cFLIP regulates death receptor-mediated apoptosis in an ovarian granulosa cell line by inhibiting procaspase-8 cleavage

More than 99% of follicles in mammalian ovaries undergo atresia, but the mechanisms regulating the strict selection process are still unclear. Granulosa cell apoptosis is considered the trigger of follicular atresia, which occurs in advance of the death of an oocyte. Cellular FLICE-like inhibitory protein (cFLIP), a homologue of procaspase-8 (also called FLICE), is an intracellular anti-apoptotic protein. It is expressed in granulosa cells of porcine ovaries, where its levels decreases during follicular atresia. We hypothesized that cFLIP regulates granulosa cell apoptosis by acting as a pro-survival factor. In the present study, to further reveal the function of cFLIP in granulosa cells, we examined the anti-apoptotic mechanism of cFLIP using KGN, a human granulosa tumor cell line. Fas-mediated apoptosis was induced by co-treatment with anti-Fas antibody (CH-11), which acts as an agonist of Fas-ligand, and cycloheximide (CHX). When cFLIP was stably expressed in KGN cells following transfection of an expression vector, the Fas-mediated apoptosis was inhibited. Suppression of cFLIP by small interfering RNA (siRNA) spontaneously induced cell death. Silencing of cFLIP promoted cleavage of procaspase-8, and the cell death caused by cFLIP siRNA was completely blocked by a caspase-8 inhibitor (Z-IETD-FMK), indicating that cFLIP regulates apoptosis in KGN cells by inhibiting cleavage of procaspase-8. In conclusion, cFLIP is an essential pro-survival factor for granulosa cells, and it prevents granulosa cell apoptosis by inhibiting procaspase-8 activation.     

Soil solution chemistry at different positions on slope in a conifer plantation forest

It is known that soil property varies along the slope. It suggests that soil solution chemistry also differs topographically. To determine the variation in soil solution chemistry within one watershed, soil solution chemistry at the different positions of the slope was investigated. Soil N transformation changed along the slope. NH₄ ⁺ ratio to inorganic N (NH₄ ⁺ + NO₃ ⁻) increased upslope. The tendency was verified by laboratory incubation. After incubation most of the mineralized N was nitrified at the lower part of the slope, while little nitrification occurred at the upper part of the slope. At the ridge and the backslope inorganic N form in soil solution was concomitant with inorganic N form by incubation. At the ridge NH₄ ⁺ was predominant form in soil solution, at that time major anion was sea salt originated Cl⁻. From this, soil solution chemistry seems to be regulated by the external nutrient source at the ridge. In the second year of lysimeter installation NO₃ ⁻ concentration increased in both sites and the ratio of NH₄ ⁺ to inorganic N decreased. It was considered due to the effect of lysimeter installation. The lag time and the magnitude of NO₃ ⁻ increase were different between the ridge and the backslope. It would be related with soil N transformation in pre-disturbance. The influence of disturbance were shown in other solute concentrations of soil solution.     

Identification of a blue photoluminescent composite material from a combinatorial library

A quaternary combinatorial masking strategy was used in conjunction with photolithography to generate compositionally diverse thin-film phosphor libraries containing 1024 different compositions on substrates 2.5 centimeters square. A parallel imaging system and scanning spectrophotometer were used to identify and characterize compositions in the library with interesting luminescent behavior. Optimal compositions were identified with the use of gradient libraries, in which the stoichiometry of a material was varied continuously. This process led to the identification of an efficient blue photoluminescent composite material, Gd3Ga5O12/SiO2. Experimental evidence suggests that luminescence in this material may arise from interfacial effects between SiO2 and Gd3Ga5O12.     

Juveniles of threadsail filefish, <Emphasis Type="Italic">Stephanolepis cirrhifer</Emphasis>, can survive and grow by feeding on moon jellyfish <Emphasis Type="Italic">Aurelia aurita</Emphasis>

We conducted feeding experiments on threadsail filefish Stephanolepis cirrhifer juveniles for 16 days to evaluate the efficacy of moon jellyfish Aurelia aurita as a prey item. Four treatments, each with 40 individuals, were prepared to compare fish growth performance. The treatments consisted of filefish that were starved (control) (S), fed only jellyfish (J), fed only krill (K), and fed both jellyfish and krill (JK). Fish in the S treatment exhibited a 50% mortality rate and reduced body weight, whereas the J treatment exhibited a zero mortality rate and increased body weight. Fish in the JK treatments showed a significantly faster growth than those in the K treatment. Filefish consumed as much as 24 and 13 times their own body weight in jellyfish per day in the J and JK treatments, respectively. This is the first report showing that growth can be sustained by feeding a marine fish only jellyfish, and indicates the potential of jellyfish as a fish prey in both nature and captivity.     

Amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis> interleukin-10 negatively suppresses host cell-mediated immunity

Interleukin-10 (IL-10) is an anti-inflammatory cytokine and negatively regulates cell-mediated immunity (CMI) induction by inhibiting cytokine production in type 1 T helper cells. IL-10 genes have been isolated from several fish, and inflammatory cytokine inhibition by IL-10 has been well examined. However, a CMI regulator of IL-10 in fish has not yet been identified. In this study, we cloned the IL-10 gene in amberjack Seriola dumerili and analyzed its function using its recombinant protein (rIL-10). In an in vitro culture experiment, gene expression of inflammatory cytokines was suppressed in leukocytes incubated with rIL-10 compared with cells that only received Nocardia seriolae stimulation. This result suggests amberjack IL-10 has conserved function as an inflammatory cytokine inhibitor. Bactericidal activity of amberjack cells against intracellular pathogen stimulation was decreased in a rIL-10 dose-dependent manner. Furthermore, a significant reduction in the T-bet/GATA-3 ratio was observed in N. seriolae living cell (LC)?+?rIL-10-injected fish. Taken together, these results suggest amberjack rIL-10 suppresses CMI induction both in vitro and in vivo. In addition, the number of IgM⁺ cells among spleen leukocytes in N. seriolae?+?rIL-10-injected fish was higher than in only N. seriolae LC, suggesting that Th2-dominant immunity was induced by adding rIL-10.     

Phytotoxin produced by the netted scab pathogen, <Emphasis Type="Italic">Streptomyces turgidiscabies</Emphasis> strain 65, isolated in Sweden

Streptomyces spp. are a highly diverse group of bacteria most of which are soil-inhabiting saprophytes. A few are plant pathogens that produce a family of phytotoxins called thaxtomins and cause significant economic losses, e.g., by reducing the marketability of potato tubers (Solanum tuberosum). In northern Europe, S. scabies, S. turgidiscabies and S. europaeiscabiei are the most common plant pathogenic species. In this study, a Streptomyces strain isolated from a netted scab lesion on a tuber of potato cv. Bintje in northern Sweden was identified as S. turgidiscabies but was found to differ in the genomic region carrying genes required for thaxtomin biosynthesis. Our results showed that the strain did not produce thaxtomin but rather phytotoxin fridamycin E, which is an anthraquinone novel to plant pathogenic Streptomyces spp. Fridamycin E was shown to reduce or inhibit sprouting of potato microtubers in vitro. While fridamycin E is known to have antibiotic activity against Gram-positive bacteria, the inhibitory activity of fridamycin E on plant growth is a novel finding.     

Two new species,Phytophthora nagaii sp. nov. and P. fragariaefolia sp. nov., causing serious diseases on rose and strawberry plants,respectively, in Japan

A new disease of rose was noticed in Chiba Prefecture of Japan in 1968, and the pathogen was initially identified as Phytophthora megasperma based on morphological characteristics. Similar Phytophthora isolates have since been collected from rose plants in Chiba, Kanagawa, and Shizuoka Prefectures. In 2005, several Phytophthora isolates were recovered from crowns of strawberry plants in Hokkaido Prefecture. These were considered to be members of a new species. In this study, we re-examined all these isolates using morphological and physiological studies and a multilocus phylogenetic analysis. The rose and strawberry isolates were mostly similar morphologically and physiologically, with some exceptions. The rose isolates differed significantly from P. megasperma sensu stricto and other related Phytophthora species. The rose and strawberry isolates had external proliferation of sporangia, characteristic funnel-shaped oogonia, predominantly paragynous antheridia, and fast growth rates of 10.5 mm/24 h at an optimum temperature of 28 °C. In the multilocus phylogenetic tree constructed using sequences from the rDNA ITS regions, rDNA LSU, and the translation elongation factor 1-α, β-tubulin and coxI genes, they formed a distinct monophyletic group in clade 7 with strong bootstrap support. The rose and strawberry isolates separated into two distinct groups. The results indicate that the rose and strawberry isolates constitute two separate species, designated here as Phytophthora nagaii and P. fragariaefolia.     

Mechanism of the rice hull‐induced germination of Monochoria vaginalis seeds in darkness

The seeds of Monochoria vaginalis require light to germinate. However, they are able to germinate in darkness in the presence of rice hulls. The mechanism of this phenomenon was investigated in this study, in which only the unsterilized seeds of Monochoria were induced to germinate by the presence of a rice hull extract. The rice hull extract was able to be replaced by either a mixture of amino acids and phosphate or a yeast extract. The culture media in which germination was observed always showed turbidity, indicating the propagation of bacteria, which were identified as Bacillus sp., Pedobacter sp., Pantoea sp. and Sphingomonas sp. Treatment with the individual bacterial species or combinations of these bacteria induced Monochoria seed germination when added to the media containing the rice hull extract. It was concluded that the rice hull extract accelerates the propagation of bacteria, which in turn digest the seed coat of Monochoria, facilitating its germination.     

Impact of the spatial and temporal arrangement of pastoral use on land degradation around animal concentration points

In mobile pastoral systems, the spatial movement of herders is tied to requirements such as water, markets and medical services, resulting in the concentration of livestock in particular areas and subsequent desertification in those areas. The spatial and temporal distributions of these requirements are subject to changes in external forces, such as political regimes and economic systems. To assess and counteract desertification requires an understanding of, and ability to predict, the spatial and temporal arrangements of such concentration points and how these arrangements cause or inhibit desertification. To this end we developed a model that explicitly simulates how animals and vegetation interact. The model has spatial settings for extensive pasture to represent the points at which animals concentrate. We found that the spatial dynamics of the interaction between animal behavior and vegetation were nonlinear and markedly affected the size of the area desertified, and that the distribution of grazing pressure was more important than total grazing pressure, which had only a limited influence on desertification. These findings indicate that application of the carrying capacity concept is not capable of preventing desertification in extensive pasture, even under equilibrium conditions. Therefore, explicit management of the spatial distribution of animals is essential to prevent desertification in extensively grazed rangelands. Copyright © 2010 John Wiley & Sons, Ltd.