6957792 research outputs found
Sort by
Threat Modelling for Active Directory
Get PDFAbstract: This paper analyses the security threats that can arise against an Active Directory server when it is included in a Web application. The approach is based on the STRIDE classification methodology. The paper also provides outline descriptions of countermeasures that can be deployed to protect against the different threats and vulnerabilities identified here
Fasting Insulin, Adiponectin, hs-CRP Levels, and The Components of Metabolic Syndrome
Get PDFABSTRACT 3, 5, 7, 7, 4
RELATIONSHIPS BETWEEN BROWSING DAMAGE AND THE SPECIES DOMINANCE BY THE HIGHLY FOOD-ATTRACTIVE AND LESS FOOD-ATTRACTIVE TREES
Get PDFAbstract ČERMÁK, P., BERANOVÁ, P., ORÁLKOVÁ, J., HORSÁK, P., PLŠEK, J.: Relationships between browsing damage and the species dominance by the highly food-attractive and less food-attractive trees. Acta univ. agric. et silvic. Mendel. Brun., 2011, LIX, No. 1, pp. 29-36 The paper analyses data on the browsing damage to Acer pseudoplatanus, Carpinus betulus, Fraxinus excelsior, Quercus spp., Tilia cordata and Fagus sylvatica. Field research was carried out in the period 2007-2010 and analysed data came from 33 transects at 10 localities with the various abundance of game in the CR (everywhere Capreolus capreolus, on several plots also Cervus elaphus, Ovis musimon or Dama dama). Trees were monitored up to a height of 150 cm in natural regeneration under stands and in plantations and the occurrence was noted of new browsing damage. Diff erences between the percentage of damaged individuals of the given species of a food-attractive species (A. p., C. b., F. e.) and the percentage of damaged individuals of all tree species on a transect as well as the proportion of these parameters correlate negatively with the given species dominance and thus, they appear to be suitable parameters for the analysis of relationships between the damage intensity and dominance. The higher the percentage proportions of highly food-attractive species and the lower the percentage of less-attractive species, the lower the relative intensity of damage to highly food-attractive species. At the same time, the higher the percentage proportion of highly food-attractive species and the lower the percentage of less-attractive species then the lower a diff erence between damage to less food-attractive species and all species. browsing, dominance, highly food-attractive species, less food-attractive species, correlation The attractiveness of tree species for ruminant consumers is highly species-specifi c, however, dama ge to particular tree species markedly diff ers at various localities, namely in absolute values and in relation to other species. Generally, the species structure of advance regeneration and the species rareness are the most mentioned factors aff ecting the preference of a species by browsing animals. That is, tree species are usually more intensively searched if they are in combination with less attractive species and relative damage is o en in relation to the species relative abundance or its dominance. In some cases, the damage increases with the relative proportion of a species, in other cases increases with the decreasing proportion of these species in advance growth both polarities of relationships being interpretabl
Between the dazzle of a new building and its eventual corpse
No full textAbstract This paper presents the development of a lightweight component model that allows user to manage the introduction and arrangement of new interactive services and devices in the home. The model is responsive to ethnographic studies of the interplay between the Space-plan or interior layout and Stuff or artefacts placed within the fabric of the home. Interaction techniques developed through user-participation enable household members -rather than designers -to configure and reconfigure interactive devices and services to meet local needs. As a result, we have developed a tabletbased editor that discovers available ubiquitous components and presents these to users as 'jigsaw pieces' that can be dynamically assembled and recombined
Generation of reflected second-harmonic light beam with inhomogeneous transversal distribution of polarization from the surface of chiral medium by normally incident Gaussian beam
