Finding of a highly efficient ZFN pair for Aqpep gene functioning in murine zygotes

The generation efficiencies of mutation-induced mice when using engineered zinc-finger nucleases (ZFNs) have been generally 10 to 20% of obtained pups in previous studies. The discovery of high-affinity DNA-binding modules can contribute to the generation of various kinds of novel artificial chromatin-targeting tools, such as zinc-finger acetyltransferases, zinc-finger histone kinases and so on, as well as improvement of reported zinc-finger recombinases and zinc-finger methyltransferases. Here, we report a novel ZFN pair that has a highly efficient mutation-induction ability in murine zygotes. The ZFN pair induced mutations in all obtained mice in the target locus, exon 17 of aminopeptidase Q gene, and almost all of the pups had biallelic mutations. This high efficiency was also shown in the plasmid DNA transfected in a cultured human cell line. The induced mutations were inherited normally in the next generation. The zinc-finger modules of this ZFN pair are expected to contribute to the development of novel ZF-attached chromatin-targeting tools.     

Consecutive monitoring for conidiogenesis by Oidium neolycopersici on tomato leaves with a high-fidelity digital microscope

Conidiogenesis by Oidium neolycopersici KTP-01 on tomato leaves was vitally monitored with a high-fidelity digital microscope. Conidiophores were initially formed 3 days after inoculation and then elongated to a maximum length within at least 12h. The apical part was split into two cells after two successive septations, accompanied by apical expansion. These cells subsequently developed into primary and secondary conidia. An additional septation at the stem portion of the conidiophores produced a generative and a foot cell. Subsequent conidiation occurred during repeated cycles of splitting of the generative cell, maturation of the apical cell into a conidium, and abstriction of the conidium. To our knowledge, this report is the first on the developmental process of conidiogenesis by powdery mildew on host leaves as revealed with the digital microscope.     

Infectivity of a Japanese isolate of <Emphasis Type="Italic">Oidium neolycopersici</Emphasis> KTP-01 to a European tomato cultivar resistant to <Emphasis Type="Italic">O. lycopersici</Emphasis>

The infectivity of a Japanese isolate of tomato powdery mildew, Oidium neolycopersici KTP-01, to tomato cultivars was examined using a resistant cultivar Grace bred in The Netherlands to O. lycopersici, which was recently proposed to be renamed O. neolycopersici. Grace was severely infected with KTP-01, and its susceptibility was similar to that on susceptible tomato cultivars Moneymaker and Ponderosa, suggesting that KTP-01 differs in pathogenicity on tomatoes from those of European and American isolates.     

Correlated responses of respiratory disease and immune capacity traits of Landrace pigs selected for Mycoplasmal pneumonia of swine (MPS) lesion

Five generations of Landrace pigs selected for average daily gain, backfat thickness, Mycoplasmal pneumonia of swine (MPS) lesion score, and plasma cortisol levels, was executed to decrease the MPS lesion score. Genetic parameters and correlated genetic responses for respiratory disease and peripheral blood immune traits were estimated in 1395 Landrace pigs. We estimated the negative genetic correlation of MPS lesion score with phagocytic activity (PA) at 7 weeks of age (‐0.67). The breeding values of PA at 7 weeks of age and 105 kg body weight and the correlated selection response of the ratio of granular leukocytes to lymphocytes at 105 kg body weight were significantly increased, and sheep red blood cell‐specific antibody production (AP) was significantly decreased in a selection‐dependent manner. Increasing of natural immunological indicators (e.g. PA) and decreasing of humoral immunological indicator (e.g. AP) were observed due to genetically decreasing MPS lesion score.     

Participatory forest conservation in Southwest lowland rainforests of Sri Lanka

Natural forest in Sri Lanka has been decreased significantly in the last few decades. The remaining natural forests especially in floristic region seven have undergone less conservation efforts in the past. Considering the capacity of the government and dependence on forest resources by local villagers, the only way for conservation of these forests in the Southwest lowland is forest management through local participation. Management plans for community-based resource management have been completed. However promotion of such management requires integrated measures, which are beyond jurisdiction of Forest Department. A holistic approach with political commitment concerning buffer zone villages and economic incentives with income generation opportunities supported by external inputs are expected to be implemented as a matter of urgency.     

