Fas-associated protein with death domain (FADD)-independent recruitment of c-FLIPL to death receptor 5

Here we show a novel mechanism by which FLICE-like inhibitory protein (c-FLIP) regulates apoptosis induced by tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and one of its receptors, DR5. c-FLIP is a critical regulator of the TNF family of cytokine receptor signaling. c-FLIP has been postulated to prevent formation of the competent death-inducing signaling complex (DISC) in a ligand-dependent manner, through its interaction with FADD and/or caspase-8. In order to identify regulators of TRAIL function, we used the intracellular death domain (DD) of DR5 as a target to screen a phage-displayed combinatorial peptide library. The DD of DR5 selected from the library a peptide that showed sequence similarity to a stretch of amino acids in the C terminus of c-FLIP(L). The phage-displayed peptide selectively interacted with the DD of DR5 in in vitro binding assays. Similarly, full-length c-FLIP (c-FLIP(L)) and the C-terminal p12 domain of c-FLIP interacted with DR5 both in in vitro pull-down assays and in mammalian cells. This interaction was independent of TRAIL. To the contrary, TRAIL treatment released c-FLIP(L) from DR5, permitting the recruitment of FADD to the active DR5 signaling complex. By employing FADD-deficient Jurkat cells, we demonstrate that DR5 and c-FLIP(L) interact in a FADD-independent manner. Moreover, we show that a cellular membrane permeable version of the peptide corresponding to the DR5 binding domain of c-FLIP induces apoptosis in mammalian cells. Taken together, these findings indicate that c-FLIP interacts with the DD of DR5, thus preventing death (L)signaling by DR5 prior to the formation of an active DISC. Because TRAIL and DR5 are ubiquitously expressed, the interaction of c-FLIP(L) and DR5 indicates a mechanism by which tumor selective apoptosis can be achieved through protecting normal cells from undergoing death receptor-induced apoptosis.     

An Approach to Design Highly Durable Piezoelectric Nanogenerator Based on Self‐Poled PVDF/AlO‐rGO Flexible Nanocomposite with High Power Density and Energy Conversion Efficiency

Till date, fabrication of piezoelectric nanogenerator (PNG) with highly durable, high power density, and high energy conversion efficiency is of great concern. Here a flexible, sensitive, cost effective hybrid piezoelectric nanogenerator (HPNG) developed by integrating flexible steel woven fabric electrodes into poly(vinylidene fluoride) (PVDF)/aluminum oxides decorated reduced graphene oxide (AlO‐rGO) nanocomposite film is reported where AlO‐rGO acts as nucleating agent for electroactive β‐phase formation. The HPNG exhibits reliable energy harvesting performance with high output, fast charging capability, and high durability compared with previously reported PVDF based PNGs. This HPNG is capable for harvesting energy from a variety and easy accessible biomechanical and mechanical energy sources such as, body movements (e.g., hand folding, jogging, heel pressing, and foot striking, etc.) and machine vibration. The HPNG exhibits high output power density and energy conversion efficiency, facilitating direct light on different color of several commercial light‐emitting diodes instantly and powers up many portable electronic devices like wrist watch, calculator, speaker, and mobile liquid crystal display (LCD) screen through capacitor charging. More importantly, HPNG retains its performance after long compression cycles (≈158 400), demonstrating great promise as a piezoelectric energy harvester toward practical applications in harvesting biomechanical and mechanical energy for self‐powered systems.     

Effect of grazing removal on aboveground vegetation and soil seed bank composition in sub-alpine grasslands of northern Iran

Background: Knowledge about vegetation and soil seed bank composition and the processes that contribute to vegetation recovery after the removal of heavy grazing is lacking in sub-alpine ecosystems.

Aims: In order to assess the effects of large herbivores on above-ground vegetation (AGV) and soil seed bank (SSB) characteristics, intensively sheep-grazed areas were compared to adjacent areas where grazing had been removed 10 years previously in a sub-alpine grassland of northern Iran.

Methods: A total of 40 4-m² (2 m × 2 m) plots were established in each treatment, and soil samples were collected from each plot within a depth of 10 cm. Plant species composition was determined for each plot during the flowering stage of herbaceous species in June 2011. The seedling emergence method was used to estimate SSB composition.

Results: A total of 45 species (23 annuals and 22 perennials) emerged from the soil samples of the grazed area, while the number of species emerged from the soil samples of the previously grazing area was 76 (37 annuals and 39 perennials). The removal of grazing led to a significant increase in species richness and seed density in the SSB. Species turnover of AGV was higher, and that of the SSB was similar for grazed areas compared with areas that were formerly grazed. Detrended correspondence analysis ordination of AGV composition showed a clearly separate structure between grazed plots and plots where grazing has been removed, while the segregation was less clear for SSB composition.

Conclusions: We concluded that restoration of locally degraded sites cannot rely on the SSB when grazing is stopped simultaneously and unvegetated gaps are colonised by vegetative growth rather than by seed.     

