Molecular basis of oocyte-paracrine signalling that promotes granulosa cell proliferation

Copyright © 2006 Company of BiologistsOocytes regulate follicle growth by secreting paracrine growth factors that act on neighbouring granulosa cells (GCs). Those factors identified to date are mainly members of the transforming growth factor-ß (TGFß) superfamily, but little is known about which specific receptor/signalling system(s) they employ. This study was conducted to determine the requisite pathways utilised by oocytes to promote GC proliferation. We used an established oocyte-secreted mitogen bioassay, where denuded mouse oocytes are co-cultured with mural GCs. Oocytes, growth differentiation factor-9 (GDF9), TGFß1 and activin-A all promoted GC DNA synthesis, but bone-morphogenetic protein 6 (BMP6) did not. Subsequently, we tested the capacity of various TGFß superfamily receptor ectodomains (ECD) to neutralise oocyte- or specific growth factor-stimulated GC proliferation. The BMP type-II receptor (BMPR-II) ECD antagonised oocyte and GDF9 bioactivity dose-dependently, but had no or minimal effect on TGFß1 and activin-A bioactivity, demonstrating its specificity. The TGFßR-II, activinR-IIA and activinR-IIB ECDs all failed to neutralise oocyte- or GDF9-stimulated GC DNA synthesis, whereas they did antagonise the activity of their respective native ligands. An activin receptor-like kinase (ALK) 4/5/7 inhibitor, SB431542, also antagonised both oocyte and GDF9 bioactivity in a dose-dependent manner. Consistent with these findings, oocytes, GDF9 and TGFß1 all activated SMAD2/3 reporter constructs in transfected GC, and led to phosphorylation of SMAD2 proteins in treated cells. Surprisingly, oocytes did not activate the SMAD1/5/8 pathway in transfected GCs although exogenous BMP6 did. This study indicates that oocyte paracrine factors primarily utilise a similar signalling pathway first identified for GDF9 that employs an unusual combination of TGFß superfamily receptors, the BMPR-II and a SMAD2/3 stimulatory ALK (4, 5 or 7), for transmitting their mitogenic actions in GC. This cell-signalling pathway may also have relevance in the hypothalamic-pituitary axis and in germ-somatic cell interactions in the testis.Robert B. Gilchrist, Lesley J. Ritter, Samu Myllymaa, Noora Kaivo-Oja, Rebecca A. Dragovic, Theresa E. Hickey, Olli Ritvos and David G. Mottershea

Binary Weighted DAC with 2-ξ Resistor Ratio

In this paper we present a new digital analog converter (DAC) design, based on the binary weighted resistor network. The proposed design ensures high conversion accuracy using low precision resistors with ±1% ±2%, ±5%, ±10% and ±20% resistor tolerance. High accuracy is achieved due to better coverage of the analog domain of the transfer characteristic. In binary weighted converters the imprecision of resistors introduces positive and negative differential nonlinearities (DNL). Positive DNL causes gap in the analog domain of the transfer characteristic and negative DNL causes non-monotonicity. In the proposed solution we change the resistor ratio of the two consecutive DAC branches from 2 to 2-ξ, where ξ is small positive number. With this change, we intentionally introduce an additional negative DNL in order to entirely avoid the positive gap. Simulation results confirm that even with resistors tolerance of up to ±10%, we can achieve a converter with maximal gap in the transfer characteristic less than or around one LSB

On Nonperturbative Calculations in Quantum Electrodynamics

A new approach to nonperturbative calculations in quantum electrodynamics is proposed. The approach is based on a regular iteration scheme for solution of Schwinger-Dyson equations for generating functional of Green functions. The approach allows one to take into account the gauge invariance conditions (Ward identities) and to perform the renormalization program. The iteration scheme can be realized in two versions. The first one ("perturbative vacuum") corresponds to chain summation in the diagram language. In this version in four-dimensional theory the non-physical singularity (Landau pole) arises which leads to the triviality of the renormalized theory. The second version ("nonperturbative vacuum") corresponds to ladder summation and permits one to make non-perturbative calculations of physical quantities in spite of the triviality problem. For chiral-symmetrical leading approximation two terms of the expansion of the first-step vertex function over photon momentum are calculated. A formula for anomalous magnetic moment is obtained. A problem of dynamical chiral symmetry breaking (DCSB) is considered, the calculations are performed for renormalized theory in Minkowsky space. In the strong coupling region DCSB-solutions arise. For the renormalized theory a DCSB-solution is also possible in the weak coupling region but with a subsidiary condition on the value of $\alpha$.Comment: 31 pages, Plain LaTex, no figures. Journal version: some discussion and refs. are adde

