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Here, we present homogeneous time resolved FRET (HTRF) as an efficient method for rapid detection of insulin secreted from cells.
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Aslanoglou, D., George, E. W., Freyberg, Z. Homogeneous Time-resolved Förster Resonance Energy Transfer-based Assay for Detection of Insulin Secretion. J. Vis. Exp. (135), e57531, doi:10.3791/57531 (2018).
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The detection of insulin secretion is critical for elucidating mechanisms of regulated secretion as well as in studies of metabolism. Though numerous insulin assays have existed for decades, the recent advent of homogeneous time-resolved Förster Resonance Energy Transfer (HTRF) technology has significantly simplified these measurements. This is a rapid, cost-effective, reproducible, and robust optical assay reliant upon antibodies conjugated to bright fluorophores with long lasting emission which facilitates time-resolved Förster Resonance Energy Transfer. Moreover, HTRF insulin detection is amenable for the development of high-throughput screening assays. Here we use HTRF to detect insulin secretion in INS-1E cells, a rat insulinoma-derived cell line. This allows us to estimate basal levels of insulin and their changes in response to glucose stimulation. In addition, we use this insulin detection system to confirm the role of dopamine as a negative regulator of glucose-stimulated insulin secretion (GSIS). In a similar manner, other dopamine D2-like receptor agonists, quinpirole, and bromocriptine, reduce GSIS in a concentration-dependent manner. Our results highlight the utility of the HTRF insulin assay format in determining the role of numerous drugs in GSIS and their pharmacological profiles.
The regulation of energy metabolism is fine-tuned by a major anabolic hormone, insulin. Insulin is synthesized and released by pancreatic beta cells in response to increased extracellular glucose levels. The released insulin triggers the uptake of glucose by insulin-sensitive tissues1,2. Physiologically, this is linked to the elevation of glucose concentration after a meal, followed by secretion of insulin to regulate glucose uptake. Disturbances in glucose homeostasis lead to metabolic impairments culminating in insulin resistance and ultimately in the onset of type 2 diabetes2,3,4.
Although insulin secretion has been extensively studied, its regulatory mechanisms remain poorly understood. A critical area of investigation has been identification of novel modulators of insulin secretion by beta cells5,6,7,8. These studies require a better understanding of the coupling relationship between glucose stimulation and insulin secretion. Therefore, the ability to accurately monitor and quantify the levels of glucose-stimulated insulin secretion (GSIS) has been essential. To date, however, only a limited number of methods were available to allow quantification of GSIS using insulin-secreting cell lines and/or pancreatic islets. One is radioimmunoassay (RIA), which utilizes radioisotope-tagged insulin and antibodies. The main limitations of this approach include safety issues due to the handling and disposal of radioactive materials. Additionally, this method is labor-intensive, involving multiple long washing and incubation steps. Enzyme-linked immunosorbent assay (ELISA) is another costly and labor-intensive approach that utilizes antibodies for insulin detection. Variation in antibody affinities and in the efficiency of recognizing insulin are limiting factors of this method and can affect the reproducibility of the results. Neither ELISA nor RIA was designed for high-throughput experiments. AlphaScreen is a homogeneous assay used for detecting and measuring levels of insulin secretion. AlphaScreen technology is based on the conversion of ambient oxygen into an excited oxygen singlet state that can react with chemiluminescent species, resulting in the generation of chemiluminescence. Because the assay is homogenous, many of the washing steps associated with RIA and ELISA are eliminated. However, the instability of the signal due to the nature of the reaction is a limiting factor that may affect the readout of the assay. (TR-PINCER, developed by Heyduk and colleagues9, is another homogenous approach to insulin measurement based on the binding of two separate antibodies to different epitopes on the insulin molecule. The antibodies are each chemically linked to double stranded DNA with short complementary single stranded DNA overhangs. Binding of the antibodies to insulin brings them together and leads to a double stranded DNA duplex. Each antibody is also associated with a respective donor or acceptor fluorophore, and the association of the DNA duplex brings together these fluorophores to generate Förster resonance energy transfer (FRET). One potential limitation of TR-PINCER, however, rests with the FRET itself. The inability to rapidly dissipate background fluorescence during the FRET reaction may lead to relatively high levels of background fluorescence and a low signal to noise ratio within the assay. Therefore, a need still exists for a reliable, robust, and cost-effective assay for quantifying GSIS in a high-throughput manner.
Recent advances in biophysics have culminated in the development of a homogeneous time-resolved fluorescence energy resonance transfer (HTRF) based assay. Specifically, while the energy transfer within the assay may be described as FRET-based, more accurately, HTRF relies on luminescence energy resonance transfer (LRET)10 which is the non-radiative transfer of energy between the donor and acceptor species11,12,13. This distinction is important, since the timing of a fluorescence or quenching based FRET interaction is much different than it is for LRET, though the same types of gating can be used for FRET and LRET. Moreover, the use of rare earth lanthanide cryptate compounds such as europium or terbium cryptate in HTRF produces long fluorescence half-lives12,14. This offers the unique advantage of the introduction of a time delay (µsec) between donor excitation and the measurement of emission from the acceptor (i.e., time-resolved assay). This time delay allows for sufficient time for the background fluorescence to dissipate prior to measurement of acceptor emission fluorescence. Consequently, the readout is free of non-specific fluorescence and thus, a high signal-to-noise ratio is achieved (Figure 1). Furthermore, the homogenous nature of HTRF eliminates the need for washing steps to wash off the unbound species, making the assay much more rapid than ELISA or RIA-based methods.
Figure 1: Schematic of the mechanism for HTRF insulin detection. Two independently generated monoclonal antibodies specifically recognize and bind to insulin at separate sites. These antibodies are conjugated to either the europium cryptate donor or the XL665 acceptor. Excitation of the donor at 337 nm results in emission at 620 nm. The resulting energy transfer causes XL665 to emit at a longer wavelength, 665 nm. Please click here to view a larger version of this figure.
Here, we provide a detailed protocol for using an HTRF-based approach to determine the levels of GSIS from INS-1E cells, a well-established insulin-secreting beta cell-derived rat insulinoma cell line15. In addition, this assay may be used for identifying the pharmacological profile of molecular regulators of insulin secretion. We apply this HTRF-based insulin assay to examine dopamine D2-like receptor regulation of GSIS. Increasing studies have revealed that the neurotransmitter dopamine is an important regulator of GSIS8,16,17,18,19,20,21,22. Dopamine affects GSIS in a negative autocrine/paracrine manner via actions on the dopamine D2-like receptors (D2, D3, D4 receptors) expressed at the surface of beta pancreatic cells8,16,19. Using this assay, we confirm dopamine's role as a negative regulator of GSIS and demonstrate that the dopamine D2-like receptor agonists bromocriptine and quinpirole also reduce GSIS.
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1. INS-1E Cells: Maintenance and Plating
- Maintain INS-1E cells in a humidified 37 ˚C/5% CO2 incubator and cultured with RPMI 1640 medium supplemented with 5% (v/v) heat-inactivated fetal bovine serum, 2 mM L-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 100 U/mL Penicillin/Streptomycin solution, 50 µM β-mercaptoethanol. Culture cells in 10 mL complete RPMI 1640 medium (per plate), until they reach 80 - 90% confluence, when they can be trypsinized and passaged or used for the insulin secretion assay.
- Day 1: Aspirate media and wash cells once with 5 mL of pre-warmed PBS. Add 0.5 mL of trypsin (0.025%) diluted 1:1 in 0.5 mL PBS to trypsinize the cells and incubate for 3 - 4 min at 37 ˚C. Deactivate trypsin by adding 9 mL complete media and transfer cells to a 15 mL centrifuge tube by pipetting.
- Pellet cells by centrifugation and re-suspend the cell pellet in approximately 5 mL fresh media.
- Take 10 µL of the re-suspended cells and mix with 10 µL Trypan Blue vital dye to check for cell viability. Count live and dead cells in 10 µL of this mixture using a hemocytometer. Viability level should be above 90%. Dilute the re-suspended cells in fresh media to 1 million cells per mL.
- Seed 0.5 mL INS-1E cells per well in a poly-L-Lysine pre-coated 24-well plate, at a density of 500,000 cells/well.
- Day 2: Remove media 18 - 24 h after plating and add 500 µL/well of fresh RPMI 1640 media. Incubate the cells for another 24 h to allow cells to fully recover from the prior passaging step. This additional time permits the cells to spread on the tissue culture plate.
2. Insulin Secretion Assay (Day 3)
- Prepare KRB buffer: 132.2 mM NaCl, 3.6 mM KCl, 5 mM NaHCO3, 0.5 mM NaH2PO4, 0.5 mM MgCl2, 1.5 mM CaCl2, and 0.001 g/mL bovine serum albumin (BSA), pH 7.4.
- Aspirate media from cells and wash twice with pre-warmed PBS.
- For the glucose starvation step, add 450 µL/well KRB (containing BSA) without glucose for 1 h at 37 ˚C/5% CO2.
- During the glucose starvation step, prepare serial dilutions of the drug(s) in KRB containing 200 mM Glucose (10x concentration).
- Prepare drug at 10x the final concentration in KRB supplemented with 200 mM glucose (also 10x the final assay glucose concentration). If the drug stock (plus the added 10x glucose) is in DMSO, make sure DMSO percentage is kept consistent throughout the assay (ideally final percentage of less than 0.1% DMSO).
- For dopamine and quinpirole treatments, use a final assay drug concentration range of 100 µM to 100 pM (from highest to lowest concentration), with the last point of the dose response containing no drug. For bromocriptine, use a final assay concentration range of 10 µM to 10 pM, with the last point of the dose response being the drug-free control.
- After glucose starvation, add drug serial dilutions to the assay to produce a dose response.
- Add 50 µL/well of each serial dilution to the corresponding wells (total assay volume 500 µL).
- For the glucose stimulation step, incubate cells with the respective drug serial dilutions (in the presence of 20 mM glucose) for 90 min at 37 ˚C/5% CO2. Include a set of control wells: (1) stimulation with 20 mM glucose alone in the absence of any additional drug and (2) cells that are neither stimulated with drug nor glucose (which provides a basal rate of secretion).
- After the stimulation step, carefully remove the supernatants (use directly or store at 4 ˚C).
NOTE: An additional 5 min gentle centrifugation step (600 x g, 1 min) may be introduced at this point to remove any remaining cells in the assay supernatants.
3. HTRF to Measure Insulin Secretion
- Dilute assay supernatants 1:10 in KRB (without BSA), preferably in clear 96-well plates.
- Prepare the insulin standard curve for the HTRF insulin assay (Table 1).
|Standard stock solution 500 ng/ml
|Serial dilutions
|Working [insulin] ng/ml
|30 µl Stock + 140 µl KRB
|30 µl STD 7 + 45 µl KRB
|30 µl STD 6 + 45 µl KRB
|30 µl STD 5 + 45 µl KRB
|30 µl STD 4 + 45 µl KRB
|30 µl STD 3 + 45 µl KRB
|30 µl STD 2 + 45 µl KRB
|45 µl KRB
|Note: STD Stock is 500 ng/ml
Table 1. Serial dilutions to make the insulin standard curve.
- Add the standard curve samples and the diluted assay supernatants to the HTRF plate. The measurement of secreted insulin by HTRF can be carried out in either a 96-well or a 384-well plate format, keeping in mind that the assay volume has to be re-adjusted. Use 10 µL/well sample in 96-well white half-area plates or 5 µL/well in a 384-well white low-volume, round-bottom plate (see Table of Materials).
- Prepare antibody mix in detection buffer (see Table of Materials) in a 1:2 donor (cryptate)/acceptor (XL-665) ratio.
NOTE: Further specific details about the HTRF assay are available from the manufacturer.
- Add antibody mix to the assay at 30 µL/well (for a 96 well-plate assay format) or 15 µL/well (for a 384-well plate assay format).
- Seal the plate and incubate at room temperature.
- Read plate after 2 h, 4 h, and/or overnight incubation with antibodies using the plate reader and the appropriate HTRF optic module (337 665 620 nm) (see Table of Materials and manufacturer's instructions). Set the integration start at 60 µs and the integration time at 400 µs. Use 200 flashes per well.
NOTE: These parameters were based on the use of our particular reader. The readout at 620 nm and 665 nm may vary between different instruments. This is one of the reasons we suggest using the 665/620 ratio. In calculating this ratio, any potential differences from reader to reader will be normalized and provide consistent values irrespective of the instrument used to measure HTRF.
4. Data Analysis and Normalization
- Calculate the insulin concentrations of the assay wells via extrapolation of ratiometric fluorescence readings (665 nm/620 nm) to a second order quadratic polynomial curve (Figure 2).
Figure 2: Insulin standard curve. Human insulin stock of known concentrations was used to generate the insulin standard curve. The resulting HTRF ratios (665 nm/ 620 nm) were plotted against the insulin concentrations. The data was best fit to a second order quadratic polynomial curve (R2 = 0.99996). This is a representative standard curve. Error bars = SEM. Please click here to view a larger version of this figure.
- With the extrapolated data as ng/mL of insulin secreted, normalize (insulin secreted in response to increasing ligand concentrations) to the average value of the % maximum insulin secretion assay wells (20 mM glucose alone condition).
- Use the curve fit (R2) from a single experiment to calculate the intraplate variation. Within the experiment, derived the individual R2 values from the intra-experimental duplicates, allowing calculation the standard error of the mean for the curve fit.
- For determining the interplate variation, use the data from at least three individual experiments to calculate the standard error of the mean for the R2 value of the collective curve.
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Representative Results
We validated our insulin HTRF assay by generating an insulin standard curve using purified human insulin standards of predefined concentrations (Figure 2). Generation of the standard curve permitted us to extrapolate the ratiometric fluorescence readings and thus to determine the secreted insulin levels in response to the drug treatments (Figure 2). Intraplate variation for curve fitting was minimal (R2 = 0.99996 ± 5.0 x 10-5) as was the interplate variation (R2 = 0.947 ± 5.5 x 10-5; n = 3 plates).
We next determined the basal levels of insulin secretion from INS-1E cells. Using a seeding density of 5 x 105 cells per well in a 24-well plate format, we found that cells secreted 54.53 ± 2.2 ng/mL of insulin in the absence of glucose over a duration of 90 min incubation in a static bath. Addition of 20 mM glucose to stimulate GSIS resulted in a significant enhancement in GSIS (112.4 ± 5.2 ng/mL insulin; p <0.0001) (Figure 3).
