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text:
Morphology control in mesoporous carbon films using solvent vapor annealing
Zhe Qiang, Jiachen Xue, Kevin A. Cavicchi, Bryan D. Vogt
Ordered mesoporous (2-50 nm) carbon films were fabricated using cooperative self-assembly of a phenolic resin oligomer with a novel block copolymer template (poly(styrene-block-N,N-dimethyl-n-octadecylamine p-styrenesulfonate), (PS-b-PSS-DMODA)) synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization. Due to the high Tg of the PS segment and the strong interactions between the phenolic resin and the PSS-DMODA, the segmental rearrangement is kinetically hindered relative to the cross-linking rate of the phenolic resin, which inhibits long-range ordering and yields a poorly ordered mesoporous carbon with a broad pore size distribution. However, relatively short exposure (2 h) to controlled vapor pressures of methyl ethyl ketone (MEK) yields significant improvements in the long-range ordering and narrows the pore size distribution. The average pore size increases as the solvent vapor pressure during annealing increases, but an upper limit of p/p0 = 0.85 exists above which the films dewet rapidly during solvent vapor annealing. This approach can be extended using mesityl oxide, which has similar solvent qualities to MEK, but is not easily removed by ambient air drying after solvent annealing. This residual solvent can impact the morphology that develops during cross-linking of the films. These results illustrate the ability to fine-tune the mesostructure of ordered mesoporous carbon films through simple changes in the processing without any compositional changes in the initial cast film.
https://doi.org/10.1021/la304915j
10.1021/la304915j
Fingerprint Dive into the research topics of 'Morphology control in mesoporous carbon films using solvent vapor annealing'. Together they form a unique fingerprint.
Carbon films Chemical Compounds
phenol-formaldehyde resin Chemical Compounds
Annealing Chemical Compounds
phenolic resins Physics & Astronomy
Vapors Engineering & Materials Science
Phenolic resins Engineering & Materials Science
vapors Physics & Astronomy
Pore size Chemical Compounds
Qiang, Z., Xue, J., Cavicchi, K. A., & Vogt, B. D. (2013). Morphology control in mesoporous carbon films using solvent vapor annealing. Langmuir, 29(10), 3428-3438. https://doi.org/10.1021/la304915j
Qiang, Zhe ; Xue, Jiachen ; Cavicchi, Kevin A. ; Vogt, Bryan D. / Morphology control in mesoporous carbon films using solvent vapor annealing. In: Langmuir. 2013 ; Vol. 29, No. 10. pp. 3428-3438.
@article{7fc1cc11791641999490022968841e41,
title = "Morphology control in mesoporous carbon films using solvent vapor annealing",
abstract = "Ordered mesoporous (2-50 nm) carbon films were fabricated using cooperative self-assembly of a phenolic resin oligomer with a novel block copolymer template (poly(styrene-block-N,N-dimethyl-n-octadecylamine p-styrenesulfonate), (PS-b-PSS-DMODA)) synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization. Due to the high Tg of the PS segment and the strong interactions between the phenolic resin and the PSS-DMODA, the segmental rearrangement is kinetically hindered relative to the cross-linking rate of the phenolic resin, which inhibits long-range ordering and yields a poorly ordered mesoporous carbon with a broad pore size distribution. However, relatively short exposure (2 h) to controlled vapor pressures of methyl ethyl ketone (MEK) yields significant improvements in the long-range ordering and narrows the pore size distribution. The average pore size increases as the solvent vapor pressure during annealing increases, but an upper limit of p/p0 = 0.85 exists above which the films dewet rapidly during solvent vapor annealing. This approach can be extended using mesityl oxide, which has similar solvent qualities to MEK, but is not easily removed by ambient air drying after solvent annealing. This residual solvent can impact the morphology that develops during cross-linking of the films. These results illustrate the ability to fine-tune the mesostructure of ordered mesoporous carbon films through simple changes in the processing without any compositional changes in the initial cast film.",
author = "Zhe Qiang and Jiachen Xue and Cavicchi, {Kevin A.} and Vogt, {Bryan D.}",
doi = "10.1021/la304915j",
journal = "Langmuir",
Qiang, Z, Xue, J, Cavicchi, KA & Vogt, BD 2013, 'Morphology control in mesoporous carbon films using solvent vapor annealing', Langmuir, vol. 29, no. 10, pp. 3428-3438. https://doi.org/10.1021/la304915j
Morphology control in mesoporous carbon films using solvent vapor annealing. / Qiang, Zhe; Xue, Jiachen; Cavicchi, Kevin A.; Vogt, Bryan D.
In: Langmuir, Vol. 29, No. 10, 12.03.2013, p. 3428-3438.
T1 - Morphology control in mesoporous carbon films using solvent vapor annealing
AU - Qiang, Zhe
AU - Xue, Jiachen
AU - Cavicchi, Kevin A.
AU - Vogt, Bryan D.
N2 - Ordered mesoporous (2-50 nm) carbon films were fabricated using cooperative self-assembly of a phenolic resin oligomer with a novel block copolymer template (poly(styrene-block-N,N-dimethyl-n-octadecylamine p-styrenesulfonate), (PS-b-PSS-DMODA)) synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization. Due to the high Tg of the PS segment and the strong interactions between the phenolic resin and the PSS-DMODA, the segmental rearrangement is kinetically hindered relative to the cross-linking rate of the phenolic resin, which inhibits long-range ordering and yields a poorly ordered mesoporous carbon with a broad pore size distribution. However, relatively short exposure (2 h) to controlled vapor pressures of methyl ethyl ketone (MEK) yields significant improvements in the long-range ordering and narrows the pore size distribution. The average pore size increases as the solvent vapor pressure during annealing increases, but an upper limit of p/p0 = 0.85 exists above which the films dewet rapidly during solvent vapor annealing. This approach can be extended using mesityl oxide, which has similar solvent qualities to MEK, but is not easily removed by ambient air drying after solvent annealing. This residual solvent can impact the morphology that develops during cross-linking of the films. These results illustrate the ability to fine-tune the mesostructure of ordered mesoporous carbon films through simple changes in the processing without any compositional changes in the initial cast film.