The quality of sliced carrots affected by modified polyethylene foil and storage temperatures

The aim of this study was to examine the influence of different additive contents for oxygen absorption (5, 10 and 15%) in low-density polyethylene (PELD) plastic foils and storage temperatures (4 and 28 °C) on the quality and shelf-life of sliced carrots during storage. Quality and storage-life of packaged carrots slices were determined by observing changes of mass, total carotenoide pigments, microbial counts (mesophilic aerobic bacteria, enterobacteria, sulphite-reducing clostridia, yeast and moulds), sensory quality and texture by the use of penetrometer. The PELD foils modified with 10 and 15% of oxygen absorber (O2, CO2 and N2 permeability at 4 °C of around 700 ml m-2d-1atm-1) were the most suitable for the storage and prevention of deterioration of minimally processed carrots. Findings indicated that in these foils the best quality and shelf-life of carrot were maintained by 6 days of storage at 4 °C, without significant changes in parameters studied. The absorber for oxygen added to the foil had no influence on the permeability to CO2, O2and N2. The permeability of foils, which were used for carrot packaging increased by the increase of storage temperature to 28 °C and decreased by decreasing the temperature to 4 °C, and was not significantly affected by the additive content either. In the same time the diffusion constants of unused and used PELD foils for carrot packaging at 4 °C and 28 °C changed according to the change of film permeability during storage at those temperatures

Multi-lingual and multi-cultural information literacy; perspectives, models and good practice

Purpose This paper reviews current approaches to, and good practice, in information literacy development in multi-lingual and multi-cultural settings, with particular emphasis on provision for international students. Design/methodology/approach A selective and critical review of published literature is extended by evaluation of examples of multi-lingual information literacy tutorials and MOOCs. Findings Multi-lingual and multi-cultural information literacy are umbrella terms covering a variety of situations and issues. This provision is of increasing importance in an increasingly mobile and multi-cultural world. This article evaluates current approaches and good practice, focusing on issues of culture vis a vis language, the balance between individual and group needs, specific and generic information literacy instruction, and models for information literacy, pedagogy and culture. Recommendations for good practice and for further research are given, Originality/value This is one of very few articles critically reviewing how information literacy development is affected by linguistic and cultural factors

Effect of processing on the drying kinetics and functional value of dried apricot

Apricots can be considered as a good source of phenolic compounds, which are beneficial for human health. Microwaves may be an alternative to the conventional sun or hot air drying techniques used to obtain dried apricot. Nevertheless, their impact on the functional compounds must be taken into account if they are to be recommended as an attractive drying option. This work compares the drying kinetics and the change in the organic acids, phenolic compounds and antioxidant activity of dried apricot when using hot air drying and microwave energy. Empirical (linear and Page) equations can be used to model the drying kinetics in air, combined air-microwave and microwave processes. From the obtained results, it can be concluded that the industrial processing of dried apricots may be improved by using microwave energy, as the drying time is considerably reduced, and the obtained fruit had a higher phenolic content, particularly of chlorogenic acid, catequin and epicatequin. Nevertheless, as the contribution of these phenols to antioxidant capacity was not significant, microwave dried samples maintained the same antioxidant capacity as the air-dried ones. When sulphite is added previous to the drying processes, care should be taken with the total phenols and the antioxidant capacity quantified as it may interfere with the results depending on the methodology used. © 2011 Elsevier Ltd.Igual Ramo, M.; García Martínez, EM.; Martín-Esparza, M.; Martínez Navarrete, N. (2012). Effect of processing on the drying kinetics and functional value of dried apricot. Food Research International. 47(2):284-290. doi:10.1016/j.foodres.2011.07.019S28429047

Release of oxidizing fluids in subduction zones recorded by iron isotope zonation in garnet

