text:
Journal of Biomedicine and Biotechnology
Volume 2012, Article ID 782642, 10 pages
Camel Milk Modulates the Expression of Aryl Hydrocarbon Receptor-Regulated Genes, Cyp1a1, Nqo1, and Gsta1, in Murine hepatoma Hepa 1c1c7 Cells
1Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, 11451 Riyadh, Saudi Arabia
2Faculty of Pharmacy & Pharmaceutical Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E1
3Department of Pharmacology, College of Medicine, King Saud University, 11461 Riyadh, Saudi Arabia
Received 6 September 2011; Revised 24 October 2011; Accepted 8 November 2011
Academic Editor: Ikhlas A. Khan
Copyright © 2012 Hesham M. Korashy et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
There is a traditional belief in the Middle East that camel milk may aid in prevention and treatment of numerous cases of cancer yet, the exact mechanism was not investigated. Therefore, we examined the ability of camel milk to modulate the expression of a well-known cancer-activating gene, Cytochrome P450 1a1 (Cyp1a1), and cancer-protective genes, NAD(P)H:quinone oxidoreductase 1 (Nqo1) and glutathione S-transferase a1 (Gsta1), in murine hepatoma Hepa 1c1c7 cell line. Our results showed that camel milk significantly inhibited the induction of Cyp1a1 gene expression by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the most potent Cyp1a1 inducer and known carcinogenic chemical, at mRNA, protein, and activity levels in a concentration-dependent manner. In addition, camel milk significantly decreased the xenobiotic responsive element (XRE)-dependent luciferase activity, suggesting a transcriptional mechanism is involved. Furthermore, this inhibitory effect of camel milk was associated with a proportional increase in heme oxygenase 1. On the other hand, camel milk significantly induced Nqo1 and Gsta1 mRNA expression level in a concentration-dependent fashion. The RNA synthesis inhibitor, actinomycin D, completely blocked the induction of Nqo1 mRNA by camel milk suggesting the requirement of de novo RNA synthesis through a transcriptional mechanism. In conclusion, camel milk modulates the expression of Cyp1a1, Nqo1, and Gsta1 at the transcriptional and posttranscriptional levels.
1. Introduction
The aryl hydrocarbon receptor (AhR), a cytosolic ligand-activated transcriptional factor, belongs to the basic-helix-loop-helix (bHLH)/Per-ARNT-Sim (PAS) family of transcription proteins, which are involved in regulation of cell differentiation and proliferation [1, 2]. Mechanistically, AhR is located in the cytoplasm bound with heat shock protein-90 (HSP90) and AhR interacting protein (AIP) forming inactive complex. Activation of AhR upon binding with its ligands, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a polycyclic aromatic hydrocarbon (PAH), causes dissociation of HSP90 and AIP from the activated ligand-receptor complex, subsequently leading to translocation of the complex into nucleus. In the nucleus, the ligand-receptor complex dimerizes with AhR nuclear translocator (ARNT), which subsequently binds to xenobiotic-responsive element (XRE) located in the promoter region of so-called AhR-regulated genes resulting in promoting its transcription and protein translation processes [3, 4].
The AhR-regulated gene batteries include phase I xenobiotic metabolizing enzymes such as the cytochrome P450 1A1 (CYP1A1), CYP1A2, CYP1B1 and phase II enzymes such as NAD(P)H:quinone oxidoreductase 1 (NQO1), glutathione S-transferase A1 (GSTA1), uridine diphosphate glucuronosyltransferase 1A6, and aldehyde dehydrogenase-3 [3, 4]. Among these genes, CYP1A1 plays a particular role in bioactivating procarcinogens into carcinogen and toxic metabolites and hence is considered as cancer-activating gene [5], whereas the NQO1 and GSTA1 catalyze reduction of several environmental contaminants and endogenous compounds that maintain endogenous antioxidants, such as ubiquinone and vitamin E, to protect tissues against mutagens, carcinogens, and oxidative stress damage [6, 7]. Accordingly, one of the strategies for protecting human cells and tissues from the toxic effects of carcinogenic and cytotoxic metabolites generally include attenuation of the carcinogen-activating genes, CYP1A1 signaling pathways, and/or enhancing the adaptive mechanisms by increasing the expression of detoxification and antioxidant genes, such as NQO1 and GSTA1 [8].
Chemoprevention by dietary constituents in the form of functional food has a well-established beneficial role in health promotion and emerged as a novel approach to control cancers [9]. Camel's milk is different from other ruminant milk, having low cholesterol, lactoferrin, low sugar, high minerals (sodium, potassium, iron, copper, zinc and magnesium), high vitamin C, B2, A, and E, low protein, and high concentrations of insulin. Recent studies have reported that camel milk is the most active milk among other species against E. coli, Staphylococcus aureus, Salmonella typhimurium, and rotavirus [10, 11]. In addition, it has been demonstrated that milk, in addition to secretory IgA and IgM, also contains numerous nonantibody components that possess antiviral activity, including lactoferrin [10].
Until recently, it is traditionally claimed that drinking camel milk may help to fight against serious diseases and cure numerous cases of cancer; however, this claim has never been exposed to scientific scrutiny investigation. Therefore, the main objective of the current study was to explore the capacity of camel milk to modulate the expression of Cyp1a1, Nqo1, and Gsta1 genes as target for cancer prevention in murine hepatoma Hepa 1c1c7 cells.
2. Materials and Methods
2.1. Materials
7-Ethoxyresorufin (7ER), Dulbecco's Modified Eagle's Medium (DMEM), protease inhibitor cocktail, sulforaphane (SNF), resveratrol (RES), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and rabbit anti-goat IgG peroxidase secondary antibody were purchased from Sigma-Aldrich (St. Louis, MO). 2,3,7,8-Tetrachlorodibenzo-p-dioxin, >99% pure, was purchased from Cambridge Isotope Laboratories (Woburn, MA). Resorufin and 100 × vitamin supplements were purchased from ICN Biomedicals Canada (Montreal, QC). TRIzol and Lipofectamine 2000 reagents were purchased from Invitrogen (Carlsbad, CA). The High-Capacity cDNA reverse transcription kit and SYBR Green PCR Master Mix were purchased from Applied Biosystems (Foster City, CA). Actinomycin D (Act-D) was purchased from Calbiochem (San Diego, CA). Dual-Luciferase Reporter Assay System was obtained from Promega Corporation (Madison, WI). Chemiluminescence Western blotting detection reagents were obtained from GE Healthcare Life Sciences (Piscataway, NJ). Nitrocellulose membrane was purchased from Bio-Rad Laboratories (Hercules, CA). Cyp1a1 goat anti-mouse polyclonal primary antibody, glyceraldehyde-3-phosphate dehydrogenase (Gapdh) rabbit anti-goat polyclonal antibody, and anti-rabbit IgG peroxidase secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). All other chemicals were purchased from Fisher Scientific Co. (Toronto, ON).
2.2. Milk Sample Collection and Preparation
Camel milk was collected aseptically from five healthy domestic camels (Camelus dromedaries). The camel milk was collected from farm and desert living animals. The collection of milk was usually conducted during the feeding time and was performed by experienced attendants. Milk was allowed to flow directly into sterile stainless steel containers and then transferred to glass vials. Camel samples were transported to the laboratory as soon as practical (<4 h) and were frozen at −80°C. Aqueous portion (fat-free, skimmed milk) was removed from the lipids (cream) as described before [12]. Briefly, aliquots of pooled milk samples were centrifuged at 1400 ×g for 30 minutes at 4°C, thereafter, the creamy layer consisting largely of fat was removed by filtration through a glass wool plug in a Pasteur pipette. Camel milk was collected and kept in −80°C freezer until use.
2.3. Cell Culture and Treatments
Murine hepatoma Hepa 1c1c7 cells (American Type Culture Collection, Manassas, VA) were maintained in DMEM, without phenol red supplemented with 10% heat-inactivated fetal bovine serum, 20 M L-glutamine, 100 IU/mL penicillin, 10 g/mL streptomycin, 0.1 mM nonessential amino acids, and vitamin supplement solution. Cells were grown in 75 cm2 tissue culture flasks at 37°C under a 5% CO2 humidified environment.
Hepa 1c1c7 cells were plated onto 96- and six-well cell culture plates in DMEM culture media for the mRNA, protein, and catalytic activity assays. In all experiments, the cells were pretreated for indicated time interval in serum-free media with various concentrations of camel milk in the presence or absence of TCDD as indicated. Stock solutions of TCDD were prepared in dimethyl sulfoxide (DMSO) and stored at −20°C, in which the concentration of DMSO did not exceed 0.05% (v/v).
2.4. Cytotoxicity of Camel Milk
The effects of camel milk (fat-free) on Hepa 1c1c7 cell viability were determined by measuring the capacity of reducing enzymes present in viable cells to convert MTT salt to formazan crystals as described previously [13]. Twenty-four hours after incubating the cells with the tested milk in the presence and absence of TCDD in a 96-well cell culture plate at 37°C under a 4% CO2 humidified incubator, the media were removed and a 100 L of serum-free medium containing 1.2 mM of MTT dissolved in phosphate-buffered-saline (PBS), pH 7.4, was added to each well. The plate was then incubated in a CO2 incubator at 37°C for 2 h. The media were then decanted off by inverting the plate, and a 100 L of isopropyl alcohol was added to each well, with shaking for 1 h to dissolve the formazan crystals. The color intensity in each well was measured at wavelength of 550 using BIO-TEK Instruments EL 312e microplate reader, Bio-Tek Instruments (Winooski, VT). The percentage of cell viability was calculated relative to control wells designated as 100% viable cells.
2.5. Determination of Cyp1a1 Enzymatic Activity
Cyp1a1-dependent 7-ethoxyresorufin (7ER) O-deethylase (EROD) activity was performed on intact living Hepa 1c1c7 cells using 7ER as a substrate [14]. After incubation of the cells with increasing concentrations of fat-free camel milk for 24 h, media were aspirated and the cell monolayers were rinsed with PBS. Thereafter, 100 L of 2 M 7ER in assay buffer (0.05 M Tris, 0.1 M NaCl, pH 7.8) was then added to each well. Immediately, an initial fluorescence measurement () at excitation/emission (545 nm/575 nm) was recorded from each well using Baxter 96-well fluorometer (Deerfield, IL). The plates were then replaced in the incubator, and additional set of fluorescence measurements of the wells were recorded every 5 min for 20 min interval. The amount of resorufin formed in each well was determined by comparison with a standard curve of known concentrations. The working solution was then aspirated, the cells were rinsed twice with PBS, and 50 l of double de-ionized water was added to lyse the cells. After placing of the cell plates at −80°C for 30 min, the cell lysates were allowed to thaw, and protein levels were determined using a modified fluorescent assay [15]. The rate of resorufin formation was expressed as pmol/min/mg protein.
2.6. RNA Extraction and cDNA Synthesis
Total RNA was isolated using TRIzol reagent (Invitrogen) according to the manufacturer's instructions and quantified by measuring the absorbance at 260 nm. RNA quality was determined by measuring the 260/280 ratio. Thereafter, first strand cDNA was synthesized using the High-Capacity cDNA reverse transcription kit (Applied Biosystems) according to the manufacturer's instructions. Briefly, 1 g of total RNA from each sample was added to a mix of 2.0 L of 10x reverse transcriptase buffer, 0.8 L of 25x dNTP mix (100 mM), 2.0 L of 10x reverse transcriptase random primers, 1.0 L of MultiScribe reverse transcriptase, and 3.2 L of nuclease-free water. The final reaction mix was kept at 25°C for 10 min, heated to 37°C for 120 min, heated for 85°C for 5 s, and finally cooled to 4°C [16].
2.7. Quantification of mRNA Expression by Real-Time Polymerase Chain Reaction (RT-PCR)
Quantitative analysis of specific mRNA expression was performed by RT-PCR by subjecting the resulting cDNA to PCR amplification using 96-well optical reaction plates in the ABI Prism 7500 System (Applied Biosystems). The 25 L reaction mix contained 0.1 L of 10 M forward primer and 0.1 L of 10 M reverse primer (40 nM final concentration of each primer), 12.5 L of SYBR Green Universal Master Mix, 11.15 L of nuclease-free water, and 1.25 L of cDNA sample. The primers used in the current study (Table 1) [17, 18] were purchased from Integrated DNA Technologies (IDT, Coralville, IA). Assay controls were incorporated onto the same plate, namely, no-template controls to test for the contamination of any assay reagents. The RT-PCR data was analyzed using the relative gene expression (i.e., ) method, as described in Applied Biosystems User Bulletin [19]. The data are presented as the fold change in gene expression normalized to the endogenous housekeeping gene (-actin) and was determined using the equation: fold change = , where and .
2.8. Protein Extraction and Western Blot Analysis
Twenty-four hours after incubating the cells with increasing concentrations of camel milk (fat-free), the cells were washed once with cold PBS and collected by scraping in 100 L of lysis buffer (50 mM HEPES, 0.5 M NaCl, 1.5 mM MgCl2, 1 mM EDTA, 10% (v/v) glycerol, 1% Triton X-100, and 5 L/mL of protease inhibitor cocktail). The lysates were incubated on ice for 1 h with intermittent vortexing every 10 min, followed by centrifugation at 12,000 ×g for 10 min at 4°C. The supernatant was then stored at a −80°C freezer for later use in the Western blot analysis.