No full textAbstract A principle possibility of second harmonic generation (SHG) from the surface of a chiral medium by normally incident focused fundamental beam has been shown earlier, and the key features of this phenomenon (forbidden in a planewave approximation) have been outlined in [N.I. Koroteev, V.A. Makarov, S.N. Volkov, Laser Phys. 8 (1998) 532-535]. In our work we have obtained analytical expressions, which describe the distributions of intensity and polarization in the cross-section of a second-harmonic (SH) light beam. It is found that the polarization state drastically changes along the cross-section of the signal light beam. The polarization effects concerning the transversal inhomogeneous polarization distribution were studied in detail. It is shown that the measurement of the polarization state in certain areas of the SH beam cross-section gives us quantitative information directly about the medium material constants
No full textABSTRACT The mouse mu opioid receptor (MOR) gene has two promoters, referred to as distal and proximal promoter. Previously our colleagues reported that a 26 bp cis-acting element of the mouse MOR gene activates MOR gene expression. Here, we report the cloning of four members of the poly(C) binding protein (PCBP) family and show that the 26 bp polypyrimidine stretch in MOR proximal promoter interacts with these PCBPs and activates MOR transcription. The PCBPs bind not only to single-stranded, but also to double-stranded DNA. The nuclear run-off assay and semi-quantitative RT-PCR show that PCBPs enhance the transcription rate of MOR gene. Further, we perform refined mapping to elucidate the core region (-317/-304) involved in mediating the PCBPs induced MOR promoter activity. Decoy oligonucleotides against the polypyrimidine stretch inhibit the PCBP induced MOR promoter activity, thereby reconfirming the role of this element in regulating MOR promoter activity. ChIP assay confirms the interaction of PCBPs with MOR promoter in vivo. In conclusion, we demonstrate that PCBPs act as a transcription factor and positively regulate MOR gene expression in NMB cells. 0 0 5 / 0 1 2 2 4 5 4 INTRODUCTION Opioids exert their pharmacological and physiological effects through binding to their endogenous receptors. There are, at least, three major types of opioid receptors, namely mu, delta and kappa (µ, δ and κ) (Kieffer, 1995). These receptors belong to the superfamily of G protein-coupled receptors (GPCR). Expression of opioid receptors is mainly limited to the central nervous system (CNS), and each receptor type has a distinct distribution pattern The mu opioid receptor (MOR) is of particular interest, as it is the receptor which mediates important opioid effects like analgesia and addiction Ever since the cloning of the mu opioid receptor silencer element (Kim et al., 2004), an AP2-like element (Ko et al., 2003) and canonical Sp binding sites (Ko et al., 1998). Recently, we reported a 26 bp polypyrimidine rich cis-element in the MOR proximal promoter located at -334/-308, very close to the transcription initiation sites (Ko and Loh, 2001). This stretch of pyrimidine rich bases is able to form single-stranded DNA like structure and can bind to single strand nucleic acid binding proteins. Later, we identified the interacting protein with the polypyrimidine rich element to be a poly(C) binding protein (PCBP) (Ko and Loh, 2005). Poly(C) binding proteins (PCBPs) belong to a family of RNA-binding proteins which are characterized by high affinity and sequence-specific interaction with poly r(C). These PCBPs have been divided into two sets in mammalian cells; hnRNP K/J The KH domains are about 70 amino acid long RNA binding modules; however, they do not provide binding specificity to poly(C) RT-PCR of MOR Gene galactosidase activities of each lysate were measured using assays kits as described by the manufacturer (Promega). In Vitro Translation 10 annealed and labeled with T4PNK. The end-labeled DNA probes were incubated with TNT reaction products of PCBP expression constructs in a final volume of 20 µ l EMSA buffer (10 mM Tris, pH 7.5, 5% glycerol, 1 mM EDTA, 50 mM NaCl, 1 mM dithiothreitol and 0.1 mg/ml poly dI-dC) at room temperature for 20 min. For oligonucleotide competition analysis, a 100-fold molar excess of cold competitor oligonucleotide was added to the