Effects of cycloheximide treatment on in-vitro development of porcine parthenotes and somatic cell nuclear transfer embryos

This study aimed to verify the beneficial effect of cycloheximide (CHX) treatment on the development of porcine somatic cell nuclear transfer (NT) embryos, which were constructed with enucleated oocytes and cumulus cells by using a single direct current (DC) pulse. In the first experiment, a single DC pulse applied to the induction of fusion and activation of NT embryos gave a high fusion rate. However, cleavage and subsequent development of fused couplets (NT embryos) to the blastocyst stage were poor. Experiment 2 was conducted to determine whether CHX treatment could enhance metaphase II (M II) oocyte activation and improve the subsequent parthenogenetic development. After giving the DC pulse and incubation with or without CHX, M II oocytes incubated with CHX showed higher cleavage and development to blastocysts compared with those incubated without CHX (P < 0. 05). Experiment 3 was carried out to verify the beneficial effect of CHX on the development of NT embryos. The NT embryos treated with the DC pulse and CHX treatment showed higher cleavage and subsequent development compared with those treated with the DC pulse alone (P < 0.05) . The present study demonstrates that CHX treatment enhances the electrical stimulus-induced activation of oocytes and NT embryos, and improves the subsequent development of parthenotes and NT embryos. The results indicate that protein synthesis inhibition treatment required for the induction of oocyte activation promotes the development of NT embryos.     

Effect of Leukemia Inhibitory Factor and Forskolin on Establishment of Rat Embryonic Stem Cell Lines

This study was designed to investigate whether supplementation of 2i medium with leukemia inhibitory factor (LIF) and/or forskolin would support establishment of germline-competent rat embryonic stem (ES) cell lines. Due to the higher likelihood of outgrowth rates, supplementation of forskolin with or without LIF contributed to the higher establishment efficiency of ES cell lines in the WDB strain. Germline transmission competency of the chimeric rats was not influenced by the profile of ES cell lines until their establishment. When the LIF/forskolin-supplemented 2i medium was used, the rat strain used as the blastocyst donor, such as the WI strain, was a possible factor negatively influencing the establishment efficiency of ES cell lines. Once ES cell lines were established, all lines were found to be germline-competent by a progeny test in chimeric rats. In conclusion, both LIF and forskolin are not essential but can play a beneficial role in the establishment of “genuine” rat ES cell lines.     

The Distribution of <Emphasis Type="Italic">Perilla</Emphasis> Species

Perilla (Lamiaceae) contains one tetraploid species, P. frutescens (L.) Britt. and three diploid species, P. citriodora (Makino) Nakai, P. hirtella Nakai and P. setoyensis G. Honda. Tetraploid species have been traditionally cultivated in Asia for their seed oil and for their fragrant leaves that are used as medicine or as a garnish for fish. The center of diversity is still obscure. To conserve the genetic resources, it is important to know the diversity of the tetraploid species. The three diploid species, which are possible parents of the tetraploid species, are all believed to be indigenous to Japan. Their distribution in China and Korea was clarified on the basis of herbarium and field surveys. The tetraploid species is assumed to have originated somewhere around the mid-to downstream area of the Changjiang River. Though Perilla is not cultivated as often in these areas as in northern China, Korea, the Himalayan region, or Myanmar, these areas should also be important for the conservation of genetic resources of tetraploid Perilla crops because of the expected high genetic diversity.     

Acoustoelastic birefringence effect in wood III: ultrasonic stress determination of wood by acoustoelastic birefringence method

Stress conditions produced in wood were analyzed by means of the acoustoelastic birefringence method. Bending load was applied against a wood beam specimen. Under loading, ultrasonic shear waves were propagated through the breadth direction of the wood beam specimen. The velocities of shear waves polarized in the longitudinal or tangential direction of the wood beam specimen were measured with the sing-around method. Bending stresses were determined by dividing the difference between the acoustic anisotropy and the texture anisotropy by the acoustoelastic birefringence coefficient. Shear stresses were also determined. These stress distributions of the beam specimen were in good agreement with those obtained by the strain gauge method and mechanical calculation.     

Analysis of phloem exudate collected from fruit-bearing stems of coconut palm: Palm trees as a source of molecules circulating in sieve tubes

Sieve tubes have been attracting widespread research interest because of their possible role in mediating physiological signals within the whole plant. However, progress in research into the function of sieve tubes has been limited by the low volume of sap available. To overcome this problem, we attempted to collect phloem exudate from tropical coconut palm trees (Cocos nucifera L. cv. Namhom). As much as 3 to 15 mL of exudate per hour was collected from the cut surface of the plant's fruit-bearing stem. Our analyses revealed that the characterized profiles of sugars (sucrose: 339 mM), amino acids (total concentration: 17.1 mM), cations (potassium: 48.3 mM), and proteins (total concentration: 0.1 /-lg /-lL-1) in the exudate were mostly consistent with those of phloem sap or phloem exudate collected from rice plants, castor bean plants, etc. This exudate was assumed to reflect the composition of the phloem sap from the source organs of coconut palm trees. The large volume of exudate collected contributed significantly to the analyses of the various compounds in the stream of sieve tubes.     

Evaluation of the effects of spores and their heat-treated residues from different Bacillus strains on the initial growth of rice plants

This study evaluated the effects of the spores and their heat-treated residues from different strains of Bacillus species (B. pumilus, B. altitudinis, and B. megaterium) on the early growth of paddy rice cultivars, including Hitomebore (short-grain japonica rice), Takanari (high-yielding indica rice), and two new lines, TULK-143-6 and LTAT-29. The spores of seven Bacillus strains positively affected Hitomebore root growth, while, the root volume of TULK-143-6 with inoculation of B. pumilus TUAT1 and JM52, and root length and root surface area of LTAT-29 with inoculation of B. megaterium MAFF301694 were increased significantly. In contrast with Hitomebore, Takanari root growth was significantly inhibited by the spores of six Bacillus strains. Surprisingly, inoculations with the spore residues from all tested Bacillus strains increased the root dry weight of Hitomebore, with the effects of four bacterial strains being significant. Furthermore, there were more Bacillus spores than vegetative cells at different time points during the initial rice growth stage, and most plant samples mainly consisted of Bacillus spores. Thus, the spores of Bacillus species likely promote rice root development.     

Glycoprotein hormone in the pituitary of hagfish and its evolutionary implications

The pituitary gland is present in all vertebrates, from agnathans (jawless vertebrates) to mammals, but not in invertebrates. Reproduction in gnathostomes (jawed vertebrates) is controlled by two pituitary gonadotropins (GTHs), luteinizing hormone and follicle-stimulating hormone, which are part of the pituitary glycoprotein hormone (GPH) family. Hagfishes, which lack both jaws and vertebrae, are considered the most primitive vertebrate known, living or extinct. Accordingly, they are of particular importance in understanding the evolution of the pituitary GPHs and their functions related to vertebrate reproduction. Nevertheless, key elements of the reproductive endocrine system in hagfish have yet to be elucidated. Our current report has revealed the first identification of a functional GPH composed of two subunits that possess gonadotropic action at the pituitary of brown hagfish. It seems most likely that an ancestral GPH gave rise to only one GTH in hagfish pituitary and that multiplicity of GPHs arose later during the early evolution of gnathostomes. This paper briefly summarizes the latest findings on the hagfish GPH from an evolutionary point of view.