Noxa, a BH3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis

A critical function of tumor suppressor p53 is the induction of apoptosis in cells exposed to noxious stresses. We report a previously unidentified pro-apoptotic gene, Noxa. Expression of Noxa induction in primary mouse cells exposed to x-ray irradiation was dependent on p53. Noxa encodes a Bcl-2 homology 3 (BH3)-only member of the Bcl-2 family of proteins; this member contains the BH3 region but not other BH domains. When ectopically expressed, Noxa underwent BH3 motif-dependent localization to mitochondria and interacted with anti-apoptotic Bcl-2 family members, resulting in the activation of caspase-9. We also demonstrate that blocking the endogenous Noxa induction results in the suppression of apoptosis. Noxa may thus represent a mediator of p53-dependent apoptosis.     

A spontaneously occurring malignant ovarian Sertoli cell tumor in a young Sprague Dawley rat

Primary ovarian tumors are generally uncommon in rats used in toxicologic studies. A malignant Sertoli cell tumor was present in the ovary of a 19-week-old female Sprague Dawley rat. Macroscopically, the mass was white and firm, 10 × 13 × 17 mm in size, and located in the right ovary. Histopathologically, the mass was composed of nests of pleomorphic cells, which formed seminiferous-like tubules separated by a thin fibrovascular stroma. The tubules were lined by tumor cells, which had basally located nuclei and abundant eosinophilic and vacuolated cytoplasm. In some areas, the tumor cells were arranged in a retiform growth pattern, mimicking a rete testis/ovarii. Disseminated metastases to the surfaces of the mesentery, spleen and liver were also present. Immunohistochemically, many tumor cells were strongly positive for vimentin, estrogen receptor α and Ki 67. Some tumor cells were positive for pancytokeratin and inhibin α. These findings closely resemble those of an ovarian-derived human malignant Sertoli cell tumor. From our review of the literature, we believe this is the first report of a spontaneous malignant Sertoli cell tumor in the ovary of a young laboratory rat. This case might provide useful historical control information for rat toxicity studies.     

Transepithelial Transport of Theasinensins through Caco-2 Cell Monolayers and Their Absorption in Sprague-Dawley Rats after Oral Administration

The aim of this study is to illustrate the in vivo and in vitro absorption of theasinensins B and A that are (-)-epigallocatechin-3-O-gallate (EGCG)-(-)-epigallocatechin (EGC) dimer and EGCG dimer, respectively, and their transport pathway across the intestinal membrane. Our animal study by a single oral administration to rats demonstrated the intact absorption of theasinensins into the blood system, which was estimated to be a >10-fold lower absorption amount than EGCG. The in vitro absorption study indicated that theasinensins can be transported across Caco-2 cell monolayers, while their permeability coefficients were also >10-fold lower than those of EGCG and EGC. Transport experiments using cytochalasin D or quercetin as a tight junction (TJ) modulator and a non-saturable permeation revealed that theasinensins were transported across Caco-2 cells in a TJ paracellular diffusion route. In conclusion, the dimers of condensed catechins, theasinensins B and A, can be absorbed intact into rat blood and transported across Caco-2 cell monolayers probably through a TJ paracellular pathway.     

Effects of management, environment and landscape conditions on establishment of hardwood seedlings and saplings in central Japanese coniferous plantations

There is an increasing need to restore natural hardwood forests in landscapes dominated by monocultural conifer plantations. A convenient restoration approach is to exploit natural regeneration processes. Natural regeneration, however, is affected by diverse interacting factors, for which better understanding is required, in order to optimize restoration programs. To identify optimal management practices for improving natural regeneration of hardwood trees in coniferous plantations, we examined the effects of multiple factors on the abundance of seedlings, small saplings and large saplings (height <0.3, 0.3-1.3 and ?1.3 m, respectively) of hardwood tree and shrub species in both line thinned (LT) and unthinned (UT) plantations of sugi (Cryptomeria japonica) and hardwood forests (HF) in central Japan. The effects of management practices (number of the times of weeding and cleaning, thinning method, years after thinning and forest age), environment (slope position, slope angle and canopy openness), and landscape conditions (distance from nearest hardwood forest, altitude and landuse before planting) on the number of hardwood individuals were examined by using the data obtained from the LT plantations. We also compared hardwood density between LT and UT plantations to examine the effect of line thinning. Finally, we examined species composition of LT plantations and HF to identify hardwood forest components in the thinned plantations. The effects on hardwood regeneration of environmental conditions, landscape factors and management practices applied in the plantations varied, depending on the size class and life form of the regenerating species. The abundance of large saplings of tall tree species was affected by several management factors, especially number of the times of weeding. Landscape conditions (distance from the nearest hardwood forest and altitude) affected the abundance of small saplings and seedlings of tall tree species, but not the other classes. Seedlings and small saplings of many tall tree species that contribute to hardwood forest canopies were less abundant in the LT plantations. The results show that numerous factors affect the establishment and abundance of naturally regenerating hardwood tree species, and suggest that successful establishment during early plantation stages can have long-lasting effects on natural regeneration of tall tree species.     

