Catalase in manipulation buffer enhances the developmental competence of DNA-injected embryos

To improve the efficiency of transgenesis, we investigated the effects of a radical scavenger during microinjection on the development to blastocysts or pups of mouse pronuclear embryos, microinjected with the enhanced green fluorescent protein (EGFP) transgene. When embryos were microinjected in medium containing 0-1,000 units/ml catalase, the developmental rate to blastocysts was significantly higher (P<0.01) in 100-units/ml catalase (81%) than those in 0 and 1,000 units/ml (56 and 65%). To investigate the ontogenetic ability of DNA-injected embryos, EGFP-injected embryos manipulated under 0 or 100 units/ml catalase were transferred separately to recipient mice. The proportion of fetuses derived from EGFP-injected embryos manipulated under 100 units/ml catalase (29%) was significantly higher (P<0.05) than that manipulated under 0 units/ml catalase (19%). Furthermore, the numbers of transgenic pups were 17 in 100 units/ml catalase and 14 in 0 units/ml catalase. The results of the present study indicate that scavenging reactive oxygen species during in vitro micromanipulation is beneficial for the development of DNA-injected embryos.     

Expression and subcellular localization of GSE protein in germ cells and preimplantation embryos

We previously identified a novel gonad-specific expression gene (Gse) and investigated its expression during gametogenesis in the mouse testis and ovary. In this study, we generated a polyclonal antibody to GSE protein and determined the profiles of the protein's expression in germ cells and preimplantation embryos in detail using immunocytochemical and immunofluorescence staining. In a Western blot analysis, the anti-GSE antibody recognized long and short isoforms (approximately 27.6 kDa and 23.1 kDa) of the protein in the mouse testis and the long isoform in the ovary. In the mouse testis, GSE protein was expressed in spermatocytes I in the pachytene stage, round spermatids, and elongated spermatids. In the mouse ovary, the protein was located in the cytoplasm and nucleus of all oocytes regardless of the stage of the ovarian follicles. In preimplantation embryos from the pronuclear to blastocyst stage, however, GSE protein was mainly detected in the nuclei of cells. At the blastocyst stage, the protein was confirmed to have accumulated in the inner cell mass (ICM), whereas it had mostly disappeared from the trophectoderm (TE). These findings suggest that GSE protein may play a role in the establishment of nuclear totipotency and may be associated with early lineage specification.     

Semiquantitative multi‐analysis of plasma obtained from Romney lambs (Ovis aries) by inductively coupled plasma mass spectrometry,and the classification according to feed type

The establishment of a classification system for domestic animals on consumed feed stuff is thought to be important from both a hygiene and market point of view. We collected plasma samples of Romney lambs (Ovis aries) which were fed one of the following: a herb‐clover mix (n = 10) which included chicory, red clover, white clover and plantain; a plant‐grass mix (n = 10) which included plantain, ryegrass and white clover; or a grass mix (n = 10) which included ryegrass and white clover. A total of 20 elements in plasma samples obtained from the lambs were analyzed using inductively coupled plasma mass spectrometry. The data were then analyzed by principal component analysis. The lambs were divided into three groups on a score plot depending on the different feed conditions. Furthermore, discriminant analyses of the elements were examined, using linear discriminant analysis with forward stepwise regression. This discriminant function correctly classified the samples from each group. The accuracy of classification of each group, as shown by 10‐fold cross‐validation, proved the effectiveness of the established discriminant function. It is concluded that using linear discriminant analysis might be a useful tool for the validation of elements from plasma in lambs grown in different conditions.     

Ozone treatment of spent medium from Auricularia polytricha cultivation for enzymatic saccharification and subsequent ethanol production

We examined the enzymatic saccharification and ethanol fermentation of the spent media (SMs) from Auricularia polytricha cultivation using wood meals of Falcataria moluccana, Shorea sp., and Tectona grandis. Although the hydrolysis weight decrease and reducing sugar yield were higher in SM of F. moluccana, the ethanol yield was higher in SM of Shorea sp. Ozone treatment of SM further increased the hydrolysis weight decrease, reducing sugar, and ethanol yields in Shorea sp. These results indicate that SM of A. polytricha is a suitable biomass material to produce fermentable sugars for ethanol production, and that ozone treatment is a suitable method for increasing the ethanol yield.     

Effects of resveratrol treatment on mitochondria and subsequent embryonic development of bovine blastocysts cryopreserved by slow freezing

This study evaluated the effects of cryopreservation by slow freezing on the mitochondrial function, DNA integrity, and developmental ability of bovine embryos and examined whether resveratrol treatment of the frozen‐thawed blastocysts improved embryonic viability. In vitro produced bovine embryos were subjected to slow freezing. After thawing, the ATP content and mitochondrial DNA integrity (mtDNA), determined by real‐time PCR targeting short and long mitochondrial sequences, was found to be lower in frozen‐thawed embryos than in fresh embryos, and mtDNA copy number was significantly reduced during the 24‐hr incubation post warming. Furthermore, immunostaining against double‐strand DNA revealed DNA damage in frozen‐thawed embryos. When frozen‐thawed embryos were incubated in the medium containing 0.5 µM resveratrol, SIRT1 expression, and survival rate of the embryos significantly improved compared with the vehicle‐treated embryos. In addition, cell‐free mtDNA content in medium was higher in case of resveratrol‐treated embryos than of vehicle‐treated embryos. In conclusion, slow freezing affects mitochondrial integrity and function in the blastocysts. In the frozen‐thawed embryos, mitochondria were removed during post‐thawing incubation and resveratrol enhanced the process, resulting in improved survivability of the embryos.     

