The developmental ability of vitrified oocytes from different mouse strains assessed by parthenogenetic activation and intracytoplasmic sperm injection

Assessment of the developmental ability of oocytes following freezing and thawing is an important step for optimizing oocyte cryopreservation techniques. However, the in vitro fertilization of frozen-thawed mouse oocytes is often inefficient because of incomplete capacitation of spermatozoa in the absence of surrounding cumulus cells. This study was undertaken to determine whether the oocyte cryopreservation efficiency of different strains of mice could be assessed from the development of oocytes following parthenogenetic activation and intracytoplasmic sperm injection (ICSI). Oocytes were collected from hybrid (C57BL/6 x DBA/2) F1 or inbred (C57BL/6J, C3H/HeN, DBA/2J and BALB/cA) strains and were vitrified in a solution containing ethylene glycol, DMSO, Ficoll and sucrose. In the first series of experiments, oocytes were activated parthenogenetically by Sr(2+) treatment after warming. The oocytes from the inbred strains, but not those of the F1 hybrid, were diploidized by cytochalasin treatment to obtain a sufficient number of blastocysts. In all strains tested, parthenogenetic embryos derived from vitrified oocytes developed into blastocysts at rates between 23 and 68%. In the second series of experiments, vitrified oocytes from each strain were injected with homologous spermatozoa after warming. Normal offspring were obtained from all strains at rates between 5 and 26% per embryo transferred. Thus, the feasibility of oocyte cryopreservation protocols can be assessed easily by in vitro development of parthenogenetic embryos or by in vivo development of ICSI embryos. Moreover, the oocytes of these four major inbred strains of mice can be cryopreserved safely for production of offspring.     

Changes in the expression of pituitary gonadotropin subunits during reproductive cycle of multiple spawning female chub mackerel Scomber japonicus

The endocrine regulation of reproduction in a multiple spawning fish with an asynchronous-type ovary remains largely unknown. The objectives of this study were to monitor changes in the mRNA expression of three gonadotropin (GtH) subunits (GPα, FSHβ, and LHβ) during the reproductive cycle of the female chub mackerel Scomber japonicus. Cloning and subsequent sequence analysis revealed that the cDNAs of chub mackerel GPα, FSHβ, and LHβ were 658, 535, and 599 nucleotides in length and encoded 117, 115, and 147 amino acids, respectively. We applied a quantitative real-time PCR assay to quantify the mRNA expression levels of these GtH subunits. During the seasonal reproductive cycle, FSHβ mRNA levels remained high during the vitellogenic stages, while GPα and LHβ mRNA levels peaked at the end of vitellogenesis. The expression of all three GtH subunits decreased during the post-spawning period. These results suggest that follicle-stimulating hormone (FSH) is involved in vitellogenesis, while luteinizing hormone (LH) functions during final oocyte maturation (FOM). Both GPα and FSHβ mRNA levels remained high during the FOM stages of the spawning cycle and increased further just after spawning. Thus, FSH synthesis may be strongly activated just after spawning to accelerate vitellogenesis in preparation for the next spawning. Alternatively, LHβ mRNA levels declined during hydration and then increased after ovulation. This study demonstrates that chub mackerel are a good model for investigating GtH functions in multiple spawning fish.     

A sustained controlled release formulation of soil nitrogen based on nitrate-layered double hydroxide nanoparticle material

Purpose

Nitrate-layered double hydroxide material (nitrate-LDH) matrix can be considered as a potential formulation of delivering nitrogen into soil in a sustained manner.

Materials and methods

The nitrate-LDH matrix was formulated by a co-precipitation technique and subsequently characterized by scanning electron microscopy, X-ray analysis, and infrared spectroscopy. The release of nitrate was monitored in different buffer mediums: buffer A as a simulated acidic soil solution and buffer B as a simulated neutral soil solution.

Results and discussion

The stability of nitrate-LDH against thermal decomposition was evaluated by thermal gravimetric analysis. The nitrate-LDH supported a sustained controlled release process of nitrate during 16 days into acidic soil at 15 °C, while the release was continued to 20 days into neutral soil at the same temperature. The increase of soil temperature slightly enhanced the release of nitrate.