AB - Ordered mesoporous (2-50 nm) carbon films were fabricated using cooperative self-assembly of a phenolic resin oligomer with a novel block copolymer template (poly(styrene-block-N,N-dimethyl-n-octadecylamine p-styrenesulfonate), (PS-b-PSS-DMODA)) synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization. Due to the high Tg of the PS segment and the strong interactions between the phenolic resin and the PSS-DMODA, the segmental rearrangement is kinetically hindered relative to the cross-linking rate of the phenolic resin, which inhibits long-range ordering and yields a poorly ordered mesoporous carbon with a broad pore size distribution. However, relatively short exposure (2 h) to controlled vapor pressures of methyl ethyl ketone (MEK) yields significant improvements in the long-range ordering and narrows the pore size distribution. The average pore size increases as the solvent vapor pressure during annealing increases, but an upper limit of p/p0 = 0.85 exists above which the films dewet rapidly during solvent vapor annealing. This approach can be extended using mesityl oxide, which has similar solvent qualities to MEK, but is not easily removed by ambient air drying after solvent annealing. This residual solvent can impact the morphology that develops during cross-linking of the films. These results illustrate the ability to fine-tune the mesostructure of ordered mesoporous carbon films through simple changes in the processing without any compositional changes in the initial cast film.
U2 - 10.1021/la304915j
DO - 10.1021/la304915j
JO - Langmuir
JF - Langmuir
Qiang Z, Xue J, Cavicchi KA, Vogt BD. Morphology control in mesoporous carbon films using solvent vapor annealing. Langmuir. 2013 Mar 12;29(10):3428-3438. https://doi.org/10.1021/la304915j

redpajama_set_name: RedPajamaCommonCrawl


text:
require File.expand_path('../boot', __FILE__)

require 'action_controller/railtie'
require 'sprockets/railtie'

require 'jquery-rails'
require '../lib/date_time_picker.rb'

module App
  class Application < Rails::Application
  end
end

redpajama_set_name: RedPajamaGithub


text:
Comparative study of two protocols for quantitative image-analysis of serotonin transporter clustering in lymphocytes, a putative biomarker of therapeutic efficacy in major depression
Raquel Romay-Tallon1,2,
Tania Rivera-Baltanas3,
Josh Allen1,
Jose M. Olivares3,
Lisa E. Kalynchuk2,4 &
Hector J. Caruncho1,2
Biomarker Research volume 5, Article number: 27 (2017) Cite this article
The pattern of serotonin transporter clustering on the plasma membrane of lymphocytes extracted from human whole blood samples has been identified as a putative biomarker of therapeutic efficacy in major depression. Here we evaluated the possibility of performing a similar analysis using blood smears obtained from rats, and from control human subjects and depression patients. We hypothesized that we could optimize a protocol to make the analysis of serotonin protein clustering in blood smears comparable to the analysis of serotonin protein clustering using isolated lymphocytes.
Our data indicate that blood smears require a longer fixation time and longer times of incubation with primary and secondary antibodies. In addition, one needs to optimize the image analysis settings for the analysis of smears. When these steps are followed, the quantitative analysis of both the number and size of serotonin transporter clusters on the plasma membrane of lymphocytes is similar using both blood smears and isolated lymphocytes.
The development of this novel protocol will greatly facilitate the collection of appropriate samples by eliminating the necessity and cost of specialized personnel for drawing blood samples, and by being a less invasive procedure. Therefore, this protocol will help us advance the validation of membrane protein clustering in lymphocytes as a biomarker of therapeutic efficacy in major depression, and bring it closer to its clinical application.
In a seminal and classical report, [1] described the plasma membrane as a lipid bilayer (which is mostly composed of phospholipids and sterols) with embedded proteins that can span the membrane (transmembrane proteins) or attach to it peripherally (peripheral proteins). The structure of the membrane was described as a "fluid mosaic", which allowed a certain degree of mobility for both lipids and proteins. The presence of tight junctions, the link of membrane proteins to the cytoskeleton, and the clustering of proteins within lipid rafts define specific membrane domains and allow for protein-protein interactions that influence their physiological roles [2]. Among the proteins that tend to be clustered within specific membrane domains are numerous neurotransmitter transporters and receptors that locate in pre- and/or post-synaptic sites, and that in many cases (e.g. transporters and G-Protein-coupled-receptors) accumulate in lipid rafts [3]. The partitioning of these molecules into different membrane compartments has important consequences for their physiological actions, to the extent that alterations in membrane protein clustering may be a marker of cellular physiology/pathophysiology.