Subduction zones are key regions of chemical and mass transfer between the Earth’s surface and mantle. During subduction, oxidized material is carried into the mantle and large amounts of water are released due to the breakdown of hydrous minerals such as lawsonite. Dehydration accompanied by the release of oxidizing species may play a key role in controlling redox changes in the subducting slab and overlying mantle wedge. Here we present measurements of oxygen fugacity, using garnet–epidote oxybarometry, together with analyses of the stable iron isotope composition of zoned garnets from Sifnos, Greece. We find that the garnet interiors grew under relatively oxidized conditions whereas garnet rims record more reduced conditions. Garnet δ56Fe increases from core to rim as the system becomes more reduced. Thermodynamic analysis shows that this change from relatively oxidized to more reduced conditions occurred during lawsonite dehydration. We conclude that the garnets maintain a record of progressive dehydration and that the residual mineral assemblages within the slab became more reduced during progressive subduction-zone dehydration. This is consistent with the hypothesis that lawsonite dehydration accompanied by the release of oxidizing species, such as sulfate, plays an important and measurable role in the global redox budget and contributes to sub-arc mantle oxidation in subduction zones

Monoubiquitination of syntaxin 3 leads to retrieval from the basolateral plasma membrane and facilitates cargo recruitment to exosomes

Syntaxin 3 (Stx3), a SNARE protein located and functioning at the apical plasma membrane of epithelial cells, is required for epithelial polarity. A fraction of Stx3 is localized to late endosomes/lysosomes, although how it traffics there and its function in these organelles is unknown. Here we report that Stx3 undergoes monoubiquitination in a conserved polybasic domain. Stx3 present at the basolateral—but not the apical—plasma membrane is rapidly endocytosed, targeted to endosomes, internalized into intraluminal vesicles (ILVs), and excreted in exosomes. A nonubiquitinatable mutant of Stx3 (Stx3-5R) fails to enter this pathway and leads to the inability of the apical exosomal cargo protein GPRC5B to enter the ILV/exosomal pathway. This suggests that ubiquitination of Stx3 leads to removal from the basolateral membrane to achieve apical polarity, that Stx3 plays a role in the recruitment of cargo to exosomes, and that the Stx3-5R mutant acts as a dominant-negative inhibitor. Human cytomegalovirus (HCMV) acquires its membrane in an intracellular compartment and we show that Stx3-5R strongly reduces the number of excreted infectious viral particles. Altogether these results suggest that Stx3 functions in the transport of specific proteins to apical exosomes and that HCMV exploits this pathway for virion excretion

Placental Vesicles Carry Active Endothelial Nitric Oxide Synthase and Their Activity is Reduced in Preeclampsia.

Preeclampsia (PE), a multi-system hypertensive disorder of pregnancy, is associated 25 with increased systemic vascular resistance. Placentae from PE patients have 26 reduced levels of endothelial nitric oxide synthase (eNOS) and thus less nitric oxide 27 (NO). Syncytiotrophoblast extracellular vesicles (STBEV), comprised of microvesicles 28 (STBMV) and exosomes (STBEX), carry signals from the STB to the mother. We 29 hypothesized that STBEV bound eNOS (STBEV-eNOS), capable of producing NO, 30 are released into the maternal circulation. Dual-lobe ex vivo placental perfusion and 31 differential centrifugation was used to isolate STBEV from PE (n=8) and normal 32 pregnancies (NP) (n=11). Plasma samples of gestational age matched PE and NP 33 (n=6) were used to isolate circulating STBMV. STBEV expressed placental alkaline 34 phosphatase (PlAP), confirming placental origin. STBEV co-expressed eNOS, but not 35 iNOS, confirmed using Western blot, flow cytometry and immuno-depletion. STBEV-36 eNOS produced NO which was significantly inhibited by L-NAME (eNOS inhibitor, 37 *p<0.05), but not 1400W (iNOS inhibitor). STBEV-eNOS catalytic activity was 38 confirmed by visualising eNOS dimerization. STBEV-eNOS was more abundant in 39 uterine vein compared to peripheral blood, indicating placental origin. STBEV isolated 40 from PE perfused placentae had lower levels of STBEV-eNOS (STBMV; *p<0.05) and 41 overall lower NO activity (STBMV, ns; STBEX, *p<0.05) compared to NP. Circulating 42 plasma STBMV from PE women had lower STBEV-eNOS expression compared to NP 43 women (**p<0.01). This is the first observation of functional eNOS expressed on 44 STBEV from NP and PE placentae, as well as in plasma. The lower STBEV-eNOS 45 NO production seen in PE may contribute to the decreased NO bioavailability in this 46 disease