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p53 inhibitors as targets in anticancer therapy
Category Archives: Hydroxylases
Supplementary MaterialsSupplementary data mmc1
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Supplementary MaterialsSupplementary data mmc1. AGD is the protozoan [find Refs. [12,13]], which may be the most phylogenetically divergent types [14]. Though normally free living, can colonise the gills and cause the disease, which is definitely characterised by multifocal white patches within the gill surface. At a histological level AGD causes hyperplasia of the epithelial and mucous cells, which can lead to lamellar fusion, generally in association with attached amoebae [15]. Cumulative pirinixic acid (WY 14643) mortalities can reach up to 50% if remaining untreated [16]. Currently, a commercial AGD vaccine is not available [17]. Though initial studies have been conducted to evaluate the effectiveness of several potential chemotherapeutants [[18], [19], [20]], at present, exposure to freshwater remains the most effective treatment [21]. One of the important difficulties to developing and evaluating new therapeutants is the availability of a cost effective ethically sound model system. systems have the potential to address these requirements, and, because of the clonal nature demonstrate less inherent heterogeneity between replicates than would be observed between live fish replicates, therefore potentially reducing the need for animal use in experiments [22]. An system to study host-pathogen connection in AGD requires the ability to isolate and grow the parasite and the ability to maintain pirinixic acid (WY 14643) suitable sponsor cells. Protocols for the isolation of from diseased fish, and tradition onto malt candida agar (MYA) are available. Under these conditions, the parasite retains its virulence and capacity to cause AGD pirinixic acid (WY 14643) in Atlantic salmon after at least 70 days of clonal tradition [12]. However, cultured has been shown to lose virulence after 3 years of repeated passage in tradition [23]. There is little published information about illness (Walbaum, 1792) gills (RTgill-W1) [25]. When cultured at an osmolarity above 700 mOsm kg?1, this system has been shown to support the growth of [observe Ref. [27]]. However, unlike requires full salinity sea water and can't be subjected to web host cells in cell lifestyle media that have lower osmolality than ocean drinking water [27]. Transwell? lifestyle inserts give a permeable support which seeded cells can connect and type confluent monolayers. By changing apical mass media with either seawater or freshwater, lifestyle conditions could be modified to determine asymmetrical systems which create a cell lifestyle environment that allows the establishment of effective polarised epithelia and even more carefully resembles the condition. This program continues to be utilized assays successfully to attempt pirinixic acid (WY 14643) chemotaxis, drug transportation, and toxicity lab tests with seafood gill principal cell civilizations [find 29 for critique]. RTgill-W1 cells can develop on the Transwell? in immediate contact with clean or saltwater on the pirinixic acid (WY 14643) apical surfaces developing tight epithelia, and also have been suggested being a sentinel model for aquatic toxicology [29], enabling the analysis of gill illnesses and may as a result be suitable for studies on system being a model to review host-interactions, utilizing the rainbow trout gill cell series RTgill-W1 seeded onto Transwell? inserts and subjected to two clones: a outrageous type clone and a lab passing attenuated one. The association of using the gill epithelium, the parasite development and the appearance of an array of genes mixed up in Atlantic salmon innate immune system response to AGD are analysed. The application of the system as an proxy to judge therapeutics to fight AGD is discussed. 2.?Material & methods 2.1. Ethics statement Animal procedures were approved by the Animal Welfare and Honest Review Body Rabbit Polyclonal to PKC delta (phospho-Ser645) (AWERB) in the Cefas Weymouth Laboratory and carried out in compliance with the Animals (Scientific Methods) Take action 1986. 2.2. isolates and tradition trophozoites were isolated from your gills of naturally infected Scottish farmed sea-cage Atlantic salmon showing standard AGD lesions as explained before [30]. Isolated amoebae were then cultured on malt candida agar (MYA: 0.01% malt, 0.01% candida, 2% Bacto agar, 0.2?m filtered sea water (SW) at 35 salinity) overlaid with 0.2?m filtered SW. Plates were incubated at 18?C and amoebae subcultured fortnightly by transfer of SW to fresh MYA plates with an additional overlay of 0.2?m filtered SW while described previously [12]. The isolation of or related varieties was confirmed by a species-specific PCR as explained below. Cell counting was performed inside a haemocytometer and in a TC20 automated cell counter (Bio-Rad, Herts, UK). In order to obtain a clonal tradition, an isolated trophozoite was separated and propagated as explained above. Two clones were used in this study: a.
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CDT2 targets proteins involved in replication licensing (CDT1), cell cycle control (p21), and chromatin modification (SET8) for destruction by the CUL4-based E3 ligase (CRL4)
CDT2 targets proteins involved in replication licensing (CDT1), cell cycle control (p21), and chromatin modification (SET8) for destruction by the CUL4-based E3 ligase (CRL4). may provide tumors with a proliferative advantage. INTRODUCTION The CHK1 protein kinase maintains genome integrity in normal cycling cells and in cells exposed to replication or genotoxic stress (1, 2). Replication stress that occurs during the normal course of DNA replication or following exposure to antimetabolites or certain DNA-damaging agents generates single-stranded DNA (ssDNA). ssDNA is also generated in the course of DNA repair and double-strand break (DSB) end resection. The CHK1 signaling pathway is engaged by checkpoints that detect ssDNA. Replication protein A (RPA) coats ssDNA, therefore recruiting a Rabbit polyclonal to ITPKB DNA damage-sensing complicated comprising ATR (ataxia telangiectasia- and RAD3-related proteins) and ATRIP (ATR-interacting proteins) (3, 4). The ATR/ATRIP module, with RAD17 as well as the 9-1-1 complicated collectively, activates CHK1 inside a claspin-dependent way on chromatin (5C9). ATR phosphorylates CHK1 on serine 317 (S317) and serine 345 (S345), which activates CHK1 by facilitating autophosphorylation on S296 (10C13). Activated CHK1 can be after that released from phosphorylates and chromatin downstream effectors to briefly halt cell routine development, stabilize Primaquine Diphosphate stalled replication forks, and regulate DNA restoration (4, 14). ATR-mediated phosphorylation activates CHK1 and in addition promotes its ubiquitin-mediated proteolysis by facilitating relationships with two specific E3 ubiquitin ligases that use CUL1 and CUL4A (15C17). These cullin proteins work as scaffolds in multisubunit complexes referred to as cullin-RING ligases (CRLs) (18). CRLs recruit substrates via adaptor protein scaffold particular for every cullin. CRL1 utilizes SKP1 (S-phase kinase-associated proteins 1), and CRL4 utilizes DDB1 (broken DNA binding proteins 1). Cullin-adaptor complexes require additional substrate receptors to recruit and ubiquitinate focus on protein often. Substrate receptors offer E3 ubiquitin ligases using the specificity necessary to focus on their varied repertoire of mobile substrates for ubiquitination. While F-box protein recruit substrates to CRL1, CRL4 frequently recruits its substrates via DCAFs (DDB1- and CUL4-connected elements) (19C21). Greater than a hundred DCAFs and putative DCAF proteins have already been identified predicated on quality motifs, including WD40 repeats, WDXR motifs, and DDB containers (19C23). The DCAF proteins CDT2 identifies substrates including a specific PCNA (proliferating cell nuclear antigen) discussion protein theme (PIP package) known as a PIP degron (24). Chromatin-bound PCNA mediates the recruitment of PIP degron-containing substrates to CRL4CDT2 (24). The F-box proteins FBX6 facilitates relationships between CHK1 and CRL1 (16), however the substrate receptor mediating relationships between CHK1 and CRL4 is not determined. Furthermore, it is unclear why two distinct E3 ubiquitin ligases mediate CHK1 degradation. Here we demonstrate that CDT2 targets the activated form of CHK1 to CRL4 using a noncanonical mechanism and that CHK1 stability is usually regulated in distinct cellular compartments by CRL1FBX6 and CRL4CDT2. We also demonstrate that CHK1 kinase activity is essential for the maintenance of G2 cell cycle arrest in CDT2-depleted cells. MATERIALS AND METHODS Cell culture, antibodies, and reagents. HeLa cells were produced in Dulbecco's Primaquine Diphosphate modified Eagle's medium (DMEM) (Life Technologies) supplemented with 10% bovine growth serum, l-glutamine, and penicillin-streptomycin. HeLa Tet-on cells (Clontech) were produced in DMEM supplemented with 10% Tet system-approved fetal bovine serum (Clontech), l-glutamine, penicillin-streptomycin, and 100 g/ml Primaquine Diphosphate Geneticin (Life Technologies). Primaquine Diphosphate 293T cells were produced in DMEM supplemented with 10% fetal bovine serum and l-glutamine. The following antibodies were purchased: CHK1 (G-4), CUL1 (H-213), CDT2 (B-8), Myc (9E10), PCNA (PC10), SKP1, and FBX6 (7B11) antibodies were purchased from Santa Cruz Biotechnology; actin, Flag (M2), and claspin antibodies were purchased from Sigma; CUL4 and CDT1 antibodies were purchased from Bethyl Laboratories; CUL4A antibody was purchased from Rockland Immunochemicals; V5, CDT2, DDB1, and tubulin antibodies were purchased from Abcam; CHK1 phospho-S296 (pS296) antibody was purchased.
Supplementary MaterialsTransparent reporting form
Supplementary MaterialsTransparent reporting form. varied processes, including zebrafish development, murine intestinal crypt and human being tumor organoids, demonstrating that Wnt transport by cytonemes and its control via the Ror2 pathway is definitely highly conserved in vertebrates. and as well as the manifestation of tissue-specific genes, and subsequently controls both?cell proliferation?and cells patterning. In the -catenin-independent Wnt/PCP pathway (Yang and Mlodzik, 2015), Wnt proteins bind to Frizzled and to?co-receptors such as the receptor-tyrosine kinase-like orphan receptor 2 (Ror2) to regulate cytoskeleton corporation by actin polymerization and cell polarity (Grumolato et al., 2010; Ho et al., 2012; Oishi et al., 2003). To this end, the?small GTPases Rho, Rac1, and Cdc42 are regulated to control the?formation of filopodia and lamellipodia, as well as cell motility and morphogenetic motions of?cells?in vertebrates. Although the PCP pathway and -catenin signaling generally take action inside a mutually repressive fashion, by competing for related hub proteins?such as the effector protein Dishevelled (Dvl) (van Amerongen and Nusse, 2009), recent evidence suggests that PCP signaling can act dependent on the context either in opposition to, in concert with, or independently of -catenin signaling. The Triethyl citrate production and secretion of Wnt ligands requires lipid modification from the acyltransferase Porcupine (Porcn) followed by binding to Evi/Wls, which serves as a Wnt chaperone and facilitates its transportation in the endoplasmic reticulum towards the plasma membrane (Bartscherer and Boutros, 2008; B?nziger et al., 2006; Yu et al., 2014). Following that, lipophilic Wnt is normally transported with the neighboring tissues to exert its long-range signaling activity. Extracellular binding protein have been recommended to improve the solubility of Wg/Wnt within the aqueous extracellular space and facilitate this activity (Mii et al., 2009; Mulligan et al., 2012). Various other studies,?however, indicate membrane-associated mechanisms of Wg/Wnt delivery, which?perform?not really compromise the signaling capacity for?Wg/Wnt?(McGough and Vincent, 2016; Basler and Port, 2010; Scholpp and Stanganello, 2016). These trafficking routes consist of Wg/Wnt proteins distribution over the plasma membrane of dividing supply cells (Alexandre et al., 2014; Farin et al., 2016) and?positively migrating cells (Serralbo and Marcelle, 2014), or the dissemination of Wg/Wnt proteins in exovesicles (Pankov et al., 2005)?(more specifically on?exosomes [Beckett et al., 2013; Gross et al., 2012; Korkut et al., 2009]). Wg/Wnt proteins and their receptors are transported in cell protrusions in a variety of tissues also. Lipid-modified Wnt protein were bought at the cell membrane of signaling filopodia ?so-called cytonemes in and zebrafish (Holzer et al., 2012; Luz et al., 2014; Stanganello et al., 2015), whereas Fzd receptor protein could be localized to filopodia in and poultry (Huang and Kornberg, 2016; Sagar et al., 2015). In zebrafish, an evaluation of cytonemes shows these are specific filopodia, with stabilizing actin bundles at their cores, which serve as a Triethyl citrate primary transport gadget for the -catenin ligand Wnt8a during neural dish patterning (Stanganello et al., 2015). Wnt8a is normally packed on cytoneme guidelines and used in the neighboring cells by immediate cellCcell get in touch with. At the get in touch with sites, Wnt8a cytonemes induce Lrp6/Fzd receptor clustering in to the Lrp6 Mouse monoclonal to ZBTB16 signalosome to activate -catenin signaling. Even though numbers and lengths of cytonemes are necessary in?determining the -catenin signaling vary during embryogenesis (Stanganello et al., 2015), it isn't?yet?apparent what mechanism handles the forming of Wnt cytonemes within a tissues. Here, we present that Wnt8a can activate both PCP pathway by connections with Ror2 as well as the -catenin pathway by connections with Lrp6. This dual function enables Triethyl citrate Wnt8a to regulate its own path of dissemination. In the foundation cells, Wnt8a binds and activates the Ror2 co-receptor accompanied by activation from the PCP pathway. Wnt8a-PCP affects convergent expansion (CE) motion and activates the tiny GTPase Cdc42, that leads towards the outgrowth of signaling filopodia. Wnt8a is normally packed onto these cytonemes, and it is transported with the tissues to bind towards the -catenin-specific co-receptor Lrp6 within the responding cells, illustrations being PAC2 seafood fibroblasts and HEK293T individual embryonic kidney cells, where it activates the -catenin pathway within a paracrine style. Activation from the -catenin pathway by Wnt cytonemes results in target-gene induction, which in.
