Sardine procalcitonin amino-terminal cleavage peptide has a different action from calcitonin and promotes osteoblastic activity in the scales of goldfish

The nucleotide sequence of a sardine preprocalcitonin precursor has been determined from their ultimobranchial glands in the present study. From our analysis of this sequence, we found that sardine procalcitonin was composed of procalcitonin amino-terminal cleavage peptide (N-proCT) (53 amino acids), CT (32 amino acids), and procalcitonin carboxyl-terminal cleavage peptide (C-proCT) (18 amino acids). As compared with C-proCT, N-proCT has been highly conserved among teleosts, reptiles, and birds, which suggests that N-proCT has some bioactivities. Therefore, both sardine N-proCT and sardine CT were synthesized, and their bioactivities for osteoblasts and osteoclasts were examined using our assay system with goldfish scales that consisted of osteoblasts and osteoclasts. As a result, sardine N-proCT (10− 7 M) activated osteoblastic marker enzyme activity, while sardine CT did not change. On the other hand, sardine CT (10− 9 to 10− 7 M) suppressed osteoclastic marker enzyme activity, although sardine N-proCT did not influence enzyme activity. Furthermore, the mRNA expressions of osteoblastic markers such as type 1 collagen and osteocalcin were also promoted by sardine N-proCT (10− 7 M) treatment; however, sardine CT did not influence their expressions. The osteoblastic effects of N-proCT lack agreement. In the present study, we can evaluate exactly the action for osteoblasts because our scale assay system is very sensitive and it is a co-culture system for osteoblasts and osteoclasts with calcified bone matrix. Both CT and N-proCT seem to influence osteoblasts and osteoclasts and promote bone formation by different actions in teleosts. © 2017 Elsevier Inc.Embargo Period 12 month

Involvement of integrin-linked kinase in capillary/tube-like network formation of human vascular endothelial cells

Angiogenesis is a complex process involving an ECM and vascular endothelial cells (EC), and is regulated by various angiogenic factors including VEGF. The ability to form a capillary/tube-like network is a specialized function of EC. Therefore, in vitro angiogenesis was assessed by a capillary/tube-like network formation assay. There are three angiogenic parameters: capillary length, number of capillaries, and relative capillary area per field. We evaluated capillary length per field in the assay. VEGF promoted capillary/tube-like network formation of EC in a type I collagen gel matrix in vitro. Moreover, we demonstrated the involvement of ILK in a VEGF signaling pathway mediating capillary/tube-like network formation of EC using dominant-negative, kinase deficient ILK. This is a straightforward assay to monitor responses of human vascular endothelial cells

Dissolved methane distribution in the South Pacific and the Southern Ocean in austral summer

[1] Oceanic methane (CH_4) was examined in the South Pacific and the Southern Ocean from December 2001 to February 2002. From the oligotrophic South Pacific (10°S) to the Subtropical Front (STF), we observed the maximum concentrations of CH_4 in the subsurface layer which ranged from 2.7 to 3.9 nmol kg^. South of the STF, higher levels of CH_4 were often detected in both the surface and the subsurface layers. Elevated surface CH_4 concentrations (3.4-6.1 nmol kg^) south of the STF tended to be higher than those north of the STF. Higher CH_4 were often concomitant with an increase in chlorophyll a levels in the Seasonal Ice Zone (SIZ). Considering that phytoplankton does not generate methane directly, the high CH_4 production probably resulted from the grazing processes of Antarctic krill and/or zooplankton fed on phytoplankton and the subsequent microbial methanogenesis in addition to the aerobic CH_4 production in the water column. Present results showed a good relation between surface CH_4 data (, r = 0.68, n = 49, p d^ between 10°S and the PF (54°S), and from 0.8 to 2.3 mol CH_4 km^ d^ south of the PF, where the sea-air CH_4 flux was largely affected by the wind speed

Pilot-Plant Scale 12 kW Microwave Irradiation Reactor for Woody Biomass Pretreatment

A simple, low reflection, and highly-efficient pilot-plant scale microwave irradiation reactor for woody biomass pretreatment was fabricated. Pretreatment is an essential process for effective bioethanol production.The fabricated reactor consists of 8 microwave irradiators which are attached to a metal pipe. The woody biomass mixture which contains water and organic acid flows through the metal pipe and is heated by microwaves at a total power of 12 kW. To design the microwave irradiators, we used a 3D Finite Element Method (FEM) simulator, which was based on the measured complex permittivity data of the woody biomass mixture.The simulation results showed that the reflection coefficient |S11| from the reactor was less than -30 dB when the woody biomass mixture temperature was between 30°C and 90°C.Finally, we experimentally confirmed that the fabricated irradiation reactor yielded a microwave absorption efficiency of 79%