Western blot analysis was performed as described previously [20]. For Cyp1a1 immuno-detection, 30 g of proteins from each treatment group were diluted with same amount (1 : 1) of 2X loading buffer (0.1 M Tris-HCl, pH 6.8, 4% SDS, 1.5% bromophenol blue, 20% glycerol, 5% β-mercaptoethanol), boiled and loaded onto a 10% SDS-polyacrylamide gel. Samples were electrophoresed at 120 V for 2 h, and the separated proteins were transferred to Trans-Blot nitrocellulose membrane (0.45 μm) in a buffer containing 25 mM Tri-HCl, 192 mM glycine, and 20% (v/v) methanol. Protein blots were blocked overnight at 4°C in a solution containing 5% skim milk powder, 2% bovine serum albumin, and 0.5% Tween-20 in TBS solution (0.15 M NaCl, 3 mM KCl, 25 mM Tris-base). Thereafter, the blocking solution was removed and the blots were rinsed three times in a wash buffer (0.1% Tween-20 in TBS). Proteins were detected by incubation with a primary polyclonal goat anti-mouse Cyp1a1 antibody for 2 h at 4°C in TBS containing 0.01% sodium azide and 0.05% Tween-20. The primary antibody solution was removed and blots were rinsed three times with a wash buffer, followed by incubation with horseradish peroxidase-conjugate rabbit anti-goat secondary antibody for 1 h at room temperature followed by washing as previously described. Antibody detection was performed using the enhanced chemiluminescence method. The intensity of Cyp1a1 bands was quantified, relative to the signals obtained for Gapdh, using Java-based image-processing software, ImageJ (W. Rasband (2005) National Institutes of Health, Bethesda, MD, http://rsb.info.nih.gov/ij/).
2.9. Transient Transfection and Luciferase Assay
Transient transfection and luciferase assay were carried out as described previously [21]. Briefly, Hepa 1c1c7 cells (3 × 104 cells per well) were plated onto 12-well cell culture plates. Each well was cotransfected with 1.5 g of XRE-driven luciferase reporter plasmid pGudLuc 1.1 and 0.1 g of the renilla luciferase pRL-CMV vector, used for normalization. The pGudLuc 1.1 plasmid was provided as a gift from Dr. Michael S. Denison (University of California, Davis, CA), while pRL-CMV vector was obtained from Promega Corporation (Madison,WI). Transfection procedure was carried out using Lipofectamine 2000 reagent according to the manufacturer's instructions (Invitrogen), the luciferase assay was performed according to the manufacturer's instructions (Promega), and luciferase activity was reported as relative light unit of firefly luciferase to renilla luciferase (Fluc/Rluc).
2.10. Statistical Analysis
All results are presented as mean ± SEM. The comparative analysis of the results from various experimental groups with their corresponding controls was performed using SigmaStat for Windows, Systat Software Inc., (San Jose, CA). One-way analysis of variance (ANOVA) followed by Student-Newman-Keul's test was carried out to assess the significant difference between different groups. The differences were considered significant when .
3. Results
3.1. Effect of Camel Milk on Hepa 1c1c7 Cell Viability
To determine the cellular toxicity effects of camel milk (fat-free) in the presence and absence of TCDD on Hepa 1c1c7, cells were treated for 24 h with increasing concentrations of camel milk (0, 15, 25, 50, 100, and 200 L/mL) in the presence and absence of TCDD (1 nM) and the cell viability was determined by MTT assay. Figure 1 shows that neither camel milk alone nor with TCDD were toxic to Hepa 1c1c7 cells at all concentrations of camel milk used, with the exception of mixture of TCDD plus camel milk (200 L/mL) that significantly decreased cell viability by approximately 25%. Based on these results, concentrations of 0, 25, and 100 L/mL of fat-free camel milk in the presence and absence of TCDD were chosen to be used in the subsequent experiments.
3.2. Effect of Camel Milk on the TCDD-Mediated Induction of Cyp1a1 Catalytic Activity in Hepa 1c1c7 Cells
To determine the capacity of camel milk to alter the induction of Cyp1a1 catalytic activity by TCDD, Hepa 1c1c7 cells were preincubated with increasing concentrations of camel milk (fat-free) (0, 25, and 100 L/mL) or RES (25 M), a positive control, for 30 min before the incubation with 1 nM TCDD for additional 24 h. The TCDD 1 nM concentration was selected from our previous concentration-response study that caused maximum induction of Cyp1a1 gene without significant cell toxicity [22]. Thereafter, Cyp1a1 activity was determined by EROD assay. Figure 2 shows that TCDD alone markedly induced Cyp1a1 enzymatic activity level by 180-fold. Furthermore, camel milk at all concentrations tested significantly inhibited the TCDD-mediated induction of Cyp1a1 activity by approximately 63% and 80% at the concentrations of 25 and 100 L/mL, respectively, (Figure 2). On the other hand, RES, a well-known AhR antagonist, significantly reduced the Cyp1a1 induction by TCDD (Figure 2). The observed inhibition of Cyp1a1 by camel milk (fat-free) particularly at the highest concretion is due to a direct effect of camel milk and not because of any cell toxicity (Figure 1).
3.3. Effect of Camel Milk on the TCDD-Mediated Induction of Cyp1a1 mRNA Level in Hepa 1c1c7 Cells
To determine whether the inhibitory effect of camel milk (fat-free) on the TCDD-mediated induction of Cyp1a1 activity (Figure 2) is attributed to a transcriptional mechanism, Cyp1a1 mRNA levels were determined in Hepa 1c1c7 cells treated for 6 h with TCDD (1 nM) in the presence and absence of different concentrations of camel milk (0, 25, and 100 L/mL) or RES (25 M) as positive control. Our results showed that TCDD significantly induced Cyp1a1 mRNA level by approximately 15-fold (Figure 3). Importantly, incubation of Hepa 1c1c7 cells with camel milk (fat-free) significantly decreased the TCDD-mediated induction of Cyp1a1 mRNA in a concentration-dependent manner (Figure 3). The maximum inhibition (90%) was observed at the highest concentration tested (100 L/mL) (Figure 3). Similar to Cyp1a1 activity, the positive control RES significantly decreased the TCDD-mediated induction of Cyp1a1 mRNA (Figure 3).
3.4. Effect of Camel Milk on the TCDD-Mediated Induction of Cyp1a1 Protein in Hepa 1c1c7 Cells
To further investigate whether the obtained inhibition on TCDD-mediated induction of Cyp1a1 mRNA levels by camel milk is translated into a functional Cyp1a1 protein, Western blot analysis was carried out. Total protein was isolated from Hepa 1c1c7 cells treated for 24 h with TCDD (1 nM) in the presence and absence of different concentrations of camel milk (fat-free) (0, 25, and 100 L/mL). Figure 4 shows that the induction of Cyp1a1 protein by TCDD (72-fold) was significantly reduced by camel milk (fat-free) by approximately 30% and 65% at concentrations of 25 and 100 L/mL, respectively. This pattern of inhibition is similar to what was observed at the activity and mRNA level (Figures 2 and 3).
Taken together, these results showed that camel milk stands prominently in its inhibition of the induction of Cyp1a1 gene expression at the activity, mRNA, and protein levels.
3.5. Effect of Camel Milk on XRE-Dependent Luciferase Activity
To explore the ability of fat-free camel milk to inhibit the AhR-dependent reporter gene expression, Hepa 1c1c7 cells were transiently cotransfected with the XRE-dependent luciferase reporter gene and renilla luciferase vector, which was used for normalization of transfection efficiency. Cells were then incubated for 24 h with TCDD (I nM) in the presence and absence of camel milk (100 L/mL). Our results showed that TCDD significantly induced XRE-dependent luciferase activity by approximately 7-fold (Figure 5). Importantly, camel milk (fat-free, 100 L/mL) completely blocked the induction of XRE-dependent luciferase activity by TCDD to its control level (Figure 5).
3.6. Effect of Camel Milk on Ho-1 mRNA in Hepa 1c1c7 Cells
The capacity of camel milk to inhibit TCDD-mediated induction of Cyp1a1 at the activity level more than at the protein levels prompted us to investigate the possible role of Ho-1, a rate-limiting enzyme of heme degradation, in this inhibitory effect. For this purpose, Ho-1 mRNA level was determined in Hepa 1c1c7 cells treated with increasing concentrations of camel milk (0, 25, and 100 L/mL) or SFN (5 M), as a positive control, for 6 h. Our results show that camel milk (fat-free) significantly increased the Ho-1 mRNA expression level by the highest concentration tested (100 L/mL) by approximately 3-fold (Figure 6). Similarly, SFN caused a 9-fold induction in Ho-1 mRNA level. These data may suggest a role for Ho-1 in camel milk-mediated effects.
3.7. Effect of Camel Milk on Nqo1 and Gsta1 mRNA Levels in Hepa 1c1c7 Cells
To further investigate whether camel milk (fat-free) is able to increase the expression of chemoprotective genes, Nqo1 and Gsta1 mRNA levels were measured in Hepa 1c1c7 cells treated for 6 h with camel milk (0, 25, and 100 L/mL) or SFN (5 M), as a positive control. Figure 7 shows that camel milk (fat-free) significantly induced Nqo1 and Gsta1 mRNA levels by approximately 2.5- and 8-fold, respectively, at the highest concentration tested (100 L/mL). A similar induction was observed with SFN, strong inducer of Nqo1 and Gsta1 (Figure 7).
3.8. Effect of Transcription Inhibitor, Act-D, on Camel-Milk-Induced Nqo1 mRNA Levels
To further investigate whether camel milk (fat-free) is able to increase the de novo Nqo1 RNA synthesis, Hepa 1c1c7 cells were treated with Act-D (5 g/mL, RNA synthesis inhibitor) immediately before the addition of camel milk (fat-free, 100 L/mL) for additional 6 h. Thereafter, Nqo1 mRNA was quantified by real-time PCR. If camel milk increases the amount of Nqo1 mRNA through a transcriptional mechanism, a decrease in the Nqo1 mRNA level would be expected. Figure 8 shows that Act-D slightly but significantly inhibited the constitutive expression of Nqo1 mRNA, whereas markedly blocked the camel milk-induced Nqo1 mRNA by approximately 70% (Figure 8).
4. Discussion
The current study provides the first mechanistic evidence of the ability of camel milk to significantly inhibit the induction of Cyp1a1, a cancer-activating gene, and to induce Nqo1 and Gsta1, cancer protecting genes in Hepa 1c1c7 cells at the transcriptional and posttranscriptional levels.
One of the strategies for protecting human cells and tissues from the toxic effects of carcinogenic and cytotoxic metabolites include attenuation of the carcinogen-activating genes signaling pathways and/or enhancing the adaptive mechanisms by increasing the expression of detoxification and antioxidant genes. Accordingly, we hypothesize that camel milk exhibits anticancer effects by inhibiting the expression of Cyp1a1 and/or inducing Nqo1 and Gsta1 genes. Therefore, to test our hypothesis we examined the capacity of camel milk to (a) inhibit the induction of Cyp1a1 at the mRNA, protein, and activity levels, (b) induce the expression of Nqo1, and Gsta1 mRNA levels in Hepa 1c1c7 cells, and (c) explore the underlying molecular mechanism.
Several lines of evidence showed that induction of CYP1A1 is strongly correlated with increased incidence of several human colon, rectal, and lung cancers [23, 24]. In addition, studies on the carcinogenicity and mutagenicity of PAHs have demonstrated a significant role for the induction of CYP1A1 in bioactivating these environmental toxicants into their ultimate carcinogenic forms [25]. Thus Cyp1a1 induction is considered a useful biomarker of exposure to carcinogenic substances [26]. Accordingly, we have first examined the potential effect of camel milk on the expression of Cyp1a1 gene after induction by TCDD using EROD as a probe for Cyp1a1 activity [27]. Our results showed that camel milk (fat-free) at all concentrations tested reduced Cyp1a1 EROD activity in a concentration-dependent manner (Figure 2). Importantly, the modulation of Cyp1a1 activity by camel milk is attributed to a decrease in transcriptional and translational regulation of Cyp1a1 gene. This was evidenced by the ability of camel milk to alter the expression of Cyp1a1 at the mRNA and protein levels (Figures 3 and 4).
The transcriptional regulation of Cyp1a1 and Nqo1 genes by camel milk (fat-free) was demonstrated by several lines of evidence. First, the inhibition of XRE-dependent luciferase assay that occurs only through the AhR activation suggests an AhR-dependent transcriptional control (Figure 5). Second, the ability of the transcription inhibitor, Act-D, to significantly block the newly synthesized Nqo1 mRNA (Figure 7) suggests a requirement of de novo RNA synthesis for the induction of Nqo1 mRNA by camel milk. Taken together, these results strongly suggest that the inhibition of Cyp1a1 and induction of Nqo1 genes by camel milk is mediated, at least in part, at the transcriptional level.
Interestingly, the observation that camel milk-mediated inhibition of Cyp1a1 induction by TCDD at the activity levels is higher than those observed at the protein levels (Figure 2), suggest that a possible involvement of a posttranslational mechanism, such as phosphorylation, proteasomal degradations, modulation of HO-1 gene expression, could be involved [22, 28]. Among these mechanisms, HO-1 gene expression, a rate-limiting enzyme in heme catabolism, has been shown to alter cellular heme, the prosthetic group of CYP450, content and hence the enzyme activity [29]. To test this hypothesis, we have examined the effect of camel milk (fat-free) on the expression of Ho-1 mRNA level. Our results showed that the camel milk increases the level of Ho-1 mRNA levels (Figure 6). Taken together, we postulated here that the capacity of camel milk to induce the expression of Ho-1 mRNA resulted in a decrease in Cyp1a1 activity levels through degrading its heme content. This postulation is supported by our previous observations that in mesoporphyrin, competitive Ho-1 inhibitor or knockdown of Ho-1 using siRNA significantly restored the inhibition of Cyp1a1 activity by heavy metals [30, 31].