Figure 3: Estimation of basal versus stimulated insulin secretion. INS-1E cells were treated in the absence or the presence of high glucose (20 mM) stimulation for 90 min. A doubling in insulin secretion levels was detected in response to high glucose stimulation relative to the unstimulated cells (basal secretion condition; **** p <0 .0001, two-tailed unpaired t-test). N = 5; all assays were carried out in triplicates. Error bars = SEM. Please click here to view a larger version of this figure.
To investigate the role of dopaminergic D2-like receptor stimulation in GSIS within our INS-1E cell system, we incubated cells in the presence of D2-like receptor agonist drugs (dopamine, quinpirole, and bromocriptine) during the 90-min stimulation period with 20 mM glucose (Figure 4).
Figure 4: HTRF detection of dopamine D2-like receptor agonist inhibition of GSIS. INS-1E cells were treated with increasing concentrations of dopamine D2-like receptor agonists for 90 min in the presence of high glucose (20 mM; 37 ˚C). (A) Dopamine inhibited GSIS with an IC50= 1.78 ± 3.9 µM (R2 = 0.4355). (B) Bromocriptine was more potent than dopamine in reducing GSIS (IC50 = 13.9 ± 2.4 nM, R2 = 0.7501). (C) Quinpirole's potency was similar to dopamine (IC50 = 3.3 ± 3 µM, R2 = 0.3617). All data were best fit to a three-parameter fit sigmoidal curve. The '0 mM Glucose' point indicates basal insulin secretion by the INS-1E cells in the absence of high glucose (20 mM) stimulation. N >3; all assays were carried out in triplicates on separate experimental days. Error bars = SEM. Please click here to view a larger version of this figure.
The efficacy and potency of these agonist drugs was determined by graphing the insulin secretion curves. We found that in the absence of stimulation with high glucose, the INS-1E cells continued to secrete insulin, albeit to a markedly lower degree (indicated by the 0 mM condition), consistent with earlier reports15. Our results demonstrated that dopamine, the endogenous agonist of D2-like receptors, inhibited GSIS in a concertation-dependent manner (DA IC50 = 1.78 ± 3.9 µM; Figure 4A). In contrast, bromocriptine, a drug approved to treat type 2 diabetes23,24, was markedly more potent than dopamine in producing GSIS inhibition (IC50 = 13.9 ± 2.4 nM; Figure 4B). Lastly, though less efficacious than bromocriptine, quinpirole exhibited similar potency to dopamine in GSIS inhibition (IC50 = 3.3 ± 3 µM; Figure 4C).
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The HTRF insulin assay described here offers a rapid, efficient system to measure insulin secretion from a cultured cell-based system. Among its most important advantages, this assay offers a low background signal due to the high signal-to-noise ratio. Additionally, we have confirmed that the HTRF signal is stable for extended periods of time (>24 h)7. Nevertheless, since the insulin-binding monoclonal antibodies quickly reach binding saturation after addition to the assay, kinetics of antibody binding may be affected by ambient temperature or variations in secreted levels of insulin. Therefore, we read the assay plate at multiple time points (2 h, 4 h, and overnight) to precisely identify when the antibodies have reached full saturation and HTRF signal is most robust7. We have also recently shown the insulin antibodies do not significantly cross-react with proinsulin or c-peptide, suggesting that the assay is quite specific for mature insulin7.
An important step in the optimization of the assay involved establishing a broad dynamic range of insulin secretion. This dynamic range is defined as the difference in detected insulin during basal (unstimulated) versus stimulated insulin secretion. We find that an initial brief glucose starvation step (0 mM glucose, 1 h) preceding high glucose stimulation expands the dynamic range by priming the INS-1E cells to secrete more insulin during GSIS. In earlier work, we showed the assay's dynamic range to be from 0.17 ng/mL (limit of detection) to as high as ~110 ng/mL insulin with a signal to noise ratio of 10.027. Additionally, the amount of secreted insulin is dependent on initial cell number plated per well. We further optimized the assay by standardizing the passaging and handling of the INS-1E cells. Maintaining consistency in passaging schedule and plating in a specific plate type (e.g., 10 cm dishes) is essential for obtaining consistent secreted insulin levels.
Among potential sources of assay variability is bubbling in the assay solutions which interferes with the HTRF readout. An important source of these bubbles is the presence of bovine serum albumin (BSA) in the various KRB-based assay solutions. To troubleshoot this potential problem, reducing or even eliminating BSA significantly minimizes bubbles in the solution and therefore improves HTRF signal reliability. This is particularly important when generating the insulin standard curve and drug dilutions. Cell passage number-dependent differences in basal insulin secretion are another source of potential assay variability (data not shown). This is consistent with earlier work by Merglen and colleagues who also suggested that optimal insulin secretion is found between INS-1E cell passage numbers ~60 - 10015. Therefore, to minimize possible variability in insulin secretion related to cell passage number, we recommend limiting secretion assays to this well-defined cell passage range.
The primary limitation of the cell-based insulin secretion assay is the variability of the basal secretion intrinsic to the INS-1E cells. This may be related to the growing insulin-secreting cells outside of the physiological context of the pancreatic islet. Indeed, the other cell types within the islet including alpha and delta cells secrete numerous factors that may modulate basal and stimulated insulin secretion from beta cells in a local, paracrine manner25. The absence of these other islet cell types may disturb the tight regulation of insulin secretion, leading to greater variability than otherwise observed physiologically. Another potential limitation is the relatively small linear range of the assay, since insulin secreted in response to 20 mM glucose was almost saturating in some cases. As a result, in the undiluted cell supernatants, effects of insulin secretagogues on stimulated insulin secretion may be out of the linear portion of the calibration curve. This is, however, obviated by diluting the insulin-containing supernatant sufficiently to allow the insulin to fall into the linear range of the calibration curve.
We validated our assay by showing that agonism of dopamine D2-like receptors via dopamine, bromocriptine and quinpirole treatments inhibited GSIS in a concentration-dependent manner. These data allowed us to determine pharmacological properties of these drugs (i.e., IC50 and efficacy) in terms of their effects on GSIS. Our results are consistent with a growing body of evidence suggesting an important role for dopamine and dopaminergic signaling outside of the central nervous system20,21,22,26,27. Notably, our findings showing that the dopamine D2-receptor agonist bromocriptine potently inhibits GSIS, yet was recently approved by the FDA to treat type 2 diabetes24, raises the important question: how can this drug treat type 2 diabetes (T2D) if it lowers insulin secretion, at least acutely? One potential explanation may be related to bromocriptine's capacity to incur "beta cell rest." According to this hypothesis, since one of the cardinal features of T2D is an overall oversecretion of insulin, that ultimately may contribute to beta cell failure28. Moreover, we suggest that insulin oversecretion during T2D may also exacerbate insulin resistance by diminishing insulin sensitivity of organs including skeletal muscle, liver and adipose tissue. Thus, blocking or diminishing GSIS may result in gradual resensitization of these target tissues, making them more responsive to insulin and thus improving insulin sensitivity. Having an assay system amenable to high-throughput screening opens the door to future work to both dissect the intracellular signaling pathways mediated by dopamine D2-like receptors in pancreatic beta cells that are responsible for paracrine/autocrine GSIS inhibition as well as to further elucidate mechanisms by which drugs like bromocriptine may improve symptoms of diabetes.
The development of a cell-based insulin secretion assay that takes advantage of HTRF technology for insulin secretion and represents a significant advance in rapid, and efficient detection of insulin at significantly diminished cost relative and/or greater safety compared to older insulin detection assays relying on RIA and ELISA-based approaches. Indeed, though ELISA and RIA insulin assays are comparably sensitive and specific to the HTRF insulin assay, the homogenous nature of the detection, eliminates numerous additional washing steps present in these other assay systems7. Furthermore, the relative stability of the HTRF signal provides another key advantage for reliable insulin measurement.
Taken together, the HTRF insulin secretion assay has the capability of rapidly discriminating between the pharmacological properties of multiple drugs in the context of cellular insulin secretion. Moreover, this assay's relatively ease of use and robust signal make it amenable for high-throughput screening.
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We thank Nicolas Pierre (Cisbio Bioassays) for helpful advice and Dr. Pierre Maechler (University of Geneva) for generously providing INS-1E cells. This work was supported by funding from the Department of Defense (grant PR141292 to Z.F.), and the John F. and Nancy A. Emmerling Fund of The Pittsburgh Foundation (to Z.F.).
The authors have nothing to disclose.
|Catalog Number
|96-well white half area plate
|Greiner Bio-One
|384-well white low-volume, round-bottom plate
|Insulin High Range Kit
|Cisbio Bioassays
|10,000 test HTRF-based insulin assay kit
|PHERAstar FSX plate reader
|BMG Labtech
|Plate reader adapted for HTRF-based assay readings
|Plate sealers
|Fisher Scientific
|To seal plate while antibodies incubate
|Hausser Scientific
|RPMI Medium 1640 (1x)
|[+] L-glutamine
|Trypsin 0.05%
|0.53 mM EDTA, (-) sodium bicarbonate
|DPBS (Dulbecco's phosphate buffered saline)
|Without calcium and magnesium
|Fetal Bovine Serum
|Sodium pyruvate
|Penicillin/Streptomycin solution 100x
|Trypan Blue Stain (0.4%)
|Dopamine hydrochloride
|Bromocriptine mesylate
|(-)-Quinpirole hydrochloride
|Bovine Serum Albumin
|DMSO (Dimethyl sulfoxide)
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- Vetere, A., Choudhary, A., Burns, S. M., Wagner, B. K. Targeting the pancreatic beta-cell to treat diabetes. Nat Rev Drug Discov. 13, (4), 278-289 (2014).
- Despres, J. P., Lemieux, I. Abdominal obesity and metabolic syndrome. Nature. 444, (7121), 881-887 (2006).
- Kahn, S. E., Hull, R. L., Utzschneider, K. M. Mechanisms linking obesity to insulin resistance and type 2 diabetes. Nature. 444, (7121), 840-846 (2006).
- Barg, S. Mechanisms of exocytosis in insulin-secreting B-cells and glucagon-secreting A-cells. Pharmacol Toxicol. 92, (1), 3-13 (2003).
- Braun, M., Ramracheya, R., Rorsman, P. Autocrine regulation of insulin secretion. Diabetes Obes Metab. 14 Suppl 3, 143-151 (2012).
- Farino, Z. J., et al. Development of a rapid insulin assay by homogenous time-resolved fluorescence. PLoS One. 11, (2), e0148684 (2016).
- Simpson, N., et al. Dopamine-mediated autocrine inhibitory circuit regulating human insulin secretion in vitro. Mol Endocrinol. 26, (10), 1757-1772 (2012).
- Heyduk, E., Moxley, M. M., Salvatori, A., Corbett, J. A., Heyduk, T. Homogeneous insulin and C-Peptide sensors for rapid assessment of insulin and C-peptide secretion by the islets. Diabetes. 59, (10), 2360-2365 (2010).
- Heyduk, T. Measuring protein conformational changes by FRET/LRET. Curr Opin Biotechnol. 13, (4), 292-296 (2002).
- Degorce, F. HTRF((R)): Pioneering technology for high-throughput screening. Expert Opin Drug Discov. 1, (7), 753-764 (2006).
- Degorce, F., et al. HTRF: A technology tailored for drug discovery - a review of theoretical aspects and recent applications. Curr Chem Genomics. 3, 22-32 (2009).
- Mathis, G. HTRF(R) Technology. J Biomol Screen. 4, (6), 309-314 (1999).
- Daijo, J. E., Sportsman, J. R. A time-resolved fluorescence immunoassay for insulin in rodent plasma. J Pharm Biomed Anal. 19, (3-4), 335-342 (1999).
- Merglen, A., et al. Glucose sensitivity and metabolism-secretion coupling studied during two-year continuous culture in INS-1E insulinoma cells. Endocrinology. 145, (2), 667-678 (2004).
- Ustione, A., Piston, D. W. Dopamine synthesis and D3 receptor activation in pancreatic beta-cells regulates insulin secretion and intracellular [Ca(2+)] oscillations. Mol Endocrinol. 26, (11), 1928-1940 (2012).
- Ustione, A., Piston, D. W. A simple introduction to multiphoton microscopy. J Microsc. 243, (3), 221-226 (2011).
- Ustione, A., Piston, D. W., Harris, P. E. Minireview: Dopaminergic regulation of insulin secretion from the pancreatic islet. Mol Endocrinol. 27, (8), 1198-1207 (2013).
- Rubi, B., et al. Dopamine D2-like receptors are expressed in pancreatic beta cells and mediate inhibition of insulin secretion. J Biol Chem. 280, (44), 36824-36832 (2005).
- Garcia-Tornadu, I., et al. Disruption of the dopamine d2 receptor impairs insulin secretion and causes glucose intolerance. Endocrinology. 151, (4), 1441-1450 (2010).
- Garcia-Tornadu, I., et al. New insights into the endocrine and metabolic roles of dopamine D2 receptors gained from the Drd2 mouse. Neuroendocrinology. 92, (4), 207-214 (2010).
- Lopez Vicchi, F., et al. Dopaminergic drugs in type 2 diabetes and glucose homeostasis. Pharmacol Res. 109, 74-80 (2016).
- Kalra, S., Kalra, B., Agrawal, N., Kumar, S. Dopamine: The forgotten felon in type 2 diabetes. Recent Pat Endocr Metab Immune Drug Discov. 5, (1), 61-65 (2011).
- Mahajan, R. Bromocriptine mesylate: FDA-approved novel treatment for type-2 diabetes. Indian J Pharmacol. 41, (4), 197-198 (2009).
- Caicedo, A. Paracrine and autocrine interactions in the human islet: more than meets the eye. Semin Cell Dev Biol. 24, (1), 11-21 (2013).
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- de Leeuw van Weenen, E. J., et al. The dopamine receptor D2 agonist bromocriptine inhibits glucose-stimulated insulin secretion by direct activation of the alpha2-adrenergic receptors in beta cells. Biochem Pharmacol. 79, (12), 1827-1836 (2010).
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Bodo Tribal Community of Assam is considered as the earliest immigrants of Assam. & plays very important role in the culture and tradition of the state.
Bodo Tribe is known to be the earliest immigrants in Assam. This ethnic community is mainly concentrated in Brahmaputra valleys. It is the largest minority group of the state. It is believed that this tribe arrived in Assam from the Tibet through Bhutan passes. As, the primitive settlers of Assam valley, the bodo community is considered to be the most traditionally and culturally rich community of the state.
Bodo people speak Bodo language. For writing they used Roman script and Assamese script. Now they have taken up the Nagari script for their writing.
Culture and Religion: Their rich culture incorporates elements like dancing, singing etc. which reflects the entire they have many religious practices and beliefs, among which Bathouism has special importance.
Food habit of Bodo tribe: This community is very fond of conventional drink called Zu mai. When people visit their house they used to offer this drink as a kind of respect. Their main food consists of non-vegetarian dish like pork and fish. Oma Bedor, Onla and Narzi are their main cuisines.
Fairs and festival of Bodo tribe "Baishagu is the main festival of Bodo community. It is celebrated during the month of April every year.
Economy of Bodo tribe: Earlier, this tribe used to practiced cultivation and farming. Among them rice farming, tea plantation, poultry and silkworm rearing were the main occupation. Bodo people are expert in Bamboo craft. Weaving is the main occupation of Bodo women.