mixture prior to adding the probe. The reaction mixtures were electrophoresed in a 4% polyacrylamide nondenaturing gel in 0.5 X TBE (45 mM Tris-borate and 1 mM EDTA) at 4°C and the gel was dried. Gel was exposed to phosphoimaging screen and scanned on PhosphoImager. Chromatin Immunoprecipitation (ChIP) Assay NMB cells transiently transfected with HA-tagged PCBP were used for ChIP assay using the protocol from Upstate Biotechnology (Lake Placid, NY). Twenty-four hrs after transfection of NMB cell with Effectene transfection reagent (Qiagen), chromatin was cross-linked by the addition of formaldehyde to the culture medium at a final concentration of 1% and incubated for 10 min at 37°C. Cells were washed twice with PBS containing protease inhibitor cocktail (Roche). Washed cells were counted and 1 x 10 6 cells were resuspended in 200 µl of SDS lysis buffer (Zhang and Dufau, 2002). We determined conditions to produce sheared chromatin of average lengths between 0.5 and 1.5 kb using a cell destructor (Ultrasonic Inc.). Each sample was sonicated using the cell destructor at 30% intensity for 6 x 10 s bursts with a gap of 30 s between each burst. Cells were maintained on ice throughout. Sheared chromatin was centrifuged for 10 min at 14,000 g at 4°C, after which the supernatant was diluted 10-fold with dilution buffer (Zhang and Dufau, 2002). As input control, 1% of each diluted supernatant was retained at this step. Chromatin was pre-cleared for 1 h at 4°C with 75 µl 50% Protein G agarose (Invitrogen), 20 µg of sonicated and sheared salmon sperm DNA, and 50 µg of BSA. Anti-HA antibodies (2 µg) (Roche) were added to pre-cleared supernatant, and the immunoprecipitates were allowed to form overnight at 4°C with end-over-end rocking. Immune complexes were incubated for 1 h at 4°C with 60 µl of 50% Protein G agarose beads and 4 µg of sonicated and sheared salmon sperm DNA, and then centrifuged gently for 1 min at 4°C. Beads were washed four times for 5 min, and eluted twice in 250 µl of fresh elution buffer (1% SDS, 0.1 M NaHCO 3 ). Samples were heated at 65°C for 4 h to reverse cross-links, and the DNA samples were purified by phenol/chloroform extraction followed by ethanol SDS-PAGE. Based on the size comparison with the molecular weight markers, the molecular weight of the translated PCBP family was found to be approximately similar to the predicted molecular weight deduced from the full-length cDNA sequence (data not shown). 0 0 5 / 0 1 2 2 4 5 13 RESULTS Cloning of Poly (C) Binding Protein Family PCBP Family Proteins Bind to the MOR Proximal Promoter In Vitro To verify the binding of PCBPs to the polypyrimidine stretch in MOR proximal promoter, we 14 performed electrophoretic mobility shift assay (EMSA) with single stranded (SS) as well as double stranded (DS) 26 nt long oligonucleotide sequence As shown in EMSA with other PCBP family members (PCBP2, PCBP3 and PCBP4) and the above probe yielded similar kind of results as in case of PCBP1 Specific Binding of PCBP Family Proteins to the MOR Promoter In Vivo Effect of PCBP Family on MOR Proximal Promoter Activity Use of Decoys to inhibit the PCBP mediated induction of proximal promoter activity We also used decoy oligonucleotide approach to confirm the PCBPs binding specifically to polypyrimidine tract in MOR promoter. For this purpose, a luciferase reporter gene construct under MOR promoter fragment containing PCBP binding region was cotransfected with various concentrations of decoy oligonucleotide in NMB cells. The decoy 17 oligonucleotide binds to the PCBPs and decreases the ability of PCBPs to bind to their cellular target sequences. Hence, decoy-induced inhibition of PCBP-mediated induction of promoter activity was measured siRNA Mediated Inhibition of PCBP Expression Our data showed that PCBPs bind directly to the MOR proximal promoter region and increase transcription activity. siRNAs have been used extensively to study or confirm the gene function in the mammalian system. We used duplex siRNAs against PCBPs individually and in combination to inhibit the expression of PCBPs family and studied their effects on MOR gene expression. There was not effect on the MOR expression level when the cells were transfected with siRNA against on type of PCBP PCBPs may mainly contribute to the regulation of MOR gene expression in NMB cells. DISCUSSION