The chemo protective effect of camel milk was further supported, in addition to inhibition of Cyp1a1, by the ability to upregulate antioxidant genes, particularly Nqo1 and Gsta1 mRNA levels in a concentration-dependent manner (Figure 7). Our results are in agreement with previous studies that reported the ability of camel milk to induce GST levels in healthy and schistosoma-infected mice [32]. Thus increased expression of Nqo1 and Gsta1 by camel milk will increase the levels of several antioxidant enzymes which prevent the formation of highly reactive oxygen radicals and hence reduce DNA adduct and cell damage [33]. In addition, overexpression of NQO1 in several human solid tumors and cancer cells has been shown to activate bioreductive chemotherapeutic agents in tumor cells that allow tumor cytotoxicity without corresponding toxicity to normal cells [34].
Although the potential mediators in camel milk involved in the downregulation of Cyp1a1 and induction of Nqo1 and Gsta1 were not examined in this study, several previous studies have reported that camel milk contains considerably higher amounts of antioxidant vitamins, such as E and C, in comparison to cow milk [35], lysosomes [11], lactoferrins [11, 36], and immunoglobulines [36]. In addition, ongoing research in our laboratory has shown the presence of several compounds in camel milk that could be involved in milk-mediated effect (unpublished data). Among these mediators, lactoferrin, an iron-binding glycoprotein, is known to exert in vitro and in vivo antitumor activity [37]. In this context, it has been recently reported that lactoferrin inhibits the development of cancer through inhibiting CYP1A1 activation in 7,12-dimethylbenz[a]anthracene- (DMBA-) induced hamster buccal pouch carcinoma model. Taken together the results obtained from our laboratory and previously published reports, we speculate that lactoferrin could be responsible for camel-milk-mediated effect. In addition, the main components of the camel milk have been already determined [38]. Ongoing research in our laboratory focuses on identifying and characterizing the most effective component of camel milk using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and one-dimensional gel electrophoresis, where several proteins were found to be relatively abundant in camel milk (data not shown).
In conclusion, the results of current study suggest that camel milk could protect against or decrease the deleterious effects of many environmental toxicants and carcinogens such as PAHs, probably through modulation of AhR-regulated genes of Cyp1a1, Nqo1, and Gsta1 at the transcriptional and posttranscriptional mechanisms. These results are of potential clinical significance to humans in that it uncovers the molecular mechanism involved and could explain the anecdotal evidence for the successful use of camel milk in the treatment and/or prevention of various medical conditions.
|Aryl hydrocarbon receptor
|Actinomycin D
|Cytochrome P450 1A1
|Dimethyl sulfoxide
|7-ethoxyresorufin O-deethylase
|Glyceraldehyde-3-phosphate dehydrogenase.
|Glutathione S-transferase a1
|(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)
|NAD(P)H:quinone oxidoreductase
|Polycyclic aromatic hydrocarbons
|Xenobiotic responsive element.
Conflict of Interests
The authors declare that there is no conflict of interests.
The authors extend their appreciation to the Deanship of Scientific Research at King Saud University for funding the work through the research group project no. RGP-VPP-141.
- J. K. Kerzee and K. S. Ramos, "Constitutive and inducible expression of Cyp1a1 and Cyp1b1 in vascular smooth muscle cells: role of the Ahr bHLH/PAS transcription factor," Circulation Research, vol. 89, no. 7, pp. 573–582, 2001. View at Google Scholar · View at Scopus
- M. L. Whitelaw, M. Gottlicher, J. A. Gustafsson, and L. Poellinger, "Definition of a novel ligand binding domain of a nuclear bHLH receptor: co-localization of ligand and hsp90 binding activities within the regulable inactivation domain of the dioxin receptor," EMBO Journal, vol. 12, no. 11, pp. 4169–4179, 1993. View at Google Scholar · View at Scopus
- J. P. Whitlock Jr., "Induction of cytochrome P4501A1," Annual Review of Pharmacology and Toxicology, vol. 39, pp. 103–125, 1999. View at Publisher · View at Google Scholar · View at Scopus
- D. W. Nebert, A. L. Roe, M. Z. Dieter, W. A. Solis, Y. Yang, and T. P. Dalton, "Role of the aromatic hydrocarbon receptor and [Ah] gene battery in the oxidative stress response, cell cycle control, and apoptosis," Biochemical Pharmacology, vol. 59, no. 1, pp. 65–85, 2000. View at Publisher · View at Google Scholar · View at Scopus
- D. W. Nebert, T. P. Dalton, A. B. Okey, and F. J. Gonzalez, "Role of aryl hydrocarbon receptor-mediated induction of the CYP1 enzymes in environmental toxicity and cancer," Journal of Biological Chemistry, vol. 279, no. 23, pp. 23847–23850, 2004. View at Publisher · View at Google Scholar · View at Scopus
- D. Ross, "Quinone reductases multitasking in the metabolic world," Drug Metabolism Reviews, vol. 36, no. 3-4, pp. 639–654, 2004. View at Publisher · View at Google Scholar · View at Scopus
- V. Vasiliou, D. Ross, and D. W. Nebert, "Update of the NAD(P)H:Quinone oxidoreductase (NQO) gene family," Human Genomics, vol. 2, no. 5, pp. 329–335, 2006. View at Google Scholar · View at Scopus
- M. Cuendet, C. P. Oteham, R. C. Moon, and J. M. Pezzuto, "Quinone reductase induction as a biomarker for cancer chemoprevention," Journal of Natural Products, vol. 69, no. 3, pp. 460–463, 2006. View at Publisher · View at Google Scholar · View at Scopus
- N. Kontou, T. Psaltopoulou, D. Panagiotakos, M. A. Dimopoulos, and A. Linos, "The mediterranean diet in cancer prevention: a review," Journal of Medicinal Food, vol. 14, no. 10, pp. 1065–1078, 2011. View at Publisher · View at Google Scholar
- C. Conesa, L. Sánchez, C. Rota et al., "Isolation of lactoferrin from milk of different species: calorimetric and antimicrobial studies," Comparative Biochemistry and Physiology B, vol. 150, no. 1, pp. 131–139, 2008. View at Publisher · View at Google Scholar
- E. I. el Agamy, R. Ruppanner, A. Ismail, C. P. Champagne, and R. Assaf, "Antibacterial and antiviral activity of camel milk protective proteins," Journal of Dairy Research, vol. 59, no. 2, pp. 169–175, 1992. View at Google Scholar · View at Scopus
- C. Saravanan, Z. Cao, J. Kumar et al., "Milk components inhibit Acanthamoeba-induced cytopathic effect," Investigative Ophthalmology & Visual Science, vol. 49, no. 3, pp. 1010–1015, 2008. View at Google Scholar
- H. M. Korashy and A. O. S. El-Kadi, "The role of aryl hydrocarbon receptor and the reactive oxygen species in the modulation of glutathione transferase by heavy metals in murine hepatoma cell lines," Chemico-Biological Interactions, vol. 162, no. 3, pp. 237–248, 2006. View at Publisher · View at Google Scholar · View at Scopus
- S. W. Kennedy, A. Lorenzen, C. A. James, and B. T. Collins, "Ethoxyresorufin-O-deethylase and porphyrin analysis in chicken embryo hepatocyte cultures with fluorescence multiwell plate reader," Analytical Biochemistry, vol. 211, no. 1, pp. 102–112, 1993. View at Publisher · View at Google Scholar · View at Scopus
- A. Lorenzen and S. W. Kennedy, "A fluorescence-based protein assay for use with a microplate reader," Analytical Biochemistry, vol. 214, no. 1, pp. 346–348, 1993. View at Publisher · View at Google Scholar · View at Scopus
- B. N. M. Zordoky, M. E. Aboutabl, and A. O. S. El-Kadi, "Modulation of cytochrome P450 gene expression and arachidonic acid metabolism during isoproterenol-induced cardiac hypertrophy in rats," Drug Metabolism and Disposition, vol. 36, no. 11, pp. 2277–2286, 2008. View at Publisher · View at Google Scholar · View at Scopus
- A. Anwar-Mohamed, O. S. Degenhardt, M. A. M. El Gendy, J. M. Seubert, S. R. Kleeberger, and A. O. S. El-Kadi, "The effect of Nrf2 knockout on the constitutive expression of drug metabolizing enzymes and transporters in C57Bl/6 mice livers," Toxicology in Vitro, vol. 25, no. 4, pp. 785–795, 2011. View at Publisher · View at Google Scholar
- A. Anwar-Mohamed, R. H. Elbekai, and A. O. S. El-Kadi, "MG-132 inhibits the TCDD-mediated induction of Cyp1a1 at the catalytic activity but not the mRNA or protein levels in Hepa 1c1c7 cells," Toxicology Letters, vol. 182, no. 1-3, pp. 121–126, 2008. View at Publisher · View at Google Scholar · View at Scopus
- K. J. Livak and T. D. Schmittgen, "Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method," Methods, vol. 25, no. 4, pp. 402–408, 2001. View at Publisher · View at Google Scholar · View at Scopus
- J. Sambrook, E. F. Fritsch, and T. Maniatatis, "Molecular Cloning," in A Laboratory Manual, N. Ford, Ed., Cold Spring Harbour Laboratory Press, Plainview, NY, USA, 1989. View at Google Scholar
- M. A. M. El Gendy, A. A. Soshilov, M. S. Denison, and A. O. S. El-Kadi, "Transcriptional and posttranslational inhibition of dioxin-mediated induction of CYP1A1 by harmine and harmol," Toxicology Letters, vol. 208, no. 1, pp. 51–61, 2012. View at Publisher · View at Google Scholar
- H. M. Korashy and A. O. S. El-Kadi, "Regulatory mechanisms modulating the expression of cytochrome P450 1A1 gene by heavy metals," Toxicological Sciences, vol. 88, no. 1, pp. 39–51, 2005. View at Publisher · View at Google Scholar · View at Scopus
- M. L. Slattery, W. Samowtiz, K. Ma et al., "CYP1A1, cigarette smoking, and colon and rectal cancer," American Journal of Epidemiology, vol. 160, no. 9, pp. 842–852, 2004. View at Publisher · View at Google Scholar · View at Scopus
- P. P. Shah, K. Saurabh, M. C. Pant, N. Mathur, and D. Parmar, "Evidence for increased cytochrome P450 1A1 expression in blood lymphocytes of lung cancer patients," Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, vol. 670, no. 1-2, pp. 74–78, 2009. View at Publisher · View at Google Scholar · View at Scopus
- T. Shimada and Y. Fujii-Kuriyama, "Metabolic activation of polycyclic aromatic hydrocarbons to carcinogens by cytochromes P450 1A1 and 1B1," Cancer Science, vol. 95, no. 1, pp. 1–6, 2004. View at Publisher · View at Google Scholar · View at Scopus
- T. D. Williams, J. S. Lee, D. L. Sheader, and J. K. Chipman, "The cytochrome P450 1A gene (CYP1A) from European flounder (Platichthys flesus), analysis of regulatory regions and development of a dual luciferase reporter gene system," Marine Environmental Research, vol. 50, no. 1–5, pp. 1–6, 2000. View at Publisher · View at Google Scholar · View at Scopus
- B. Hasspieler, D. Haffner, M. Stelljes, and K. Adeli, "Toxicological assessment of industrial solvents using human cell bioassays: assessment of short-term cytotoxicity and long-term genotoxicity potential," Toxicology and Industrial Health, vol. 22, no. 7, pp. 301–315, 2006. View at Publisher · View at Google Scholar · View at Scopus
- V. Werlinder, M. Backlund, A. Zhukov, and M. Ingelman-Sundberg, "Transcriptional and post-translational regulation of CYP1A1 by primaquine," Journal of Pharmacology and Experimental Therapeutics, vol. 297, no. 1, pp. 206–214, 2001. View at Google Scholar · View at Scopus
- G. Kikuchi, T. Yoshida, and M. Noguchi, "Heme oxygenase and heme degradation," Biochemical and Biophysical Research Communications, vol. 338, no. 1, pp. 558–567, 2005. View at Publisher · View at Google Scholar · View at Scopus
- I. E. A. Amara, A. Anwar-Mohamed, and A. O. S. El-Kadi, "Mercury modulates the CYP1A1 at transcriptional and posttranslational levels in human hepatoma HepG2 cells," Toxicology Letters, vol. 199, no. 3, pp. 225–233, 2010. View at Publisher · View at Google Scholar · View at Scopus
- A. Anwar-Mohamed and A. O. S. El-Kadi, "Arsenite down-regulates cytochrome P450 1A1 at the transcriptional and posttranslational levels in human HepG2 cells," Free Radical Biology and Medicine, vol. 48, no. 10, pp. 1399–1409, 2010. View at Publisher · View at Google Scholar · View at Scopus
- A. S. Maghraby, M. A. Mohamed, and A. M. Abdel-Salam, "Anti-schistosomal activity of colostral and mature camel milk on Schistosoma mansoni infected mice," Asia Pacific Journal of Clinical Nutrition, vol. 14, no. 4, pp. 432–438, 2005. View at Google Scholar · View at Scopus
- M. Mohora, "Role of Nad(P)h: quinone oxidoreductase in the regulation of intracellular redox state," Romanian Journal of Internal Medicine, vol. 38-39, pp. 33–50, 2000. View at Google Scholar · View at Scopus
- S. Danson, T. H. Ward, J. Butler, and M. Ranson, "DT-diaphorase: a target for new anticancer drugs," Cancer Treatment Reviews, vol. 30, no. 5, pp. 437–449, 2004. View at Publisher · View at Google Scholar · View at Scopus
- Z. Farah, R. Rettenmaier, and D. Atkins, "Vitamin content of camel milk," International Journal for Vitamin and Nutrition Research, vol. 62, no. 1, pp. 30–33, 1992. View at Google Scholar
- G. Konuspayeva, B. Faye, G. Loiseau, and D. Levieux, "Lactoferrin and immunoglobulin contents in camel's milk (Camelus bactrianus, Campus dromedarius, and Hybrids) from Kazakhstan," Journal of Dairy Science, vol. 90, no. 1, pp. 38–46, 2007. View at Google Scholar · View at Scopus
- A. Roseanu, P. E. Florian, M. Moisei, L. E. Sima, R. W. Evans, and M. Trif, "Liposomalization of lactoferrin enhanced its anti-tumoral effects on melanoma cells," BioMetals, vol. 23, no. 3, pp. 485–492, 2010. View at Publisher · View at Google Scholar · View at Scopus
- M. S. Y. Haddadin, S. I. Gammoh, and R. K. Robinson, "Seasonal variations in the chemical composition of camel milk in Jordan," Journal of Dairy Research, vol. 75, no. 1, pp. 8–12, 2008. View at Publisher · View at Google Scholar · View at Scopus


