Morphological reproductive characteristics of testes and fertilization capacity of cryopreserved sperm after the Fukushima accident in raccoon (Procyon lotor)

Since the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident, we have established an archive system of livestock and wild animals from the surrounding ex-evacuation zone. Wildlife within the alert zone have been exposed to low-dose-rate (LDR) radiation for a long continuous time. In this study, we analysed the morphological characteristics of the testes and in vitro fertilization (IVF) capacity of cryopreserved sperm of racoons from the ex-evacuation zone of the FDNPP accident. The radioactivity of caesium-137 (¹³⁷Cs) was measured by gamma-ray spectrometry, and the measured radioactivity concentration was 300–6,630 Bq/kg in the Fukushima raccoons. Notably, normal spermatogenesis was observed in the seminiferous tubules of the testes, with the germinal epithelium composed of a spermatogenic cell lineage with no evident ultrastructural alterations; freeze-thawing sperm penetration ability was confirmed using the interspecific zona pellucida-free mouse oocytes IVF assays. This study revealed that the chronic and LDR radiation exposure associated with the FDNPP accident had no adverse effect on the reproductive characteristics and functions of male raccoons.     

Effect of simvastatin on cell proliferation and Ras activation in canine tumour cells

Statins are inhibitors of the mevalonate cascade that is responsible for cholesterol biosynthesis and the formation of intermediate metabolites, farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP) used in the prenylation of proteins. Although statins are widely used in the treatment of hypercholesterolemia, recent studies suggest that they also inhibit proliferation of tumour cells by reducing prenylation of small GTP‐binding proteins, such as, Ras. This study aimed to evaluate the effect of simvastatin on cell proliferation and Ras activation in various canine tumour cell lines, including hemangiosarcoma (HSA), melanoma, and lymphoma cell lines. Simvastatin inhibited cell proliferation of all cell lines tested in a concentration‐ and time‐dependent manner, but the susceptibilities were different amongst the cell lines. Simvastatin induced apoptotic cell death via activation of caspase‐3 and cell cycle arrest. The cytotoxic effects of simvastatin were attenuated by GGPP and FPP. Simvastatin decreased the amount of prenylated Ras and GTP‐bound Ras in HSA and melanoma cell lines, but not in lymphoma cell lines. These results indicate that simvastatin induces cytotoxic effects through the depletion of GGPP and FPP in a variety of canine tumour cells, whereas multiple mechanisms are involved in the effects. Further study is required to elucidate the underlying mechanisms of simvastatin‐induced cytotoxic effects in a variety of canine tumour cells.     

Nuclear and mitochondrial DNA polymorphisms suggest introgression contributed to garden beet (Beta vulgaris L.) domestication

Genetic Resources and Crop Evolution - Garden beet is the ancestor of fodder beets and sugar beets, but the origin of garden beet’s genetic potential to evolve novel beet types is debatable....     

Evaluation and mapping of relative sensitivity of the terrestrial ecosystems to acidic deposition in Fujian

Based on the semi-quantitative approach, four environmental factors of sites (i.e. bedrock lithology, soil type, land use, and rainfall) were categorized, weighted and combined to determine and assess the relative sensitivity of the terrestrial ecosystems to acidic deposition in Fujian Province. Then the factors have been digitized and combined to assign an overall value for each mesh square (16.77 km×18.39 km) by using the geographic information system (GIS) The results indicated that the most sensitive area in Fujian was mainly located in the southeast, and the least: ensitive area was distributed sporadically in the east along the coast. Due to slow weathering rate of siliceous rocks, acid to weakly acid reactions of the soils, along with the greater percent of coniferous forests, more than 80 percent of the total area exhibits higher sensitivity classes (4–7). This project is jointly sponsored by Forestry Administration of Fujian Province and Japan International Forestry Promotion and Cooperation Center (k99066). Fan Houbao, male, born in 1965, associate professor of Fujian Forestry College, Nanping 353001, P.R. China. Responsible editor: Zhu Hong     

Differential kinetic activities of glycerol kinase among African trypanosome species: phylogenetic and therapeutic implications

African trypanosome species are causative agents for sleeping sickness in humans and nagana disease in cattle. Trypanosoma brucei can generate ATP via a reverse reaction with glycerol kinase (GK) when alternative oxidase (AOX) is inhibited; thus, GK is considered to be a crucial target for chemotherapy combined with AOX. However, the energy metabolism systems of African trypanosome species other than T. brucei are poorly understood. Thus, GK genes were surveyed from genome databases and cloned by PCR from T. vivax and T. congolense. Then, recombinant GK proteins (rGK) of T. vivax, T. congolense and T. brucei were expressed and purified. Kinetic analysis of these rGK proteins revealed that the K(m) values of T. congolense rGK for ADP and G-3-P substrates were lower than those of T. vivax and T. brucei. The expression level of GK molecules was highest in T. congolense cells and lowest in T. vivax cells. Based on these results, effective combination dosages of ascofuranone, a specific inhibitor of AOX, and glycerol, an inhibitor of the GK reverse reaction, were determined by using in vitro-cultured trypanosome cells.     