In a series of recent reports, we have proposed that alterations in membrane protein clustering in peripheral lymphocytes may represent a novel biomarker of depression: We observed that the pattern of clustering of both the serotonin transporter (SERT) and serotonin 2A receptor on the plasma membrane of lymphocytes from naïve depression patients identified two subpopulations of patients that showed a different therapeutic response after 8 weeks of conventional antidepressant treatment [4,5,6,7]. We concluded that analysis of membrane protein clustering in lymphocytes could predict which patients are more likely to respond to antidepressant treatment. This issue is quite important as more than 1/3 of patients fail to show a proper response to antidepressant medication, and this response can only be determined after several weeks of antidepressant use (see [8]). If the pattern of protein clustering in peripheral lymphocytes can accurately predict therapeutic responsivity in patients with depression, this would offer a significant advancement over the hit and miss approach often used in the clinic to map out a course of treatment for individual patients.
One of the challenges associated with the collection and proper preservation of blood samples from the depression patients used in our previous experiments was that both patients and clinical personnel found the drawing of blood samples and isolation of lymphocytes to be both cumbersome and expensive. For research purposes, the traditional drawing of blood samples requires the presence of a trained professional, which increases the cost and complexity of the research project. And for the eventual clinical application of this approach for evaluating potential treatment options for patients, one needs to have a simple technique that can be implemented in any medical clinic. Therefore, we developed a new protocol for the study of SERT clustering that can be done using blood smears (from human and rodents) rather than whole blood samples. In this manuscript, we describe the blood smear protocol in detail and compare the results of SERT labelling in blood smears with those from the analysis of isolated lymphocytes in laboratory rats, and in control human subjects and depression patients.
Immunocytochemistry of SERT in whole blood isolated lymphocytes
The analysis was carried out on a total of 10 healthy subjects and 5 naïve depression patients. All the procedures were carried out in compliance with the Code of Ethics of the World Medical Association (Declaration of Helsinki) and approved by the Alvaro Cunqueiro Hospital Ethics Committee (Spain), which includes a consent form signed up for all participants. The blood was drawn by a trained nurse with the use of a 10 ml BD-Vacutainer ® glass whole-blood tube containing 1.5 ml ADC solution [tri-sodium citrate (22 g/l), citric acid (8 g/l), and dextrose (24.5 g/l)]. Samples were diluted 1:1 in phosphate buffered saline (PBS) prior to lymphocyte isolation by centrifugation at 20 °C in a gradient of Ficoll-Paque Plus at 400 g for 30–40 min. The band containing the lymphocytes was extracted and rinsed with PBS. Subsequently, cells were centrifuged at 1000 g for 10 min. This step was repeated three times. Finally, cells were fixed with 1% paraformaldehyde solution in 0.1 M phosphate buffer for 1 min. We tried different fixation conditions and ascertained that these parameters were the ones that worked best. Fixed lymphocytes could be stored at 4 °C for up to 1 week. For longer periods of time, they were stored at -80 °C in a cryoprotectant solution (Dimethyl-sulfoxide 10%; Fetal bovine serum 10% and RMPi-1640).
A total of 11 naïve male Long Evans (Charlers River) have been used in this analysis. All the procedures were conducted under a protocol approved by the Committee on Animal Care and Supply, Animal Research Ethics Board of the University of Saskatchewan (Canada). To obtain peripheral lymphocytes from rats, blood was drawn either by heart puncture (up to 3 ml in total, when animals were sacrificed for other research purposes) or directly from the tail vein (up to 1 ml). Blood was collected in 5 ml Eppendorf tubes containing blood and ADC solution in a 1:7 proportion. Centrifugation of samples, extraction and fixation of lymphocytes was carried out as explained above.
For SERT immunolabeling, samples were re-suspended in PBS and brought to room temperature, followed by centrifugation at 100 g for 10 min. To block unspecific staining, we performed a pre-incubation step for 10 min with 3% human IgG, or 3% rat IgG (I2511 and I4131, respectively, Sigma, St Louis, Missouri), and 1% bovine serum albumin (BSA, 3059-100Gr, Sigma, St Louis, Missouri) in PBS. For labeling of SERT, we used a polyclonal primary antibody raised in rabbit that specifically recognizes epitopes of both human and rat SERT [anti-serotonin transporter: AB10514P (Millipore, Billerica, Massachusetts)], and proceeded to incubate the lymphocytes with the primary antibody diluted 1:250 in 1% BSA in PBS at 4 °C overnight. After incubation with primary antibody, the samples were rinsed with PBS by centrifuging at 100 g for 10 min (repeated 3 times). After rinsing samples were then incubated for 1 h at room temperature with a 1:200 solution of the secondary antibody [Alexa Fluor 568, goat anti-rabbit: (A-11008, Molecular Probes, Eugene, Oregon)] in 1% BSA in PBS. This step is light sensitive, and therefore was carried out in the dark. After 3× rinsing with PBS (by centrifuging 3 times, 10 min each, at 100 g) the samples were re-suspended in 100-200 μl PBS. Sets of 20 μl from the samples were deposited on Super-Frost Plus slides (Menzel-Glaser, Braunschweig, Germany), coverslipped, and stored in the freezer at -20 °C.
Immunocytochemistry of SERT in blood smears
Smears were prepared from blood collected by a quick prick on the tip of the forefinger in 6 human control subjects and a prick of the lateral tail vein in 19 male Long Evans untreated rats. Thin smears were spread on Super-Frost Plus slides, dried at room temperature for at least 1 hr., and then stored at 80 °C until used for immunocytochemistry.
We tested different fixation conditions: formalin 10% incubated for a minute, methanol incubated for 2 min, precooled acetone incubated at -20C for 10 min and at 4C for 2 h; the quality of the immunolabeling was not as good (see Discussion). We obtained the best results if the smears were fixed with 0.5 ml of 1% PFA for 5 min at room temperature. The fixation step was performed on dry slides, which were then washed and dried before being stored at -80 °C. We also examined the results if fixation was done in unfixed frozen slides brought back at room temperature and allowed to dry. There were no differences in the data obtained with either of these protocols.
