Inhibition of host protein synthesis in B95a cells infected with the HL strain of measles virus

The shut-off of host protein synthesis in virus-infected cells is one of the important mechanisms for viral replication. In this report, we showed that the HL strain of measles virus (MeV-HL) as well as other field isolates, which were isolated from human blood lymphocytes using B95a cells, induce the shut-off in B95a cells. Since the Edmonston strain of MeV failed to induce the shut-off in B95a cells, the ability to induce the shut-off was considered to be dependent on virus strains. Although, the modification of eukaryotic translation initiation factors (eIF) including eIF4G, eIF4E, and 4E-BP1 was reported for shut-off by various viruses, the involvement of these eIFs was not observed in MeV-HL-infected B95a cells. Instead, the accumulation of phosphorylated eIF2alpha was found to coincide to the decrease of host protein synthesis, suggesting the involvement of phosphorylation of eIF2alpha in inhibition of translation as one of the mechanisms of the shut-off.     

Spontaneous Adenosquamous Carcinoma with Rapid Growth and EMT-like Changes in the Mammary Gland of a Young Adult Female BALB/c Mouse

This study histopathologically and immunohistochemically investigated a spontaneously occurring single mass subcutaneously located in the left lower abdomen of a female BALB/cAJcl−nu/+ mouse at 10 weeks of age. The mass was about 20 × 15 × 10 mm in size after formalin fixation; nevertheless, it was not detected by clinical observations at 9 weeks of age. H&E staining revealed the tumor origin was epithelial and probably arose from the mammary gland, and the tumor cells demonstrated a squamous, acinar or polyhedral/basal pattern. A cell kinetics analysis revealed that many of the tumor cells of the squamous, acinar or polyhedral/basal component were positive for PCNA and cyclin D1, although there were a few of TUNEL-positive tumor cells in all of the components. An epithelial/mesenchymal analysis demonstrated that most of the tumor cells of the squamous and acinar components contained keratin and E-cadherin; however, most of the tumor cells of the polyhedral/basal component were less or very weakly positive for these markers. The tumor cells of the squamous component were negative for vimentin and SMA; however, many of the tumor cells of the polyhedral/basal component exhibited vimentin. In addition, expression of SMA was confirmed in some tumor cells of the acinar and basal components. Based on the microscopic and immunohistochemical characterizations, the tumor was diagnosed to be adenosquamous carcinoma that originated from the mammary gland with rapid growth, and the tumor cells demonstrated epithelial-mesenchymal transition-like changes.     

Growth of juvenile chub mackerel Scomber japonicus in the western North Pacific Ocean: with application and validation of otolith daily increment formation

The growth of juvenile chub mackerel Scomber japonicus collected in the western North Pacific Ocean in 2007 and 2009 was examined based on the evidence of otolith daily increment formation in captive specimens. There was a significant difference in the relationship between known age and number of increments in the frontal and sagittal planes. Repeated markings on the otolith using Alizarin complexone and the coefficient of variation in number of increments suggest that the increments in the frontal plane of the otolith are more suitable for age estimation than those in the long and short axes of the sagittal plane. The increments in the frontal plane formed daily, and the first ring was usually deposited 3 days after hatch. Age of wild juveniles ranged from 24 to 211 days after hatch based on the frontal plane method. The estimated hatching periods of specimens ranged from February to June, but the April-hatched specimens were collected throughout the sampling periods of 2007 and 2009. The Gompertz growth model showed a difference in growth pattern in specimens between 2007 and 2009. The juveniles in 2009 appeared to grow more quickly than those in 2007 until summer, but thereafter the 2009 specimens seemed to grow more slowly.     

Expression and characterization of the recombinant swine interleukin-6

The swine interleukin-6 (SwIL-6) cDNA was cloned by RT-PCR and each expression system of recombinant SwIL-6 in Escherichia coli, insect cells, and mammalian cells was developed. Recombinant SwIL-6 produced in bacteria was applied for generation of the polyclonal antibodies. The rSwIL-6 was purified from supernatant of insect cells with a Q-sepharose or anti-SwIL-6 monoclonal antibody based immunoaffinity column. The antibodies showed that the molecular weight of rSwIL-6 was approximately 26kDa in E. coli, 25, 26, 30kDa in insect cells, and 26 and 30kDa in mammalian cells. These variations of molecular weight were probably due to the different modifications of glycosylation. All these recombinant proteins retained the antigenicity and biological activity on 7TD1 mouse cells.     

Birth of piglets through the non-surgical transfer of blastocysts produced in vitro

In this study, we attempted to produce piglets by non-surgically transferring blastocysts produced in vitro, using a flexible catheter as the transfer instrument. Cumulus-oocyte complexes (COCs) were aspirated from the follicles of ovaries obtained at a local slaughterhouse. They were then matured in modified North Carolina State University (NCSU)-37 medium for 44-46 h and fertilized in porcine gamete medium (PGM). Ten hours after in vitro fertilization (IVF), presumptive zygotes were removed from the cumulus cells and cultured in porcine zygote medium (PZM)-5. Blastocysts were cultured for five days after IVF and, using a catheter for deep intrauterine insemination without sedation, they were transcervically transferred into the uterine horn of six recipients (45-50 blastocysts/recipients) whose estrous cycles were synchronized, at 5 days after human chorionic gonadotropin (hCG) injection. Of the six recipients, one sow became pregnant and farrowed seven piglets (four live piglets) 119 days after hCG injection. The body weight at birth of the newborns ranged from 0.8 to 1.4 kg. These results indicate that it is possible to obtain piglets by transcervically transferring blastocysts produced by IVF and in vitro cultures in chemically defined media.     