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Kirkland, Wash – Kirkland Police Chief Cherie Harris announced this morning that the department has initiated a formal investigation to determine whether officers that responded to a call for suspicious activity at a Menchie's frozen yogurt franchise in the Totem Lake neighborhood of Kirkland acted in accordance with department protocol.
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The City of Kirkland will issue additional information after the internal investigation has been completed. The police report from the incident is available online, as is the 911 call that was placed by the owner of the Kirkland Menchie's.
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Lehigh Valley Higher Ed Web Design Team Partners with Albright College
Higher ed web design team KDG helped Albright College launch its brand-new website.
Allentown, PA, December 21, 2017 --(
)-- Nestled in the heart of Reading, PA, Albright College is a liberal arts school where students are free to discover and explore their interests both in and out of the classroom. However, when the college embarked on a new brand and marketing strategy to highlight this, a new website was also in order. With a Wordpress design already in place, Albright turned to KDG for help in migrating more than 3,000 pages of old web content to its new site.
"There was a lot of content that needed to be organized, shortened, and migrated in a way that fit the design, matched the school's vision, and also made it easier for audiences to navigate," explains Dan Stover, UI/UX Manager at KDG.
Both Albright and KDG's higher ed web design team quickly discovered that the migration project would be no easy task. A new sitemap needed to be created and a series of spreadsheets were used to schedule an extensive "migration strategy." During this process, old web pages were matched up with new pages and all old content had to be cut, condensed, and edited so it could fit into the new sitemap. By the end of the process, 3,000 pages had turned into 400.
"Our new site was a long time coming and was totally worth the wait," says David Markowski, Web Manager at Albright. "It looks great, functions beautifully and responsively, and is easier to maintain than our old site."
In addition to content migration, Albright and KDG ensured that the site was both accessible and user-friendly with several other features. Sign-up and application forms were migrated, alt tags and other ADA regulations were implemented, and device testing was completed to make sure the site worked well for all users.
"The meat of the design was there at the onset, but KDG added some extra touches that enhanced the user experience even more," continues Markowski.
Now, visitors to Albright's site have an informative, responsive, and engaging place to discover all that Albright has to offer.
About KDG: KDG (formerly the Kyle David Group) is a leading provider of web development for higher education. With over 15 years of experience in using technology to help education clients improve enrollment, campus climate, and alumni engagement, KDG has developed a reputation for being able to see and respond proactively to changing markets. Most recently, KDG won wide acclaim for developing unique and engaging inbound marketing for higher education crowdfunding campaigns. Learn more at http://www.kyledavidgroup.com/.
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Price: $20.00
Quantity: 1 available
Book Condition: Fine
FINE condition. SOFTCOVER. 1988. UNCORRECTED PAGE PROOFS as issued by publisher for the hardcover edition, April 1988. AS NEW, bound in white coated wraps, square and tight, UNREAD. As issued, two slits on front cover to hold a promotional letter opener, not included. Serial killer in Jerusalem.
Title: BUTCHER'S THEATER
Categories: HERE'S THE PROOF!, AUTHOR J~K~L,
Edition: Uncorrected Proof
Publisher: NY, Bantam: 1988
Binding: Soft Cover
Book Condition: Fine
Jacket Condition: Original Wraps
Item: 1.00 Item
Seller ID: 000349
Keywords: NOISBN, Uncorrected Proof, Jerusalem, Mystery, Detection, Scene Of The Crime