Immunocytochemistry was performed directly on the smears using a protocol that was slightly modified from the one used on isolated lymphocytes. Smears include a whole blood sample: red blood cells, platelets and white blood cells, this causes more unspecific background with the immunolabelling. To avoid this, we have performed a pre-incubation step for 1 h at room temperature with 10% human IgG, or rat IgG diluted in 1% BSA and PBS. In the same way as the described protocol for isolated lymphocytes, samples were incubated overnight at 4C with primary antibody (same antibody as indicated above). After incubating, smears were rinsed with PBS for 10 min (3 times). The samples are subsequently incubated with fluorescent secondary antibody for 2 h at room temperature (same secondary antibody specified previously). We rinsed the excess of antibody by rinsing the sample with PBS 3 times 10 min each. To facilitate the identification of the lymphocytes, we used a nuclear marker [Hoechst 33,258 (Invitrogen, Molecular Probes, Carlsbad, California), applied at 2 μg/ml for 15 min at room temperature]. After nuclear labeling, we rinsed the smears with PBS for 10 min, let them dry, and coverslipped them. The slides were then stored in the freezer at -20 °C.
Imaging processing and quantification
We analyzed SERT membrane protein clustering in isolated lymphocytes and in smears from both human and rat samples We obtained pictures at 100× magnification from 100 individual lymphocytes per sample using a fluorescence microscope. The pictures were subsequently analyzed using ImageJ software (1.48v, NIH, USA). We need to set appropriate conditions to accurately analyse the size of membrane protein clusters in both smears and isolated lymphocytes. ImageJ performs the measurements in pixels, so the first step must be the calibration of the program to get real values in our measurements (i.e. number and size of the SERT clusters). This process implies introduce a known distance to the program to establish equivalence in pixels to the distance given in μm. In a second step, we must adjust the background conditions in ImageJ to facilitate the analysis of the clusters. We will go to menu Process → Substract background and in Rolling Ball Radius we will indicate the size of the smallest particle which is not background (approximately 5–10 to isolated lymphocytes, and 1–3 in smears). When working with smears, an additional step is needed, Remove Outliers: Radius 1 and Threshold 50, in this way we can remove small dots that might interfere in the analysis and give us an overestimation of the number of clusters or an underestimation of the size. Following this step, a binary image is created where the clusters are easily quantified. However, if two clusters appear too close, the software could interpret as only one, and so provide an underestimation of the number of clusters and an overestimation of their size. To avoid this effect, we will add one more editing in menu Process → Binary → Watershed. In a third step, we will proceed to measure and quantify the clusters. By Analyze → Measure, the software gives us the measurement area, and by Analyze → Analyze Particles, the program provides a number and measure of the clusters. It is necessary indicate the minimum size of particle that the program will analyze to avoid a mistaken measurement. We have set that 0.05-Infinite provides an accurate measure of the clusters. As an additional setting to gather an accurate measure will be indicate analyze Pixel Units.
Importantly, once we established a set of useable conditions in ImageJ, we kept them constant to make the results comparable across conditions. From the binary image of each lymphocyte we obtain two sets of measurements: the number of SERT expressing clusters, the size of those clusters. We average the measurements from each lymphocyte within each sample, and used to compare results between analysis done in smears and in isolated lymphocytes.
For the statistical analyses, we used SPSS 20 (IL, Chicago, USA). We compared the number and size of SERT protein clusters in lymphocytes analyzed from each protocol using two-tailed Student's T-test in both human and rodent samples. The criterion for statistical significance was set at p < 0.05.
We compared two different methods to analyze SERT protein clustering in lymphocytes obtained from human and rat peripheral blood samples. Panel 1 shows low magnification images of isolated rat lymphocytes (Fig. 1a), and rat blood smears (Fig. 1b) that were immunostained for SERT. Although logically the number of lymphocytes per microscope field is higher in images from isolated lymphocyte preparations, the presence of lymphocytes is also easily seen in blood smears (arrow in Fig. 1b), where they are shown interspersed with red blood cells (asterisk), and platelets (arrowhead). Platelets also show SERT immunoreactivity, but are easily distinguished from SERT immunoreactive lymphocytes.
Images of lymphocytes immunostained for SERT in preparations of isolated lymphocytes extracted from whole rat blood samples (a), and in rat blood smears (b). Note than in rat blood smears lymphocytes can be easily identified (arrow), and are interspersed with numerous SERT immune-negative red blood cells (asterisk), and platelets that also show inmmunoreactivity for SERT (arrowhead). Calibration Bar: 10 μm
Once we determined the feasibility of SERT immunolabeling in lymphocytes within blood smears, we proceeded to carry out a quantitative study of SERT clusters on the plasma membrane of lymphocytes, in both isolated lymphocyte preparations and in blood smears obtained from rats and humans. In Fig. 2, we show an example of SERT clustering quantification in one isolated lymphocyte from a rat (A), and after converting the image to a binary colour and subtraction of background (B), the image was automatically analyzed by the Image J software to quantify the number of clusters per lymphocyte and average size of SERT clusters (C). The same composition of the analysis of one lymphocyte from a rat blood smear is shown in Fig. 2 d-e. In a similar way, Fig. 3 portrays images of the analysis of SERT clustering in isolated human lymphocytes (A-C), and in human blood smears (D-E).