Supplementary MaterialsSupplementary informationSC-010-C9SC04151F-s001
Supplementary MaterialsSupplementary informationSC-010-C9SC04151F-s001. two ligands. Incorporation of a MK-1064 di-2-picolylamine binding unit into the ligand design provided efficient intracellular zinc uptake, resulting in metallochaperone capability for both LI and LH. The ability of LI to lessen mutant p53 aggregation leads to increased repair of p53 transcriptional function and mediates both caspase-dependent and -3rd party cell loss of life pathways. We show that LI displays minimal toxicity in non-cancerous organoids further, and that it's well tolerated in mice. These outcomes demonstrate that iodination of our ligand platform restores p53 function by getting together with and inhibiting mutant p53 aggregation and shows LI as the right candidate for extensive anticancer preclinical assessments. Intro Amyloidogenic proteins are inclined to endogenous misfolding and prion-like transformation from a soluble, folded protein into alternative fibrillar and oligomeric set ups.1 Proteins vunerable to this technique include amyloid-, tau, TDP-43, SOD1, and -synuclein and donate to an array of diseases including Alzheimer's disease and ALS.2 These protein show toxic gain-of-function (GoF) results by self-propagating and performing as seed products to start aggregation.3 Just like neurodegenerative diseases, latest studies possess demonstrated that proteins misfolding and aggregation are likely involved in cancer advancement through misfolding from the tumour suppressor proteins p53.4 Several reviews possess highlighted that p53 aggregation not merely leads to lack of function, but that it could co-aggregate with homologous protein p63 and p73 to create amyloid fibrils and oligomers.4a,5 p53 takes on a critical part in managing the cell routine by initiating apoptosis, DNA fix, and cell routine arrest of damaged cells.6 The core DNA-binding domain of p53 (p53C) consists of an individual Zn2+ ion that's needed for proper proteins folding and function.7 However, p53 is mutated in over 50% of tumor diagnoses, the most frequent mutations affecting the protein's tertiary framework and frequently producing a reduction or alteration of Zn-binding at the primary site.8 This may lead to proteins unfolding and improved aggregation because of publicity of amyloidogenic parts of the proteins (residues 251C257).8a,9 Kinetic research indicate that happens a two-step approach wherein the first requires relatively decrease unfolding of MK-1064 p53C to expose the aggregation nucleus accompanied by a second, rapid step aggregation.10 Interestingly, apo p53C (zinc-free) escalates the aggregation approach nucleation with zinc-bound p53C and plays a part in lack of protein function.11 The normal hotspot mutant p53-Y220C destabilizes the protein's tertiary structure because of an exposed cavity at the top of proteins. This can lead to lack of Zn2+ and causes accelerated proteins aggregation.7,8,10a,12 While study regarding repair of p53 function has centered on stabilization of mutant p53C largely,13 repopulating the metal-depleted site metallochaperones14 has been proven to revive function to common p53 mutants.13a,14aCc,15 More broadly, targeted metal ion chelation and redistribution shows utility both as an anticancer strategy16 and in modulating amyloidogenic protein aggregation.17 Furthermore, several small molecule/peptide inhibitors of p53 aggregation have already been developed18 and a cell-penetrating peptide (ReACp53) developed by Eisenberg and co-workers rescued p53 function in high-grade serous ovarian carcinomas and led to decreased tumour proliferation in xenograft models.9 Given the increased propensity for Rabbit Polyclonal to CYSLTR2 aggregation and possible zinc loss in the common mutant p53-Y220C, we used this as a model for testing compounds targeted to modulate mutant p53 aggregation. Herein, we describe two novel bifunctional ligands, LI and LH (Fig. 1), designed to reactivate p53 by inhibiting mutant p53 aggregation and restore zinc-binding using a metallochaperone approach. With reports showing that Zn-free p53 exhibits accelerated protein aggregation,5b,8a,11 the incorporation of a zinc metallochaperone unit to remetallate apo-p53 in combination with an aggregation-targeting moiety could provide advantages over reported single-target compounds. Furthermore, multifunctional agents are advantageous due to their ability MK-1064 to act on multiple targets, resulting in.
Background Fluoranthene (FR) is a common environmental pollutant that exists inside a complex mixture with additional polycyclic aromatic hydrocarbons (PAHs)
Background Fluoranthene (FR) is a common environmental pollutant that exists inside a complex mixture with additional polycyclic aromatic hydrocarbons (PAHs). exposure. Conclusion The exposure of BM-MSCs to FR induced impressive CNT2 inhibitor-1 alterations in cellular biology and the proteome, allowing for identification of novel biomarkers for FR exposure. Furthermore, AHR antagonists might be able to prevent cellular damage due to FR exposure. cell line studies, cells were cultured and maintained in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum and different concentrations of FR (25, 50, 100, 150, and 200 M). The medium was replaced every 24 hours to avoid chemical decay of FR. Reagents and antibodies FR (98%) was purchased from Sigma (Sigma-Aldrich, St. CNT2 inhibitor-1 Louis, MO, USA) and dissolved in dimethyl sulfoxide (DMSO) (Sigma-Aldrich) to produce a stock solution at a concentration of 100 mM. A working solution was obtained with dilution of the stock solution using cell culture medium. Cell viability measurement For the assay of viable cell numbers, 2103, 4103, and 5103 cells per well were seeded in triplicate into a 96 well plate and subjected to an Enhanced Cell Viability Assay Kit (EZ-CyTox, Daeil Lab Service Co., Seoul, Korea), according to the instructions of the manufacturer. The tetrazolium-based colorimetric assay evaluates viable cell numbers, taking advantage of NADPH-dependent cellular oxidoreductase enzymes in viable cells to reduce the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to insoluble formazan, which has a purple color. The absorbance at 450 nm of each well was measured using a VERSA Max microplate reader (Molecular Devices, Sunnyvale, CA, USA). DNA fragmentation assay DNA fragmentation assays were performed with the DNeasy mini kit Rabbit polyclonal to ANXA8L2 (Qiagen, Valencia, CA, USA) according to the manufacturer's protocol. MSC cells had been treated with 150 M and 200 M FR for 72 hours, and genomic DNA was extracted. Up coming, DNA fragments had been separated by gel electrophoresis on the 1.5% agarose gel. DNA was visualized using ethidium bromide staining and photographed under ultraviolet light. Recognition of mitochondrial membrane potential Mitochondrial membrane potential was assessed using MitoTracker Crimson CMXRos (Molecular Probes Inc., Eugene, OR, USA) based on the guidelines provided by the maker. Quickly, after 72 hours of treatment, MSC cells had been washed double with phosphate buffer saline (PBS) and packed with 100 nm pre-warmed CMXRos for thirty minutes in serum free of charge culture moderate. Subsequently, cells had been washed double with fresh moderate and transferred to a mounting chamber on the microscope stage. Images of red MitoTracker Red CMXRos fluorescence were captured using a 568-nm excitation light from the argon/krypton laser, a 560-nm dichroic mirror, and a 590-nm long pass filter. Western blot analysis Protein expression was determined in BM-MSCs using the western blotting technique according to standard procedures. Briefly, total protein from untreated or treated cells was extracted in RIPA lysis buffer, and an equivalent amount of protein (50 g) from each group was separated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto a polyvinylidene fluoride (PVDF) membrane (Bio-Rad Laboratories, Hercules, CA, USA). After blocking with sodium azide at room temperature for 90 minutes, each membrane was incubated with a specific primary antibody (1:1,000) at 4 overnight. After three washes in washing buffer (20 mM Tris-HCl, 500 CNT2 inhibitor-1 mM NaCl, and 0.1% Tween 20), each membrane was incubated with the appropriate secondary antibody at room temperature for 90 minutes. Specific protein bands were visualized using a chemiluminescence detection system (Amersham ECL system, London, UK). Cellular proteome analysis Proteomic analysis was performed by a commercial laboratory (Yonsei Proteome Research Center, Seoul, Korea; www.proteomix.org). Briefly, the FR-treated cultured cells (5106/mL) were washed extensively with ice-cold PBS. The cells were centrifuged at 600 G for 5 minutes at 4 then. The cells had been resuspended in 1.0 mL of just one 1 cytosol extraction buffer containing dithiothreitol and protease inhibitors from a industrial mitochondria/cytosol fractionation kit CNT2 inhibitor-1 (Pierce Biotechnology). Purification from the cytosolic small fraction was carried out, and samples had been ready for and operate on two-dimensional gel electrophoresis (2-DE), that was performed using published protocols [22] previously. Coomassie.
Data Availability StatementThe datasets used and/or analyzed through the current research are available through the corresponding writer on reasonable demand
