Serum- and glucocorticoid-induced kinase 1 (Sgk1) regulates many ion channels and transporters in epithelial cells and promotes cell survival under stress conditions. In this study we demonstrate that Sgk1 is a short-lived protein regulated by the endoplasmic reticulum (ER)-associated degradation system and subcellular localization to the ER. We identified a hydrophobic motif (residues 18-30) as the signal for ER localization and rapid degradation by the ubiquitin (Ub)/proteasome pathway in both yeast and mammalian cells. Deletion or reduction of hydrophobicity of the motif redistributes Sgk1 to the cytosol and nucleus and markedly increases its half-life. We determined that the Ub-conjugating UBC6 and UBC7 and the Ub ligase HRD1 are the ER-associated Ub enzymes that mediate degradation of Sgk1; thus, Sgk1 has been identified as a cytosolic substrate for mammalian HRD1. Compartmentalization of Sgk1 controls the functional and spatial specificities of Sgk1-mediated signaling pathways, whereas rapid protein turnover provides a means to rapidly adjust Sgk1 abundance in response to different hormonal and external stimuli that increase Sgk1 gene transcription.
The efficient amplification of genomic libraries, cDNA libraries and other complex mixts. of genes by PCR is impeded by two phenomena: firstly, short fragments tend to be amplified in preference to larger ones; and, secondly, artifactual fragments are generated by recombination between homologous regions of DNA. Recombination in this case occurs when a primer is partially extended on one template during one cycle of PCR and further extended on another template during a later cycle. Thus, chimeric mols. are generated, the short ones of which are then preferentially amplified. A variety of PCR protocols have been proposed to minimize these problems, most of which rely on high template concns. and low nos. of PCR cycles. Clearly, however, such an approach is not viable if little template DNA is available. Here we describe a protocol for amplifying complex DNA mixts., based on the compartmentalization of genes in a water-in-oil (w/o) emulsion. Template fragments are segregated in the minute aq. droplets of the emulsion and amplified by PCR in isolation. This approach alleviates the problems described above while enabling the use of small amts. of template DNA and high nos. of PCR cycles. Box 1 described an alternative method for generating very stable emulsions for emulsion PCR using the surfactant ABIL EM 90. [on SciFinder(R)]
Sturgeons are known throughout the world as the source of black caviar. Their declining populations in their native habitats, mainly the Caspian Sea, due to over-fishing for meat and caviar production, destruction of their spawning grounds and water pollution, have led to their introduction into aquaculture in areas with suitable conditions, including Israel. Recently, we noticed an unusual phenomenon in these normally gonochoristic species. Several 5-year-old female sturgeons were found to have one or more testicular sections in each of their two gonads, forming an intersexual gender. Further examination of other fish from the same age group revealed 14% fish with intersex gonads among a population of 5000 fish that had been pre-selected as females. This phenomenon has not been found however in other age groups of Russian sturgeons, cultured at the same facility. Sturgeons are a generally gonochoristic species, and hermaphroditism is only very infrequently observed under natural or normal breeding conditions. Moreover, these rare cases have all been from polluted habitats. The present work is the first description of fish containing intersex gonads in Russian sturgeon (Acipenser gueldenstaedtii). We describe the phenomenon anatomically and histologically, and examine plasma steroid levels and pituitary gonadotropin gene expression by comparing fish with intersex gonads with normal females and males of the same age group. Intersex gonads were typical female ovaries with one or more white testicular components embedded in each. The testis components were not uniform in size or location among the two gonads of each fish or among different fish, and they showed marked differences in distribution. The ovarian component of the intersex gonad was at the pre-vitellogenic stage as in normal females, and the testis component contained spermatids and mature spermatozoa as in normal males of the same age. However, in terms of estradiol and 11-ketotestosterone plasma levels, as well as of pituitary gonadotropin (betaLH and betaFSH) gene-expression levels, the fish with intersex gonads were more similar to the normal males than to the normal females, even though the testis part of the intersex gonad was smaller than the ovarian part. To examine the possibility that the fish containing intersex gonads were hybrids, phylogenetic trees were constructed from the consensus sequences of Cytochrome b and control region (D-loop) genes. Results indicated no differences between the fish with intersex gonads and normal males or females of the same age group. However, statistically significant differences were found between different age groups of Russian sturgeon, as well as of white sturgeons (A. transmontanus), grown under the same culture conditions.