Example of analysis of SERT protein clusters in one rat lymphocyte isolated from whole blood samples (a-c), and one lymphocyte from rat blood smears (d-f). Figs. a and c show the microscope image of SERT immunostaining, while Figs. b and e represent the binary image as obtained in Image J, and Figs. c and f show the actual counts of the number and size of cluster as performed by the image analysis system. Calibration Bar: 10 μm
Example of analysis of SERT protein clusters in one human lymphocyte isolated from whole blood samples (a-c), and one lymphocyte in blood smears from human control subjects (d-f). Figs a and c show the microscope image of SERT immunostaining, while Figs. b and e represent the binary image as obtained in Image J, and Figs. c and f show the actual counts of the number and size of cluster as performed by the image analysis system. Calibration Bar: 10 μm
Figure 4 shows the quantified number and size of clusters in both rat (A-B) and human lymphocytes (C-D) from isolated lymphocyte preparations and blood smears. Our statistical analyses of these data revealed no significant differences in the number of clusters and their size when we compare the two protocols in rats [Number: t (28, .910); p = .371; Size: t (28, −.278); p = .783] or humans [Number: t (14, 1.366); p = .193; Size: t (14, .253); p = .804].
Histograms of the counts of SERT clusters number and size in isolated lymphocytes and lymphocytes in blood smears from rat and human control samples. Note that after protocol optimization the results obtained in the analysis of SERT clusters and size are quite similar
Figure 5 shows the quantitative comparison of number and size of SERT clusters in isolated lymphocytes or blood smears from depression patients. There are no significant differences in the number and size of clusters when comparing the two protocols [Number: t (14, 1.032); p = .324; Size: t (14, .631); p = .541].
Histograms of the counts of SER clusters number and size in isolated lymphocytes and lymphocytes in blood smears from naïve depression patients. Note that after protocol optimization the results obtained in the analysis of SERT clusters and size are quite similar
Figure 6 shows the quantitative analysis of SERT clustering in lymphocytes from human control subjects and depression patients, when using isolated lymphocytes or blood smears. Statistical analysis reveals no alterations in SERT numbers for both protocols; and an increase in SERT clusters which is similarly shown in both protocols [SERT Number blood smears: t (14, 1.687); p = .143; SERT number isolated lymphocytes: t (14, 1.639); p = .119; SERT size blood smears: t (14, −4.858); p = .002; SERT size isolated lymphocytes: t (14, −2.810); p = .012].
Histograms of the counts of SER clusters number and size in isolated lymphocytes and lymphocytes in blood smears from human control subjects and depression patients. Note that after protocol optimization the results obtained in the analysis of SERT clusters and size are quite similar for both protocols, indicating in both cases a lack of significant changes in the number of SERT clusters, and an increase in SERT clusters size in naïve depression patients
The identification of biomarkers to support a proper diagnosis, prognosis, and evaluation of therapeutic efficacy of antidepressant response is of the utmost importance [8]. In this experiment, we evaluated the feasibility of analysing membrane protein clusters in lymphocytes from blood smears, as this would provide a much easier and cheaper way to obtain appropriate samples in a clinical setting and it would also facilitate the study of lymphocytes in a research environment. Our results indicate that when the smears protocol is optimized, the data obtained from this approach are comparable to the data obtained when using isolated lymphocytes.
Our previous studies have been performed with isolated lymphocytes from peripheral blood samples, and we had adjusted the fixation, immunocytochemistry protocols, and image analysis settings to provide an accurate and replicable measurement of the number and size of membrane protein clusters (i.e., SERT and serotonin 2A receptors) in lymphocytes [5,6,7]. In this experiment, we developed a new protocol for analysing membrane protein clustering in lymphocytes in blood smears and optimized it to make it comparable to the analysis of protein clustering in isolated lymphocytes. We found that the major challenges associated with making the protocols comparable were related primarily to fixation and immunocytochemistry. For the fixation and immunolabeling of isolated lymphocytes, we use centrifugation (to obtain a pellet) and shaking the samples in a vortex (to re-suspend the samples) in every step, whereas we found that in the case of blood smears, we could conduct the fixation and immunolabeling in one spread layer of cells dried onto a coverslip. In these conditions, we determined after trying different fixation protocols, with different concentrations of PFA, at different temperatures, and for different periods of times, that 1% PFA for 5 min provides the most accurate results for immunolabeling of SERT clusters. This time is longer than the time required for fixation of isolated lymphocytes (i.e., which is 1 min), but it ensures that the quality of the immunolabeling in smears is similar to immunolabeling obtained in isolated lymphocytes (see Figs. 2 and 3). Our preliminary experiments also asserted that similar fixation characteristics are valid when using other anti-SERT antibodies or when labeling other membrane receptors and transporters (Romay-Tallon, personal communication).
Immunolabeling of SERT in smears also requires an increase in incubation time with at almost each step of the process. This is primarily because with smears the cells are adhered to the slide and therefore the labeling of clusters in the cell membranes requires a longer period of exposure to antibodies than what would normally be necessary for immunolabeling re-suspended cells, as it is the case for isolated lymphocytes. We have also discovered that the temperature at which incubation with the primary and secondary antibodies occurs is vital both for smears and isolated lymphocytes, as other temperature settings during these steps (e.g., incubation at room temperature) results in an increase in size and decrease in number of SERT clusters.
Finally, it is important to define a set of conditions for the image analysis in ImageJ software (e.g. background clearing conditions, define the minimum size of a particle that would be quantified) that offer a realistic representation of the lymphocytes which provides a more accurate analysis of the membrane protein clustering. It is crucial to keep these settings for all subsequent analyses to compare the results across all the analyzed groups.