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Servando Cabrera Moreno
Servando Cabrera Moreno (1923 – 1981) Cuban
Servando Cabrera Moreno born in 1923 died in 1981. Servando Cabrera he studied painting at the Fine Arts Academy of San Alejandro, where he graduated in 1942. In September, 1943 he presented his first solo show at the Havana Lyceum. In the years that followed he exhibited works related to his academic experience at the annual salons of the Circle of Fine Arts and other institutions. In 1946 he travelled to the United States and took a course at the Art Students' League in New York. Among other artists, he discovered Picasso, who was to become – according to the painter himself – the greatest influence in his work throughout his life. In 1949 he travelled to Europe, visited museums and attended the Grande Chaumière in Paris. His first rupture occurred in those years with oil paintings from 1950 and 1951 in which a geometrical steadiness of cubist origin led him close to abstraction. Afterwards, the influence of Miró and Klee dominated his brief but intense abstract experience (1951-1954), whose results he exhibited in Spain and France. Since 1959 the themes of the Revolution became part of his painting. Works of his great epic cycle reached their climax with the series Héroes, jinetes y parejas. In 1970 he left aside the violent expressionism and began with the styling of torsos, parts of the human body and couples, which are part of the cycle of eroticist painting in which he was to work from that moment on. Works of his are in numerous museums, among them in Sofia, Bucharest, Bogota, the University of Tampa, Venezuela, Peru, the United States, Spain, Italy, Mexico, England, and in institutions and private collections in Cuba.
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Car Crafting: 10 Toys That Made America Great!
10 Toys That Turned Kids Into Car Crafters
Things sure have changed in the half century since I was a kid. When I think what kids do today versus what we did back in the day, it becomes clearer why attitudes are so different now when you consider the toys we had. There are a ton of toys that shaped a generation of hot rodders that, because they accelerated the evolutionary process too quickly, are frowned upon today. Here's my Top 10:
Bottle rocket. Essentially a high-speed mortar round filled with water and air and powered by a hand-operated pump, these high-impact plastic projectiles were lofted skyward with breathtaking force. Easily 10 times more dangerous than a BB gun, one of these could put your sister in the hospital.
Chemistry set. Any kid getting a chemistry set in 2020 is likely a candidate for child protective services, but in 1970 it was the equivalent of getting a good STEM education. The Rutherford-Bohr model of the atom, balancing equations, and molar quantities are still important to grasp, but so is keeping the house from burning down. I wasn't a chemistry nut, but I did like
Firecrackers and cherry bombs. These were already considered dangerous at the time, but you could still get 'em if you knew where to find 'em. A decent-sized box could be had for about the cost of your allowance, and they were most fun when used to blow stuff up. My preference was putting them in plastic ship models and then floating them down the creek. My combustion education got a shot in the arm.
Wood-burning set. I got one, and actually hated it. The idea is to use a 700-degree electric soldering iron (no danger in that!) to burn designs into plywood. When used as advertised, it caused impenetrable boredom. Its main usefulness was to learn what different substances smelled like when burning.
Model kits. Beyond the obvious built-in hot rod education, model kits gave us up-close and personal experience with model glue, paint, and other VOCs. And not just car models, either. The best were balsa-wood models that used a glue called "dope," and, well, it was.
Cap gun. The cap gun was for little kids (you "moved up" to a BB gun, then bullets ), and used a roll of percussion caps made from potassium perchlorate, sulfur, and antimony sulfide. After shooting a half roll of these with about a 50 percent success rate, you'd ditch the "gun," and just pound the caps with a big rock.
Real steam engine. Yes, they actually let kids play with these, and my buddy Skip had one in the second grade. (Best show-and-tell ever.) Add water to the boiler, put a carbon fuel puck in the furnace, oil the bearings, light it, wait for pressure to build, and let it rip!
Radio set/circuit kit. My dad is an electronics geek and I am not so much, which is why I think I'm related to the milkman. Like all good dads in 1971, mine took me to RadioShack and got me the then-ubiquitous 100-in-1 Electronic Projects kit. While I never learned how to build a proper RLC circuit (milkman, remember?), I did learn that electrons can bite. I successfully scaled this experiment up 10 years later when I welded my hand to a screwdriver while working on a Volkswagen voltage regulator.
Mini-bike. There is perhaps no other toy in the young man's or woman's arsenal that foreshadows life in Car Craft land more than the mini-bike. (The Richie McRichersons of the world got go-karts.) You were large and in charge if you were 11 and had a mini-bike with gas in the tank.
Anything with a 0.09ci two-stroke glow engine. Don't let its small size belie its importance! Where on earth could you combine three of the world's most dangerous things in one toy and have it be legal? If you do it right, you'll have high-speed flying apparatus with virtually no control that runs on both nitromethane (a high explosive) and castor oil (containing the nerve agent ricin). I had a Cox Sopwith Camel but no nitro. Later, I discovered Mom read the directions after Dad bought it and vetoed the fuel!


