Genetic diagnosis of band 3 deficiency and sexing in bovine preimplantation embryos

Band 3 deficiency with hereditary spherocytosis and hemolytic anemia in Japanese black cattle, band 3(Bov.Yamagata), is caused by a total lack of band 3 protein with an autosomal dominant inheritance. Genotyping for band 3 deficiency and sexing were successfully achieved in biopsied embryo cells with efficiencies of 98.4% and 97.4%, respectively. Transfer of the embryo that was determined as homozygous for the mutant allele into a recipient cow resulted in the production of a fetus exhibiting the genotype and red cell phenotypes characteristic of band 3(Bov.Yamagata). These results demonstrate that our procedure is reliable and applicable to produce animals free from or homozygous for the mutant allele by breeding carrier animals.     

Heavy-ion-induced mutants in sweet pepper isolated by M1 plant selection

We examined the effects of heavy-ion bombardment on mutagenesis in sweet pepper (Capsicum annuum L.). Dose–response studies indicated that 10 Gy irradiation of ¹²C or ²⁰Ne ions on dry seeds is suitable for inducing mutations in plants. From ²⁰Ne-irradiated M₁ plants, putative mutants included two dwarf plants and one plant whose pericarp was yellow were isolated. Phenotypes of their M₂ progeny were similar to those of the M₁ plants and did not segregate. F₁ plants resulting from reciprocal crosses between the mutants and wild-type plants showed the wild-type phenotype, but phenotypes of F₂ and BC₁F₁ segregated at 1:3 (mutant:wild) and 1:1, respectively. These crossing experiments indicate that the three mutants have monogenic recessive mutations in nuclear genes. In light of these data, we discuss the effectiveness of using heavy-ion bombardment to mutate sweet peppers.     

Pigments in green tea leaves (Camellia sinensis) suppress transformation of the aryl hydrocarbon receptor induced by dioxin

Environmental contaminants such as dioxins enter the body mainly through diet and cause various toxicities through transformation of the aryl hydrocarbon receptor (AhR). We previously reported that certain natural flavonoids at the dietary level suppress the AhR transformation induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). In this study, we identified lutein and chlorophyll a and b from green tea leaves as the novel antagonists for AhR. These active compounds suppressed AhR transformation dose-dependently with the 50% inhibitory concentration (IC(50)) values against 0.1 nM TCDD-induced AhR transformation at 3.2, 5.0, and 5.9 microM, respectively. (-)-Epigallocatechin gallate, which is the most abundant flavonoid in green tea leaves, also showed stronger suppressive effects than did other major tea components, with the IC(50) value of 1.7 microM. Thus, these pigments of green tea leaves have the potential to protect from dioxin toxicity through the suppression of AhR transformation.     

Suppressive effects of ethanolic extracts from propolis and its main botanical origin on dioxin toxicity

Suppressive effects of ethanolic extracts prepared from propolis group 12 and its main botanical origin (leaf bud of Baccharis dracunculifolia) on transformation of the aryl hydrocarbon receptor (AhR), the initial action of dioxin toxicity, were investigated. It was found that suppressive effects of propolis on AhR transformation were relatively higher than those of resins of its botanical origin in cell-free system and in Hepa-1c1c7 cells. When the composition of chemical ingredients was measured, propolis contained slightly higher amounts of flavonoid aglycones as compared with its botanical origin with the same characteristics. Moreover, antiradical activity, one of the typical biological activities of flavonoids, in propolis was also slightly higher than that in its botanical origin. These results indicate that not only propolis but also its botanical origin contains high amounts of flavonoid aglycones and that both of them are useful dietary sources for flavonoids with a potency to prevent dioxin toxicity.     

Molecular identification of Candida parapsilosis from crop mucosa in a cockatiel.

A 2-month-old cockatiel was evaluated for diarrhea, dyspnea, and death. Histologic examination of lesions in the crop mucosa revealed hyperkeratosis and the presence of blastoconidia and hyphae. Positive immunohistochemical staining of the organisms was achieved with an antibody directed against Candida spp. Polymerase chain reaction amplification of DNA from crop lesion material with internal transcribed spacer 2 (ITS2) primers yielded fragments of approximately 300 bp, which demonstrated 95% DNA homology with the corresponding sequence from a strain of Candida parapsilosis deposited in the GenBank data base. The Candida species in the lesion of the crop mucosa was therefore identified by DNA sequence analysis as C. parapsilosis.     