Data Availability StatementThe datasets used and/or analyzed through the current research are available through the corresponding writer on reasonable demand. and tumor necrosis aspect (TNF)- had been also discovered using ELISA products to measure the inflammatory response. Finally, traditional western blotting and invert transcription-quantitative PCR had been utilized to investigate the appearance degrees of linked protein and mRNAs. Ad-G-DEC1 RNA interference markedly decreased DEC1 expression levels. In addition, following the downregulation of DEC1 expression, the infarct size, CK, LDH, Toll-like receptor (TLR)4, NF-B, IL- and TNF- levels were all significantly decreased. In conclusion, the results of the present study suggested that this downregulation of DEC1 may decrease the inflammation by suppressing the TLR4/NF-B signaling pathway, which may represent a therapeutic target for MIRI. (12) reported that DEC1 modulated access to food and water. Small interfering RNA (siRNA) interference The adenovirus (RNA interference) made up of siRNA against DEC1 was synthesized and generated with a pBHGlox_E1,3Cre plasmid (Microbix Biosystems) using the AdMax system (Microbix Biosystems) according to the manufacturer's protocols. A scrambled sequence (500 l; Sangon Biotech Co., Ltd.) was used as the unfavorable control. 293T cells (cat. no. CRL-1573; American Type Culture Collection; 4×105 cells/ml) were used to package and amplify the adenoviruses (500 l) with Lipofectamine? 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) in DMEM (Gibco; Thermo Fisher Scientific, Inc.), supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37?C in a 5% CO2 incubator for 10-15 days. After the majority of 293T cells exhibited common cytopathic effects, samples were frozen-thawed at -70?C/37?C three times and centrifuged at 7,000 Dimethyl phthalate x g at 4?C for 5 min. Computer virus supernatants were subsequently collected. The final computer virus concentration was 1×1011 plaque-forming models, as using the endpoint dilution method (21). Establishment of MIRI model rats The 48 healthy male SD rats were divided into four groups (n=12/group): The sham group (Sham); I/R group (I/R); adenovirus expressing green fluorescent protein Control group (Ad-G-Control); and DEC1-targeting RNA interference group (Ad-G-DEC1). The rat MIRI model was prepared in the I/R, Ad-G-Control and Ad-G-DEC1 groups. Briefly, sodium pentobarbital (30 mg/kg; intraperitoneal) was used to anesthetize the rats and open the chest. A volume of 80 l Ad-G-Control or Ad-G-DEC1 diluted in saline (1×1010 PFU/ml; 10 occasions diluted) was injected into the heart wall of 8-week-old rats. A total of 4 times afterwards, the Dimethyl phthalate rats had been re-anesthetized, the rats' chests had been reopened as well as the still left anterior descending artery (LAD) was ligated utilizing a 6-0 silk suture. Pursuing 30 min of ligation, PRHX the blood flow was restored for ~4 h by detatching the silk suture; when the rat's limbs confirmed slight motion (~70 min intervals), the rats had been re-anesthetized to maintain them under anesthetic in this treatment. Post-reperfusion, the rats had been sacrificed by jugular vein shot of potassium chloride (75 mg/kg). Loss of life was confirmed utilizing a regular body component II-lead electrocardiogram. Both bloodstream (5 ml) and area of the anterior wall structure of the still left ventricular myocardium close to the cardiac apex had been subsequently harvested through the rats. The sham groupings offered as the control no occlusion from the LAD was performed. Perseverance of myocardial enzymes Harvested bloodstream was centrifuged at 500 x Dimethyl phthalate g for 5 min at 4?C to acquire serum, that was utilized to measure myocardial enzyme amounts subsequently, including creatine kinase (CK; kitty. simply no. QS1107) and lactate dehydrogenase (LDH; kitty. simply no. QS1001) using commercially obtainable biochemical products (Beijing Solarbio Research & Technology Co., Ltd.), based on the producers' protocols. The info obtained are shown as U/l. Recognition of myocardial infarct region (IA)/area in danger (AAR) Evans Blue/triphenyltetrazolium chloride (TTC) staining was utilized to look for the IA pursuing MIRI. In short, the LADs from the rats had been immediately ligated pursuing 4 h of reperfusion and ~1 ml Evans blue (Sigma-Aldrich; Merck KGaA) was intravenously injected to tell apart non-ischemic and risk areas. Next, rats had been sacrificed by injecting 75 mg/kg potassium chloride in to the jugular vein, and hearts had been removed, iced at -20?C for 10 min and sliced into 2 mm heavy sections. Areas were stained with 1 subsequently.5% TTC (Sigma-Aldrich; Merck KGaA) for.
Supplementary MaterialsSupplementary information 41467_2019_10460_MOESM1_ESM
Supplementary MaterialsSupplementary information 41467_2019_10460_MOESM1_ESM. approaches focusing on the senescence state using senolytic drugs. The combination of PARPi and a senolytic is effective in preclinical models of ovarian and breast cancer suggesting that coupling these synthetic lethalities provides a rational approach to their clinical use and may together be more effective in limiting resistance. mutations and have high rates of copy number anomalies23C26. In particular, OV4453 carries a mutation that is likely responsible for PARPi sensitivity4,23. Real-time imaging confirmed dose-dependent Olaparib-mediated inhibition of cell proliferation in which higher concentrations were required for two cell lines and IC50 were consistent with those obtained using clonogenic assays (Fig.?1a, Supplementary Fig.?1A). Interestingly, live-cell imaging revealed that inhibition of cell proliferation was not accompanied by significant cell detachment. This was confirmed by correspondingly small increases in total cumulative cell death/apoptosis, as only 20C40% of cells were cumulatively AnnexinV and/or DRAQ7 positive 6 days after treatment initiation, even at the highest Olaparib concentrations (Fig.?1b, Supplementary Fig.?1B). However, real-time images revealed treatment-associated changes in cell morphology, including cell enlargement that started at day 3 and became more pronounced at day 6 (Supplementary Fig.?1C), suggesting a senescence cell fate response. Open in a separate window Fig. 1 Olaparib induces a senescence-like phenotype in HGSOC RO4927350 cell lines. a Cell proliferation Rabbit Polyclonal to BST1 curves of HGSOC H2B-GFP cell lines exposed to increasing concentrations of Olaparib. b, c HGSOC dead cells analyzed by flow cytometry (b) and SAgal positive HGSOC cells (c) following 6 days treatment with selected Olaparib concentrations (Supplementary Fig.?1A). d HGSOC cell morphology analyzed by flow cytometry following 6 days of treatment with Olaparib IC50 concentrations (see Supplementary Fig.?1A, E for details). e, f Levels of IL-6 (e), IL-8 (f) were measured by ELISA assay following 6 days treatment with Olaparib IC50 concentrations. g Number of -H2AX foci per nucleus in HGSOC cells lines pursuing 6 times of treatment with Olaparib IC50 concentrations. h, i Evaluation of 8-h (h) or 24-h (i) EdU pulse after 6 times publicity of HGSOC cells to Olaparib IC50 concentrations. j Movement cytometry evaluation of cell routine populations pursuing 6 days publicity of HGSOC cells to Olaparib IC50 concentrations. Data in (a) are representative curves of at least three 3rd party experiments. For all your data, the mean??SEM of three individual tests is shown. Data had been examined using the two-tail College student check. *Denotes mutant position22, that was verified for HGSOC cells with this research23C26. Therefore, improved degrees of the immediate p53 transcriptional focus on p21 are unpredicted. However, p53-3rd party activation of p21 continues to be reported during embryonic- and oncogene-induced senescence33 and pursuing overexpression from the Chk2 DDR kinase in epithelial tumor cells34. To check whether a Chk2-p21 pathway regulates PARPi-induced proliferation arrest in HGSOC cells likewise, we confirmed the Chk2 (test. *Denotes test. *Denotes test. * Denotes RO4927350 test. * Denotes mutations in this type of malignancy40. Olaparib doseCresponse curves for mutant triple unfavorable breast cancer (TNBC) MDA-MB-231 cells41 revealed a concentration-dependent inhibition of cell proliferation that was in a IC50-intermediate range when compared to HGSOC cells (Fig.?6a, IC50: 2.92??0.17?M). As in HGSOC cells, Olaparib induced a senescence-like phenotype in MDA-MB-231 cells, including a very low cumulative cell death rate even at concentrations above the IC50 (Fig.?6b, Supplementary Fig.?11A), a significant increase in SAgal positive cells (Fig.?6c, Supplementary Fig.?11B), and a clear cell enlargement even at a RO4927350 lower concentration (2.5?M) (Supplementary Fig.?11C, D). Short and long EdU pulse-labeling assays revealed a dose dependent decrease in DNA synthesis at day 6 in Olaparib-treated TNBC cells (Fig.?6d), indicating an.
Astrocytes have long been considered the supportive cells from the central nervous program, but over the last years, they have got gained a lot more attention for their dynamic involvement in the modulation of neuronal function
Posted on August 9, 2020 by
Astrocytes have long been considered the supportive cells from the central nervous program, but over the last years, they have got gained a lot more attention for their dynamic involvement in the modulation of neuronal function. migration. The coordination between astrocytes Rabbit Polyclonal to OR10R2 takes place generally through Connexin (Cx) stations, by means of immediate cell-cell get in touch with (difference junctions, GJs) or get in touch with between your extracellular matrix as well as the astrocytes (hemichannels, HCs). Reactive astrocytes raise the expression degrees of many proteins involved with astrocyte migration, such as for example v3 Integrin, Syndecan-4 proteoglycan, the purinergic receptor P2X7, Pannexin1, and Cx43 HCs. Proof provides indicated that Cx43 HCs are likely involved in regulating astrocyte migration through the discharge of small substances towards the extracellular space, which in turn activate receptors in the adjacent AZD7762 irreversible inhibition or same cells to keep the signaling cascades necessary for astrocyte migration. Within this review, the conversation is normally defined by us of astrocytes through Cxs, the function of Cxs in astrocyte and irritation migration, and discuss the molecular systems that regulate Cx43 HCs, which might provide a healing window of possibility to control astrogliosis as well as the development of neurodegenerative illnesses. by freeze-fracture electron microscopy (Brightman and Reese, 1969; Dermietzel, 1974). Afterwards, in 1991, Cx43 was discovered to be among the main Cx subtypes in astrocytes (Dermietzel, 1991). The pivotal function of Cxs in astroglial connection was showed with Cx43/Cx30 dual knockout (KO) mice, where intercellular conversation was dropped (Dermietzel, 2000). Nevertheless, the first romantic relationship between Cxs and astrocyte migration was uncovered in Cx43 KO mouse fetuses, using organotypic human brain slice civilizations that demonstrated an abnormal distribution of astrocytes (Perez Velazquez, 1996). Significantly, this finding resulted in the basic proven fact that Cx43 played another role in regulating astrocytic mobility. Since then, many studies have got reported that Cxs have an effect on astrocyte migration (Homkajorn et al., 2010; Mayor and Kotini, 2015; Lagos-Cabre, 2018). The concentrate of the critique will be on the power of Cxs to create HCs in astrocytes, specifically Cx43 HCs, and exactly how they control astrocyte migration by launching small molecules towards the extracellular AZD7762 irreversible inhibition space. These substances activate receptors in the adjacent or same cells, AZD7762 irreversible inhibition which continue the signaling cascades necessary for astrocytes to go then. We may also compare the functions of HCs and GJs in cell communication and the interplay between these two cellular channels in the rules of cell migration. Astrocytes and Cell Communication Astrocytes possess a characteristic star-like shape that distinguishes them from additional non-neuronal cells of the glial family; however, despite the fact that astrocytes outnumber neurons and the additional glia (i.e., microglia and oligodendrocytes) in rodents, their AZD7762 irreversible inhibition important role has always been undermined by neurons (Sosunov, 2014; Allen and Eroglu, 2017). In the human brain, there are many different types of astrocytes that can be identified from the combination of unique cell markers, such as CD44, EAAT1, EAAT2, Aquaporin, and GFAP (Sosunov, 2014). The number of astrocytes in the human brain seems to vary according to the region, from 20C50%, and the exact percentage of total glial cells to neurons, although controversial, seems to be closer to one (von Bartheld et al., 2016). The previous conception of astrocytes as being mere assisting cells for neurons is definitely no longer valid. Today, it is known that astrocytes surround the pre- and post-synaptic membranes, therefore forming the tripartite synapse (Allen and Eroglu, 2017), and achieving practical integration and physical proximity to stimulate and regulate the activity of chemical synapses. Astrocytes also support and enhance the delivery of substrates required by neurons and take action, for example, like a highway for glucose (Muller et al., 2018). Notably, and because astrocytes function primarily by anaerobic glycolysis, they can survive in low oxygen environments AZD7762 irreversible inhibition much longer than neurons. Astroglial Cx30 and Cx43 allow the diffusion of energy metabolites such as glucose and lactate and therefore,.
Supplementary MaterialsSupplementary Information 41467_2018_4863_MOESM1_ESM
Supplementary MaterialsDataSheet1
Epithelia are polarized levels of adherent cells that will be the blocks for appendage and body organ buildings throughout pets
Supplementary Materialscells-08-00570-s001
H+-ATPase
H+, K+-ATPase
M2 Receptors
MAGL
Mannosidase
Matrix Metalloprotease
Matrix Metalloproteinase (MMP)
Matrixins
MBOAT
MC Receptors
MCH Receptors
Mcl-1
Melanin-concentrating Hormone Receptors
Melanocortin (MC) Receptors
Melastatin Receptors
Melatonin Receptors
Membrane Transport Protein
Membrane-bound O-acyltransferase (MBOAT)
Metastin Receptor
Methionine Aminopeptidase-2
mGlu Group II Receptors
mGlu Group III Receptors
mGlu Receptors
Mineralocorticoid Receptors