Enzyme-linked immunosorbent assay with monoclonal and single-chain variable fragment antibodies selective to coplanar polychlorinated biphenyls

Coplanar polychlorinated biphenyls (Co-PCBs) consisting of non-ortho and mono-ortho-chlorinated PCBs are dioxin-like compounds and cause wide contamination in the environment. To monitor Co-PCB residues, it was attempted to establish an enzyme-linked immunosorbent assay (ELISA) with monoclonal and recombinant antibodies selective to Co-PCBs. When 3,3',5,5'-tetrachlorobiphenoxybutyric acid (PCBH)-keyhole limpet hemocyanin conjugate was immunized into mice, two monoclonal antibodies, Mab-0217 and Mab-4444, were obtained. 3,3',5,5'-Tetrachlorobiphenyl (PCB80) was determined with an IC(50) value of 2.6 and 0.46 ng mL(-1) in ELISA based on Mab-0217 and Mab-4444, respectively. Mab-4444 cross-reacted with Co-PCB congeners, except for PCB77 and PCB81. Mab-0217 reacted with PCB80 and cross-reacted with PCB111. A single-chain variable fragment (scFv) antibody derived from Mab-4444 was produced in recombinant Escherichia coli cells. The scFv antibody showed nearly the same sensitivity toward PCBH as the parent monoclonal antibody in ELISA. These results clearly suggested that Mab-4444 and its scFv antibodies were suitable for monitoring the representative congeners of Co-PCBs.     

Identification of novel genetic markers and evaluation of genetic structure in a population of Japanese crested ibis

Japanese population of the Japanese crested ibis Nipponia nippon was founded by five individuals gifted from the People's Republic of China. In order to exactly evaluate genetic structure, we first performed development of novel genetic makers using 89 microsatellite primer pairs of related species for cross‐amplification. Of these, only three primer pairs were useful for the genetic markers. Additionally, we sequenced allelic PCR products of these three markers together with 10 markers previously identified. Most markers showed typical microsatellite repeat units, but two markers were not simple microsatellites. Moreover, over half of the markers did not have the same repeat units as those of the original species. These results suggested that development of novel genetic markers in this population by cross‐amplification is not efficient, partly because of low genetic diversity. Furthermore, the cluster analysis by STRUCTURE program using 17 markers showed that the five founders were divided into two clusters. However, the genetic relationships among the founders indicated by the clustering seemed to be questionable, because the analysis relied largely on a small number of triallelic markers, in spite of the addition of the three useful markers. Therefore, more efficient methods for identifying large numbers of single nucleotide polymorphisms are desirable.     

Systemic spindle-cell proliferative disease in broiler chickens

The major organs and tissues of 24 broiler chickens (70 or 71 days old) suspected of spindle-cell proliferative disease (SPD) because of showing the tumorous lesions distributed throughout the body at meat inspection were collected for histopathological and immunohistochemical examination. Macroscopically, liver, spleen and cecal tonsil showed severe enlargement and white nodules or plaques were observed in the liver, spleen, kidney, intestine and bone marrow of the femur. All chickens were diagnosed with SPD based on the histopathological examination. The lesions of SPD were observed in the liver, spleen, kidney, heart, lung, pancreas, proventriculus, gizzard, duodenum, jejunum, ileum, rectum, cecal tonsil, bursa of Fabricius, bone marrow of the femur and skin. Hemangioma was observed in the lung of 1 bird. Eight 1-day-old specific pathogen-free chicks were inoculated intraperitoneally with 0.25 ml of a 20% homogenate of the affected spleens of three naturally occurring cases. One inoculated bird, necropsied at 10 weeks of age, macroscopically had a white nodule in the kidney and histopathologically had spindle-cell proliferative lesions, a pattern similar to that seen in the naturally occurring cases, in the liver, spleen, kidney, heart, lung, pancreas, proventriculus, duodenum, cecal tonsil and bone marrow of the femur, and was diagnosed with SPD. Immunohistochemically, significant positive reactions with a rabbit antiserum against avian leukosis virus antigens were detected in all spindle cells in the proliferative lesions of all examined SPD cases and in tumor cells of the hemangioma of a field case.