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Wednesday, 06 January 2010 01:42 PM
Morgan Stanley has won a legal battle with former subsidiary Discover Financial Services over nearly $800 million that Morgan Stanley says it was owed by the credit card company.
New York state Supreme Court Justice Barbara Kapnick has ruled Discover Financial must pay Morgan Stanley a special dividend the pair negotiated when Morgan Stanley spun off Discover in 2007.
At the time of the spinoff, the two agreed to share money received from an antitrust lawsuit Discover had pending against Visa and MasterCard. Discover settled the antitrust case for $2.75 billion.
However, Discover refused to share the settlement with Morgan Stanley, claiming it interfered in the Visa and MasterCard settlement.
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Eric Ludwig for
Provo District 4
Why am I running?
My name is Eric Ludwig, and I am running for Provo City Council to represent District 4. I first moved to Provo in 2012 to attend school at Brigham Young University. In the seven years since I moved to Provo, the city has become home. I am running for City Council because I believe I can bring a unique perspective to our local government.
How can you help?
Donate to my campaign by sending a venmo to @eric4provo. Please put your address in the notes as city law requires an address to be associated with each donation.
Get in Touch
Eric Ludwig
Eric Ludwig for District 4


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More Horror Prompts
Here are a few more horror prompts for anyone having trouble with their horror stories. Our current challenge is a short (1000-word) haunted house piece, so perhaps not all of the prompts will suit—but, then again, if you're in the mood to challenge yourself even further, it could be fun to see what kind of story you can craft from one of these prompts within the limitations already set. Also, plot generators, anyone? They're so much fun to play with. This first one is specifically horror and VERY involved—perhaps better suited to a longer work, such as a novel or a … Continue reading More Horror Prompts


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London, Wandsworth. 2016
With its mix of new homes, modern office space and active retail frontages, Upper Richmond Road improves the streetscape and enlivens the heart of London's SW15 to provide a catalyst for urban regeneration in the area.
Composed of a cascading series of interlocking volumes containing residential accommodation above retail and commercial opportunities the two blocks are connected via a six storey link block.
Together the two phases provide 113 double aspect apartments and an array of balconies, winter gardens and terraces.
The building's stepped dovetailing design language is expressed coherently at multiple scales, from the overall massing through to the relief brick texture.
Five AHMM projects have been shortlisted for this year's RIBA Regional Awards. Weston Street, Upper Richmond Road, Hawley School and White Collar Factory are among the 93 buildings shortlisted within the London region, while Liverpool's Royal Court Theatre has been shortlisted in the RIBA North West region. Shortlisted projects will be visited over the next month, and winning projects will be announced in May. A RIBA Regional Award is given to a UK building for its regional importance as a piece of architecture; winners are considered for RIBA National Awards, announced later in the year. Find out more about the RIBA Awards and see past winners here.
A total of nine projects by AHMM have been shortlisted for this year's awards in the RIBA's London, East, North West and South regions. In London, The Library at Willesden Green, Grand Union Studios/The Ladbroke Grove, New Scotland Yard, 1 King William Street, Upper Richmond Road and Google 6 Pancras Square have been visited by regional judges; Liverpool's Royal Court was shortlisted in the North West region; Alconbury Weald Club in RIBA East; and Barns Place in RIBA South. Regional winners will be announced at ceremonies in May.


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Posts by tag
2 posts
Raags to Riches: Globalism and Vernacular in Contemporary Pakistani Rap
Languages such as Punjabi and Pashto offer a versatile slang to many a Pakistani street. And the streets are where lyrics overwhelmingly situate rap. The introduction of Punjabi-American rap into Pakistan invigorated its rappers by furnishing them with a vernacular familiar with their experiences. Far from a shadow of its American progenitor, contemporary Pakistani rap thrives in engaged dialectic with local and global trends in the genre.


text: These three spiraling hearts form a perfect row just beneath your neckline and are crowned by a stunning high quality labradorite cabochon stone hand set in 18K yellow gold with a silver back. There are hidden six pointed stars as a texture on each heart, and the silver has been oxidized to accentuate the textures. It is a substantial necklace– as a nice piece of jewelry should be, but it is not too heavy. Each labradorite stone is hand picked and may vary slightly. This piece is a one-of-a-kind, but custom orders are available. It has been worn for special occasions, and every day. As a gift for yourself, for your wedding day, Mother's Day, Hanukkah or birthday, it is sure to be treasured and worn. It comes with a sturdy sterling silver chain in your choice of length, and gift boxed. Enjoy!


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Those who claim to know Buhari very well, praised him for his Spartan nature, saying he remains the same man that he was many years ago despite presiding over billions of Nigeria's cash.
Details of how President Muhammadu Buhari applied and got bank loan of N260, 000 to build his first and only house in Kaduna, have emerged, with some Nigerians questioning whether his Spartan life of 1984 when he served as head of state is still the same as the current Nigeria's president.
The bank document, which Saturday Vanguard got exclusively, shows clearly that Buhari had resorted to taking a bank loan to be able to build his first house in Kaduna, having not been able to raise enough cash for such a project through all other means.
After applying for the cash, which came with interest, Buhari, a Major General in the army at the time, accepted all the terms and conditions imposed by Union Bank, which provided the N260,000 loan to him and laid out the conditions attached to the offer.
In the loan document dated June 1, 1984, and addressed to Major General Muhammadu Buhari, Head of State and Commander-in-Chief, Dodan Barracks, Lagos, the bank made it clear that the loan was approved for the building of his house.
The letter signed by one M. D. Yusif, who was the manager of Union Bank at the time, the bank said: "Dear sir, we are pleased to advise that the bank has agreed to renew your facility as follows: Building Loan-N260,000, has been marked for review on the 2nd of March 1985 but in accordance with the normal banking practice; it is repayable on demand and subject to review or withdrawal at any time the bank's option.
"Interest will be charged for the time being, at the rate of 11 percent per annum with monthly rents. Commission on turnover will be charged at 20 percent.
"The facility has been renewed subject to the following conditions: All the security items have been holding before now continue to support this lending and ordinary account should be kept strictly on creditor's basis while loan outstands.
"Kindly signify your acceptance of the terms and conditions of the renewal by signing and returning the duplicate copy of this letter," M. D. Yufif, said.
But as it appeared that Major Gen. Buhair was desperately in need of the loan, he quickly signed and returned the letter to Union Bank, which immediately released the cash to him for the building project in Kaduna.
Although this gesture has given out Buhari as a man of Spartan nature, many Nigerians are unconvinced if he would have take the loan today, given that he has made a lot of money from being the president of Nigeria for almost four years and surrounding himself with people who are said to be money-conscious and ready to do anything to get rich quick.
However, those who claim to know Buhari very well, praised him for his Spartan nature, saying he remains the same man that he was many years ago despite presiding over billions of Nigeria's cash.
A Kaduna-based politician, Aliyu Musa, who claims to have know Buhari from childhood, told Saturday Vanguard in Kaduna on Friday that Buhari remains incorruptible despite all the posts he has held and the passage of time given his austere background and fear of God not to dip his hands into public till.
"We, who are Buhari's friends, are proud of him and we will continue to vouch for him as an honest, contented and sincere leader who is not driven by ill-gotten wealth and that is why he went for commercial bank loan to build his first house instead of looting the public treasury as most leaders would have done.
"We know that despite being Nigeria's president at a time when everybody wants to steal and enrich themselves, Buhari will remain steadfast to his creed not to steal a kobo from the peoples treasury," Musa stated.
But second republic lawmaker, Junaid Mohammed, told Saturday Vanguard that Buhari might not be corrupt by taking money from the public till but has thrown himself into the ring by surrounding himself with questionable elements, who do not believe in his anti-corruption war.
"We want Buhari to do away with nepotism and retainership of a cabal that does not work in the interest of Nigeria because corruption extends beyond stealing public funds to conferring undue advantage on a particular set of people and tribe at the expense of other groups," Junaid said.
Source: Vanguard


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The search found 45 results in 0.01 seconds.
We propose a synthesis of two lines of sociological research on boundary spanning in cultural production and consumption. One, research on cultural omnivorousness, analyzes choice by heterogeneous audiences facing an array of crisp cultural offerings. The other, research on categories in markets, analyzes reactions by homogeneous audiences to objects that vary in the degree to which they conform to categorical codes. We develop a model of heterogeneous audiences evaluating objects that vary in typicality.
This study examines the relationship between interreligious contact and negative attitudes toward the religious outgroup among minority Christians and majority Muslims in Indonesia. It answers two research questions: Does interreligious contact reduce negative outgroup attitudes equally for minority Christians and majority Muslims? Are mediation by perceived group threat and moderation by perceived discrimination equally important for religious minorities and majorities?
This article offers a theoretical comparison between field and ecology, as developed by Pierre Bourdieu and the Chicago School of sociology. While field theory and ecological theory share similar conceptualizations of actors, positions, and relations, and while they converge in their views on structural isomorphism, temporality, and social psychology, they are quite different on several other scores: power and inequality, endogeneity, heterogeneity, metaphorical sources, and abstraction.
Social scientists have drawn on theories of embeddedness to explain the different ways legal, political, and cultural frameworks shape markets. Often overlooked, however, is how the materiality of nature also structures markets. In this article, I suggest that neo-Polanyian scholars, and economic sociologists more generally, should better engage in a historical sociology of concept formation to problematize the human exemptionalist paradigm their work upholds and recognize the role of nature in shaping markets and society.
Debates about science and religion—whether they conflict and how they factor into public opinion, policies, and politics—are of longstanding interest to social scientists. Research in this area often examines how elites use science and religion to justify competing claims. But, how do members of the public more generally incorporate science and religion into their worldviews? The assumption that science and religion inherently conflict with one another has come under increasing scrutiny and recent studies reveal that science and religion are more compatible than previously assumed.
Contemporary social research often has located religion's public influence by focusing on individual or collective religious actors. In this unitary actor model, religion is a stable, uniform feature of an individual or collectivity. However, recent research shows that people's religious expression outside religious congregations varies by context.
For many Americans, safety pins have suddenly appeared everywhere: Pinned to shirts, posted to Facebook, or worn by celebrities. When I started wearing one a handful of strangers asked "what the heck are these safety pins all about?" This is the challenge of new symbols. Before they can work people need to know what they mean.
C'est avec grand plaisir que je vous acceuille dans mon bout de pays, "La Belle Province." That we meet in Montréal to debate "Culture, Inequality, and Social Inclusion across the Globe" is particularly fitting as these very topics have been at the center of the construction of the Canadian community since 1608, in the context of multiple ethno-national and colonial conflicts. Today, many perceive Canadian society as exemplary when it comes to collective wellbeing, immigration policy, and multiculturalism.
More than 5,500 sociologists will convene in Montreal this August to explore scientific research relating to social inequality and many other topics, as part of the American Sociological Association's 112th Annual Meeting. This year's theme, "Culture, Inequalities, and Social Inclusion across the Globe," draws attention to the nexus of culture, inequalities, and group boundaries in order to promote greater social inclusion and resilience, collective well-being, and solidarity in Canada, the United States, and globally.