I'm launching my 1st Official Online Radio Show Tonight at 7:30 PM Eastern!!
You have to Tune in to RatedStarsRadio.com Tuesday, January 28, 2014 at 7:30 PM Eastern for Talent Tuesday!
We are bring you the best music and info from the hottest people on the come up.
Featuring Hotshot, #TeamPlush, and PrinceSaloEnt!
Plus Upcoming music artists and other talent, Thought Provoking Topics, Gossip and much more entertainment!
Call (347) 237-4368 to speak with the host at the time of the show! And Save the RatedStarsRadio.com Call in number in your phone!


On one hand, my nickname sounds like Art and (busy) Bee. Which makes me part of the 'Beehive Clan' together with my friends Shell Bee, Wood Bee, Can Bee and Angel B. We had a lot of fun, doing graphics projects together and organizing workshops on 3D Graphics and Photography. And I do like some honey colored accents on my site, too.
On the other hand, my nickname sounds like my initials R t B, for Ronald ter Burg, the Netherlands, 1958. For about ten years I am consultant and interim manager on Business and Project Controls. Mainly outdoor infrastructure, like roads and tracks. Before that, I wandered around in telecoms (ten years too), and before that I worked on ICT projects and teached classes on design methods and applied math (another ten years). Before that, I studied physics, and graduated as well.
Since those early days I liked the graphics part of computing most. From the green monochrome Tektronics terminals, the character graphics for the Hack game (now: NetHack), the 8-bit colored piecharts in Lotus 123, the 16-bit colored sprites for the Asteroids and Racing games, to the capabilities of 3D Studio (for DOS!) and the still exiting POV (Persistence of Vision).
High end PC's were required to run the first versions of Bryce (2 and on), Vue d'Esprit (2 and on) and Poser (1 and up). Some of them evolved to great hights, like Vue, Poser and Carrara. Bryce lagged more and more behind, while others didn't make it (MojoWorld, as in The Day After Tomorrow) or struggle for life (Terragen). From 3D Studio I migrated to 3DS MAX easily especially when the Windows NT machines kicked in, and I saw high end packs decimating their pricing schedules to keep up with serious candidates "from below the line" like Cinema4D in which I did some price-winning shorts (well, local competition that was).
Okay, so here I am, after many years of strolling around in the Computer Graphics arena, following developments and making some websites, images and shorts. But without real focus. Until now, from where I'll take the route to merge Graphics techniques and Photograpic principles.
And this is my website. Enjoy.
click the image below to view the movie.
When you've trouble viewing the Windows Media Video (WMV) then go [here] to get the browser plugin, and reopen your browser after installation.