Natural antibiotic function of a human gastric mucin against Helicobacter pylori infection

Helicobacter pylori infects the stomachs of nearly a half the human population, yet most infected individuals remain asymptomatic, which suggests that there is a host defense against this bacterium. Because H. pylori is rarely found in deeper portions of the gastric mucosa, where O-glycans are expressed that have terminal alpha1,4-linked N-acetylglucosamine, we tested whether these O-glycans might affect H. pylori growth. Here, we report that these O-glycans have antimicrobial activity against H. pylori, inhibiting its biosynthesis of cholesteryl-alpha-D-glucopyranoside, a major cell wall component. Thus, the unique O-glycans in gastric mucin appeared to function as a natural antibiotic, protecting the host from H. pylori infection.     

Epigenetically immature oocytes lead to loss of imprinting during embryogenesis

Loss of imprinting (LOI) is occasionally observed in human imprinting disorders. However, the process behind the LOI is not fully understood. To gain a better understanding, we produced embryos and pups from mouse oocytes that lacked a complete methylation imprint using a method that involved transferring the nuclei of growing oocytes into the cytoplasm of enucleated fully grown oocytes following in vitro fertilization (IVF). We then analyzed the imprinting statuses. Our findings show that the incomplete methylation imprint derived from growing oocytes results in epigenetic mosaicism or a loss of methylation imprint (LOM) at maternal alleles in embryos. In some embryos, both hypo- and hypermethylated maternal Kcnq1ot1 alleles were detected, whereas either hypo- or hypermethylated maternal Kcnq1ot1 alleles were detected in others. Such tendencies were also observed at the Igf2r and Mest loci. Gene expression levels of imprinted genes were linked with their methylation statuses in some but not all embryos. Possible explanations of the inconsistency between the data from DNA methylation and gene expression include epigenetic mosaicism in embryos. Pups were successfully produced from growing oocytes at a quite low frequency. They exhibited an obese phenotype and LOI with respect to Igf2r, Snrpn and Mest. Our finding suggests the possibility that LOI/LOM at maternal alleles in human concepti could be derived from epigenetically immature/mutated oocytes.     

Production and characterization of interspecific hybrids between Asparagus kiusianus Makino and A. officinalis L.

Interspecific hybridization is a useful technique to introduce characteristics from wild species into crops. Garden asparagus (Asparagus officinalis, 2n = 2x = 20) is an economically important vegetable that is native to the Mediterranean region but widely cultivated in many countries. The genus Asparagus is comprised of over 100 species. Asparagus kiusianus (2n = 2x = 20) is a wild asparagus species endemic to Japan. This species occurs on the coast and is likely to be a salt-resistant species. Although the geographic distribution of these two species is not close, molecular phylogenetic analysis demonstrated that these two species are closely related. In this study, a reciprocal cross between A. officinalis and A. kiusianus was carried out by hand pollination, and progeny were obtained from both crossings. These progeny exhibited a morphologically intermediate phenotype in terms of flower shape, and restriction fragment length polymorphism analysis confirmed that these were indeed interspecific hybrids. The interspecific hybrids were fertile, and backcross progeny with garden asparagus was also generated. These interspecific hybrids are expected to supply novel traits to garden asparagus.     

Ordered metal nanohole arrays made by a two-step replication of honeycomb structures of anodic alumina

A highly ordered metal nanohole array (platinum and gold) was fabricated by a two-step replication of the honeycomb structure of anodic porous alumina. Preparation of the negative porous structure of porous alumina followed by the formation of the positive structure with metal resulted in a honeycomb metallic structure. The metal hole array of the film has a uniform, closely packed honeycomb structure approximately 70 nanometers in diameter and from 1 to 3 micrometers thick. Because of its textured surface, the metal hole array of gold showed a notable color change compared with bulk gold.