text:
Senator are bad at picking winners, but losers are prettty good at picking Senators...since 2004!
Thursday, April 10, 2014
Guess who wasn't an economics major?
"I came up as a wrestler and I can tell you I worked just as hard as any football player in the country, as any basketball player, in fact I would say I worked harder than those guys,"
"The fact is we have student-athletes in all sorts of sports that, if you apply any form of value to their labor, you cannot pay football players and not pay gymnasts just because the football player has the blessing of an adoring public. That's the only difference. There are a lot of student athletes that are worthy."
Well maybe he did take a class in Marxian econ and fell in love with the labor theory of value!
Of course a simple glance at the real world (as opposed to wherever the hell Bowlsby lives) refutes his thesis.
Just change the context and see if you buy it: "you cannot pay movie stars more than teachers just because the movie star has an adoring public. Teachers work harder than those guys".
Look people, the hard truth is that football and basketball players have been cross-subsidizing tennis players, swimmers, rowers, golfers, you name it, for a very long time. Now that the handwriting is on the wall for that (to me at least) blatantly unfair system, there are going to be big adjustments.
No more cross country air travel for the golf / tennis / field hockey teams. Heck maybe even no more scholarships for them either.
6 comments:
- Ryan Long said...
There's more to it than that. Student athletes are prevented by the NCAA from working over 20 hours per week (at a part-time job). More popular sports may generate some revenue for the less popular sports, but the less popular athletes are often forced to "volunteer" to help the football team as a way of reducing football's operating costs.
For example, as an NCAA cross-country athlete, I was required to work the parking lots during football and basketball games.
I think we probably agree about the ultimate conclusions here - reduce the power of the NCAA and don't fund any sport that fails to generate a profit - but this issue looks very different from the athlete's perspective. Being forced to work for the football team at the expense of working for myself to pay my own bills, having 40% of my available credit hours sucked away by PE classes, and thus being forced to pay an additional 2 years of tuition fees after my NCAA eligibility is up...
These are the reasons I quit the team after one year. Either let kids pay their own way, or pay them. Fair is fair.
- 9:44 AM, April 10, 2014
- Jeremy Wells said...
I'm sure he'd agree that professors work harder than conference commissioners and therefore deserve more pay as well...
- 1:18 PM, April 10, 2014
How much more do you think the commissioner of the BIG 12 makes than the commissioner of the Great Lakes Valley Conference?
Shoot, aren't they all just working for student athletes, who we've already established are equals?
- 5:26 PM, April 10, 2014
- dismal_scientist said...
Cross sensitization is mostly a myth. See Rod Fort and Jason Winter's 15 sports myths and why they are wrong, myth number 1.
- 6:50 PM, April 10, 2014
- Mirandah Farre said...
Nice read! Very informative. Did you know that Chinese lead in online shopping? Full story here: http://on.fb.me/1gRKzis
- 3:55 AM, April 11, 2014
What also complicates the picture is athletes of some big revenue programs receiving non-monetary forms of compensation that overall cost the taxpayer and the people far more than the profit they make. How much is the police covering up a rape worth? Several million dollars? What about more mundane criminal activity, like drug dealing?
The problem of course is that when you add up the individuals receiving such compensation it's not necessarily according to their performance as athletes. It might not always be the star player getting these benefits even when they accrue to the football team as a whole.
- 1:36 AM, April 13, 2014