Best Eye Doctors in Denver, CO with Verified Reviews – Book Online!
Need to make an eye doctor appointment this week? Use Zocdoc to find eye doctors in Denver who take your insurance. It's simple, secure and free.
She is very nice and professional. She took the time to explain each step in the eye exam process. I got a prescription for bifocal computer glasses when I left. The entire appointment took less than an hour.
I'm so glad I took the recommendation of a friend to come to Central Denver Eyecare. Everyone was friendly and efficient. A great experience overall.
I am very satisfied with my visit.


What is Content and why should I (or my business) be creating it?
I'm a commercial photographer, based in Kent and I specialise in creating dynamic, eye-catching images for my clients. My images are created to add value to every message conveyed across the spectrum of digital and offline platforms alike, including websites, brochures, blogs, social media channels, case studies and any other medium as required. My clients range from BAFTA award winners, multi-national building firms, television personalities and heads of state to local businesses, respected artists and ambitious entrepreneurs. I was born and raised in Little Rock, Arkansas (U.S.A.), and after a stint as a lab technician followed by time out to travel, I began a career as a production rigger, working primarily in television and film. This experience allowed me to become accustomed to pressurised working environments and a demanding clientele! In 2011, I began studying at the London Film Academy, which eventually led to me devoting my time and training to becoming a commercial photographer. Whether you have a specific project in mind or you're looking for a complete brand identity overhaul, I'd love to hear from you. You can email me at kevan@smithstudios.co.uk or give me a call on +44 77 6666 1253.
When using social media, there's no better way to communicate professionalism than by using professional images. However, many growing businesses forego the expense of professional photography and look to create their own images. As such, I've compiled a few tips for creating your own professional looking images, which will hopefully hold you over until you can afford to give me a call!
Perhaps the most important point in creating your own images is to introduce your customers to the faces of the business. This is your opportunity to bring a personable touch to your business, and you should leave customers with a feel for the way that you operate.
Your actual photos should be a feeling/concept as opposed to a thing. Make sure to leave space for branding and copy, which will help to create the emotion/feeling which will compel your customers to act. Also, make sure to include the company logo or colour scheme when possible for added impact.
Company headshots and stock images should reflect the company's brand. A consistent set of images communicates quality and it helps instil trust and brand recognition.
Amateur lighting is perhaps the most obvious tell when a company chooses to create their own images.
Lighting is also the most difficult pointer to give without going into extensive detail. The simplest way to describe it is that a bigger light source will produce a softer image with less contrast. Almost all commercial images look better with soft gradients and less contrast. A flashlight is a small and contrasty light source. A grey, cloudy day is a very large and soft light source with little contrast. A bright, sunny day at high noon counts for a small light source. Look to create your images in spaces that offer large, balanced light sources for higher quality images.
Soft, uniformed lighting is the key to capturing greater detail in your images. Appropriate lighting will allow your camera to focus properly, especially when using a mobile device. Along with appropriate lighting, a good photographic technique will minimize camera shake; another telltale sign (blurred image) of amateur image craft.
This image to the left is EXACTLY what the client requested. The client wanted a minimal, original stock image that depicted one of their typical clients. Whilst there are many things that are positive about this concept and potential application, I felt that a little creativity would add impact and flexibility to their set of images.
I suggested that we change the generic magazine to a branded booklet. I also asked the client to bring in one of their signs which I simply sat balancing on the chair to the left. Having already placed a branded cup in the frame, the end result was a highly impactful image that could easily endure as a signature image for the company.
Images and content are essential for most businesses in terms of growing the business online. Whilst it is possible to create your own high-quality images for your business, there is simply no substitute for the quality and consistency of using professional images.
If you would like to grow your business by investing in professional bespoke stock photography, email me at kevan@smithstudios.co.uk for more details.
And if you've found this article to be of interest I would be grateful if you could please "like" & "share" this article with your network.
Kevan Smith I'm a commercial photographer, based in Kent and I specialise in creating dynamic, eye-catching images for my clients. My images are created to add value to every message conveyed across the spectrum of digital and offline platforms alike, including websites, brochures, blogs, social media channels, case studies and any other medium as required. My clients range from BAFTA award winners, multi-national building firms, television personalities and heads of state to local businesses, respected artists and ambitious entrepreneurs. I was born and raised in Little Rock, Arkansas (U.S.A.), and after a stint as a lab technician followed by time out to travel, I began a career as a production rigger, working primarily in television and film. This experience allowed me to become accustomed to pressurised working environments and a demanding clientele! In 2011, I began studying at the London Film Academy, which eventually led to me devoting my time and training to becoming a commercial photographer. Whether you have a specific project in mind or you're looking for a complete brand identity overhaul, I'd love to hear from you. You can email me at kevan@smithstudios.co.uk or give me a call on +44 77 6666 1253.


is a web site dedicated to educating people about issues of poverty and human rights worldwide, and encouraging action to help those in need. The site was launched by Mr. Elia Van Tuyl in March, 2005. The show is an example of how tutorials can be created. Slide narration is used extensively. Timing slide timing for narrated slides is done automatically by the software .Click here for an example of the important work being done by the Common Wisdom Project and here for more examples of how StillMotion software is being used.


text:
Former champions recall BJO memories
Donna Helmer2019-01-04T21:40:43+00:0030th Dec 2018|Categories: 2019, Features, NEWS|Tags: Ali Farag, Camille Serme, Emily Whitlock, Mohamed ElShorbagy, Nour El Sherbini, Olivia Blatchford|
By Mark Sleightholm
As hundreds of talented juniors prepare to descend on Birmingham next week for the 93rd edition of the Dunlop British Junior Open (BJO), several past champions including Ali Farag, Nour El Sherbini and Camille Serme share their memories and pass on their pearls of wisdom to today's aspiring youngsters.
Two-time World Champion and current World No.2 Nour El Sherbini won five titles between 2007 and 2012, and playing at the BJO year-after-year provided some unforgettable experiences. She said:
"Every BJO tournament has a special memory. Winning my first ever international tournament when I was 11 is one I will never forget. I was proud to end my junior career with five BJO titles.
"It's a huge opportunity to play this tournament, gain experience, meet new friends from different countries, and most importantly it's just the start, not the end."
Sherbini's experience is echoed by that of fellow Egyptian Ali Farag who won the Under 19 title in 2011. He said:
"Every top player of every nation used to compete at the BJO, so to win you have to be the best in the world. It was by far the biggest thing you can achieve as a junior."
"My fondest memory was definitely winning the Drysdale Cup. To reach a final for the first time, beat Amr Khalifa who'd taken away my dream in the final of the World Champs six months earlier, and in straight games is by far my biggest achievement to date."
The opportunity to play against the best in the world also made the British Junior Open special for Olivia Blatchford, who won the Under 15 title in 2007.
"To know that you were playing with the best is one of the most incredible experiences there is to have as a young person. It gives you such an opportunity to grow and find yourself in a way that's so powerful. I'd go home every year a better squash player and a wealth of experiences richer," says Blatchford.
For two-time champion Camille Serme, playing in the British Junior Open was a real privilege.
"The BJO was our kind of world championships, everyone was playing it and it was really prestigious. It was an honour to compete at this event and even crazier to win it," Serme recalls.
It also meant the world to two-time champion Emily Whitlock of England who said:
"I won my first title in 2011 which was insane then my second title in 2013 – it was my last ever junior match. It meant a lot to win," recalls Whitlock.
Meanwhile, the British Junior Open offered five-time champion and current World No.1 Mohamed ElShorbagy, the opportunity to progress as a player.
"Whenever I went to these top junior events, I went to learn. The main target was never to be the best junior player in the world, it was to be the best player in the world," says Elshorbagy
Last Words :
Camille Serme's advice to today's young hopefuls is to learn from the experience.
"Enjoy your squash, take as much as experience as you can get and don't worry, you can get chopped in the first round and still win it one day," she said.
Blatchford, meanwhile advises youngsters to stand back and take stock.
"Don't be afraid to slow down, take a moment to remind yourself where you are, how blessed you are to be here and how capable you are to leave everything on the line," she explains.
Finally, Farag encourages youngsters to keep going whatever happens.
"Give it your 110%, but always keep in mind that it's not the end of the world if you don't win it. Always remember, Amr Shabana never got past the semi-final stages of the BJO, and he's probably the GOAT (after Jansher and Jahangir)," he said.
The 2012 and 2007 Presentations, featuring many of the world's top players of the following decade
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Top seeds prevail to set up final showdowns in Birmingham

redpajama_set_name: RedPajamaCommonCrawl


ERP systems can assist with several aspects of the supply chain management process and provide real-time operational information and insights across multiple departments.
Let's take a brief look at the various functions in which ERP supports the creation and management of supply chains.
An ERP system provides a flexible and easier way to establish and modify all variables required in the supply chain implementation. Planning includes product stock and inventory, production strategies, and selection of appropriate marketing channels for distribution.
Product supply and procurement methods are vital links in the supply chain. An ERP system is capable of offering cross-platform visibility on various aspects affecting procurement, including manufacturing, transportation, and warehousing resources.
Supply chain management is only competitive and cost-effective with the ability to monitor and perform changes when and where needed. Using an ERP system ensures accurate monitoring and maintenance of all supply chain activities.
Lastly, the supply chain process generates multiple activities and workflow systems. An ERP solution is needed to aggregate and analyse all operations generated through this process and measure the output against projected targets to ensure compliance and growth.


2/10/2011�� Thank you SO much for the 2215 views. Love you! These Demos are like Let's Plays, but just for The Sims 3 Pets Create a Pet Demo. However, I will be creating The Sims 3 �... I think that the horses in the Sims 3 Pets PC should have more accessories. So far all that I have heard is that they will be able to wear a saddle and bridle. So far all that I have heard is that they will be able to wear a saddle and bridle.
Mod The Sims Where can I find good horse cc?
Take, for example, this romance on horseback pose pack where your sims can take a photo with someone on a horse. For updates and news on The Sims � sims 4 how to get rid of vampire perks Horses are not included in Sims Pets, the only way to have them in your game is to download them from websites such as Mod the Sims, just search for horses and follow the inst � ructions on how to properly install it into your game.
Equestrian related CC for The Sims 3 Pets. Please do NOT re-upload any of my creations & claim them as your own, always link back to my site. Please do NOT re-upload any of my creations & claim them as your own, always link back to my site.


text:
"the light" st. luke's church bulletin for june 27, 1954 (2)
college news (1)
page eight (1)
sand-spur: officers, editorials (1)
&lt (4)
br&gt (4)
advertising -- florida -- winter park. (2)
college student newspapers and periodicals -- florida -- winter park. (1)
college student newspapers and periodicals. (1)
commencement ceremonies -- florida -- winter park. (1)
concerts -- florida -- winter park. (1)
glee club -- societies and clubs. (1)
grand army of the republic -- newspapers. (1)
illustrations -- drawings. (1)
Description: announcements
Sandspur, Vol. 23 No. 02, October 21, 1921.
"The Light" St. Luke's church bulletin for June 27, 1954
By 1954, St. Luke's was publishing its Sunday bulletins in a "bilingual format." One page was written in Slovak and referenced the 10:30am Slovak language service, as well as announcements primarily of interest to the older Slovak-speaking members....
Sandspur, Vol. 17 No. 04, May 1912
St. Cloud Tribune Vol. 12, No. 32, April 1, 1920.
Newspaper for St. Cloud (Fla.) from 1920.
Sandspur, Vol. 11 No. 01, 1904


text:
Women Discussing The Women In Game Of Thrones [VIDEO]
This Exists... Because of A Lady
By Jill PantozziMar 27th, 2012, 5:46 pm
There are a tremendous amount of strong, interesting women characters in George R.R. Martin's A Song of Ice and Fire series. And not all of them get to use sword. Apparently this has caused some strife within the fandom so a group of women got together to talk about three in particular that get a lot of flack: Sansa Stark, Catelyn Tully-Stark, and Cersei Lannister.
The YouTube videos, uploaded by "elsabette" give this description of the series:
A three part series that looks at the (most hated) women in A Song of Ice and Fire: Catelyn, Cersei, and Sansa. Each video will discuss the featured character in depth and analyze the social location of that character and the way her status as a woman has effected her social mobility, health, and overall well-being. These videos are meant to address the issue within fandom of pitting one female character against another and deeming one better due to more masculine behavior. Trigger warning for discussion of abuse.
There's lot of really interesting analysis in these. Here's the one on Sansa Stark. Spoilers through A Clash of Kings.
And her mother, Catelyn. Spoilers through A Storm of Swords.
And finally, Cersei Lannister. Spoilers through A Feast for Crows.
P.S. I love all the women in A Song of Ice and Fire. Sue me.
(via io9)
A Song of Ice and FireGame of ThronesGeorge R.R. Martinliterary Figuresliterature
Jill Pantozzi - Editor in Chief
Jill Pantozzi is a pop-culture journalist and host who writes about all things nerdy and beyond! She's Editor in Chief of the geek girl culture site The Mary Sue (Abrams Media Network), and hosts her own blog "Has Boobs, Reads Comics" (TheNerdyBird.com). She co-hosts the Crazy Sexy Geeks podcast along with superhero historian Alan Kistler, contributed to a book of essays titled "Chicks Read Comics," (Mad Norwegian Press) and had her first comic book story in the IDW anthology, "Womanthology." In 2012, she was featured on National Geographic's "Comic Store Heroes," a documentary on the lives of comic book fans and the following year she was one of many Batman fans profiled in the documentary, "Legends of the Knight."
More Stories by Jill Pantozzi