Effects of delipidation and oxygen concentration on in vitro development of porcine embryos

The effects of delipidation and the oxygen (O(2)) concentration in the atmosphere during culture on in vitro development and H(2)O(2) content were investigated in porcine in vivo fertilized embryos and embryos after in vitro maturation and in vitro fertilization (IVM/IVF embryos). There was no significant difference in the developmental rates to the blastocyst stage between the intact and delipidated IVM/IVF embryos. However, the mean number of cells in blastocysts derived from delipidated IVM/IVF embryos (19.8 +/- 0.8 cells) was significantly smaller than that from intact embryos (24.2 +/- 1.2 cells). Although there were no significant differences in the developmental rates to the blastocyst stage of intact and delipidated IVM/IVF embryos between the cultures under 5% O(2) and 20% O(2), the developmental rate of intact IVM/IVF embryos cultured under 5% O(2) (27.1%) was significantly higher than that of the delipidated embryos cultured under 20% O(2) (19.3%). On the other hand, there was no difference in the developmental rate to the blastocyst stage between in vivo fertilized embryos cultured under 5% O(2) and 20% O(2). Hydrogen peroxide (H(2)O(2)), one of the reactive oxygen species (ROS), is thought to cause damage to embryos. The H(2)O(2) content per embryo derived from oocytes cultured under 5% O(2) (in vivo fertilized, 58.0 +/- 2.5 pixels; IVM/IVF, 79.6 +/- 3.2 pixels) was significantly lower than that (in vivo fertilized, 100.2 +/- 3.8 pixels; IVM/IVF, 103.9 +/- 3.2 pixels) under 20% O(2). Furthermore, the level of H(2)O(2) in delipidated IVM/IVF embryos (94.7 +/- 3.9 pixels) was significantly lower than that in intact embryos (103.9 +/- 3.2 pixels) cultured under 20% O(2). The present results indicate that the delipidation of porcine IVM/IVF embryos and reduction of the O(2) concentration decreased the H(2)O(2) level rather than the in vitro developmental rate to the blastocyst stage.     

A time-course study of flavonoids in the sprouts of tartary (Fagopyrum tataricum Gaertn.) buckwheats

The evolution, from 1 to 10 days after germination, of flavonoid content in sprouts of tartary buckwheat (Fagopyrum tataricum Gaertn.), grown in a greenhouse under low light conditions (16 μmol m⁻² s⁻¹), was investigated. Chlorogenic acid and flavonoids including C-glycosylflavones (orientin, isoorientin, vitexin, isovitexin), rutin and quercetin were separated from the sprouts by HPLC and quantified with their commercial standards. Rutin content in the edible portion of the sprouts (mean 20 and 37 mg g⁻¹ DW in ‘Hokkai T 8’ and ‘Hokkai T 10’, respectively) was 3- to 31-fold greater than that in the roots or pericarp. The free radical scavenging activity of seed sprouts was assessed through the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. From 6 to 10 days after sowing, the free radical scavenging activity in the edible portions rose significantly from 1.52 to 2.33 μmol Trolox equiv. g⁻¹ DW in ‘Hokkai T 8’ and from 1.46 to 2.09 μmol Trolox equiv. g⁻¹ DW in ‘Hokkai T 10’, but differences between ‘Hokkai T 8’ and ‘Hokkai T 10’ were not significant. As the results, the sprouts of tartary buckwheat, particularly those of ‘Hokkai T 10’ are strongly recommended as new high rutin food.     

Estimation of aboveground biomass in mangrove forests using high-resolution satellite data

Mangroves play important roles in providing a range of ecosystem services, mitigation of strong waves, protection of coastlines against erosion, maintenance of water quality, and carbon sink in the context of global warming. For trees in mangrove forests in southern Ranong Province, Thailand, we investigated the allometric relationship between crown area derived from high-resolution satellite data and stem diameter and used the resulting model to estimate aboveground biomass. We used QuickBird panchromatic and multispectral data acquired for the study area on 15 October 2006 as the high-resolution satellite data. Individual tree crowns were extracted from the satellite image of panchromatic data by using the watershed method, and the species were identified by using the maximum-likelihood method for the multispectral data. Overall classification accuracy for species identification was 88.5 %. The biomass derived from our field survey was plotted against aboveground biomass in the sample plots, estimated from the QuickBird data. The regression line through the origin between the satellite-estimated biomass and biomass based on the field data had a slope of 1.26 (R ² = 0.65). Stand aboveground biomass estimated from the high-resolution satellite data was underestimated because of a lack of data on the biomass of suppressed trees and inappropriate segmentation of crowns of large trees into two or more trees.     

Groundwater Deterioration Caused by Induced Recharge: Field Survey and Verification of the Deterioration Mechanism by Stochastic Numerical Simulation

Our field survey showed that thequality of shallow groundwater around the KatsuraRiver in the Kyoto Basin was strongly affected by theinfiltration of river water. Furthermore, that thedeterioration of the groundwater in the southern areato the west of the Katsura River may be related to theincrease in groundwater extraction. To clarify themechanism of groundwater deterioration, we havedeveloped a stochastic method to simulate groundwaterflow. The results showed that there was a largereduction in the groundwater level where groundwaterextraction was intense and recharge flowed from theKatsura River to the high extraction areas in thesouthern region. Another simulation showed that if thegroundwater extraction was 10% of the present removalrate, there would be little recharge from the KatsuraRiver into the groundwater and the quality of thegroundwater would be improved. Thus, we conclude thatthe cause of groundwater deterioration is probably dueto the induced recharge of deteriorated river waterfrom the Katsura River.