Assessing sexual maturity of feminized Japanese eel Anguilla japonica by measuring eye size

For efficient production of Japanese eel Anguilla japonica eggs, knowledge of the status of the sexual maturity of potential broodstock females is important because this status directly influences the time required to produce mature eggs by successive hormone doses. Here we apply an eye index (relative eye size) to evaluate the gonadal status of feminized A. japonica, which were induced by administration of estradiol-17β. Examination of gonad somatic and eye indexes of 267 feminized eels, cultivated for 12–56 months, revealed a significant correlation between these two indexes. Artificial maturation of 35 feminized eels showed that the number of hormone injections administered before ovulation was significantly lower as the eye index score increased, indicating availability of this noninvasive method of predicting sexual maturity of feminized eel. There was no correlation between eye index and fertilization rate, hatching rate, or larval survival rate, suggesting that sexual maturity before hormonal treatment does not affect egg quality.     

Observation of three‐dimensional flow structures and effluent transport around fish cages using a towed ADCP and free‐fall multi‐parameter profiler

A field campaign was conducted around salmon cages, using a combination of a towed ADCP and a free‐fall multi‐parameter profiler, in order to investigate flow structures and the possible distribution of effluent materials. Two transect observations showed that hydrographic conditions changed dramatically within 5 days, from highly stratified open water conditions to weak stratification. Three‐dimensional observation revealed that flow was blocked behind the cages and that the blockage was reduced as the distance from the cages increased. The flow speed was positively correlated with the intensity of the backscattering signal. The R² value was high immediately behind the cages and decreased with distance from the cages. The flow time series behind the cages exhibited a k^–2 power law spectrum that was consistent with a typical internal wave spectrum. This suggests that eddies shed from the cages were highly influenced by stratification. We estimated the rate of kinetic energy dissipation, , from YODA Profiler data based on the Thorpe scale approach. We also estimated the eddy diffusivity coefficient, . Both and followed a lognormal probability density function. The mean was consistent with a one‐dimensional diffusion model assessed from the R² values of flow speed and backscatter intensity.     

Commercialized European bumblebee can cause pollination disturbance: An experiment on seven native plant species in Japan

Non-native pollinator species are now widely utilized to facilitate pollination of agricultural crops. Evaluation of the ecological risk of alien pollinators is necessary because they could have a large impact on native ecosystems through disturbing native plant-pollinator interactions. We conducted a greenhouse experiment to examine the impact of the non-native commercialized European bumblebees, Bombus terrestris, on the pollination success of seven Japanese bumblebee-pollinated plant species. Plants were exposed to three groups of bumblebees: native bumblebee(s) only (NATIVE treatment); the alien bee only (ALIEN) and a mix of the two (MIX). ALIEN treatment had negative effects on fruitset and/or fruit quality of five plants, including self-incompatible and compatible, herb and woody, and queen- and worker-pollinated species. The negative effects were caused by a decrease in legitimate flower visitation due to (1) physical inaccessibility to nectary in deep-corolla flowers by the alien bee with insufficient tongue length and, (2) biased flower preference between short-corolla flowers. Fruitset tended to decrease drastically for the self-incompatible species while fruit quality decreased moderately for the self-compatible species. Effects of MIX were not intermediate between NATIVE and ALIEN in most plant species, and caused pollination success to vary in an unpredictable manner amongst plant species, probably due to interaction between native and alien bees. This non-linear relationship between plants’ pollination success and the relative density of the alien suggests that the alien bee can disturb pollination of a plant species even when only representing a small fraction of the total pollinator community.