Madani won the vacant WBFed jr middle title on scores of116-111, 115-112 and 115-114. Madani was down in the third.
De Donato won the vacant Italian title on scores of 97-92 twice and 96-93. Siciliano took a standing eight count in the sixth.
Allakhverdiev won the IBO jr welter title.


text:
Q: why spring boot yaml have to be called application.yml or it can not load successfully I want to map yml data to Map object. Below is my yml data(wikiuser.yml)
spring:
    profiles: default

server:
    wikiuser:
        foo: foo@bar.com

below is Config object
@Component
@ConfigurationProperties(prefix="server",locations="wikiuser.yml")
public class WikiUserConfig {
    private HashMap<String, String> wikiuser;
    //......
}

When start my project, it throws NullPointException, because wikiuser is null.
Now changed wikiuser.yml to application.yml , everything is ok! Why have to be named application even I have specified another name(locations="wikiuser.yml") explicitly?

A: Because by default spring boot looks for a property or yaml file named application. Of course you can override this file name.
If you don't like application.properties as the configuration file name you can switch to another by specifying a spring.config.name environment property. You can also refer to an explicit location using the spring.config.location environment property (comma-separated list of directory locations, or file paths).
java -jar myproject.jar --spring.config.name=myproject

or
java -jar myproject.jar --spring.config.location=classpath:/default.properties,classpath:/override.properties

For more details, consult the spring boot documentation.
Update: Add classpath to your locations parameter value, like:
@ConfigurationProperties(prefix="server",locations="classpath:wikiuser.yml")

redpajama_set_name: RedPajamaStackExchange


I upgraded from Beta 16 to Beta 17.
These are the only two error messages I get, ES still runs stable, and when I go back to Beta 16, I have no error messages. Something must have changed in the latest beta.
I initially raised this on the .net forum and it was suggested this was a SCT issue, not an ES issue. As I stated on the .net forum, the syntax of these lines is identical to the lines above and below.
I just wanted to make the UK Sectorfile developers aware, but also ask, will the changes I've made break something else?
I've popped these in github. Though, these error messages should've still appeared with previous versions of EuroScope, since they don't relate to anything new introduced in this beta!
Your amendments make the lines as they should be.


Dr. Norin has been with the The Hearing and Speech Agency since June 2009. She holds her Doctorate in Audiology, as well as two Bachelor of Science degrees in Speech Pathology and Audiology and Mass Communications from Towson University.
Her clinical expertise includes behavioral and evoked potential diagnostics of hearing loss and hearing related disorders for both pediatric and adult patients, programming and dispensing of hearing aids, aural rehabilitation, central auditory processing disorder assessment, cochlear implant mapping, and hearing conservation. During her doctoral studies, Dr. Norin's research focused on the impact of hearing protection devices on speech communication in noise, which was presented at the 2009 national convention for the American Academy of Audiology and published in Ear & Hearing (2011).
Dr. Norin is licensed by the State of Maryland, and she is certified by the American Speech-Language-Hearing Association and the American Board of Audiology. She is a fellow of the American Academy of Audiology, the Maryland Academy of Audiology, and she is a member of the Academy of Doctors of Audiology.
Where can I buy a Soundhawk? Has the company gone out of business?
If removing earwax does not improve my hearing, what is the next step?
Yes, the microphones in the hearing aids serves as the mic for the phone, so there is no need to hold the phone to your mouth.
What's the difference between the Phonak Brio and the Brio 2?
Is it possible to upgrade my current hearing aid without purchasing a new device? I wear Phonak.
Does the Oticon streamer send stereo Bluetooth to the Alta 2 Pro miniRITE?
How do I pair the Starkey Halo 2 i2400 with the iPhone 7? I am having trouble.
Does the Linx2 61 have FM capabilities?


text:
One more thing: Can you copyright porn?
By Kate Solomon 2012-02-07T16:13:00.37Z World of tech
Plus Edward Norton's new phone and iPads for all
Today's pressing issues
We've gone all Hollywood today with our indepth look at Edward Norton's career highs and, well, mainly lows, and a really relevant look at the new trailer for The Amazing Spider-Man.
Never fear, we've managed to shoe-horn in some actual tech snippets too, including a philosophical debate on the copyrightability of porn and whether the UK's government should splash out £250,000 on iPads for all its MPs. Short answer: no, it shouldn't.
Can't copyright this – Can you copyright porn? As philosophical questions go, it's not exactly up there with does a tree falling in the woods make the sound of one hand clapping, but it's the defence being used by one woman being sued by adult movie maker Hard Drive for allegedly torrenting a blue movie named Amateur Allure Jen. [TorrentFreak]
Sleb endorsement of the day – Edward Norton's back and he's brought an LG Prada 3.0 phone with him. Apparently he and Daria Werbowy who's, like, a model or something, embody the core values of the handset - namely stylish looks and smart functionality. Yes, we've often thought that Edward Norton's ability to run apps and wirelessly connect to the internet have been a real boon to his Hollywood Career. Gratuitous photo below. [PR]
Zing!-a – what do allegedly spying on players and allegedly skewing game play for the sake of profit have in common? They're both allegedly things that Zynga allegedly partakes in, as alleged by a disgruntled ex-employee answering questions about the Farmville-maker anonymously on Reddit. [Reddit]
United Kingdom of Apple – Slow clap for the Conservatives, whose Sir Alan Haslelhurst has actually gone on record saying that all MPs should be given an iPad to save the country money. He says it's because of "all the work MPs have to do" and will also "reduce the use of paper", while he added baselessly, "There is a pretty sure case to say the supply of these will lead to an overall saving for the public purse." Sure it will. [Metro]
MIND CONTROL – File this under terrifying: neuroscientists are positing that brain-machine interfaces (BMIs) could be used to allow soldiers to control weapon systems by thought alone. Knowing how our minds wander, we're quite concerned for the future of the planet. [Guardian]
Mythical city remains mythical – a Google Ocean mapping error that got wispy myth lovers of a certain ilk all excited about the prospect of Atlantis actually existing has been corrected. As you were. [MSNBC]
Image credit: MSNBC
New news we thought was old – Amazon has overtaken HMV and is now the UK's top entertainment retailer according to Kantar's research. Huh. Who knew HMV had been hanging on in there for so long. Depressingly, Tesco, Asda and Sainsbury's come in third, fourth and fifth respectively, followed by Play.com and iTunes. [Retail Gazette]
Ambidextrous headphones – You know what we hate about headphones? Having to spend one second checking we've got the buds in the correct ears. Luckily there's needless tech for that, in the form of these proximity-sensing earphone prototypes that adjust the audio depending on which ear it detects that the earphone is in. Thank god. We're not sure how much longer we could have gone on without these. [BBC]
Forgotten but not gone – Facebook photos you thought had been deleted from the site are still accessible up to three years later. Tsk tsk Facebook. Why's that then? It's because these pictures are stored on an old system but soon all photos will be stored on a new system that will delete pics in a month and a half. But then, that's only speedy when compared to the three-year alternative. [Ars Technica]
My spidey senses are tingling – The trailer for The Amazing Spider-Man is giving us butterflies in our tummies. Okay, this isn't technically tech but there is, y'know, science in it (see 0.58) and it's being shot entirely in 3D so… Oh leave us alone and just watch it. [Total Film]
See more World of tech news
Amazon Prime Day 2019: all the best tech deals in Australia
Why I bought AirPods on Amazon Prime Day even though I don't 'need' them


text:
Music / News / RIP
Survivor Vocalist Jimi Jamison Dies At Age 63
Posted by Kristyn ClarkePosted 5 years ago September 2, 2014
(PCM) It has been revealed via the Survivor Facebook page that the lead vocalist for the 80′s band, Jimi Jamison, has passed away at the age of 63.
Jamison's immediate cause of death is not known at this time and his fellow band mates shared their sadness and memories of Jamison via social media saying "The entire Survivor family is very shocked and saddened by the passing of our brother Jimi Jamison. Our thoughts, love and prayers go out to his family and friends."
Survivor are best known for their hit song "Eye Of The Tiger" which was featured on the "Rocky III" soundtrack and have also garnered success with tracks such as "Burning Heart", "I Can't Hold Back" and "Is This Love?".
It also happens to be vocals from Jamison that you hear during the theme song for the television series "Baywatch" with "I'm Always Here".
The group officially disbanded in 1989, but have more recently recorded some new tracks for a greatest hits compilation. Jamison joined up again with the band back in 2000.
Our sincere condolences go out to Jamison's family and friends during this difficult time.
The post Survivor Vocalist Jimi Jamison Dies At Age 63 appeared first on Unskinny Pop.
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The mission of the Kentwood Arts Commission is to act as an agent of the City to review and decide upon proposals for works of art on private properties in the City of Kentwood.
The Kentwood Arts Commission was established in December 2017 to review proposals for the outside display of art on private property visible from the public right of way, within the guidelines of the City of Kentwood Zoning Ordinance.
There is currently an open position on the Arts Commission. To serve, you must be a resident and/or work in the City of Kentwood and have an interest in art. To be considered, please complete this application and email it to info@kentwood.us.