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I have 6 Free General Admission Ticket for the Georgia Quilt Show. The first 6 who respond to the blog with your Name and Address will get One free Pass to the Show.
Hmmmm, here I am and I know I can't go, already have a commitment those days, however, our guild is hosting a quilt camp this weekend and I would be happy to offer them to other quilters attending. Whatcha think?
Claudia Taylor
P O Box 3662
Chattanooga, TN 37404


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Featured Interview
DBT 311: Randy L. Schmidt – Carpenters: An Illustrated Discography
This is one of the most beautiful books I have seen in a while, and you learn so much about The Carpenters and the music. The color pictures are stunning, to say the least. Randy Schmidt is so … [Read More...]
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MICHAEL LEARNED- OLIVIA WALTON FROM THE WALTONS- How exciting to talk to this wonderful actress.. she is appearing soon in a theatrical version of DRIVING MISS DAISY, and i would love to see it. … [Read More...]
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Wow, what a cute small book filled with so many interesting stats about the top 1000 greatest players of all time. Now keep in mind, this is the opinion of Derek Gentile, and you might disagree with … [Read More...]
DBT 0221: Reggie Leach – The Riverton Rifle
David talks with Reggie Leach, who had a 14-year career in the NHL and won back-to-back Stanley Cup championships. Reggie speaks candidly about his struggles with alcohol, which ended in 1988 When … [Read More...]
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DBT 313: Robert Dugoni – A Cold Trail (Tracy Crosswhite Book 7)
Robert Dugoni is one of my favorites. This is the seventh Tracy Crosswhite book to come out, and all are just so great. Dugoni really writes so well, and Tracy really jumps out of the page into your … [More...]
DBT 312: Bonnie Kistler – House on Fire
What a nice lady to interview. I met Bonnie Kistler years ago in Pennsylvania, but she has since changed her name. What a powerful book this is! A wonderful story, with characters that come to life, and one of … [More...]
DBT 310: Stephanie Evanovich – Under the Table
I don't usually interview a romance writer, so this was unusual. But I found myself enjoying this book, and Stephanie Evanovich is very outspoken and likable. Yes, Stephanie is related to Janet Evanovich, and we … [More...]
DBT 309: Mick Finlay – Arrowood (Book #1, The Arrowood Series)
What a great new series, set during the Sherlock Holmes era, but Arrowood, the lead detective, hates Sherlock Holmes and that is what makes this series so much fun. Mick Finlay is a major new talent with MIRA … [More...]
DBT 308: Alan Bradley – Golden Tresses of the Dead (A Flavia de Luce Novel)
Alan is such a great talent, so intelligent and a fun guy to interview. This is book number 10, it is hard to believe. It all started with THE SWEETNESS AT THE BOTTOM OF THE PIE, and somehow FLAVIA has totally … [More...]
DBT 0307: Charles Todd – The Black Ascot (An Inspector Ian Rutledge Mystery)
It is always a pleasure to talk to Charles and Caroline Todd, the mother / son writing duo behind the Ian Rutledge mystery series. This is book number 21 in the series, and it is hard to believe there have been … [More...]
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DBT 0304: Chris Mooney – The Snow Girls
This is one of the most haunting books I have read in a while, with a HUGE shock at the end. I hope you get to read it. Please listen to this interview. Chris is so articulate, smart and fun to talk to. A regular … [More...]


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EPSRC and BBSRC Centre for Doctoral Training in Synthetic Biology
In the following table, contact information relevant to the page. The first column is for visual reference only. Data is in the right column.
|University of Oxford
|Partner Organisation:
|University of Bristol
|Partner Organisation:
|University of Warwick
|CDT, University of Oxford
|Related theme:
The EPSRC Centre for Doctoral Training in Synthetic Biology will combine leading expertise in engineering and the physical and life sciences at the Universities of Oxford, Bristol and Warwick to create the next generation of industrial and academic leaders in this important new field. Synthetic Biology provides opportunities for revolutionary advances in fundamental science and industrial technology. The four-year programme of research and training has strong industrial links and will be highly multi-disciplinary, accepting students from a wide range of scientific backgrounds and focusing on the application of engineering principles to the design of biologically based parts, devices and systems.


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47 samurai in Sengakuji temple
Japanese culture is as strong as a magnet. The high-speed trains running on 500km/h on the magnetic rails form the extraordinary unity with the temples of 500 years old. History and future dances rhythmically in Japan. Millions of people come to the Land of the Rising Sun every year to witness this dance.
One of the most visited places in this country is located in Tokyo. Tokyo is a megacity. Residents and the tourists visiting the city make the mega chaos consisting of sounds and motion. Guests who want to hear the inner voice visit Shinagawa station in the south part of Tokyo.
After a 15-minute walk from this station, you will reach the Buddhist temple Sengakuji. There is a magical silence on the walls of this temple. You will forget all the hassle of the world while sitting on one of the benches nearby. The walls of the ancient temple, trees, and the spirit of history will embrace you.
Sengakuji is not like other temples. There are tombs of 47 Samurai in the Akō-rōshi area of the temple. The Japanese call the Samurai a ronin. Ronin is the word used to describe a leaderless samurai. Probably, you also have seen the ronin in the Hollywood movie starring with Keanu Reeves.
Let's go back to 318 years ago. We should go to Edo Castle next to the temple. The story of Ōishi and his Samurai, the love of all the Japanese people, begins in this castle. In the Edo castle, Asano, the leader of Ōishi, lost his temper and attacked a nobleman named Kira with a dagger. He wounds Kira's face. Asano's anger was not groundless. Kira repeatedly insulted him in front of his men and tried to discredit. Asano could restrain himself no longer and wanted to kill Kira, who had a higher rank. According to the rules of the shogunate, no one in Edo Castle has the right to draw a sword to a state official. So, Asano had to kill himself by seppuku for his deed. His lands were confiscated from his family. And his 300 samurai were made a ronin. Sengakuji people were furious because of all these. Everyone condemns
Kira. According to the Samurai internal code, Asano must be revenged. However, any Samurai who will take revenge will be executed according to the shogunate rules.
Leaderless Sengakuji and 300 samurai became dispersed. Ōishi, the leader of the Samurai and 46 ronin banded together, swearing a secret oath to avenge their master.
On December 14, 1702, Kira was attacked by ronin at his mansion. Ōishi cut off his head and brought to Sengakuji together with his ronin. The Samurai were very happy. Asano's revenge was taken, and his honor was restored. The Samurai could perform their duties again. Ōishi and his people waited a year and six months to eliminate Kira's suspicions so that they could take revenge.
The news of this incident was spread all over Japan. According to shogunate rules, ronin who were supposed to be executed as the criminals were allowed to honorably commit seppuku. Ōishi and his men wrote the history.
Every year on December 14, thousands of people from different parts of the country go to the Akō-rōshi cemetery in the courtyard of the temple to commemorate 47 samurai.
The place where 47 samurai sleep is a combination of honor, dignity, loyalty, and the sanctity of being a samurai.
If you visit Tokyo in December, absolutely go to Shinagawa Station. Step into the temple, which is a 15-minute walk from the station at 8 am in the morning. Sit down on the bench and give your soul a rest.
Then, buy 47 pieces of "Senko" sticks and burn them on the head of the samurai tombs. Senko smoke will make you feel weird. There is also a single-room museum in the temple. There are the armors and letters of Samurai in the room called Akogishi Kinenkan. You will have to pay 500 yen to enter the room.
The festival has a sweet end. Visitors of the temple are offered various sweets such as okonomiyaki and takoyaki.
Briefly, your day starts with tasting history and silence and ends with tasting sweets.
This article also ends.
But life continues on the mega streets of Tokyo.
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Joshgun Karimov is an author and a public speaking coach.
He wrote KVAN, which is one of the best-selling personal development books in Azerbaijan. This book was personally endorsed by world's leading personal development coach and author Brian Tracy.
Joshgun does training and mentoring for Azerbaijan Public Speaking Competition and Social Innovation Lab. He delivered trainings on communication and presentation skills to such corporate clients as Four Seasons Hotel, Fairmont Hotel, McDonalds, Formula 1 (Baku City Circuit Academy), Kapital Bank.
He won at startup competitions such as "Yeni Fikir" and "Seedstars-Baku" for the presentation of mobile application for book readers - Kvotter.
He has received Global Influencer Day Award for a positive social media impact.