When complying with all these conditions, the analysis of SERT protein clustering in cell membranes from lymphocytes in blood smears is as accurate as the analysis done in isolated lymphocytes, and provides similar results (Fig. 4).
Our previous studies focusing on the pattern of SERT labeling in isolated lymphocytes from control subjects and depression patients allowed us to differentiate two subpopulations of drug naïve depression patients that respond differently to antidepressant drugs. More specifically, we identified a group called D-I patients, which had more SERT clusters per lymphocyte but smaller SERT cluster size than the second group called D-II patients [5]. The D-I group showed very little response to antidepressant drugs after 8 weeks of treatment, but the D-II group achieved a full remission of symptoms after 8 weeks of treatment. We concluded from these observations that the pattern of SERT protein clustering in lymphocytes could be a putative biomarker of therapeutic responsivity for antidepressant medication [5,6,7]. Although these findings are important, one of the difficulties inherent in translating these data to a clinical setting relates to the fact that most naïve depression patients are diagnosed, assessed, and get their prescriptions in a family clinic setting, where appropriate conditions for routinely drawing blood samples for experimental evaluation of this biomarker may not be present. To circumvent this problem, we focussed on developing an alternative protocol by adapting our methodology to work with blood smears that are easily obtained and can be collected in an inexpensive way. In fact, the present results indicate that alterations in SERT clustering in blood smears between human control subjects and naïve depression patients are similar to those observed when using isolated lymphocytes, thereby validating the use of blood smears for quantitative analysis of membrane protein clustering in the context of further development of this technology as a biomarker of depression.
The development of this protocol will allow us to quickly advance the validation of SERT membrane protein clustering in lymphocytes as a biomarker of therapeutic efficacy in major depression, and facilitate the implementation of this type of analysis in clinical settings.
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Rivera-Baltanas T, Olivares JM, Calado-Otero M, Kalynchuk LE, Martinez-Villamarin JR, Caruncho HJ. Serotonin transporter clustering in blood lymphocytes as a putative biomarker of therapeutic efficacy in major depressive disorder. J Affect Disord. 2012;137:46–55.
Rivera-Baltanas T, Olivares JM, Martinez-Villamarin JR, Fenton EY, Kalynchuk LE, Caruncho HJ. Serotonin 2A receptor clustering in peripheral lymphocytes is altered in major depression and may be a biomarker of therapeutic efficacy. J Affect Disord. 2014;163:47–55.
Rivera-Baltanas T, Agis-Balboa RC, Romay-Tallon R, Kalynchuk LE, Olivares JM, Caruncho HJ. Serotonin transporter clustering in blood lymphocytes predicts the outcome on Anhedonia scores in Naïve depressive patients treated with antidepressant medication. Ann Gen Psychiat. 2015;14:45. doi: 10.1186/s12991-015-0085-8.
Penn E, Tracy DK. The drugs don't work? Antidepressants and the current and future pharmacological management of depression. Ther Adv Psychopharmacol. 2012;2(5):179–88.
The authors wish to acknowledge the people who anonymously donated blood samples for this study.
This work was funded by a Saskatchewan Health Foundation Establishment Grant, and a NSERC Discovery Grant awarded to HJC, and a second NSERC Discovery Grant awarded to LEK.
Please contact author for data request.
College of Pharmacy and Nutrition, University of Saskatchewan, 107 Wiggins Road, Saskatoon, SK, S7N 5E5, Canada
Raquel Romay-Tallon, Josh Allen & Hector J. Caruncho
Neuroscience Cluster, University of Saskatchewan, Saskatoon, Canada
Raquel Romay-Tallon, Lisa E. Kalynchuk & Hector J. Caruncho
Department of Psychiatry, Alvaro Cunqueiro Hospital, Vigo, Galicia, Spain
Tania Rivera-Baltanas & Jose M. Olivares
College of Medicine, University of Saskatchewan, Saskatoon, Canada
Lisa E. Kalynchuk
Raquel Romay-Tallon
Tania Rivera-Baltanas
Jose M. Olivares
Hector J. Caruncho
RR-T, TR-B, JA performed the experiments and analysed the results; RR-T, JMO, LEK, HJC designed the experiments, analysed the results, and wrote the manuscript. All authors read and approved the final manuscript.
Correspondence to Hector J. Caruncho.
All the human procedures were carried out in compliance with the Code of Ethics of the World Medical Association (Declaration of Helsinki) and approved by the Alvaro Cunqueiro Hospital Ethics Committee (Spain), which includes a consent form signed up for all participants. All the animal procedures were conducted under a protocol approved by the Committee on Animal Care and Supply, Animal Research Ethics Board of the University of Saskatchewan (Canada).
This manuscript does not include and data from individual persons and thereby there is no need to obtain individual consent for publication.
All authors declare that they have no competing interests.
Romay-Tallon, R., Rivera-Baltanas, T., Allen, J. et al. Comparative study of two protocols for quantitative image-analysis of serotonin transporter clustering in lymphocytes, a putative biomarker of therapeutic efficacy in major depression. Biomark Res 5, 27 (2017). https://doi.org/10.1186/s40364-017-0107-6
Blood smears
Lymphocytes
Membrane protein clustering


After considering some factors, I had choosen HDFC C2P and applied for it.
My medical test has been performed in 4-5 days. Few days later I got notification from HDFC that my proposal is postposned because of 'elevated level in my blood sugar'.