Ferromagnetism in LaFeO3-LaCrO3 superlattices

Ferromagnetic spin order has been realized in the LaCrO3-LaFeO3 superlattices. Ferromagnetic coupling between Fe3+ and Cr3+ through oxygen has long been expected on the basis of Anderson, Goodenough, and Kanamori rules. Despite many studies of Fe-O-Cr-based compounds, random positioning of Fe3+ and Cr3+ ions has frustrated the observation of ferromagnetic properties. By creating artificial superlattices of Fe3+ and Cr3+ layer along the [111] direction, ferromagnetic ordering has been achieved.     

Value-per-recruit analysis of bighand thornyhead <Emphasis Type="Italic">Sebastolobus macrochir</Emphasis> caught off the Pacific coast of northern Honshu,Japan

The yield-per-recruit (YPR) analysis has been used to assess growth overfishing of exploited fish stocks. Since the ex-vessel price per weight widely depends on the size of the fish, the result of the YPR does not imply the revenue. The value-per-recruit (VPR) analysis, which is an economic expansion of the YPR analysis, was conducted for bighand thornyhead Sebastolobus macrochir caught off the Pacific coast of northern Honshu, Japan. The results showed that, for the current age at first capture (t _c = 3), the current fishing mortality (F = 0.12) was close to the optimum level. However, for the current F, optimum t _c in the VPR model was 8 years and would lead to a 47% increase compared to the current revenue. The maximum YPR was obtained at t _c = 6 years, but the revenue at t _c = 6 only gave a 20% increase in the maximum revenue. The difference was caused by the size dependency of the ex-vessel price per weight. The VPR model would generate fisheries management plans with both economical and biological consideration and would help in making general consensus of fisheries management strategies.     

Quality Characteristics of Waxy Hexaploid Wheat (Triticum aestivum L.): Properties of Starch Gelatinization and Retrogradation

The viscoelastic properties and molecular structure of the starch isolated from waxy (amylose-free) hexaploid wheat (WHW) (Triticum aestivum L.) were examined. WHW starch generally had lower gelatinization onset temperature, peak viscosity, and setback than the starch isolated from normal hexaploid wheat (NHW). Differential scanning calorimetry (DSC) showed that WHW starch had higher transition temperatures (T_o, T_p, and T_c) and enthalpy (ΔH) than NHW starch. However, when compared on the basis of amylopectin (AP) content, ΔH of WHW starch was almost statistically identical to that of its parental varieties. Typical A-type X-ray diffraction patterns were observed for the starches of WHW and its parental varieties. Somewhat higher crystallinity was indicated for WHW starch. WHW starch was also characterized by having greater retrogradation resistance. The high-performance size-exclusion chromatography (HPSEC) of amylopectin showed that each amylopectin yielded two fractions after debranching. Although WHW amylopectin had somewhat long B chains, little difference was observed in the ratio of Fr.III/ Fr.II between WHW and its parental varieties.     

Propagation of Asian isolates of canine distemper virus (CDV) in hamster cell lines

Backgrounds

The aim of this study was to confirm the propagation of various canine distemper viruses (CDV) in hamster cell lines of HmLu and BHK, since only a little is known about the possibility of propagation of CDV in rodent cells irrespective of their epidemiological importance.

Methods

The growth of CDV in hamster cell lines was monitored by titration using Vero.dogSLAMtag (Vero-DST) cells that had been proven to be susceptible to almost all field isolates of CDV, with the preparations of cell-free and cell-associated virus from the cultures infected with recent Asian isolates of CDV (13 strains) and by observing the development of cytopathic effect (CPE) in infected cultures of hamster cell lines.

Results

Eleven of 13 strains grew in HmLu cells, and 12 of 13 strains grew in BHK cells with apparent CPE of cell fusion in the late stage of infection. Two strains and a strain of Asia 1 group could not grow in HmLu cells and BHK cells, respectively.

Conclusion

The present study demonstrates at the first time that hamster cell lines can propagate the majority of Asian field isolates of CDV. The usage of two hamster cell lines suggested to be useful to characterize the field isolates biologically.     

Distribution and stem growth patterns of mangrove species along the Nakara River in Iriomote Island, Southwestern Japan

We examined variations in stand structure and tree growth of a mangrove forest along two gradients: from the river mouth to upper stream and from the riverside to inland, along the Nakara River, Iriomote Island, Japan. Bruguiera gymnorrhiza (L.) Lamk. occurred throughout the intertidal area, though Rhizophora stylosa Griff. and Kandelia candel (L) Druce did not occur upstream. Basal area and maximum tree height of B. gymnorrhiza decreased downstream. The growth rate in stem diameter of B. gymnorrhiza decreased downstream in relation with the soil salinity. The basal area and the maximum tree height of B. gymnorrhiza increased with the distance from the riverside. R. stylosa increased in stem diameter and growth rate toward the riverside.