subset: commoncrawl


text:
Featured News - Current News - Archived News - News Categories
Palm Restaurant Group, Cosmopolitan, Major League Soccer, Epic Records, Zynga, Live Nation, Gilt and Viceroy Hotel Group are each featured in their own episode searching for their next outstanding employee
The series premiere of the new CBS reality competition series "The Job" will debut Friday, Feb. 8 (8 p.m.), with five candidates vying for an assistant manager position with the Palm Restaurant Group. The fine-dining steakhouse has been owned and operated by members of the Bozzi and Ganzi families for 87 years and has 25 locations across the continental U.S., as well as international outposts in Mexico City and London.
The second episode, airing Friday, Feb. 15 (8 p.m.), will feature five new candidates competing for an editorial assistant position with Cosmopolitan. The world's largest women's magazine with 18 million readers each month, Cosmo is the biggest-selling monthly magazine on U.S. newsstands and a leader in the digital, social media and tablet space.
The executives representing Palm Restaurant Group in the premiere will be Co-Chairman and Co-Owner Wally Ganzi, Executive Vice President Bruce Bozzi Jr., and Palm Too General Manager Janice Sheil. The guest companies, who can also compete for the candidates, will be Westville, which serves eclectic American cuisine in a relaxed atmosphere at their four Manhattan locations; YB & Co. by Chef Julian Medina, which features six upscale Mexican and Pan-Latino restaurants in New York City; and Maison Privé Chefs, a boutique catering company serving New York City and the surrounding areas that unites chefs, farmers and seasonal menus.
Information on the candidates for the assistant manager position with The Palm:
For the second episode, the executives representing Cosmopolitan will be Editor-in-Chief Joanna Coles, Executive Editor Joyce Chang and Senior Editor Jessica Knoll. The guest companies will be Fashionista, one of the largest independent fashion news sites; ArchetypeMe, an innovative website that redefines the concept of "search" by filtering content and community based on who you are; and Who What Wear, a leading online fashion magazine providing access to the latest celebrity styles, runway trends and beauty secrets.
Information on the candidates for the editorial assistant position with Cosmopolitan:
Hosted by Emmy Award nominee Lisa Ling, each episode of "The Job" will feature five candidates participating in several rounds of elimination challenges before a panel of executives as they compete for their dream job. In addition to the episode's featured company, representatives from three guest companies from related industries will have the opportunity to make an on-the-spot offer to one of the candidates who must decide if they will accept the offer or remain in the running for the highlighted job.
The featured companies for the additional episodes will be, in no particular order:
Major League Soccer: The top-flight professional soccer league in the U.S., MLS was founded in 1996 after the country hosted the 1994 FIFA World Cup. The league is composed of 19 teams - 16 in the U.S. and three in Canada.
Epic Records: A premiere record label boasting a vast catalog comprised of some of the most important recordings in history as well as the home for the next generation of superstars.
Zynga: The world's leading provider of social game services with 311 million monthly active users playing its games, which include "FarmVille," "FarmVille 2," "Words With Friends" and "Draw Something."
Live Nation: The world's leading live entertainment and eCommerce company, Live Nation is comprised of four market leaders: Ticketmaster.com, Live Nation Concerts, Front Line Management Group and Live Nation Network.
Gilt: A leading online shopping destination with more than 7 million members, Gilt offers daily sales at noon ET for women, men, kids, home décor and local experiences. Founded in 2007, the company has pioneered the flash sale, granting members special access to a carefully curated selection of the best brands for up to 60 percent off.
Viceroy Hotel Group: Viceroy delivers one-of-a-kind luxury experiences that bring together provocative design and intuitive service in sought-after international locations such as Abu Dhabi, Beverly Hills, New York, Anguilla and Miami.
Air dates, guest companies and candidates for additional episodes will be included in subsequent releases.


text:
- This event has passed.
Cat Volunteer Orientation
December 7, 2017 @ 10:00 am - 11:30 am
Ask for Kelly. New volunteers – this is a hands-on orientation – dress to get furry! You must be 18 years of age or older to volunteer. Please note we do not provide community service hours. If you have questions prior to orientation, please call 513-378-0300 or email firstname.lastname@example.com.
Orientation starts promptly. Please arrive a few minutes early to complete our volunteer application.


text:
Suzannah Yip joined Bonhams in August 2007 to create and head the Japanese Department, which has become Europe's leading auction house for Japanese art. Under her leadership, the department in London achieved the most valuable sales ever for Japanese art in Europe 2011, totalling nearly £10M.
She started her career at Sotheby's London in 1999 as a graduate Junior specialist in the Japanese department, where she was responsible for the cataloguing and successful dispersal of some outstanding print collections including: The Huguette Berès Collection, Part 1 & 2 (Paris 2002 & 2003) and The Adolphe Stoclet Collection of Important prints, paintings and illustrated books (London 2004). Although her main area of academic focus and interest lies in the pictorial arts of Japan, she also handles other diverse areas of Japanese art, in particular, Netsuke and works of art from the Meiji period. Working closely with Neil Davey, one of the World's leading experts on Netsuke, she co-catalogued the renowned Katchen Collection of Netsuke, part 1&2 (London 2005&2006) and more recently, she co-catalogued and researched the Edward Wrangham collection of Japanese art part I, II III & IV as well as the famous Harriet Szechenyi collection of Japanese art which achieved a total of £4.6M - Europe's most valuable auction to date of Japanese art. During the seven years she has been at Bonhams, she has sourced important works of art which have obtained world record prices at auction in London. She is fluent in Chinese (Cantonese and Mandarin), French and Japanese and holds a Masters Degree in Art and Archaeology (specialising in the arts of Japan) from SOAS, the University of London.


text:
I have a friend who is thinking about starting a kind of English studio near Shin-Yurigaoka, she has a spare house (nice problem) she has had converted into rooms suitable for acttivies, a large room with a mirrored wall (yoga classes and mom & child things etc...)and various other rooms for English lesson/lectures.
She is looking for people who can teach. I know nothing of pay etc... Just that she is looking people for possibly; Thursday Friday and Saturday... If you want to know more I can put you in touch with her.
PM me for more info.
(I want to point out, this isn't my business and I am not out to financially gain from this. Not that it would make a difference.)


text:
Alfresco Continues to Foster Growing Partner Base with New Solution Showcase
Leader in Open Source ECM Offers Online Resource for Partners to Promote Sharing of Enterprise-Level Business Solutions
LONDON – September 29, 2009 – Alfresco Software, the leader in open source enterprise content management (ECM), today announced the launch of its Partner Solution Showcase, an online content application resource that enables partners to promote solutions built with Alfresco. The Partner Solution Showcase offers a selection of solutions developed using Alfresco's content platform which enable leading companies from a range of industries to solve critical business problems.
Tweet this: #Alfresco launches Partner Solution Showcase to Promote Range of Content Applications
With an open standards and open source architecture, Alfresco software provides partners with an ideal platform for building content applications or application extensions. By supporting open standards, including the upcoming content management interoperability standard (CMIS), partners are able to easily build content applications using Alfresco and deliver portable, cost-effective business solutions . Alfresco's Partner Solution Showcase was created in response to growing interest from both customers and partners to share best-of-breed Alfresco solutions.
"Micro Strategies has been delivering high-quality, ECM solutions for more than 10 years – and we have clearly reached the next level with Alfresco's enterprise-class platform. Now more than ever, our clients must be considerably more efficient at a lower total cost of ownership, and Alfresco enables us to achieve both objectives," said Adam J. Storch, vice president of business solutions, Micro Strategies. "We are very excited about Alfresco's Partner Solution Showcase, as it provides us with a great place to market our solutions and reach a much larger audience."
In addition to achieving a reduction of costs and offering ease-of-integration, Alfresco's Partner Solution Program enables its partners to:
• Add content management, workflow and collaboration features and capabilities to each solution;
• Safely embed Alfresco content services under a commercial license that fits each OEM's needs;
• Receive support and training from Alfresco experts.
"ParaScale partnered with Alfresco to help deliver on-demand scalability, cloud-wide network security and cost efficiency to users," said Mike Maxey, director of product management, ParaScale. "Our partnership, which integrates the Alfresco ECM solution with the ParaScale cloud storage platform on a single hardware device, has allowed us to bring the manageability and performance benefits of a true cloud architecture to our combined customers."
"Our Partner Solution Showcase was designed to promote how our partners are truly driving innovation and solving business problems for leading companies using solutions built with Alfresco," explained John Powell, CEO, Alfresco Software. "The Showcase is also further proof that many now view Alfresco as a content platform that can be used to build an eco-system of Alfresco applications and add-ons. We look forward to welcoming new partners and extending the range of business critical applications available."
For more information about Alfresco's Partner Solutions Showcase, please visit http://www.alfresco.com/partners/solutions/ or contact email@example.com.


text:
So,this month it is going to be dessert recipes...
Chocolate Pumpkin Pie Bar sent by Anu's Healthy Kitchen....wow..is nt it cool??..:)
happy hosting pranati
thanks dear..:)
awesome event..will try to send some recipe..
sure will send it to you...thnks for letting me knw...
SYF&HWS - Cook With SPICES" Series - CARDAMOM (Dec 5th to Jan 5th)
100 Friends/Followers & GIVEAWAY
Happy hosting Pranathi. I have added mine Eggless Cooker Vanilla Cake.
Hi Happy hosting. I have sent you few of my entries. :)
Hi Pranathi,I somehow missed you and finally reached here today.Nice work dear.All the best for your event.Do you think my today's post can be linked here ? would love to see you soon.Thanks,
thanks friends for linking the beautiful & delicious sweets recipes in the event..:)
Hello dear, linked my recipe for your lovely event, happy hosting.
thanks for sharing sending some recipes
linked up my recipe dear..thnks for letting me knw...
Thanks for stopping by my blog and bringing this event to my notice. I would love to participate in this event and will send my recipe soon. You have a great space here.
Lovely event! Will be posting my recipes soon :) Happy Hosting Pranati :)
thanks for letting me know. linked lava cake! :)
hi..thanks for the invite..I've linked a yummy cheesecake ;)
Thank you for letting me know about the vent. I've linked in two entries. :)
Happy Hosting!
Thanx for letting me know abt the event. Have linked my entry. Happy hosting dear.
Hi there...first time here..I have linked here....Happy hosting...Merry Christmas.
Thank you for dropping by and inviting me to the event! I linked up my cranberry cake!
Hi! Thanks for visiting & inviting me to the event. Linked up my entry. Happy hosting!
Thanks for letting us share our sweets. Looks like you have quite a list.
wow this is nice event...first time here...Happy New year.....Loved your space and this event..i would like to send some of my entries tooo...
Wonderful event dear !! first time here...u have excellent space!!
Wonderful event dear.I linked up my Eggless sponge cake with curd.Happy to follow u..drop at my blog too.
Happy New Year and Prosperous life.
Ongoing Events:
Foodies Nest
Milk Shake~Event
Fasting Day Menu #1
nice event... will send my entries soon...
nice event. linked fruit salad
Thanks friends for the participation and I totally loved all the entries.
These are very informative
Good to see the
I would like to say that this blog really convinced me, you give me best information!
Thanks, very good post.
Browse the latest free classified ads in your local area...