The Commission of la Malvarrosa Falla makes official the hiring of the artist Antonio Verdugo make big Falla 2014.
The work carried out by Antonio Verdugo has not gone unnoticed by the Malvarrosa Falla that has placed their trust in him and makes it debut in special section. Executioner is a young rising value even though it has not yet planted in special section, planted in 2013, in Valencia, in Bishop Amigó-basin and sculptor Pastor-Grabador Jordan Fallas.
As for the child Falla Miguel Hernández continues after the success achieved in 2013 to achieve the seventh award of special section.


Today we attended the Royal Project Fair that celebrates the King of Thailand's support for sustainable agriculture as well as the hilltribe peoples' cultures and self-sufficiency. There we found some beautiful phaa ngung gee e-ow. The best news is that it was being sold by the Karen weaver herself with assistance from 2 bilingual (Karen/Thai) young women.
We bought 2 of these pieces in red, constructed in a traditional way with 3 long, narrow pieces sewn together. Each strip was 15" wide and almost 2 yards long. Each finished piece makes a stunning textile that traditionally is wrapped around the hips or simply tailored into a tunic top. These beautiful cotton weavings can also be used as to create contemporary fashions or home décor: wall hangings, table coverings, cushion covers and other upholstery uses.
I'm not fond of going to language classes but I love talking with people at markets – especially with tribal peoples whose mother tongue is not the one we're speaking – especially when we're talking about weaving or food.
My "Word of the Day" was gee e-ow. The young woman who taught me gee e-ow apologized that she didn't know the English translation. So I taught it to her. "Backstrap," I said pointing to my lower back while saying the Thai word for that body part. Then I fell into another vocabulary void, so I mimed a strap going around my hips.
While I may not have known the word, I did know that Karen (aka Kariang) women are renowned for their skill at backstrap weaving. The Karen people are often described as a nomadic "hilltribe" people who have migrated from China. This is literally true but many Karen settled in valleys north of Chiang Mai long before it was part of the nation state called Thailand.
While backstrap looms have largely been replaced by stationary floor looms, some traditional cultures still create beautiful textiles with this deceptively simple technology in which the weaver becomes part of the loom.
The huipiles of Guatemala are perhaps the most widely recognized example of backstrap weaving. However, every year the women who excel at this art number fewer as these traditional cultural practices are lost.
Today everyone was excited with our purchases, which also included several unbleached cotton scarves with a lovely texture. We told them about our business in Canada and they invited us to visit their village weaving group to learn more and, perhaps, to make a special order. This type of exchange often marks the beginning of a longer, fair trade relationship that is based on mutual benefit, learning and respect.


Doc Savage: The Man of Bronze is coming to your living room... and he's bringing a few thousand of his friends with him. The 1975 action film based on the classic pulp hero is just one of many movies that Warner Brothers is making available via a first-of-its-kind "on-demand DVD" service that's launching today.
By visiting warnerarchive.com you can select from many films that have previously not been made available in the DVD format. You pay twenty bucks and a week later a custom-made DVD - complete with nice case - comes in the mail. I just scanned through the catalog (only a fraction thus far of the complete library that Warners aims to make eventually available) and there are some real gems in there, including Abe Lincoln in Illinois and Countdown, the 1968 pre-Apollo 11 flick starting James Caan as an astronaut caught up in a mad race between the United States and the Soviet Union about which country will be the first to land on the Moon. There's even 1982's Yes, Giorgio starring Luciano Pavarotti (wait a sec... Pavarotti made movies too?!?).
This is one site that I am going to be watching with great interest. I think this is a very cool business model that Warners is experimenting with, and hopefully it will succeed enough for other studios to consider doing likewise.
Categories: business, DVD, movies, warner bros.


The New York Times, my favorite newspaper, has won two 2012 Pulitzer Prizes, one for its reporting on Africa (INTERNATIONAL REPORTING: Jeffrey Gettleman) and another for an investigative series on obscure tax code provisions that allow wealthy corporations and citizens to avoid paying taxes, (EXPLANATORY REPORTING: David Kocieniewski).
The prizes, celebrating achievement in newspaper and online journalism, literature, nonfiction and musical composition, were announced at Columbia University in New York. Given annually since 1917, they are awarded in 21 categories.
Here are some of this year's other winners. For a complete list, click here.


text:
HBFOL NEWSLETTER
2014 HOLIDAY BOOK SALE
SATURDAY, NOVEMBER 15th
9 A.M. TO 12 NOON
1309 BARD STREET, HERMOSA BEACH
1/2 BLOCK WEST OF THE LIBRARY
START YOUR HOLIDAY SHOPPING HERE!
Come early for our selection of beautiful coffee table books: subjects range from art and photography to a very special collection of baseball history and memorabilia. The Biography table includes books on Martha Washington, Clarence Thomas, and several Elizabeths––QEs 1 & 2, plus ET & RB (Elizabeth Taylor and Richard Burton). For Secret Santa gifts, browse and buy in the Trivia section (Do Penguins Have Knees? When Do Fish Sleep?). Also stocking stuffers large and small, and many books on holiday traditions and entertainment ideas. The hardcover fiction shelves are alphabetically arranged, constantly replenished, and all the books have dust jackets. Come early for the best selection: most paperbacks and DVDs are a spectacular bargain at 50¢, most hard covers $1.
SPECIALS FOR CHILDREN, YOUNG ADULTS, ANIMAL LOVERS, AND SPORTS FANS
The front left-hand corner of the Bard Street Book Store, always appealingly arrayed and organized, provides many gift-giving ideas: picture books and cookbooks for younger children, novels for young adults, books on animals and sports, including biographies of famous athletes, and individual children's encyclopedia volumes at just 10¢ apiece.
REAL BOOKS: HERE TO STAY
In January of this year, the New York Times Book Review posed the question to authors and readers: How do e-books change the reading experience? The majority of responses weighed in on the side of books that we can hold in our hands and store on our physical shelves. There's no denying the convenience of carrying around a virtual library on an e-reader while commuting or traveling, but feeling the weight of an actual book, turning the pages, watching the progress of a bookmark as it
travels from beginning to middle to end, are physical pleasures that enhance the reading experience. No wonder the immense success of The Last Bookstore in downtown Los Angeles has transformed it into a cultural institution and visitors magnet in just a few years. The Bard Street Book Store is the last book store in Hermosa Beach. Help us to keep it, and our Hermosa Beach Library, alive and well by patronizing our sales and donating your gently used real books for the benefit of our reading
FOR MORE BOOK SALE INFORMATION
Hermosa Beach Library: 310-379-8475
Carol Lawson, Book Sale Chairman: 310-372-2610
MONDAY BOOK STORE HOURS: 9:00 - 12:00 A.M. (Closed on all Monday holidays)
Large book donations may be brought to the Bard Street address during these hours. Call Carol Lawson at 310-372-2610 to arrange for delivery at another time. Book donations and FOL membership dues can also be left at the Hermosa Beach Library during open hours.
REMAINING SATURDAY SALE DATES FOR 2014: November 15 and December 20 .
YOUR BOOK SALE & MEMBERSHIP DOLLARS AT WORK
Since the beginning of summer, the Hermosa Beach FOL has donated over $1,000 to buy books and supplies for the children's section; most recently it sponsored and provided treats for the Halloween program featuring juggler David Cousin. HBFOL members also judged submissions in the 2014 Children's Bookmark Contest and will be providing gift certificates for winners in each category. From July through October, purchases of books and DVDs for adults with HBFOL funds total nearly $3,000;
an additional $2,800 paid for periodicals and other subscriptions.
2015 ELECTION OF OFFICERS
The following slate will be voted on at the meeting on November 19:
President: Bob Luthardt
Vice-President: Dorothy Yost
Secretary: Diane Craig
Treasurer: George Ceman
Membership: Carol Lawson
THE NEXT HERMOSA BEACH FRIENDS OF THE LIBRARY MEETING WILL BE HELD ON WEDNESDAY, NOVEMBER 19TH, AT 2 P.M. MEETINGS TAKE PLACE AT THE BARD STREET BOOK STORE. ALL MEMBERS ARE WELCOME. -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
PLEASE RENEW YOUR HBFOL MEMBERSHIP FOR 2015: YOUR DUES ARE TAX-DEDUCTIBLE AND WILL HELP TO PAY FOR PRINTING & MAILING OUR NEWSLETTER. DUES RECEIVED THROUGH DECEMBER 31 WILL EXTEND YOUR MEMBERSHIP THROUGH DECEMBER OF 2015.
(Check the mailing label for your current membership status.)
Deliver this form with your check to the library, or mail to HBFOL, P.O. Box 515, Hermosa Beach CA 90254.
NAME___________________________________ ADDRESS____________________________________________________________________________
PHONE________________________________________
STUDENT/SENIOR: $5, FRIEND: $10, SPONSOR/FAMILY: $25, PATRON(FAMILY): $50, LIFETIME: $250 (1) or $500 (2)
E-mail___________________________________________________________________OK to send newsletter via e-mail?_______________


text:
<?php

namespace App\Http\Requests\Company;

use Illuminate\Foundation\Http\FormRequest;

class StoreUpdateCompanyRequest extends FormRequest
{
    /**
     * Determine if the user is authorized to make this request.
     *
     * @return bool
     */
    public function authorize()
    {
        return true;
    }

    /**
     * Get the validation rules that apply to the request.
     *
     * @return array
     */
    public function rules()
    {
        return [
            'name' => 'required|alpha_spaces',
            'street' => 'required',
            'street_number' => 'required|numeric',
            'postal_code' => 'required|digits:4',
            'country_id' => 'required|integer|exists:countries,id',
            'vat_number' => 'required',
            'user_id' => 'required|integer|exists:users,id',
        ];
    }
}

redpajama_set_name: RedPajamaGithub


text:
IX. István (), (9. század vége – 942. október) került a keresztény egyház élére a történelem folyamán 129.-ként 939. július 14-én. Amint az már megszokottá vált, István pápasága is Róma hercegének kényétől függött. II. Alberik tette meg pápának és később egy végzetes hiba miatt ő űzte el a trónról.