Conclusions

We offered a potential management strategy of soil nitrogen leaching process. The nitrate form of layered double hydroxide material was used as a nitrogen fertilizer in order to monitor the release of nitrate anion into soil at different conditions.     

A dinuclear Ni(mu-H)Ru complex derived from H2

Models of the active site in [NiFe]hydrogenase enzymes have proven challenging to prepare. We isolated a paramagnetic dinuclear nickel-ruthenium complex with a bridging hydrido ligand from the heterolytic cleavage of H2 by a dinuclear NiRu aqua complex in water under ambient conditions (20 degrees C and 1 atmosphere pressure). The structure of the hexacoordinate Ni(mu-H)Ru complex was unequivocally determined by neutron diffraction analysis, and it comes closest to an effective analog for the core structure of the proposed active form of the enzyme.     

Regulation of CD8+ T cell development by thymus-specific proteasomes

Proteasomes are responsible for generating peptides presented by the class I major histocompatibility complex (MHC) molecules of the immune system. Here, we report the identification of a previously unrecognized catalytic subunit called beta5t. beta5t is expressed exclusively in cortical thymic epithelial cells, which are responsible for the positive selection of developing thymocytes. Although the chymotrypsin-like activity of proteasomes is considered to be important for the production of peptides with high affinities for MHC class I clefts, incorporation of beta5t into proteasomes in place of beta5 or beta5i selectively reduces this activity. We also found that beta5t-deficient mice displayed defective development of CD8(+) T cells in the thymus. Our results suggest a key role for beta5t in generating the MHC class I-restricted CD8(+) T cell repertoire during thymic selection.     

Tyrosinase inhibitory activity of proanthocyanidins from woody plants

Flavanol values, molecular weight distributions, polyphenolic patterns, and tyrosinase inhibitions of proanthocyanidins (PACs) from karamatsu (Larix sp.) bark, acacia Morishima (Acacia mearnsii) bark, and commercial quebracho (Schinopsis lorentzii) extracts were examined to investigate the relation between the structures of PACs from woody plants and tyrosinase inhibitory activities. Aqueous acetone (70%) extracts of acacia showed high flavanol contents and strong tyrosinase inhibition. On the other hand, the quebracho extracts inhibited tyrosinase activity slightly despite its high flavanol content. Phenyl nucleus analysis by the nucleus exchange reaction provided information that karamatsu, quebracho, and acacia extracts consist of procyanidin, profisetinidin, and prorobinetinidin, respectively. The relation between the phenolic hydroxylation pattern and tyrosinase inhibition suggested that the PACs with a 5,7-dihydroxyphenyl structure in the A-ring and a 3,4,5-trihydroxyphenyl structure in the B-ring have potent tyrosinase inhibitory activity.Part of this paper was presented at the 49th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1999     

Wichtige forstschädliche Insekten Japans

SummaryO. Eichhorn and K. Kanamitsu: Important forest insect pests of Japan The paper deals with the life histories, the damage and the limiting factors (parasites, diseases) of a number of forest insects which are considered to be severe pests in Japanese forest.Special regard is devoted to the pine shoot moth complex and to a group of bark and wood boring insects, which cause severe damage in the pine forests of southern Japan.

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RésuméO. Eichhorn et K. Kanamitsu: Importants insectes nuisibles aux forêts du Japon Ce travail traite du cycle biologique, des dégâts et des facteurs qui limitent la multiplication (parasites, maladies) d'un grand nombre d'insectes qui sont considérés comme très nuisibles aux forêts japonaises.Une attention spéciale est accordée au complexe des tordeuses des pousses du pin, et à un groupe d'insectes creusant l'écorce et le bois, qui produisent d'importants dégâts dans les forêts au sud du Japon.

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Den Herren Professor M. Nitto und Dr. C. Nishiguchi (Institute of Forest Zoology, Tokyo University) und K. Katagiri (Government Forestry Experiment Station Shikoku) danken wir für die Durchsicht des Manuskriptes.

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Den Herren Professor M. Nitto und Dr. C. Nishiguchi (Institute of Forest Zoology, Tokyo University) und K. Katagiri (Government Forestry Experiment Station Shikoku) danken wir für die Durchsicht des Manuskriptes.