transmitted outside the shielding by an optical fiber with radiation-proofing and the electrical circuits were removed from the beamline shielding. The lifetime of the motor encoder possibly meets the requirement for the maintenance period of the T0 chopper.     

Phonon Dispersion Measurements of YBa2Cu3O6.15 and YBa2Cu3O6.95 by Time-of-Flight Neutron Spectroscopy

We measured the phonon dispersions of YBa₂Cu₃O_6.15 and YBa₂Cu₃O_6.95 by time-of-flight inelastic neutron scattering. The in-plane bond-stretching modes in the metallic phase showed a distinct a-b plane anisotropy beyond what is expected for structural origin. Such anisotropy in the longitudinal optical modes, which is absent in the TO, suggests strong in-plane anisotropy in the underlying electronic structure. Apical oxygen bond-stretching modes showed a large frequency change between the insulating and the metallic phases. This large softening also is beyond structural origin, and suggests the effect of local electronic environment.     

Purification and characterization of recombinant human granulocyte colony-stimulating factor (rhG-CSF) derivatives: KW-2228 and other derivatives

Various derivatives of recombinant human granulocyte colony-stimulating factor (rhG-CSF) have been overproduced in Escherichia coli with the strong, inducible trp promoter. A derivative designated as KW-2228 in which the amino acids were replaced at five positions showed more potent granulopoietic activity and stability than those of wild-type both in vitro and in vivo. The purification involved a sequential renaturation process and three-step chromatography. Refolding succeeded in very high yield using a urea system. The purity of KW-2228 was greater than 99% as measured by SDS-PAGE and HPLC analysis. According to circular dichroism and nuclear magnetic resonance spectroscopy, rhG-CSF and KW-2228 have very similar conformations. This suggests that the substitution of five amino acids does not appreciably change the conformation of hG-CSF. KW-2228 ([Ala1, Thr3, Tyr4, Arg5, and Ser17]-hG-CSF) and derivative A ([Ala1, Thr3, Tyr4, Arg5]-hG-CSF) are easily crystallized and they show similar in vitro activity. On the other hand, neither rhG-CSF nor derivative B ([Ser17]-hG-CSF) are crystallized under the same conditions. Thus, the four amino acid substitution (Ala1, Thr3, Tyr4, Arg5) of the N-terminal sequence may facilitate crystallization. The change of Cys17 to Ser may not influence the stability and activity of hG-CSF.     

The role of ATP-sensitive potassium channels in striatal dopamine release: an in vivo microdialysis study

Inactivation of the retinoblastoma (RB) gene is known to be implicated in the pathogenesis of several types of human cancers. Since structural alterations of the RB gene have not been well examined in human bladder cancer, we looked for mutations in the entire coding region of this gene using polymerase chain reaction (PCR) and single-strand conformational polymorphism analysis of RNA. We also examined allelic loss of the RB gene using PCR-based restriction fragment length polymorphism analysis. Of 30 samples obtained from patients with bladder cancer, eight (27%) were found to have RB gene mutations. DNA sequencing of the PCR products revealed five cases with single point mutations and three cases with small deletions. These mutations included one (10%) of ten low-grade (grade 1) tumours, four (50%) of eight intermediate-grade (grade 2) tumours and three (25%) of 12 high-grade (grade 3) tumours. Likewise, mutations were found in four (21%) of 19 superficial (pTa and pT1) tumours and four (36%) of 11 invasive (pT2 or greater) tumours. In 15 informative cases, loss of heterozygosity at the RB locus was shown in five cases (33%), three cases with RB mutations and two without them. These results suggest that RB gene mutations are involved in low-grade and superficial bladder cancers as well as in high-grade and invasive cancers.     

Study on local immune response in diabetic mice, in which bactericidal capacity of the neutrophils had been damaged--Escherichia coli induced experimental urinary tract infection

An experimental ascending urinary tract infection was induced by transurethral instillation of Escherichia coli in streptozotocin (STZ)-induced diabetic mice, in which bactericidal capacity of the perineal exudating neutrophils had been damaged. The local immune response in uninfected diabetic mice and the response time fluctuation of local immune responses in diabetic mice following infection were compared with those in normal mice, and the nature of induced changes in the local immune response was determined. The diabetic model was prepared by a single administration of 280 mg/kg of STZ after it had been fasted for 15 hours. The mouse was then fasted an additional 2 hours to induce the diabetic state. Compared with the normal mice, the diabetic mice prepared using this method were found to have a significant suppression of the bactericidal capacity of the perineal exudating neutrophils, 3 weeks after induction of diabetes. The study of the cellular distribution in uninfected urinary tract tissue demonstrated a reduction in CD4-positive T cell distribution in bladder submucosa of diabetic mice at 3 weeks after induction of diabetes (diabetic group), compared with that in the normal mice (control group). In the diabetic group, probably in order to compensate for the reduction in CD4-positive T cell distribution in the bladder submucosa, the distribution of macrophages was increased in the bladder epithelium, compared with the control group. This finding suggested that diabetic mice are thought to have protective mechanisms against infection different from those of normal mice in the uninfected state. The study of the response time fluctuation of local immune response at the infected sites demonstrated infiltration of macrophages was the same grade as that of neutrophils in the control and the diabetic group. However, in the diabetic group, a marked infiltration of macrophages was recognized when compared with the control group in the early phase of infection. These findings suggested that macrophages aid in protection against infection when damage to the bactericidal capacity of neutrophils results in insufficient elimination of microorganisms. On the other hand, infiltration of T and B cells was weaker in the diabetic group, than in the control group.     

Field emission monotron for THz emission

Small-signal negative conductance of a field emission monotron was calculated, which was considerably higher than that of a thermionic monotron due to strong bunching of emitted electrons in a field emission. The amplification constant of 28dB/mm is achieved at THz wave region by applying the monotron to a distributed amplifier and is sufficient to overcome power loss in a transmission line. In addition, a wide band frequency tunable electromagnetic wave source may be developed at THz wave region by providing a low loss microstrip line resonator with field emission array due to the gate voltage dependent phase constant of the line.