It was a shocking news for me because I am a healthy person and never had any problem (touch wood).
I called hospital people (where my medical check-up was conducted) and after request they informed me that my sugar level count is 173 (normaly it should be between 70-100).
I still not belived that and went for medical check-up again by myself and found suger level count is 72(which is normal).
Then I decided to go for AVIVA i-life plan and applied for the policy.
Fortunetly/Unfortunetly, my medical examination was scheduled at same hospital, where it was conducted before for HDFC.
This time I carried my medical report (which I had done by myself) and asked hospital about the reason for differences in both reports.
According to their report my count was 173 & according to my report count was 75.
They checked again and found that it was a TYPING MISTAKE, actual count was 73 but they wrote it 173 by mistake.
Now, because of this mistake HDFC postponed my application & also my insurance records gets affected.
Hospital GM apologies for the mistake and sent clarification note to HDFC about the mistake.
Currently, my AVIVA application is under processing.
Lesson Learnt & Message: DON't trust on any medical report if it looks surprized for you.
Please double check it with mutiple doctors or Hospital if you think it is not correct and challenge it.
Dear Nitin, I'm also holding Aviva I-life & yes al types of deaths are covered. I have already cross checked it with Aviva itself. Relax.
Dear Investor, thanks for sharing your own experience. It seems, whenever there is a prem. loading due to medical test reports, the policy seeker should cross check it first as a standard practice.
if there is any foul, the course of action is defined there what dear Nitin & you have opted.
Last year i applied online for HDFC Click2protect and paid the premium.
My medical was scheduled with my permission and choice of hospital and it was done .
After few days, i was told that My premium is revised as my medical was not clear…..on more inquiry i came to know that MY HbA1C is reported high ………..that test means i have a high blood sugar.
I am a Dentist by profession and i was sure that i have never got high blood sugar.
I immediately rushed to other standard lab and go that test done at my expense and result was with my expectation……….HbA1C was quite normail. Actually HbA1C is a average value of blood sugar in last three months ,so there no question of day to day variation .
I complained to hdfc but as usual they didnt hear to me………but on repeated complaints they excepted my lab report and i was also asked to do one more Blood surgar test ……..at my own expensed……………. I argued that why should i pay ? And i will pay i need test reports but i was not given that also.
Finally , They excepted that I didnt have any high blood sugar and my premium was retained.
Being Medical profession, i had some surity about my health ,so i fought with them………….But Customer care service in this matter was not at all satisfactory.
They say, all medical reports will be kept online in My account after policy issuance…….But they keep most unimportant test reports like Urine analysis, fasting bl sugar……..When i demanded for all my test reports, they simply denied.
I own HDFC policy since last year.
Finally I got my policy document from AVIVA. However while going through the policy documents, I haven't find anything related to types of death cover under i-life plan. I already sent a mail to customer service about this and waiting for the reply.
During my initial discussion with customer care, they told that this plan will cover all kind of deaths including Terriorist activity, nature disasters. Even Suicide also but from 2nd year of the policy.
Does anybody have information about the type of death cover?
Thanks for the guidance as i was just about to go for a term plan.
Yes, I am also thinking about logging a complaint in consumer court against that hospital.
If you have some information regarding this then please share.
Dear Nitin, please accept my salute for your constant efforts & at the end of the day, your hard work has paid off. My take away from your query, I smell a RAT here. You may call it a nexus between the Insurers & the pathology labs (medical centers) that issue a wrong report & based upon that increase the prem.
If you are ready to fight it out more, my advice 'll be to report this matter to insurance ombudsman as well as IRDA also. Even you may file a consumer court case also for deficiency in services by this medical centre.
My medical examination for AVIVA ilife plan happened in same hospital (where I had for HDFC C2P plan).
After few days, I got information from AVIVA that they have raised my premium by 50% because of 'Increased liver enzymes' in my body and waiting for the balance payment to issue a policy.
It's again a shocking news for me, since I didn't expect it this time. But the good thing is that AVIVA has shared my medical report with me.
I checked with 2-3 doctors about my report and according to them this increase in liver enzymes is possible for those people who drink alchohal or having fever in last 1 months.
But for my case, I don't take alchohal nor having any fever in last months, so doctor suspect that this time also report may be not correct.
So again, I went for a medical test (only for Liver part this time) by myself and found that my liver report is NORMAL (as expected).
I informed AVIVA about the same and also shared my NORAML medical report.
After 2-3 days, finally got mail today that they have issued my policy without any extra preimum.
Hmmmmm……buying a online term insurance looks like winning a war for me.
Nitin, Thanks for sharing. Bizarre is the only word that comes to mind!
In July 2014, even I was asked for enhanced premium due to increased liver enzymes by Aviva. I did not accept increased premium as I was convinced that medical report was flawed. I asked Aviva to refund my entire premium. They deducted cost of medical test. Despite protests on not to deduct cost of medical test, they were adamant and they never refunded entire premium. After seeing your comments above this seems to be common practice followed by Aviva and one needs to complaint to IRDA.
Dear Nitin, yes you may keep the letter of that hospital error as proof.
@Karthikraja : HDFC gave full refund of my premium paid because of postponement from their side.
What should you suggest for me, since my insurance record is already affected because of postpond of policy (though mistake is from hospital side), what should I do so that in further my new insurance doesn't got affected due to this?
Should I ask hospital to give clarification letter to me about their mistake and kept it as a proof for further reference?
Thanks for sharing this. We all try to get second opinion from Doctors for the planned procedure ( operation/surgery) except Emergency. But even we should get second opinion for the medical lab test is really new one and we should remember it.
