Altered microviscosity of in vivo lipid-manipulated membranes inTetrahymena pyriformis: A fluorescence study

Summary By determination using fluorescence polarization measurements with 1,6-diphenyl 1,3,5-hexatriene, ergosterolreplaced pellicle and microsome membranes ofTetrahymena cells become less fluid, whereas those membranes from chimyl alcohol-fed cells are more fluid, when compared with the control native membranes.Acknowledgments. We wish to express thanks to Central Scientific Commerce, Inc., Tokyo for permission to use an Elscint Microviscosimeter MV-Ia. This work was in part supported by grant from the Ministry of Education.     

Regenerative medicine of cornea by cell sheet engineering using temperature-responsive culture surfaces

Recently, regenerative medicine has been focused on as next-generation definitive therapies for several diseases or injuries for which there are currently no effective treatments. These therapies have been rapidly developed through the effective fusion be- tween different fields such as stem cell biology and biomaterials. So far, we have proposed ＂cell sheet engineering＂ through our core technology that simply applies alterations of the temperature which allows regulation of the attachment or detachment of living cells on the culture surfaces grafted with the temperature-responsive polymer ＂poly（N-isoproplyacrylamide）＂. This tech- nology enables us to construct bioengineered sheet-like tissues, termed ＂cell sheets＂, without the need of using biodegradable scaffolds and protease treatments that are traditionally used. Therefore, our carrier-free cell sheets not only are independent of the issues observed in conventional methods, but also showed further advantages in the reconstruction of the corneal epithelium or endothelium （e.g. improvement of optical transparency and cell-cell interactions to host stroma in reconstructed tissues）. Moreo- ver, our approach does not have issues such as immune rejection or limited number of donors, due to the use of cell sheets derived from autologous limbal （in patients with unilateral disease） or oral mucosal epithelial cells （in patients with bilateral disorders）. Indeed, we have successfully performed the clinical application for the reconstruction of ocular surfaces through the transplanta- tion of our carrier-free corneal epithelial cell sheets, as evidenced by improved visual acuity as well as long-term maintenance of postoperative health conditions on ocular surfaces in all patients. We have also proposed a novel approach for the reconstruction of the corneal endothelium using corneal endothelial cell sheets by demonstrating its successful application. Thus, our cell sheet engineering technique provides a breakthrough in the field of regenerative medicine applied to the cornea.     

Airglow observation by Fabry-Perot interferometer

An all-sky and a scanning Fabry-Perot interferometers (FPI) have been developed for observations of vertical winds and two-dimensional atmospheric motions in the low-thermosphere and the mesosphere region by the measurement of Doppler shift and broadening of airglow emission lines. The system of FPI is described and the method of data analysis is proposed, an example of observation is also given. Ai Yong: born in 1958, Ph.D graduate student     

Essential role for de novo DNA methyltransferase Dnmt3a in paternal and maternal imprinting

Imprinted genes are epigenetically marked during gametogenesis so that they are exclusively expressed from either the paternal or the maternal allele in offspring. Imprinting prevents parthenogenesis in mammals and is often disrupted in congenital malformation syndromes, tumours and cloned animals. Although de novo DNA methyltransferases of the Dnmt3 family are implicated in maternal imprinting, the lethality of Dnmt3a and Dnmt3b knockout mice has precluded further studies. We here report the disruption of Dnmt3a and Dnmt3b in germ cells, with their preservation in somatic cells, by conditional knockout technology. Offspring from Dnmt3a conditional mutant females die in utero and lack methylation and allele-specific expression at all maternally imprinted loci examined. Dnmt3a conditional mutant males show impaired spermatogenesis and lack methylation at two of three paternally imprinted loci examined in spermatogonia. By contrast, Dnmt3b conditional mutants and their offspring show no apparent phenotype. The phenotype of Dnmt3a conditional mutants is indistinguishable from that of Dnmt3L knockout mice, except for the discrepancy in methylation at one locus. These results indicate that both Dnmt3a and Dnmt3L are required for methylation of most imprinted loci in germ cells, but also suggest the involvement of other factors.     

Mice deficient in protein tyrosine phosphatase receptor type Z are resistant to gastric ulcer induction by VacA of Helicobacter pylori

The vacuolating cytotoxin VacA produced by Helicobacter pylori causes massive cellular vacuolation in vitro and gastric tissue damage in vivo, leading to gastric ulcers, when administered intragastrically. Here we report that mice deficient in protein tyrosine phosphatase receptor type Z (Ptprz, also called PTP-zeta or RPTP-beta, encoded by Ptprz) do not show mucosal damage by VacA, although VacA is incorporated into the gastric epithelial cells to the same extent as in wild-type mice. Primary cultures of gastric epithelial cells from Ptprz+/+ and Ptprz-/- mice also showed similar incorporation of VacA, cellular vacuolation and reduction in cellular proliferation, but only Ptprz+/+ cells showed marked detachment from a reconstituted basement membrane 24 h after treatment with VacA. VacA bound to Ptprz, and the levels of tyrosine phosphorylation of the G protein-coupled receptor kinase-interactor 1 (Git1), a Ptprz substrate, were higher after treatment with VacA, indicating that VacA behaves as a ligand for Ptprz. Furthermore, pleiotrophin (PTN), an endogenous ligand of Ptprz, also induced gastritis specifically in Ptprz+/+ mice when administered orally. Taken together, these data indicate that erroneous Ptprz signaling induces gastric ulcers.