Élete
Rómában született és később a papi hivatást választva évek hosszas munkájával sikerült elérnie a bíborosi rangot. A Szent Szilveszter és Márton-templom kardinálisa volt. Amikor elődje, VII. Leó meghalt, a város ura Istvánt jelölte a trónra. Leó halálának másnapján, 939. július 14-én már fel is szentelték Istvánt hivatalába. Alberik nem várt tőle semmit, sőt pontosan azt akarta, hogy a pápa vonja ki magát a politikai életből. Ezért István pontifikátusa alatt igazán nem beszélhetünk pápai döntésekről, hiszen helyette Alberik uralkodott az egyházban is. István minden erejével támogatta a clunyi reformokat és a monostorizmust.
Azonban a napjait a Lateránban tengető egyházfő egy idő múlva megunta a tétlenkedést, és megkísérelt elszakadni Alberik elnyomó uralmától. A herceg távollétében kihirdette, hogy a Római Birodalom császárának a Szentszék a Karoling-dinasztia sarját, Tengerentúli Lajost ismeri el. Azt aki nem a francia uralkodót támogatja kiközösítéssel fenyegette meg. A hazatérő Alberik dühöngött István hűtlenségén, és katonáival azonnal elfogatta a pápát, majd büntetésül úgy megcsonkította István testét, hogy az sebeibe pár napon belül belehalt 942 októberében.

Művei

Jegyzetek

Források
Gergely Jenő: A pápaság története. Budapest, Kossuth Könyvkiadó, 1982.
P.G. Maxwell – Stuart: Pápák krónikája. A pápaság története pápák uralkodása szerint Szent Pétertől napjainkig. Móra Könyvkiadó, 2007.
Szántó Konrád: A katolikus egyház története. Budapest: Ecclesia. 1987.
Harald Zimmermann: A középkori pápaság. A középkori pápák története a historiográfia tükrében. Gondolat Kiadó, 2002.

Online források
Magyar Katolikus Lexikon

Kapcsolódó szócikkek
Pápa
Római pápák listája
Római pápák listája grafikusan

Pápák
10. századi püspökök
942-ben elhunyt személyek

redpajama_set_name: RedPajamaWikipedia


Natalie West thought her ability to manipulate fire was a genetic aberration straight out of a Marvel comic. Orphaned at age three, she's spent her life hiding what she can do. But like any secret superhero, she also uses her powers for good, investigating fires for the San Jose Fire Department.
When a series of arsons marked with pentagrams brings her into Andre Romero's sphere, however, Nat discovers she's not as alone as she thought. Who is this man who also wields fire? An ally? Or one of the arsonists? As Nat delves into a world of elementals, witches, and magic, learning more about her heritage in the process, she'll have to watch her back or risk getting burned.
Aileen Donovan wants nothing more than recognition as a scientist by her supernatural community. What better way to do that than to solve a mystery involving a power-hungry alchemist, hallucinogenic coral, and a homicidal sea dragon? The hard part will be working with Coast Guard officer Colin Benson–until the tides turn, and Aileen realizes that love and duty may not be mutually exclusive.
Aileen Donovan is an elemental with magical control over water. When she finds a mummified body at a scientist convention, she suspects supernatural foul play. As more bodies turn up, Aileen's amateur sleuthing not only pits her against a serial killer, but also a vampire lord, and an elemental agent with ulterior motives. Can she stop a murderer, or is she out of her supernatural league?
Nita Young doesn't know if she has a future with college sweetheart Keenan Donovan–two star-crossed lovers of opposing elements–but she invites him up to Alaska to see if Earth and Water can rekindle their old flame.
When a series of wild animal attacks strike the inhabitants of Yakutat, Nita has to put her romantic plans on hold. Mangled bodies are turning up, and a mysterious black panther has been spotted in the woods. Fur, scales, and a venomous bite suggest the cat is supernatural in origin–and evidence indicates that someone not only summoned it, but is using it to target those Nita cares about. It's the perfect murder weapon: no fingerprints, no evidence. And in a town this small, the killer is someone she knows.
Nita's strength will be put to the test as she faces losing her friends, her town, and the man she loves.
Nita Young is getting ready to marry Keenan Donovan when an unnatural earthquake crashes the wedding. As the only elementals in the area, they are sent to investigate and discover an archaeology team has unearthed an ancient and powerful artifact. After Keenan is accidentally infected with its magic, Nita must unlock the relic's secrets before its consuming powers tear her fiancé apart.
Air elemental Sylphie Seaton has the perfect job–chasing twisters. Every year she and her team of elemental scientists hit the road during tornado season to track the elusive weather phenomena in order to collect new data.
Things are about to get complicated when Eric Gage, an extreme photojournalist, comes along for the ride. He's human, and completely ignorant about the supernatural world. Sylphie can't decide whether to find the man charming or infuriating, a tension that's tested when they go up against one of Nature's most volatile forces.
Sylphie Seaton's life is about to get turned upside-down when her brother ends up wanted for murder. Framed and dying from exposure to a genetically engineered super serum, Josh turns to Sylphie for help clearing his name and finding a cure. But the conspiracy may go deeper than either of them ever imagined, and not even an acclaimed storm chaser can outrun this tempest.


Do you ever have one of those weeks when everything seems to go wrong? When it's not just a question of your train being delayed (because they couldn't find a driver – thank you so much First Great Western's so-called 15.00 Plymouth to Taunton service) or a document disappearing from your file server (everyone agrees that it shouldn't be possible, but it was) – but also of people wilfully misinterpreting your comments and choosing to take offence … offence which they then spell out in emails running to several sides of A4 paper, in carefully numbered points (1-11, go on, count them), and copied to just about every senior manager in the South West.
Despite all that, I still managed to finish off the cuff (again, casting off following the knit 2/purl 2 ribbing sequence), which looks quite nice when it's folded back; and, after a few days staring at the pullover like a cat sizing up an indolent emu – and knocking back a beer for Dutch courage – I gritted my teeth and picked up the stitches all round the other sleeve.
What a grim task it is! I approach it with all the jollity of a surgeon forced to amputate his own arm. And yet, the funny thing is, it's never as bad as I think it's going to be (is that true? Well, possibly): this time it took me just over 45 minutes, which is pretty good going really.
Well, I daresay it's close enough for jazz, as the saying is.
No time to pursue the exciting question of identifying drowned sailors this week – that'll have to wait till next week. See you then!


The Very Special Person award is given annually to a member who has given notable service to the Guild. The intention of the VSP Award is to honor the members of the Guild who do things for the Guild, often quietly and behind the scenes, not recognized by thank you blocks or other forms of acknowledgement. The following Very Special People are one of the reasons the Guild is so special.


text:
Proctor struggles to explain why he also took two cans of beans, a beer, 60 dollars, and a bath
Despite his name and appearance, Dave Scott insists he is just "a human" and not "some kind of proctorbot."
Otago University proctor Dave Scott has landed himself in hot water this week after entering a student flat without permission and stealing three bongs.
Scott, who has publicly apologised today and acknowledged he made a mistake, says he was just trying to protect the students from possible police action, and was doing it out of concern for them.
But at a 3pm press conference this afternoon, he failed to coherently explain to media why he also took two cans of beans, a couch cushion, a Playstation 4, a Rush Hour DVD set, a beer, 60 dollars, and had a bath before leaving.
"Look, everything I did, I did with the students' well-being in mind," he said. "Did I cross a line at times? Absolutely, but for one, there were too many beans in that flat. You don't need sixteen cans, I know you buy them all because they keep and you think at some point you're going to use them, but I think most students understand that those cans just get moved from flat to flat to flat and then eventually into a family home until you finally get the courage to put them straight in the bin, and there are families that could use beans that you're not using; my family, for example.
"The cushion? Uh, look, didn't fit the couch, wasn't even from the same couch that it was on. Just trying to find that a home.
"The Playstation 4, study distraction.
"Rush Hour? Kind of a racist franchise."
Scott could not explain why he took the beer and 60 dollars, but said that taking a bath was "not a criminal act."
Legal experts spoken to about the matter agreed that taking a bath is not a crime.
Students living in the flat were sympathetic to Scott, telling media today that he's a "good guy" and they didn't want to see him fired, even though it was "really weird that he had a bath here."


text:
Download our Johannesburg Schools Guide
Purchase the complete Johannesburg City Guide
Purchase the complete Expat Arrivals Johannesburg Guide (PDF)
A ddress: Johannesburg Campus: Bryanston, 2021, Johannesburg
Pretoria Campus: Midrand, 1682, Pretoria
Website: www.aisj-jhb.com
Email: info@aisj-jhb.com
Gender: Co-educational
Curriculum: American and International Baccalaureate
Ages: 3 to 18
Founded in 1982, today the American School has over 1,000 students on a sprawling campus in Bryanston which caters for students from Pre-Kindergarten to Grade 12. A second campus in Pretoria caters for Pre-Kindergarten to Grade 8. The school's facilities include science laboratories, computer laboratories, libraries, a gym, a swimming pool, sport-specific fields and courts, a fine arts centre and a cafeteria. There is a wide range of extra-curricular activities on offer.
"At AISJ, we are committed to providing strong academic support for our high school students. Our ongoing professional development assists teachers to differentiate their instruction and enables them to work effectively with a range of student learning styles and needs. Our students come from a variety of backgrounds, with over 100 nationalities represented in our student body. We are truly an international school."
In elementary, middle and high school, students follow a hybrid curriculum based on American and international best practices. Grade 11 and 12 students study toward either the American High School Diploma or the International Baccalaureate Diploma, which is internationally recognised.
The Johannesburg campus of the school is located in Bryanston, near Sandton. There is a second, smaller campus in the nearby city of Pretoria.
For the 2020/2021 academic year, annual fees range from 333,396 ZAR for Pre-Kindergarten to 598,464 ZAR for Grades 11 and 12. There is also a non-refundable capital fee of 11,025 USD for new students.
From the school director
"I am very fortunate to inherit the leadership of a school founded over 30 years ago in the best traditions of American education but a school that is also truly international in practice and demographics. I am also fortunate to lead a highly committed staff of local and international teachers, administrators, and support staff who make the school such a warm and welcoming environment for students and parents. The teachers – many recruited from around the world – add to the diverse richness of what really makes the school and South Africa such an exciting place."
– Dr Andy Page-Smith
School Director
► International Schools in Johannesburg
► Private Schools in Johannesburg
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Interview with Heloise – a Dutch expat living in Johannesburg
Interview with Srinath - expat advice on life in South Africa
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Offbeat and arty things to do in Johannesburg
Interview with Heather - an American expat living in Johannesburg
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