text:
After reading Thomas MacEntee's post at the Geneabloggers blog called "Blogger Announces New Pages Feature", I knew what I needed to do to improve this blog.
Over the weekend I added 4 pages to the top of this blog. They are:
I encourage you to take a look at Thomas's blog post and learn more about adding pages to your blog. I think that you will find this feature very helpful. Let me know what you think about my pages as well. I would love your feedback as a comment or email to email@example.com.
Monday, February 8, 2010
Adding Blogger Pages to My Blog
Posted by Tina Lyons at 11:00 AM
3 comments:
Oh, my, we had the same idea this weekend!ReplyDelete
I'm so glad GeneaBloggers could help be your muse on for pages! And they look great - I think your readers will like the new feature.
Great! I haven't even had the chance to take a look at it yet, but I have to do that soon! :)ReplyDelete
Subscribe to: Post Comments (Atom)


text:
Care in Ghana by Hannah Simmons
After a year of preparation, I arrived in Ghana with two of my best friends, Anna and Catriona, with extremely vague expectations. I was hit first by the humidity and chaos of Accra but found I felt extremely welcomed by the Ghanaians the moment I got off the plane. We were met at the airport by Nyame who was the first of many friendly and enthusiastic Projects Abroad staff who we were to meet during our two weeks in Ghana and we spent the first night in Accra. When I was first offered a sachet of water, I mistook it for an ice pack! The idea of drinking from a plastic bag seemed quite bizarre but we soon got used to it, along with our daily snacks of corn and sugar cane. I never imagined that I would miss such insignificant things in the months after I returned.
Waking up at 5am was not the easiest of things after an exhausting day of travelling before, but we were all excited to arrive in the Akuapem Hills and amused our taxi driver with our western amazement and enthusiasm to be immersed in this new culture. Our welcome at the office in the Hills was one I shall never forget – Emmanuel, the regional coordinator singing his 'I want to be your friend' song and Henry grinning away assured me that the next few weeks would be more than simply memorable. I found you very quickly adapt to a simpler lifestyle and before long I had adjusted not only to the toilets but to the refreshing bucket showers (a lovely wake up after a day's hard work) and brushing my teeth by torchlight in the back garden.
Anna, Catriona and I all shared a room at Diana's house and I have to say that I could not have been made any more welcome by Diana, her family and the other volunteers who lived with her. As all three of us love to sing and I had brought my ukulele to Ghana we were invited by Hannah, Emmanuel and Samuel (the children at the house) to play to them in return for some dance lessons! We spent many afternoons playing games, dancing and singing with the kids – I even taught Emmanuel to play Leonard Cohen's 'Hallelujah' on the ukulele and we had everyone, including Diana's two year old neighbour, joining in! Having such a wonderful host family really made this a precious experience.
The work we carried out in our two weeks in Ghana was varied which made everyday new and exciting. We began by painting Mamfe Methodist School. There were about 12 of us altogether and we all got on extremely well. There was never a dull moment at the school as the pupils were so friendly - we were blessed to be surrounded by the singing, dancing and enthusiasm of the children and they were always keen to help out.
We also had the opportunity to assist in the work at the Projects Abroad Community Farm, and our other job placements included working at Adom Day Care Centre, just a short walk from Diana's house. Here we tried our hand at building cement bricks which certainly tested our strength, as well as building and plastering a new urinal to replace what was previously a pile of bricks. I loved spending our breaks giving piggybacks, playing games and generally being clambered on by the children. We also had the chance to collect water from a well in what can only be described as a small jungle. I had a great amount of respect for the men and women who do this on a day to day basis as by the time we made it back up the hill, my arms were killing me!
During our two week programme we also visited Trinity Orphanage on several occasions. It was emotional to see so many children aging from 2 weeks to 16 years fighting for our attention but I am extremely grateful that we got the opportunity to deliver fun activities such as mask-making. The image of masked faces smiling up at me is one that I hope will remain with me forever.
The evenings, as with our days in Ghana, were never mundane. We attended dance lessons at the Projects Abroad office where we picked up some useful skills, and tried our hand at the local drumming. The office also played host to the weekly quiz nights where we could get to know the other volunteers. Everyone was really friendly and it was amazing to meet people from all over the world, not just the locals. While we were out there, Ghana celebrated 49 years since becoming a republic – a night which called for much celebration! We went to bar/restaurant with some local friends we had made and spent the evening dancing away Ghanaian style. I would definitely recommend experiencing the local nightlife, it was great fun!
At the weekend we visited the bead market as well as Cape Coast. There, we took a trip to the slave fort which taught us a lot about the horrific history of the slave trade, and also went on the canopy walk in Kakum national park. We had spectacular views of the forest and saw some incredible trees, spanning the length of more than five of us holding hands.
Going to Ghana was the best experience of my life to date. I would definitely recommend volunteering abroad because you will meet some of the friendliest and accommodating people as well as find yourself immersed in an entirely different culture, bringing back more than just photos. The trip has assured me of my future career ambition to work for charities in developing countries and has opened my mind to the vast possibilities that await us in the world.
Hannah Simmons
+44 (0) 1273 007230


text:
Gabby SNL
E!'s Marc Malkin is reporting that Gabourey Sidibe will be hosting Saturday Night Live before the end of the season. Love this choice. And not because Gabby's so great. It's also because Gabby is fresh. Gabby doesn't necessarily have to be "nice". And so Gabby can afford to be a little renegade which hopefully means the writing will be strong.
There are certain artists who commit to the show who come with a list of conditions. I won't do skits about this. I don't want to mock that. Well then f-ck. How will you make us laugh?
Over the last several weeks, Gabby's weight has become the subject of a huge debate. Howard Stern said that she was too "fat" to be successful. Many blogs criticised that she's a bad example. Gabby so far has * mostly* declined to answer the hate. Perhaps she'll be game to tackle the issue on the show.
Gabby will be closing out the year with some other very accomplished ladies including Tina Fey and Betty White. Malkin also reports that Ryan Phillippe will take the stage one night too. For why? Why Is Ryan Phillippe?
This is Gabby out and about in NY this week.
Photos from INFphoto.com


text:
f246632 necesitaba un nuevo diseño de página web y lanzó un concurso de diseño en 99designs.
Un ganador ha sido elegido entre 46 diseños de 13 diseñadores freelance.
We transform photos and digital images to canvas prints ready to display on wall.
http://www.canvasondemand.com - great ordering system
http://www.canvaspeople.com - great ordering system , simple :), too colorful :(
http://www.zartprints.com/ - gallery and gallery ordering system (same page)
1.Main page - presentation of product, order button, testemonials from customers with image of thier canvas (under main image , or left from image)
Main buttons ( must be big and simple) - Home, Options, Prices, Testemonials, Gallery, Help
Main goal: to keep customer on site and direct him to find out more about product, to inspire him to put something he likes on canvas!
Would allways like to see light and dark colored layout ! Or atleast light colored one.
2.Gallery page with categories ( photo, ilustrations ...ETC) - gallery with art and photos that we will sell trough our website as canvas prints for commisioning artists, whitin gallery page - artists can register and login , and then upload thier images with descriptions
When gallery image is clicked - enlarged image appear , with artist photo-icon with name under it , and on right side buyer can pick one of sizes and options ..( similar on http://www.zartprints.com/?view=product&id=17738
Main goal: to present artistic and other images that can be easily ordered onsite as canvas prints
3.Prices/ order page - configurator tool - dimensions, options, effects,
(take a look at http://www.canvasondemand.com/prices-and-sizes/ )
Main goal: simple , easy to use , quick checkout <---USE Canvasondemand.com as example!
Other, on each page:
Header- Contact, Help/faq, Cart, pro-login
Main buttons : Home, Options , Testemonials, Gallery, Help
Footer (sitemap) - Home , Options , Customer Showcase, Gallery, Artists, Prices etc...
Top- big phone number and text
"10087 people like Slika " - facebook widget
Slika- word for picture, image or painting on croatian
http://www.slikenaplatnu.hr - means http://www.imagesoncanvas.com :)
Summary: We need simple and beautiful website with nice presentation of canvas as product and with simple ordering steps before checkout. Design must be flexible to language changes.
Summary 2: SIMPLE , beautiful, its here to sell tons of canvas :)
We want clean design, logo is colorful but dont make site colorful, we want images to standout, not design.
When we choose design , we will supply all texts for links on croatian language to the designer , so you will deliver us Croatian and English version of designs .
UPDATE: expect updates soon with more details etc.
UP1- You can try give a solution where Slike na platnu ( IMages on canvas) - we have http://www.slikenaplatnu.hr is main "logo"


text:
Director Joe Wright's vision for the Leo Tolstoy classic "Anna Karenina" has been called "bold," "daring" and "ambitious" — words that could also be used to describe the effect the director and his leading lady, Keira Knightley, have on each other.
"Keira is quite fearless, and she encourages me to go to places I wouldn't necessarily be brave enough to go to myself," says Wright, who proceeded with the film only after Knightley, whom he'd directed to an Oscar nomination in 2005's "Pride and Prejudice," had agreed to take on the lead role.
Still, his gutsy new vision for the Tolstoy classic — it's set mostly on the premises of a decaying theater, including a horse race scene that thunders across a proscenium stage — was a surprise he dropped on the actress a mere 10 weeks ahead of filming.
VIDEO| The Envelope Screening Series: 'Anna Karenina'
"He said, 'Come around to my office. I need to explain something to you,'" remembers Knightley, seated recently beside Wright over tea at the Chateau Marmont in Los Angeles. "I went in and it was covered with drawings of this old stage, and I went, 'Oh ..."


