Maintenance of mouse trophoblast stem cells in KSR-based medium allows conventional 3D culture

Mouse trophoblast stem cells (TSCs) can differentiate into trophoblast cells, which constitute the placenta. Under conventional culture conditions, in a medium supplemented with 20% fetal bovine serum (FBS), fibroblast growth factor 4 (FGF4), and heparin and in the presence of mouse embryonic fibroblast cells (MEFs) as feeder cells, TSCs maintain their undifferentiated, proliferative status. MEFs can be replaced by a 70% MEF-conditioned medium (MEF-CM) or by TGF-ß/activin A. To find out if KnockOut^TM Serum Replacement (KSR) can replace FBS for TSC maintenance, we cultured mouse TSCs in KSR-based, FBS-free medium and investigated their proliferation capacity, stemness, and differentiation potential. The results indicated that fibronectin, vitronectin, or laminin coating was necessary for adhesion of TSCs under KSR-based conditions but not for their survival or proliferation. While the presence of FGF4, heparin, and activin A was not sufficient to support the proliferation of TSCs, the addition of a pan-retinoic acid receptor inverse agonist and a ROCK-inhibitor yielded a proliferation rate comparable to that obtained under the conventional FBS-based conditions. TSCs cultured under the KSR-based conditions had a gene expression and DNA methylation profile characteristic of TSCs and exhibited a differentiation potential. Moreover, under KSR-based conditions, we could obtain a suspension culture of TSCs using extracellular matrix (ECM) coating-free dishes. Thus, we have established here, KSR-based culture conditions for the maintenance of TSCs, which should be useful for future studies.     

Transformation processes involving mercury species in the atmosphere — results from a literature survey

Mercury is released into the atmosphere from natural and anthropogenic sources. Once in the air, Hg species will be dispersed, chemically and physically transformed, and transported over varying distances (regional, continental and global scales) while concurrently being subjected to removal processes. Modeling of the aerial transport and deposition of Hg to receptor sites offers an attractive approach for integrating existing experimental data and represents an important tool for advancing our understanding of environmental Hg pollution. Comprehensive modeling efforts of the atmospheric pathways and fate of this heavy metal require knowledge of its atmospheric chemistry. To update and consolidate available information relevant to the atmospheric chemistry of Hg, a computer-assisted search was undertaken of the primary scientific literature published during the last ten years. Selected results from this literature survey are presented in this paper.     

Cryopreservation diluents for spermatozoa of Sakhalin taimen <Emphasis Type="Italic">Hucho perryi</Emphasis>

To develop a suitable cryopreservation diluent for spermatozoa of the endangered Sakhalin taimen Hucho perryi, all possible combinations of cryoprotectants (glycerol, dimethyl sulfoxide [DMSO], methanol) and extenders (fetal bovine serum [FBS], 300 mM glucose solution [GS], artificial seminal plasma for masu salmon) were examined by observing sperm motility 10 s after thawing. Spermatozoa cryopreserved with diluents such as mixtures of 10% glycerol plus 90% FBS, 10% DMSO plus 90% FBS, and 10% methanol plus 90% GS showed the highest motility. The maximal post-thaw motility was observed at 10% among all concentrations (0, 5, 10, 15 and 20%) of these three cryoprotectants. No significant difference among three diluents was observed in motility at 10 s. Mixtures of 10% glycerol plus 90% FBS, 10% DMSO plus 90% FBS, and 10% methanol plus 90% GS are suitable cryopreservation diluents for Sakhalin taimen spermatozoa.     

Effect of oral administration of a single bolus of six different protein sources on digestive physiology of red seabream Pagrus major juveniles

Fish Physiology and Biochemistry - To reveal direct effects of various protein sources on digestive physiology of red seabream, Pagrus major (38.5 ± 0.4&nbsp;g), six different...     

Culture and climate change: Japanese cherry blossom festivals and stakeholders’ knowledge and attitudes about global climate change

Most global climate change models predict serious ecological and social problems. In Japan, biologists have found climate change is affecting species and ecosystems, including the earlier flowering time of cherry trees which are an important cultural symbol in Japan. Cherry blossom festivals are also important to local economies. This study explored the perceptions of Japanese residents regarding climate change impacts on culturally significant events such as flower timing of cherry trees. We conducted interviews of stakeholders of three cherry blossom festivals, including sixteen organizers of festivals and 26 managers of festival-dependent businesses, to understand their awareness, attitudes and behaviors toward global climate change and impacts on cherry blossom festivals. Most organizers of the festival in Kakunodate were concerned about global warming and its impact on cherry blossom times while organizers of festivals in Nakano and Komoro felt it was unimportant if flower timing affected the festival schedule. Most (92%) managers of festival-dependent businesses mentioned that global warming is occurring and affecting the flower timing of cherry trees, but there were diverse perceptions of global warming impacts on their business. Managers more dependent on income from cherry blossom festivals indicated greater concern for the effects of climate change.     

Active expression of matrix metalloproteinase-13 mRNA in the granulation tissue of equine superficial digital flexor tendinitis

The DNA microarray analysis for matrix metalloproteinase (MMP)-related mRNA expression in equine superficial digital flexor tendinitis indicated that mRNA level of MMP-13 was apparently up-regulated in the tendinitis as compared to normal tendon. In situ hybridization also revealed that fibroblastic cells proliferated in the granulation tissue generated in the tendinitis actively expressed MMP-13 mRNA. On the other hand, in normal tendon, a few fibroblastic cells and vascular components lied in the endotenon barely expressed its mRNA, but other cellular components in the tendon bundle were not positively hybridized. As mentioned above, MMP-13 but not other collagenases or gelatinases, may play an important role in tendon injuries in the racehorses.