Our group previously reported that a 26 nucleotide long cis-element rich in polypyrimidine residues in MOR proximal promoter can adopt ssDNA structure and interact with the nuclear proteins. In this study, we demonstrated that this polypyrimidine tract interacted with a family of PCBPs (Ko and Loh, 2005) . Despite their recognized function as RNA-binding proteins, PCBPs appeared to also function to directly activate gene transcription of the MOR gene. The proximal promoter of the mouse MOR gene is known to be regulated by various cis-elements and trans-factors, all of these elements are important in regulating proximal promoter activities. These features are unique in the proximal promoter of mouse MOR, as compared to the promoter of other opioid receptor genes (kappa and delta). Thus, the proximal promoter regulation of mouse MOR gene is under a fine and tight control (Ko and Loh, 2001; Ko et al., 2003; PCBPs have been involved in a spectrum of posttranscriptional controls. First insights into the functions were obtained from investigation of human α-globin mRNA stabilization (Weiss and Liebhaber, 1995). Stabilization of α-globin mRNA is tightly linked to formation of a binary complex between a single molecule of PCBPs and pyrimidine-rich region within the α-globin. We performed nuclear run-off assay in PCBP transfected NMB cells and determined et al., 1995). In a recent study using PCBP2, it was shown that PCBP2 can interact with 160 different types of mRNA and regulate a number of cellular processes (Waggoner and Liebhaber, 2003)
Clinical Study Metastatic Behavior in Melanoma: Timing, Pattern, Survival, and Influencing Factors
Get PDFMetastatic melanoma (MM) is a fatal disease with a rapid systemic dissemination. This study was conducted to investigate the metastatic behavior, timing, patterns, survival, and influencing factors in MM. 214 patients with MM were evaluated retrospectively. Distant metastases (82%) were the most frequent for patients initially metastatic. The median and 1-year survival rates of initially MM patients were 10 months and 41%, respectively. The median time to metastasis for patients with localized disease was 28 months. The timing of appearance of metastases varied minimally; however, times to metastases for distant organs varied greatly. For the first metastatic pathway, more than half of the primary metastases were M1A (57%). These findings were in contrast to the results compared with those with metastatic in diagnosis (P < 0.001). The median and 1-year survival rates of all patients were 12 months and 49%, respectively. Outcome was higher in M1A than visceral metastases (P < 0.001). In conclusion, the fact that over half of all recurrences/metastases occurred within 3 years urges us to concentrate follow-up in the early time periods following diagnosis. Because the clinical behavior of MM is variable, the factors for survival consisting of site and number of metastases should be emphasized
Undernutrition during pregnancy in mice leads to dysfunctional cardiac muscle respiration in adult offspring
Get PDFSummary Statement We show that in utero undernutrition is associated with impaired cardiac muscle energetics and increased plasma short-chain acylcarnitines in adult mice. Findings suggest that in utero undernutrition is associated with maladaptive programming processes that have negative effects on the heart. Synopsis Intrauterine growth restriction is associated with an increased risk of developing obesity, insulin resistance, and cardiovascular disease. However its effect on energetics in heart remains unknown. In this study, we examined respiration in cardiac muscle and liver from adult mice that were undernourished in utero. We report that in utero undernutrition is associated with impaired cardiac muscle energetics, including decreased fatty acid oxidative capacity, decreased maximum oxidative phosphorylation rate, and decreased proton leak respiration. No differences in oxidative characteristics were detected in liver. We also measured plasma acylcarnitine levels and found that short-chain acylcarnitines are increased with in utero undernutrition. Results reveal the negative impact of suboptimal maternal nutrition on adult offspring cardiac energy metabolism, which may have lifelong implications for cardiovascular function and disease risk