text:
Wednesday, March 14, 2012
Super Bass
10 comments:
Wow! It looks really nice...ReplyDelete
As usual, I love your work!
I like the detail in his vest.
- AnonymousMarch 15, 2012 at 6:28 AM
I wish I had that tee! You should make it on one of those random t-shirt making websites.ReplyDelete
The tee shirt is pure genius and the guy is....BADABOOM BOOM ! :)ReplyDelete
- AnonymousMarch 15, 2012 at 8:35 PM
hey~nickie! haven't seen you on blogger for a long time, really miss you here:)ReplyDelete
Those eyes!ReplyDelete
Wow! finally sthg new for your blog! Been following it and your facebook for awhile. Really enjoy your drawings. When will you upload the hi rez pictures of the Bicycle and Valentines Leather? they are awsome, I want to own them so bad!ReplyDelete
nickie- your art is fantastic. i love the clean no-nonsense lines and moody shading. i am an artist, too, and i love finding others' work that stimulates me as much as yours does. take care-- razeReplyDelete
Thank you very much dear folks for your nice comments. :) This guy is probably one of my big faves that I did so I'm mighty glad you guys like him too.ReplyDelete
May I use this for a Facebook post? How do you want me to credit you? Thanks! (Sorry if you received multiple comments from me about this. I tried it from my phone and it was glitching)ReplyDelete
Indian girls Hot Desi 3x Sex Xxx 3gp mp4 HD Video Free DownloadReplyDelete
»------------ European Sex (137)
»------------ Cumshot Sex (107)
»----------- Blowjob Sex (311)
»------------ Hot Sites (Ads)
»------------ Sex In Office (762)
»------------ Teen Sex (277)+7
»------------- Pornstar Sex (910)+10
»------------- Lesbian Sex (223)+22
»------------ Hardcore Sex (386)
»------------- European Sex (137)
»------------ Cumshot Sex (107)
»------------- Blowjob Sex (311)
»------------- Big Tits Sex (334)
»------------- Hot Sites (Ads)
»------------- Sex In Office (762)


text:
Watch how to make this recipe.
Preheat the oven to 350 degrees F.
Heat the butter to melt in a medium skillet over medium heat. Saute the apples, celery, and onions seasoned with a bay leaf, salt, and pepper, until very tender, 10 minutes. Add the stuffing cubes and moisten with 1 to 1/1 2 cups of stock; season with poultry seasoning. Cool completely, then combine in a bowl with ground turkey, salt, pepper, half the parsley, and egg. Form 4 oval-shaped loaves a couple of inches thick. Arrange on a rack over baking sheet or parchment-lined baking sheet and coat with a little oil to glisten. Roast until brown and firm, 45 minutes, depending on thickness.
Reheat the meatloaves in medium hot oven to warm through. Place them in a shallow baking dish and cover with foil. Cook's Note: To keep the loaves from drying out - add a splash of stock, this will keep the loaves super moist.
In a saucepan, whisk the oil and 1/4 cup flour over medium heat 10 to 15 minutes, until the color of peanut-butter. Add lots of coarse pepper to the roux, then whisk in a couple of cups of stock and thicken, season with salt to taste. Cool completely and store in the refrigerator. Reheat the mixture in a saucepan over medium flame.
While the meatloaves bake, boil the potatoes in enough water to cover. Drain, then return the potatoes to the hot pot and mash with milk, herb cheese, mixed herbs, remaining parsley, salt and pepper. Cool completely and store in the refrigerator.
Reheat the potatoes in a saucepan over medium-low heat, adding a little more milk as the potatoes warm to loosen them up a bit.
Serve the meatloaves, whole or sliced, with mashed potatoes and gravy.


text:
Might have a problem with the upload attachment since the forum problem on the 15th. I went to upload pictures of my pups on my dogs for sale ad. When I went to preview it, It was someone elses pictures(not even a picture I have in my files) that came up---so I didn't post it. Today I'm looking at other people's posts and I see a picture of one of my pups on a post that is supposed to be another person's adult dog (Molly). Any one else having problems with other people's pictures showing up when they try to upload a picture I see a couple more of my puppy pictures showed up in Grey Dawgs brag post
I see another of my pup pictures showed up in one of ckgsp dogs for sale ads. I see 2 more of my puppy pictures showed up in mlinnane's lewllen pupy dog for sale ad.


text:
Which programming language to choose for children
Many children, having got acquainted with the computer world, are excited by desire to become programmers. But where should we start? This question is asked not only by the children, but also by their parents. After all, it depends on their advice and farewell whether the child will love programming or not.
To answer the main question, where to start learning programming for children, we first turned to the network. On requests "where to start learning programming" or "what programming languages are in the most demand on the market", one can conclude that today the most popular languages are Java, Python, C #, C ++ or Cocoa. It is clear that these languages are really important in the modern world. For example:
- C ++ is needed to develop cross-platform games and high-performance computing
- Python is a statistical calculation
- Java is needed for those who plan to work in a large company
- PHP - creating complex websites and online stores
But all this is a difficult way, which can also alienate the young programmer from further development in this area. Therefore, before proceeding to the study of one of these languages, it is best to get the basic knowledge of the programming structure based on an easier and more intuitive language.
IT Academy
If we talk about programming courses for children and about fashion IT academies, there, most likely, your child will be offered to study web-design, teach basic knowledge of using PC, tell about the graphics and give information about HTML \ CSS for creating websites. All this knowledge is useful and can help you to decide in the future direction. But if your child said that he wants to learn programming, then the best option is to offer him a Basic programming language (by the way, he was taught in schools) or Scratch - a specially developed system for teaching children the basics of creating simple games or animations.
How to attract a child to the IT field?
Show the child that programming is not boring, but very exciting. This can be done just in the Scratch language example. In it, you can quickly get the finished result - a game or animation - without prior preparation and long training.
Give the child time to ensure that he has mastered this environment, understood what he is interested in. Do not push or try to force the child to learn programming languages that are used by professionals. They require a long and hard work, as well as basic knowledge of other subjects that your children will get only in high school.
Try to present the idea of programming for the child as a game. And Scratch is the simplest and most understandable way.
How the program is created
To create a working program, you need to think through and develop a number of components, without which you cannot get the finished result.
- Objects
- Logic
- Events
- Functions \ Operations
- Memory and Data
- Appearance of the program (GUI)
- Framework
The characters in the program (Game Hero, Picture, Sound or Data, for example, the alarm timer).
How the program will behave from the very beginning. This is a game script, an alarm clock script.
How the program will interact with the User - clicking the mouse, typing text from the keyboard. How objects will interact with each other - collision, appearance on the screen, disappearance.
What Objects can do at all: shoot, jump, fly, ring.
To the hero of the game had 3 lives and did not die with the first shot, we must make sure that he remembered how the shoots. To the alarm does not ring all day and was able to remind of a missed signal - it must remember that it already rang once. Such Data is stored in memory (in variables) or on a disk.
Graphical User Interface or whatever it looks like. Buttons, inscriptions on the screen, pictures.
Interrelation of all components of the program and adjustment of their harmonious work. That part of all programs, the creation of which immediately repels children and adolescents due to their complexity and monotony of writing code.
For a child who is just starting his way in programming, it will be difficult to immediately keep all 7 points in his or her head. That's why we recommend refusing from the programming for adults and choosing Scratch, one of the easiest languages for children.
What is Scratch and how to use it?
Scratch is a language \ programming environment, through which you can quickly (get the finished result) create a working program. This is the development environment for programs without the framework, which means that it is as simple and accessible to children and adolescents.
Without any basic training, your child will be able to write a simple game, create an animation, a calculator or a stopwatch with a beautiful design. Scratch allows you to understand in the game form the basic principles of programming and create a knowledge base, through which you can easily move to the next level of programming.
For example, in Scratch, all items are called sprites, sprites can be programmed and assign sounds. Dragging blocks from the "Scripts" tab to the "code" field, the child creates an understandable picture of the multi-colored blocks:
- Blue - responsible for the movement
- Lilac - responsible for appearance
- Pink - responsible for sound
- Green - responsible for the pen
- Orange - responsible for sprite data
- Brown - responsible for events
- Yellow - responsible for managing the sprite
- Blue - responsible for sensors
- Light green - responsible for operations
- Purple - own blocks
In the "Costume" tab, you can select the image of the sprite.
"Sounds" tab allows you to select a sound from the library or from your uploaded sound files and put on a sprite or background.
As a result, you can get a small game or a cartoon.
Try programming on Scratch will be also to parents who have never dealt with programming. This can be your new hobby, as well as help to understand what attracts your children so much in this area. A simple and intuitive Scratch interface will help you and your children to discover the world of programming and get the necessary knowledge to grow and develop in IT.
And to KidLogger users, we remind you that if you notice Scratch in the report "sites and applications", do not rush to scold your child for hours spent online. It's a chance to learn something new!
Here you can find tutorials, articles and announcements about KidLogger SAS: what new features we've implemented, how to install Kidlogger, and how to set up user monitoring for different platforms like Windows, Mac, Android, Symbian, BlackBerry and iOS.
We're happy to hear any suggestions you might have about improving KidLogger.
Compare Parental Controls in Windows 7, Mac and Kidlogger.net
"Never leave kids and employees unsupervised."
- 29 Jan
- 18 Jan
- 24 Dec
- 13 Nov
- 18 Jul
- 30 Jun
- 29 Jun
- 27 Jun
- 23 Jun
- 01 Jun


text:
Our Fitness Blog
The Calorie Games - How to Burn Fat Like A Champion
The calorie counters of the world know this pain - food tastes delicious, but it can be so bad for you! Many believe that the solution is to simply to exercise a little more...but do you know how long it would take you to work off the calories of your favorite meal or snack? This infographic is visual proof that diet is an important part of staying fit, too! Next time you look at a burrito, think about how it will take you nearly two and a half hours to work it off - or imagine that big bag of chips as three hours in the ring with Floyd Mayweather. Also take note - kickboxing is one of the best calorie burners around! So put down the sandwich and get active!
Click to view the full-size infographic


text:
Find the perfect place from the worlds best selection of holiday rentals on HomeAway.
Our 2019 accommodation listings offer a large selection of 30 holiday lettings near Paseo San Pedro. From 16 Condos/Apartments to 15 Studios, find the best place to stay with your family and friends to discover Paseo San Pedro area.
Yes, of course. HomeAway has 16 Condos/Apartments near Paseo San Pedro. Our other popular types of holiday rentals near Paseo San Pedro include:
Yes, you can select your preferred holiday accommodation with pool among our 5 holiday rentals with pool available near Paseo San Pedro. Please use our search bar to access the selection of self catering accommodations available.
Yes, HomeAway offers a selection of 30 holiday homes to book directly online and 13 with instant booking available near Paseo San Pedro. Don't wait, have a look at our self catering accommodations via our search bar and be ready for your next trip near Paseo San Pedro!