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Annie can't get through to the Town Hall and stop Alf from resigning. Alan rings Maggie from London and tells her that Elsie is making progress but they're keeping her in for the time being. Hilda continues to upset the neighbours with her insinuations about Elsie. Len isn't bothered about Alf's problems. At the Town Hall, Alf is kept waiting for Vernon Fletcher, the clerk to the council. Len has to go to a council meeting about a trip to France and offers to give Alf the good news. Mr Fletcher can't be found in the building. Ray moans in The Kabin about Len's trip to France. Rita is furious that she was kept in the dark. Mr Fletcher arrives at his office. Alf resigns and tells his story to him as Len strolls nonchalantly to the Town Hall. Mr Fletcher understands Alf's reasons though he wished he'd given him the chance to help him. He tells Alf there are ten minutes before the meeting at which he has to proffer his resignation. Norma has taken time off to have her hair done before a date at the ballet with Ken but Maggie worries that he'll hurt her. Fletcher alerts the press to a statement that will be issued later. Rita looks for Len. Jerry starts to lose his patience with Ray at the state of the office at the yard and suggests they need a secretary. Rita is livid about Len and insists on closing the shop early. She breaks down as Mavis tries to comfort her. They agree to go out on the town together. Alf returns to the Rovers with the news that Vernon rang the police superintendent himself and heard about the guilty plea from Norman Leach. The regulars tell him that Len should have passed on the news. Alf is livid at the betrayal and Rita overhears him.
Regular cast
- Bet Lynch - Julie Goodyear
- Betty Turpin - Betty Driver
- Hilda Ogden - Jean Alexander
- Stan Ogden - Bernard Youens
- Annie Walker - Doris Speed
- Maggie Clegg - Irene Sutcliffe
- Ena Sharples - Violet Carson
- Minnie Caldwell - Margot Bryant
- Len Fairclough - Peter Adamson
- Alf Roberts - Bryan Mosley
- Ray Langton - Neville Buswell
- Jerry Booth - Graham Haberfield
- Mavis Riley - Thelma Barlow
- Rita Littlewood - Barbara Mullaney
Guest cast
- Rovers Return Inn - Public and hallway
- Corner Shop
- The Kabin
- Fairclough and Langton - Yard and office
- Weatherfield Town Hall - Council clerk's office
- Canal Street
- TV Times synopsis: Will Len be in time to save the Mayor from shame?
- Viewing Figures: First UK broadcast - 6,950,000 homes (2nd place).
Notable dialogue
Bet Lynch: "It's our role in life is that - making fellers perfect for ourselves."
Hilda Ogden: "There's days I could leave you."
Stan Ogden: "An' there's days when I'd flippin' let you!"
|August 1973 episodes
|← Wed 1st • Mon 6th • Wed 8th • Mon 13th
Wed 15th • Mon 20th • Wed 22nd • Mon 27th • Wed 29th →


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- This event has passed.
See Your Future Vision Board Workshop with Bernadette Petrie.
February 22 @ 10:00 am - 1:30 pm£75.00
Join me Bernadette Petrie, The Soul's Life Coach, and Creator of The Barefoot Sanctuary for The Creative Play shop, on Saturday 8th February 2020 at The Barefoot Sanctuary, where you will create a powerful vision/vibration board to bring you back to the real you and take you into a future that feels more authentic. This is a powerful process and I invite you to join as I dive into this Life and Soul enhancing experience. Diving into the principles that I write about in my first book Permission To Shine, the workshop can be a huge catalyst for those taking part. Each participant will also receive a signed copy of that book.
Not sure? Take a peek at a video I made before a previous workshop in which I give you a taster of why I believe tapping into your Core Soul Desires is essential so that you can thrive on your Yellow Brick Road and read what some previous participants have had to say about their experience:
What others have to say…
"I guess it was a week or so after the see your future session that I had my first interview of the year. My 3 words were Free, Hope, Real and I had this great feeling after the interview that i was able to be completely real. Totally myself without the corporate stuff and it worked! And now I'm feeling so free with the massive weight of financial stress lifted. And of course as we are feeling much better things are picking up faster for Sealblades! Thanks for helping me find my renewed focus on creating the life I love."
Sorcha O'Connor, Sealblades Edinburgh.
"Bernie's workshop was truly more than I bargained for. I was challenged emotionally and personally to confront and then set aside those negative "voices" that hold me back. I felt recharged, as though my 'reset' button had been pressed. And, a bonus was being amongst a group of lovely ladies. Thank you Bernie, and ladies, for a really great day! I look forward to doing it all over again…"
Catherine Amis
"I love this place! From the moment you enter the space at the Barefoot Sanctuary you are met with thee biggest and warmest of welcomes. The See Your Future workshop totally nourished, refocused and helped me find my inner balance, it's just like a reset button! You meet some lovely new people, share your experiences, have quiet time and get creative too. Result for me – clarity in abundance! Thank you Bernie x."
Lisa Johnson.
"Thank you so very much for the wonder you have shared with me! The workshop on Saturday felt like a really cleansing, and it was a joy to be there. I enjoyed the energy of being with other like minded souls, and felt a real calmness and peace. It's really given me a lot to think about and motivated me make some change, which is just what I was looking for. I would certainly recommend it, and have already told a colleague to look out for a date in the new year. Thank you so much!"
Veronica Vaughan.
"What a beautiful workshop! I have tried to create a vision board on my own before, but I definitely created in from my head. Bernadette's guided meditation was so powerful, my new board has been created from my heart and inner wisdom and feels completely different to my previous attempt.It was amazing to get rid of my inner critic and just dream big! I am so excited to see the stuff on my board coming true. Bernadette is an Earth Angel, a morning spent in her company is an absolute joy – I recommend anyone to attend to simply spend some time in Bernadette's gorgeous energy."
Julie Begbie
"Thank you for Saturday as it was such a positive day. The inner wisdom / inner critic work was very powerful for me and each time I look at my vision board I can re-feel that positive energy that was buzzing on Saturday. The process of making it was so simple but the outcome still amazes me in the ideas that I picked out and stuck on.
Combining that and the evening of meditation and talk last night has put me into such a positive place in my mind that I can see a future that makes me smile in spite of the crazy world around us."
Joan Mann
It was so, wonderful to hear this feedback and see such happy faces. This is why I believe this experience is so valuable.
– You deserve this experience too….
After a guided visualisation, into your future, a 1-1 with your inner critic and your inner wisdom, you will discover your true desires and we will use these desires to create your very own future self board. This is vision board but with a whole depth of difference.
There will be fun music and laughter, as we create these boards together in the Sacred space that is The Barefoot Sanctuary.


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Compare reviews, ratings and up to 4 quotes for HomeBuyer Reports or Building Surveys from RICS Chartered Surveyors covering St Albans.
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If you're looking for a cost-effective quote for a surveyor in St Albans, you don't need to look any further. Here at reallymoving.com we understand that finding a quality surveyor can be stressful, which is why we want to ensure that your service remains expert and dedicated to your personal needs. Our surveyors are regulated by the Royal Institution of Chartered Surveyors so you'll know you're getting a professional service. Simply fill in the form provided on our site for up to 4 quotes for surveyors in your area and let us do the hard work for you.
11,241 user reviews
As a first time buyer and completely new to the home buying process, it was really easy to get a selection of Chartered Surveyors and make my selection
John, June 2018
4.9 out of 5.0 based on 93 reviews
Able Surveyors Limited provide a professional surveying service in the London and Home Counties area. We ensure that all our clients get the most from purchasing a property.Read more
4.8 out of 5.0 based on 103 reviews
Located only 4 miles from the M11, our RICS Regulated Hertfordshire branch, your local surveyor is ideally positioned to provide surveying and valuation services throughout parts of Cambridgeshire, Hertfordshire and Read more
5.0 out of 5.0 based on 8 reviews
Chas R Lowe and Co are located in Barnet, ideally located to cover South East England, including Hertfordshire, Bedfordshire, Essex and London.Read more
4.8 out of 5.0 based on 18 reviews
Chawton Hill Associates Ltd are a passionate, dedicated team of Chartered Building Surveyors offering an efficient, effective service providing Structural Surveys (Building Surveys), Party Wall Awards, measured surveys Read more
4.6 out of 5.0 based on 208 reviews
An Established RICS Chartered Surveying practice with surveyors operating across the country. Our surveyors are all RICS accredited chartered surveyors and registered valuers who are at the top of their field.Read more
4.8 out of 5.0 based on 33 reviews
A multi-disciplined Surveying practice regulated by the RICS. We undertake Property valuations, Homebuyer surveys and Building Surveys and can also provide expert advice on planning matters such as loft conversions and Read more
4.6 out of 5.0 based on 40 reviews
Nationwide specialists in providing Independent Structural/Building and Home Buyer Surveys, with 5 star ratings on Trustpilot/Google.Read more
4.7 out of 5.0 based on 92 reviews
We are located in Central London, at our new offices on Chancery Lane. We are strategically placed to service London and the South East. We are independent, regulated by the RICS with over 35 years surveying experience.Read more
4.6 out of 5.0 based on 89 reviews
Hugh James Surveyors aim to offer a wide range of professional property services through our specialist team of Chartered Surveyors.Read more
4.8 out of 5.0 based on 5 reviews
Jeremy Wise Surveyors are a RICS regulated firm of Residential Property Surveyors and Valuers. We have over 20 years' experience of providing HomeBuyer Reports and Building Surveys in the St Albans area.Read more
4.7 out of 5.0 based on 83 reviews
Leo Horsfield Surveying is Brighton Based. We are an experienced small practice of Building Surveyors who have extensive experience in undertaking Building Surveys and Home Buyer Reports on all types of property from Read more
5.0 out of 5.0 based on 91 reviews
Navah Consulting offers residential building surveys across London, Hertfordshire, Essex, Cambridgeshire, Kent, Surrey and Sussex.Read more
5.0 out of 5.0 based on 3 reviews
Roger Cameron has been practicing as a Surveyor for approximately 25 years. He worked as a Loss Adjustor for structural damage such as subsidence and heave before moving to valuation and survey work.Read more
4.8 out of 5.0 based on 21 reviews
Westside Services Limited are dynamic and forward thinking Chartered Construction and Property Consultants.Read more
4.6 out of 5.0 based on 184 reviews
We understand how stressful it can be to buy a new property with solicitors, the mortgage and removals …. we can help take away some of that stress and worry, by looking at your new home for you.Read more
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|091 - Strange Glasses
|092 - Cave Cover-Up
|093 - The Mirror Maze
- US Version
A couple of rapscallions have hidden some stolen goods in a cave. To protect their stash, they're planning to seal off the entrance with some wood. To do so, they need to cut the board shown below into two equal pieces.
Where should the thieves make their cut?
Draw the answer with your stylus.
- UK Version
Two criminals want to use this board to seal off the cave where they store their smuggled goods.
If they cut the board into two pieces of identical size and shape, they'll be able to seal the cave entrance completely.
Draw lines on the board to indicate where it needs to be cut.
Click a Tab to reveal the Hint.
- US Version
Too bad!
The cut they need to make isn't all that complicated. Simplify your thinking.
- UK Version
Bad luck.
It's not a particularly complicated cut that's required here.
- US Version
Nice cut!
You're exactly right.
To seal the cave, the thieves flipped one of the piece upright as shown above.
- UK Version
Good job!
The board needs to be cut as shown above.
As you can see, the criminals have flipped one of the pieces and sealed the cave entrance tight.
|Professor Layton and the Unwound FuturePuzzles in
See here for weekly puzzles.
|Hidden Door
|Puzzle Index: CV · DB · UF · LS · MM · AL · VS
A big thanks to http://professorlayton3walkthrough.blogspot.com


text:
Frustrated Takhiantia residents want to know when the much-delayed rebuilding of Takhiantia-Wat Sangkapiew Road will be completed.
The project to widen and repave 950 meters of the roadway on top of a new sewer system was scheduled for completion on Aug. 11, but a week later large holes remained in the new concrete surface and all the sewers remained open.
Resident Nattaya Wongpan said roadwork that began in February has really inconvenienced motorists and everyone is anxious for the work to finish.
In addition, she said, contractors have not put barriers or lights around many of the open sewers, posing real danger to people at night.
Sub-district officials have not given residents any notice about when the work will be completed.