Differential expression of genes encoding neurotrophic factors and their receptors along the septal‐temporal axis of the rat hippocampus

The hippocampus plays a key role in learning and emotional regulation. The hippocampus’ function varies along its septotemporal axis, with the septal pole being more frequently involved in spatial learning and memory, and the temporal pole playing a greater role in emotional behaviors. In this study, we present findings aimed at checking the expression level of the genes encoding neurotrophins and their receptors, including nerve growth factor (NGF), brain‐derived neurotrophic factor (BDNF), neurotrophin‐3 (NT‐3), and their receptors (TrkA, TrkB and TrkC) in the hippocampus along the septotemporal axis. Using real‐time PCR, several different expression patterns were observed. Remarkably, the expression of both NT‐3 and TrkA genes in the septal hippocampus was higher than in the middle and temporal hippocampus. Higher expression of NT‐3 and TrkA may implicate active neurogenesis in the dentate gyrus (DG) of the septal hippocampus because more neurogenesis occurs in the septal than the temporal DG of rats. Finally, the results obtained in this study emphasize the importance of choosing the hippocampal portion along its septotemporal axis for any hippocampal molecular and biochemical experimental studies.     

A cytochrome C oxidase model catalyzes oxygen to water reduction under rate-limiting electron flux

We studied the selectivity of a functional model of cytochrome c oxidase's active site that mimics the coordination environment and relative locations of Fe(a3), Cu(B), and Tyr(244). To control electron flux, we covalently attached this model and analogs lacking copper and phenol onto self-assembled monolayer-coated gold electrodes. When the electron transfer rate was made rate limiting, both copper and phenol were required to enhance selective reduction of oxygen to water. This finding supports the hypothesis that, during steady-state turnover, the primary role of these redox centers is to rapidly provide all the electrons needed to reduce oxygen by four electrons, thus preventing the release of toxic partially reduced oxygen species.     

The Process and Development Mechanism of Age-related Fibrosis in the Pancreatic Islets of Sprague-Dawley Rats: Immunohistochemical Detection of Myofibroblasts and Suppression Effect by Estrogen Treatment

The mechanism of spontaneous islet fibrosis in Sprague-Dawley rats was investigated. Using sections of the pancreas in naive males aged 26 to 102 weeks old and 26-week-old males injected with β-estradiol 3-benzoate (EB), the incidence of lesions and histological scores of fibrosis were examined in conjunction with immunohistochemistry for α-smooth muscle actin (α-SMA), platelet-derived growth factor receptor-α (PDGFRα) and estrogen receptor-α (ERα). The incidence of islet fibrosis increased in 78-week-old animals compared to the 26-week-old animals, and the incidence of atrophy in the fibrotic islet increased in animals over 52 weeks old. α-SMA and PDGFRα were positively stained mainly in fibrotic/inflammatory islets, and the histological score of α-SMA in the fibrotic islet decreased age-dependently. Notably, α-SMA and PDGFRα were co-expressed in inflammatory islets with a high score at all ages. The positive index of ERα in the EB-treated group increased when compared with that of the naive group. However, it was independent of the existence of fibrosis. In contrast, the score of α-SMA and PDGFRα decreased in the EB-treated group. In conclusion, it was clarified that a part of age-related fibrosis in islets became atrophy with age, and α-SMA-positive myofibroblasts were considered to contribute to the development of fibrosis. Strong PDGFRα stainability in fibrotic/inflammatory islets may imply that myofibroblasts were stimulated by PDGF to produce an extracellular matrix. Although estradiol has been known to suppress fibrosis/inflammation in the islet, nuclear-located ER-dependent signaling was considered not to be involved in the suppression mechanism. EB possibly affected the inhibition of the appearance of myofibroblasts.     

Habitat management of little terns in Japan’s highly developed landscape

The little tern (Sterna albifrons) is a species of shorebird that nests in colonies on sandy beaches and riverbanks with little vegetation cover. In Japan, the natural breeding habitats of the terns have decreased drastically, and the species is listed under the category “vulnerable” in the Red List of the Government of Japan. To reduce the effects of habitat loss on the terns, conservationists are attempting to create and manage artificial colony sites in highly developed landscapes, such as reclaimed lands located in Tokyo Bay, central Japan. We present the factors that contribute to breeding success for this species and the habitat characteristics related to nest-site selection in the artificial colony sites along Tokyo Bay. Our data show that around 3 ha of a building rooftop without vegetation cover could sustain more than 2000 little tern nests. We found a strong positive relationship between colony size and hatching success, observing that a colony size of more than 100 lowered predation rates of eggs. On sites built upon white crushed-concrete, nest densities were high and predation rates of eggs were low. We also found that feeding rates were affected by foraging habitats, of which the best types were the shoreline of sandy beaches and mud flats. The 40 km dispersal range of the breeding terns, inferred from distribution data, should be considered when establishing a network of the multiple colony sites.