A dialog with Asher Peres regarding the meaning of quantum teleportation is briefly reviewed. The Braunstein–Kimble method for teleportation of light is analyzed in the language of quantum wave functions. A pictorial example of continuous variable teleportation is presented using computer simulation.
Cofilin/ADF, beryllium fluoride complex (BeFx), and phalloidin have opposing effects on actin filament structure and dynamics. Cofilin/ADF decreases the stability of F-actin by enhancing disorder in subdomain 2, and by severing and accelerating the depolymerization of the filament. BeFx and phalloidin stabilize the subdomain 2 structure and decrease the critical concentration of actin, slowing the dissociation of monomers. Yeast cofilin, unlike some other members of the cofilin/ADF family, binds to F-actin in the presence of BeFx; however, the rate of its binding is strongly inhibited by BeFx and decreases with increasing pH. The inhibition of the cofilin binding rate increases with the time of BeFx incubation with F-actin, indicating the existence of two BeFx-F-actin complexes. Cofilin dissociates BeFx from the filament, while BeFx does not bind to F-actin saturated with cofilin, presumably because of the cofilin-induced changes in the nucleotide-binding cleft of F-actin. These changes are apparent from the increase in the fluorescence intensity of F-actin bound epsilon-ADP upon cofilin binding and a decrease in its accessibility to collisional quenchers. BeFx also affects the nucleotide-binding cleft of F-actin, as indicated by an increase in the fluorescence intensity of epsilon-ADP-F-actin. Phalloidin and cofilin inhibit, but do not exclude each other binding to their complexes with F-actin. Phalloidin promotes the dissociation of cofilin from F-actin and slowly reverses the cofilin-induced disorder in the DNase I binding loop of subdomain 2.
Apoptotic cells were shown to induce dendritic cell immune tolerance. We applied a proteomic approach to identify molecules that are secreted from apoptotic monocytes, and thus may mediate engulfment and immune suppression. Supernatants of monocytes undergoing apoptosis were collected and compared using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and differentially expressed proteins were identified using tandem mass spectrometry. Thrombospondin-1 (TSP-1) and its cleaved 26-kDa heparin-binding domain (HBD) were identified. We show that TSP-1 is expressed upon induction of monocyte apoptosis in a caspase-dependent pattern and the HBD is cleaved by chymotrypsin-like serine protease. We further show that CD29, CD36, CD47, CD51, and CD91 simultaneously participate in engulfment induction and generation of an immature dendritic cell (iDC) tolerogenic and phagocytic state. We conclude that apoptotic cell TSP-1, and notably its HBD, creates a signalosome in iDCs to improve engulfment and to tolerate engulfed material prior to the interaction with apoptotic cells.
Anti-proliferative and pro-apoptotic activities of fractions of Pleurotus ostreatus were examined using HT-29 colon cancer cells in vitro. A hot-water-soluble fraction of the mycelium of the liquid cultured mushroom was partially isolated and chemically characterized as a low-molecular-weight alpha-glucan. HT-29 cells were exposed to the different isolates and significant inhibition of proliferation was obtained in a dose-dependent manner. Proliferation inhibition was shown to be the result of apoptotic induction because the pro-apoptotic molecules Bax and cytosolic cytochrome-c were upregulated. Fluorescence-activated cell sorter analyses of polysaccharide-treated HT-29 cells showed a high percentage of Annexin-positive cells. Here, we describe a newly identified low-molecular-weight alpha-glucan with promising anti-tumorigenic properties, and demonstrate its direct effect on colon cancer cell proliferation via induction of programmed cell death.
The invention relates to a method for prepg. an aq.-based dispersion of metal nanoparticles comprising: (a) pre-reducing a metal salt suspension by a H2O sol. polymer capable of metal redn. to form a metal nuclei; and (b) adding a chem. reducer to form metal nanoparticles in dispersion. The invention further relates to aq.-based dispersions of metal nanoparticles, and to compns. such as inks comprising such dispersions. [on SciFinder(R)]
This book deals with two main questions: what are the barriers to the equality and the development of the Arabs in Israel? and how can the effect of these barriers be reduced?
The picture presented by the book is that despite certain improvements in the social area, especially in the area of education, the inequality between Jews and Arabs remains as it was. The book offers a preliminary background for the analysis of the fundamental barriers preventing equality between Arabs and Jews in Israel.