text: |COCONUT OIL 3000 mgEXTRA VIRGIN NATURAL DIETARY SUPPLEMENTManufactured for PL NutritionMade in USA2 Bottles 120 capsulesBenefits:Strengthen your immune systemImprove your digestionPrevents premature aging of your skinPromotes healthy weight loss Protects against many degenerative problemsDetoxifies and heals your body Beautifies your hair and skinNature's ultimate germ fighterReduces Blood pressureDescriptionThe coconut tree is rightly considered as the 'tree of life'. Each and every part of the tree is used for one thing or the other. Its uses range from cosmetics to food and it is part of the diet in the tropic and subtropic regions. The fibrous husk is used to make coir ropes and mats, the shell is used for charcoal and the fruit is used for oil as well as in diet. Coconut water is one of the healthiest drinks on the planet. This sweet liquid is present in the young, green coconut. Coconut is a drupe and it has three layers: endocarp, mesocarp and exocarp. According to different studies, coconut oil is among the healthiest food on earth. It is often described as a superfood as it has many health benefits. Coconut oil is rich in healthy fatty acids like capric acid and lauric acid. It also contains healthy saturated fat. The fat in coconut oil is easier to digest, has anti-fungal and anti-microbial properties and are not stored as fat readily. Coconut oil is used mainly in tropical countries but is now gaining popularity across the world. In addition, coconut oil also contains linoleic acid, oleic acid, phenolic acid, myristic acid, Vitamin E, Vitamin K, and iron.To reap the benefits of coconut oil, it is essential to choose organic unrefined coconut oil as it is extracted from fresh coconut meat and not from dried coconut.Basic benefits of coconut oilProtects liver and cares kidney infection and UTI. The medium-chain fatty acids (MCFAs) present in coconut oil are known to have antibiotic properties. It kills the bacteria by destroying the lipid coating on it. Coconut oil helps fight the kidney and urinary tract infection. It also protects the liver.Helps burn fat and lose weightThe MCFAs in coconut oil help in burning more calories and losing weight. According to a research, coconut oil helps in reducing the abdominal fat in women. It also boosts metabolic rate of the body by promoting the function of the pancreas and this helps in burning more calories.Improves brain functionAccording to a study conducted in 2004 and published in the journal of Neurobiology of Aging, MCFAs in coconut oil boost memory in older people. MCFAs are readily absorbed by the body and they fuel the cells of the brain, helping them to function efficiently.Boosts immunityLauric acid in coconut oil helps in fighting bacterial infection. Lauric acid creates an environment in the body that is toxic for the viruses. You can use coconut oil as your natural fuel instead of sugar when you are not well. The healthy fats in coconut oil support the thyroid and adrenal glands which can lower cortisol to naturally balance hormones. Coconut oil helps to suppress inflammation while supporting healthy metabolism, two things that are essential for balanced hormones.Improves digestionCoconut oil is used in cooking and it helps in preventing digestion related health issues and stomach problems. It improves digestion. It helps in the absorption of other nutrients like mineral and vitamins.Good for your skin and hairCoconut oil can be used as cleanser, sunscreen and moisturizer. The antibacterial properties of the oil will take care of any skin disorder you might have. It moisturizes your skin and reduces skin inflammation. It improves skin texture and is a great cure for all kinds of skin conditions.Helps in preventing bones problemsCoconut oil is rich in antioxidants, which fight the free radicals. One of the main causes of osteoporosis is free radicals and coconut oil is a natural cure for the condition. Coconut oil also improves the calcium absorption by the body.Coconut oil and heart healthThe main saturated fat in coconut oil is lauric acid. This type of saturated fat (SF) is shorter in length than other saturated fats. Lauric acid can be considered a medium chain triglyceride (MCT) which acts differently in the body than longer SF. Coconut oil, like other plant-based oils, does not contain cholesterol. The vast majority of the fats in coconut oil are medium chain triglycerides. These medium chain fatty acids found in coconuts have tremendous health benefits and are easy for the body to burn as fuel for energy. Coconut oil also contains antiviral, antifungal, antibacterial, and antimicrobial properties.Coconut oil and weight lossMCT's are absorbed quickly from the small intestine are sent to the liver to be used for energy. MCT's are less likely to be stored as fat and are more likely to be used for energy. Longer SF can aid in transporting cholesterol, be used for energy or stored in fat cells.Supplement FactsServing size: 3 softgelsServings per container: 20Top-Grade Virgin Coconut Oil 3000 mgCalories 26Calories from Fat 41Total Fat 3 g 5%Saturated Fat 2,6 g 12%Transfat 0 g 0%Monosaturated fat 0,075 gCholesterol 0 gTotal Carbohydrate 0 gProtein 0 gDaily Value not establishedOther ingredients: Gelatine and Purified water*These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent any disease.


text:
To keep me motivated, here are some inspiring photos I would like to see influence my home decor in 2013.
And, while we're on the subject of lighting, I would like to add sconces to the mix. And, lots of them.
I am trying to stay away from trends, but I am still in love with Kartell's ghost chair. I love the chair paired with the zinc table. And, I wouldn't mind that pine bookcase, either!
And, have lots of shelf space for family pictures, mementos, family board games and books:
I am looking forward to a 2013 (with less change) and more focus on our home and family. I hope you all had a wonderful holiday and are looking forwards to a new year!


text:
2010 Buddy Davis 70 Sportfisherman
- Description
- View Full Specifications
- Year:
- 2010
- Hull Material:
- Fiberglass
- Length:
- 70'
- Engine/Fuel Type:
- /
Davis Yachts celebrates this athletic performance with lines of unrivaled beauty. Like an artist with a huge canvas, Davis was truly inspired to create this tour de force. Admired from every angle, her signature bow flare, curved tumblehome and sweeping sheer will move even the non-boater to heartfelt emotion. Davis infuses every aspect of the Buddy Davis 70 Sportfisherman with his passion for elegance. The Buddy Davis 70 Sportfisherman is at home wherever turbulent waters are found. Smooth. Soft. Safe. Dry. All the attributes of the legendary Davis ride, only bigger. Faster. More powerful. The secret to this ride is it's ingeniously designed bottom. Deep and narrow, it will part seas in near biblical style. Yet straight-line performance alone does not measure a true sportfisherman's ultimate capability. Take the helm and put this boat through her paces. She's agile. Nimble. Especially for a boat of her size. Able to back down on that marlin. And easily maneuver into that tight slip.
Additional Specs, Equipment and Information:
Hull Shape: Semi-Displacement
LOA: 73 ft 6 in
Beam: 19 ft 6 in
Maximum Draft: 4 ft 6 in
Headroom: 6 ft 8 in
Freeboard: 7 ft 2 in
Dry Weight: 100000 lbs
Engine Options
Engine Option 1: 1x
Fresh Water Tanks: (300 Gallons)
*This price is based on today's currency conversion rate.
Email Us
Please enter a valid name!
Please enter a valid email address!
RE: Buddy Davis 70 Sportfisherman
Message cannot be empty!


text:
Thanks to the heartfelt donations of thousands of people -- many who never have, and never will, put a foot down into the red soil of the Sooner State -- Porter is at the launching point of providing 100 in-ground, concrete storm shelters for survivors of the May 19-20, 2013 storms.
Porter, disaster relief director for the Baptist General Convention of Oklahoma, hopes planting those concrete shelters will also plant seeds of hope for the future for so many who have been through so much.
The peak winds of 210 miles-per-hour that accompanied the twister on May 20 almost a year ago claimed 25 lives. Despite following the similar track of the even-deadlier May 3, 1999 Bridge Creek-Moore tornado, very few homes and neither of the two Moore schools destroyed by the 1.3-mile-wide May 20 tornado had storm shelters.
Through a partnership with the North American Mission Board (NAMB), the Oklahoma convention has a goal of installing 100 storm shelters during this year, Porter said. About half of them will be located at sites where the convention has provided, or will be providing, mobile homes for families who lost their residences in the storm.
"We will install a storm shelter for all of the families who have received a mobile home," Porter explained. "That will be a great psychological lift for them, and in a way, provide a sense of security, especially for those who have ridden out a storm in their home.
"So, for the 50 mobile homes we are going to put in, that's where half of these 100 storm shelters will go, we already know that."
The Oklahoma convention has already put in place 23 mobile homes, and has purchased another 20 to be refurbished.
"We have a total of at least 100 shelter sites identified through pastors, other agencies we work with and community leaders," Porter said.
Eight shelters will go into Steelman Estates, west of Shawnee, as community shelters, Porter indicated. They will be scattered out and be no more than one block away from any residence.
"Before the storm, there were 90 mobile homes in that area, and the majority of them were blown away by the tornado," he said.
Directing the project as long-term recovery coordinator is Sonya Saffa, an Oklahoma native who moved back to the state from Florida last year just after the storms hit. She has been involved in a significant way ever since. Saffa has extensive disaster relief experience with NAMB and helps national disaster relief staff set up disaster relief sites when the need arises. She also has owned her own ministry-related businesses and began and coordinated a ministry for orphans in Russia for seven years.
"She will be the field person tracking the project, making the contacts and coordinating the teams doing the installations," Porter said.
Saffa also has been helping coordinate the placement of the mobile homes for tornado survivors working with three other state disaster relief leaders.
Porter said, "We have also purchased a backhoe to support the project, and then we'll roll it over into our disaster relief fleet."
The Oklahoma convention currently has two skid-steer loaders, which aren't large enough to handle the heavy job of installing the storm shelters. The shelters' outside dimensions are just over 6 feet-by-8 feet. The walls are 6 inches thick, and each shelter has the capacity to accommodate between 12-14 people.
"We'll use the backhoe for storm shelter installation and also for cleaning up debris," Porter said. "With the tornado effort here, we were able to take down a lot of damaged homes once they were declared a total loss, but we had to go rent either track hoes or backhoes to knock the houses down.
"It's so dangerous to do that with a Skid Steer, because it's so much shorter, and the roofs and walls of the structures could fall back over on it and the operator. We will now be able to use the new, much larger backhoe in fire and tornado recovery; it will be a valuable asset for us."
The existence of the new backhoe opens up a new avenue of service for volunteers, Porter said.
"We are seeking several laymen with back hoe operating skills to join us on a team to install these across the May 2013 storm track," he said.
"Those individuals interested in assisting do not have to be members of the disaster relief team. We just want several individuals who want their lives and skills to make an eternal difference in someone's life."
Interested persons may contact Porter at (405)942-3800, ext. 4337.
Bob Nigh is managing editor of the Baptist Messenger. Get Baptist Press headlines and breaking news on Twitter (@BaptistPress), Facebook (Facebook.com/BaptistPress) and in your email (baptistpress.com/SubscribeBP.asp).
Copyright (c) 2014 Southern Baptist Convention, Baptist Press www.BPNews.net


text:
At Jefferson College, we welcome new faculty and staff year-round. Many of these new members of the JCHS family relocate to the beautiful Roanoke Valley from other areas for their new positions. That means there may be spouses coming with them who will need to find employment of their own.
We want to provide resources to help these "trailing spouses" find work easily in our corner of the world. The NewVA Corridor Technology Council (NCTC)—a nonprofit group seeking to develop technology in areas ranging from Roanoke to Blacksburg—has created a resource just for this purpose and we want you to have access!
"Trailing Spouse Resources" webpage on the NCTC website.
The site includes links to Job Boards, Employment Agencies and many other important resources. To learn more about the NCTC, visit their website at www.thetechnologycouncil.com/.


text:
Nice work! Enjoy the show!
Link Copied!
One year on, Brexit talks finally begin
Opening Sequence
Trump ramps up pressure on Venezuela's military
Protesters demonstrate against Trump emergency order
Former acting FBI director decries Trump 'attacks'
VERBATIM: McCabe recalls Trump saying 'I believe Putin'
Kurdish forces seek help with Islamic State prisoners
The 'slave labor' behind Kim Jong Un's new utopia
Auto tariff report already sparking backlash
Fiascos litter the road to this year's Oscars
Closing Sequence


text:
So after a few weeks of stubbornly using pen and paper, I finally broke down and decided to give digital plein air painting a shot. This was done in an area found by my friend Rustam, and we went painting there last weekend.
I painted through the computer's battery life, so something like 1 hour 45 minutes??