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It's been 34 years since a white player was selected first overall. We're not blaming the University of Indiana's Kent Benson directly, but his underwhelming career combined with the legends drafted behind him (Bernard King, Jack Sikma, Norm Nixon) certainly didn't help. Things ain't looking so good for you going No. 1 this year Jimmer.
Also Watch


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sunroof on car opened a fraction today
and the delicious smell of rain & fields
caressed my hair
Wednesday, 12 January 2011
Posted by Mark Sargeant at 20:40
Labels: aros
3 comments:
- Yvonne said...
I enjoy reading your small stones.
- 12 January 2011 at 21:38
just wait until the weather gets warmer, you can open the sunroof wider and let all the smells of summer come in!
- 12 January 2011 at 21:41
- Mark Sargeant said...
Thanks for the comment, Yvonne, I'm glad someone is enjoying reading the stones I'm sending out into the blogosphere...
@Anonymous - I keep looking up through the sunroof at the sky, in eager anticipation of a warm sunny day...
- 12 January 2011 at 22:37
Subscribe to: Post Comments (Atom)


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| 10°C Dublin
A facilities management company with its head office in Co Wexford has won a major national award.
AirconMech, which is based out of Kilcannon Business Park on the old Dublin Road, in Enniscorthy, won the 'Specialist FM Service Provider of the Year' award at this year's national Facilities Management Awards.
The facilities management sector is worth an estimated €1.2bn and has experience considerable growth in recent years. The awards, which have been going since 2014, were set up to acknowledge and recognise companies that strive for excellence within the sector.
The prestigious awards ceremony was held in the Ballsbridge Hotel where the Wexford company beat two other finalists in their category to secure the title.
In addition to the award it won the company was also shortlisted for the 'Overall Excellence in Facilities Management' award.
Speaking about the company, the MC on the night, Colm O'Regan, said it was obvious to the judges from the testimonials presented to them that customer service and satisfaction is placed at the forefront of the company's operations.
'They wanted to reward all of the hard work and dedication that went into making the company what it is,' said Mr O'Regan.
Speaking about the award the joint managing directors of the company, John Dempsey and Jim Mullett, said: 'The award is the result of a considerable team effort'.
They also said they were 'extremely proud' of everyone involved in making the company progress for over two decades.
Wexford People


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High Five Solar Powered Panorama & Garden Lights
Choosing outdoor lights can seem like a troublesome process with so many finishes, sizes and styles to choose from. Faced with the varied options and particular person strengths, conveniences and issues I've discovered many purchasers will select to install ventless fuel logs in their custom outdoor hearth. Not often will someone build a custom outdoor fire because they want a great useful heater on the patio.
Many consumers look to the weave for magnificence but check it to make sure a tightness that can enable resilience when the furniture is used outdoors whereas staying tight. So the witches, skeletons, zombies, or whatever scary characters you wish to add, are accentuated by the cooperation of the fall season which provides its rich colours to help us in our outdoor Halloween decorations. When you've got a deck or patio, the usage of several totally different lighting choices will let you use the identical space for different activities. In case you spend a great deal of time outdoors in the course of the Summer months, it makes good sense to make your outdoor dwelling area as comfy and classy as your inside areas. You'll wish to have gentle lighting on the floor for the protection and pleasure of your visitors.
You should make sure that there's an ample quantity of lighting, by using lanterns and hanging lighting supplied for the camp website, and within the form of flashlights for using your campers as they travel via the campground during night hours.
The addition of the lighted star on a pole and the wooden manger makes these yard decorations stand out. Enjoy a scenic outdoor entertaining area with our eclectic selection of outdoor furnishings sets, outdoor patio furnishings sets, seating, and decor. The very best quality (and most expensive) makes use of pure aluminum …Read More


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Cyclops Brawlbucklers are a variant of enemy Cyclops that appear later in Skylanders: Swap Force. They carry shields which protects them from attacks, until they drop their guard to strike the Skylanders.
- In Skylanders: Imaginators these, along with Coldspear Cyclopses, are in the audience in BattleBrawl Island arena challenges.
Community content is available under CC-BY-SA unless otherwise noted.


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Latest News
Visit Shropshire launches new free membership to support tourism industry. (14th May 2020) Read More...
Visit Britain Advice Hub (1st May 2020) Read More...
What's On
The Ginger and Spice Festival
World famous festival takes place 11th -13th September
World famous festival takes place 11th -13th September
Preparations are well underway for the 21st Ludlow Food Festival, which takes place Friday 11th to Sunday 13th September in and around Ludlow Castle. Nearly 200 food producers from across Shropshire and The Marches will be attending the event.
The castle will be brimming with dedicated artisans and vendors selling everything from soft shell crab to chilli plants. With cheeses, meats, wine, preserves, bread, cakes and confectionary to name a few, there will be something to suit everyone's taste buds. Luckily there will also be a shopping crèche where you can safely stow your purchases for a small donation to charity.
The festival is once again attracting top chefs and food personalities all eager to cook for the crowds expected over the three days. On Friday celebrity chef Marcus Bean has pulled together a team of culinary superstars including Cyrus Todiwala OBE and Jon Gay, Head Chef at Jimmy's Farm.
The festival will also see the very welcome return of Shaun Hill, the legendary chef who helped put Ludlow on the food map. As well as rising stars such as Suree Coates who has just been included in the new 2016 Good Food Guide.
With 'Bake Off Fever' sweeping the nation once more you can get your fix by seeing not one but two previous favourites, last year's finalist Luis Troyano and 2013's Howard Middleton. Both will be meeting fans and signing copies of their new books.
There will be lots to see, do and of course taste across the three days with the famous ale, sausage and bread trails certain to sell out quickly.
Ludlow is also lucky enough to have a world-class exhibition in town during the food festival. The Pink Lady Food Photographer of the Year exhibition can be seen at the Ludlow Assembly Rooms a minute's walk from the castle and entry is completely free.
Children will love the festival with lots of free activities just for them. Always popular is the circus skills tent while they can get hands on with cookery sessions, fun workshops and learn more about farming with the NFU. Children will also be able to pick up fun activity booklets and Let's Sow Seeds pack so visitors can try their hand at growing a variety of salad leaves in their own home.
Adults too will be spoilt for choice. When you've finished browsing the stands you can enjoy the free Talks and Tastings area or Brompton Cookery School's free pop-up taster sessions. If you want to immerse yourself in a more hands-on culinary experience then the great value master classes and Taste Workshops start from just £5.00 per person.
Full details including the downloadable 2015 programme can be found on the food festival's website www.foodfestival.co.uk You can also buy your tickets in advance with adult prices from £8.50 and family tickets £20.00.


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- Home »
- See&do In Krk Island »
Šilo info
Šilo info (Krk island)
In the northeast of Krk, you will find a small town Šilo. If you are looking for a place to relax, away from the daily hustle and noise, then Šilo is ideal for you. You can find accommodation in the camp, or in local family apartments, houses or rooms, and you will be kindly welcomed by very approachable and friendly hosts. Although it is a small town, Šilo is proud of its beaches that are very neat and clean, which is confirmed by the Blue Flag that flies proudly over the sandy Pećine beach. Along with resting, you can have fun on various recreational programs that are held on the local beaches.
Šilo 51515, otok Krk
30 min
GPS coordinates: 45.146297, 014.663566 (45°08'46.7″N, 014°39'48.8″E)
Address: Šilo 51515, otok Krk
Šilo-Malinska (15km)
Šilo-Njivice (19km)
Šilo-Baška (30km)
Šilo-Rijeka (40km)
Šilo-Pula (150km)
Šilo-Zagreb (175km)
Šilo-Split (350km)
Šilo-Dubrovnik (540km)
Šilo is located on the island of Krk
If you are driving with a car show this translation to the taxi driver:
Molim Vas da me odvezete do mjesta Šilo koje se nalazi na otoku Krku. Hvala!
You must also show to the taxi driver that he must turn on the taximeter:
Molim Vas uključite taksimetar. Hvala!
Additional translation:
Please can you tell me how to find this sight:
Molim Vas da me uputite prema mjestu Šilo koje se nalazi na otoku Krku. Hvala!


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This is the second half of my book-buying post. The first half can be found here. I haven't yet read any of the books on this list, so this post will be more of a book haul than a recent reads, but I hope it'll be of interest anyway. I always like hearing about… Continue reading Books I've Bought Recently – Part Two
Tag: reading
Consuming Stories: Different Media Ranked
Humans have an absolute obsession with stories. They are a tradition that has been carried on throughout our very earliest history, since our first attempts at sophisticated language, and we continue to love stories to this day, both real and fictional. They are our purest form of entertainment. It is something innate in us, so… Continue reading Consuming Stories: Different Media Ranked