Cellular Uptake of Pyrroloquinoline Quinone in Its Intact and Derivatized Forms from the Cell Culture Medium of 3T3-L1.

Following a growing interest in the physiological effects of pyrroloquinoline quinone (PQQ), more cell culture experiments have begun to elucidate its mechanism of action. However, to our knowledge, no reports have used instrumental analysis, such as liquid chromatography-tandem mass spectrometry (LC-MS/MS), to study cellular uptake of PQQ. In addition, despite the propensity of PQQ to react with amino acids and other compounds, only a handful of cell culture experiments have been conducted on PQQ derivatives.

Determination of pyrroloquinoline quinone by enzymatic and LC-MS/MS methods to clarify its levels in foods.

Pyrroloquinoline quinone (PQQ) is believed to be a new B vitamin-like compound, and PQQ supplementation has received attention as a possible treatment for diseases including dementia and diabetes. However, the distribution of PQQ in foods is unclear, due to the difficulty in analyzing the compound. Therefore, in this study, enzymatic and LC-MS/MS methods were optimized to enable an accurate analysis of PQQ in foods.

Establishment of oct4:gfp transgenic zebrafish line for monitoring cellular multipotency by GFP fluorescence.

The establishment of induced pluripotent stem (iPS) cell technology in fish could facilitate the establishment of novel cryopreservation techniques for storing selected aquaculture strains as frozen cells. In order to apply iPS cell technology to fish, we established a transgenic zebrafish line, Tg(Tru.oct4:EGFP), using green fluorescent protein (GFP) expression under the control of the oct4 gene promoter as a marker to evaluate multipotency in iPS cell preparations.

Identification, cDNA cloning, and mRNA localization of a zebrafish ortholog of leukemia inhibitory factor.

Leukemia inhibitory factor (LIF) maintains embryonic stem cells in an undifferentiated state. To establish stable cultures of zebrafish embryonic stem cells, recombinant zebrafish LIF (zLIF) is needed because the LIF sequence varies greatly between species. In the current study, we identified the zebrafish (Danio rerio) and pufferfish (Tetraodon nigroviridis) orthologs of lif from genomic databases, and we isolated a cDNA encoding zLIF.