Effects of 9-ene-tetrahydrocannabinol on expression of β-type transforming growth factors, insulin-like growth factor-I and c-myc genes in the mouse uterus

Effects of cannabinoid on expression of β-type transforming growth factors (TGF-β1, -β2 and -β3), insulin-like growth factor-I (IGF-I) and c-myc genes in the uteri of adult ovariectomized mice were examined using Northern blot hybridization. Mice were exposed to 9-ene-tetrahydrocannabinol (THC) alone or in combination with an injection of estradiol-17β (E₂) and/or progesterone (P₄), and uteri were analyzed at various times thereafter. TGF-β isoform messenger RNAs (mRNAs) persisted in ovariectomized uteri and their levels were not altered after THC treatment, whereas an injection of E₂ caused a modest increase in TGF-β1 and -β3 mRNA levels at 24 h. Imposition of THC treatment advanced the stimulatory effects of E₂ by changing the timing for the peak of TGF-β3 mRNA levels to 12 h. In comparison, E₂ treatment substantially elevated the levels of TGF-β2 mRNA at 6 h, and THC potentiated this E₂ response without affecting the timing for the response. Imposition of P₄ treatment did not antagonize any of these responses. P₄ treatment alone or with THC had insignificant effects on mRNA levels for these TGF-β isoforms. Uterine levels of IGF-I and c-myc mRNAs were low in ovariectomized mice and THC did not alter these mRNA levels. In contrast, E₂ treatment induced a rapid, but transient, increase in IGF-I and c-myc mRNAs, and THC antagonized the rapid c-myc mRNA response and altered the timing of the IGF-I mRNA response. P₄ treatment alone also caused the transient induction of these mRNAs, but THC failed to antagonize these effects. An injection of P₄ plus E₂ resulted in further modest increases in IGF-I and c-myc mRNA levels as compared to E₂ or P₄ treatment alone. However, THC did not antagonize these transient stimulatory effects of the combined ovarian steroids. The data suggest that THC should not be classified as estrogenic or antiestrogenic. However, this compound can modulate (potentiate, antagonize and/or alter timing) the effects of ovarian steroids on uterine gene expression.     

Role of embryonic oestrogen in rabbit blastocyst development and metabolism

Rabbit morulae were grown for 24 h in Ham's F12 medium supplemented with BSA. CI-628 citrate (1.5 micrograms/ml), a specific oestrogen antagonist, significantly inhibited the transformation of morulae to blastocysts. This inhibition was reversed with oestradiol-17 beta (1 micrograms/ml) but not oestradiol-17 alpha (1 micrograms/ml) added to the culture medium. The specific activities of phosphofructokinase, lactic dehydrogenase, malate dehydrogenase and alkaline phosphatase in blastocysts grown in vitro for 24 h in medium TC 199 + BSA showed significant elevation with blastocyst growth and expansion, while that of acid phosphatase revealed no change, and leucine aminopeptidase activity declined significantly. These changes were markedly inhibited by CI-628 citrate (2 micrograms/ml) and were reversed by oestradiol-17 beta (0.4 micrograms/ml) but not by oestradiol-17 alpha (0.4 micrograms/ml). Our findings suggest a role of oestrogen present in the rabbit morula and blastocyst in the triggering of embryonic differentiation and metabolic functions.     

Calmodulin binding to the dehydrogenase domain of NADPH oxidase 5 alters its oligomeric state

Superoxide generated by NADPH Oxidase 5 (Nox5) is regulated by Ca²⁺ through the interaction of its self-contained Ca²⁺ binding domain and dehydrogenase domain (DH). Recently, calmodulin (CaM) has been reported to enhance the Ca²⁺ sensitivity of Nox5 by binding to the CaM-binding domain sequence (CMBD), in which the interaction between CaM and Nox5 is largely unclear. Here, we used the CMBD peptide and truncated DH constructs, and separately studied their interaction with CaM by fluorescence, calorimetry, and dynamic light scattering. Our results revealed that each half-domain of CaM binds one CMBD peptide with a binding constant near 10⁶ M^-1 and a binding enthalpy change of ?3.81 kcal/mol, consistent with an extended 1:2 CaM:CMBD structure. However, the recombinant truncated DH proteins exist as oligomers, possibly trimer and tetramer. The oligomeric states are concentration and salt dependent. CaM binding appears to stabilize the DH dimer complexed with CaM. The thermodynamics of CaM binding to the DH is comparable to the peptide-based study except that the near unity binding stoichiometry and a large conformational change were observed. Our result suggests that the oligomeric states of Nox5, mediated by its DH domain and CaM, may be important for its superoxide-generating activity.     

The Combined Efficiency of Dietary Isomaltooligosaccharides and Bacillus spp. on the Growth,Hemato-Serological,and Intestinal Microbiota Indices of Caspian Brown Trout (Salmo trutta caspius Kessler, 1877)

Probiotics and Antimicrobial Proteins - The combined effects of a commercial probiotic, BetaPlus®, and a prebiotic, isomaltooligosaccharides (IMOS) on the growth, survival rate, intestinal...