So you will loose the Premium paid money if it is beyond the look up period or incur the test expenses.
Thanks for sharing that ! .. it was an eyeopenor .. Its always a good idea to note down few things which you can at the time of medicals itself , like atleast weight and height information .
Dear Nitin, my congratulations to you for the over all things & your fight for your cause.
Good that the hospital acknowledged their mistake. Your hardship not withstanding this kind of error is actually rare. What is more common is small differences. Most machines are not calibrated periodically and if a persons actual value is 95 it could come out as 105. This puts one in the danger zone. difficult to disprove this. Repeating this may or may not help.
Must as we like to blame hospitals it is actually quite a common occurrence to see people who consider themselves perfect healthy to find out that they are in the pre-diabetic stage.
Completely agree with your central point though: Recheck.
A lesson for all the readers. This implies how serious they were at the time of testing.


text:
Let'S Talk About Love by Claire Kann
Alice is secretly asexual, and that's the least important thing about her. She's a college student, has a great job, amazing friends, and is fine being single - nope, that's a lie. Alice wants rom com-grade romance: feels, cuddling, kissing, and swoons galore - as long as it doesn't lead to having sex.
When her last relationship ends, Alice swears off relationships for good. Stick a fork in her, she's done. Everyone Alice tries to date is so sure love and sex have to go together, and there doesn't seem to be any way to convince them otherwise.
But when Alice meets Takumi, who she can't stop thinking about, she doesn't know what to do. If Alice tells him the truth, it can only end in heartache. But there's something about Takumi that makes him worth the risk.
The Song of Achilles : Bloomsbury Modern Classics by Madeline Miller
Pride and Prejudice (Barnes & Noble Flexibound Classics) by Jane Austen
Call Me By Your Name by Andre Aciman


Many travellers are searching for What To Do In Bangkok In The Morning,Bangkok can well be described as a city of humble beginnings. In fact around 200 years ago, it was simply a small trading center and port community. Despite the fact that the city had grown rapidly and become renowned for being a modern and vibrant place, it has impressively retained its splendid and enchanting glory. That's why a visit to Bangkok should be on everyone's bucket list. You can find What To Do In Bangkok In The Morning travel information.
Also called Wat Chetuphon, this is a temple that was built for King Rama in Bangkok a huge selection of years back. It is actually thought to be a location where people can receive feelings, and was considered one of the primary pharmacies in all of Bangkok. Additionally it is the location of the first university which had been started by way of a descendent of King Rama, a place where you can now get not just traditional medicine but a foot massage at very inexpensive prices. It can be known mostly for the Temple in the Reclining Buddha and that is a Buddha that is laying perched on his arm. It is enormous, roughly 15 m high by 45 m long, and might just be viewed in sections due to its size. There are lots of beautiful precious stones that have been in lead from the soles of the feet, an overall amount of 108 stones total. This really is a number related to procession, the wobble of your earth, an issue that is constructed into most of the temples throughout Thailand.
It is an absolutely beautiful structure which has quite a bit of history, and it is the house of what exactly is called Wat Phra Kaeo. It is here how the emerald or jade Buddha can be obtained, in this particular palace internal 1782, the residence of your royalty that had been in control of the country at the time. This palace includes 2 hours of not only the palace itself but also the Wat Phra Kaew Tour. It is recommended that you start out early each day to take full advantage of everything that you may be able to see, or perhaps perform a half day sightseeing tour to see either one.
You can observe a wonderful representation from the ancient battles that employed to occur between Burma and Siam. This is actually a mixture of Royal Palace and Chapel, and if you can to climb to the top prior to sunset, you will notice the sun setting in to the Chao Praya River, an attractive sight to behold.
Looking at what is called Wat Suthat is really what the locals call the giant swing. It really is a 27 m high structure, use for religious ceremonies, as well as its purpose is to pass through a ritual that requires teams of three people going to the top and walking across towards the narrow board. There was a significant amount of fatal accidents back into the 1930s which make them not make use of this being a ritual anymore. However, this is a beautiful sight to see, especially facing Wat Suthat.
This can be literally something from the movies, a genuine floating market that is about 1 hour away from Bangkok. It is actually considered by some to be the Venice from the East, just like how the people in that component of Italy achieve this many things in the water. Additionally it is a tourist trap as a result of how people are looking to check this out market that is completely floating on boats manned by people selling their goods. However, when you do go, you may be treated to many of the most delicious food that is absolutely fresh, served in a authentic way.
When you have time, you are able to pay a visit to Lumpini Park which is a literal oasis in the middle of the architecture and buildings of Bangkok. There are several lawns you could hang out at, Chinese pagodas, and you will discover a small lake that you could require a boat out on. Is good for relaxing for the reason that park is indeed tranquil, despite all of the skyscrapers which are surrounding it in every directions. The park has become a place where antigovernment protests have occurred, so just be certain that if you decide to look there, it's just people chilling out to unwind.


text:
Sports\Stars
Brady super stoked on China
By Murray Greig | China Daily | Updated: 2017-06-20 07:00
Legendary QB harbors dream of playing here
If telephone booths were still ubiquitous in the United States, Tom Brady might be tempted to duck into one to don his New England Patriots uniform every Sunday during the NFL season.
You know - the same way mild-mannered reporter Clark Kent changed into his Superman tights, before the booths became extinct.
The only difference is that Brady has never needed a cape to be super.
The 39-year-old quarterback and two-time league MVP is one of only two players to win five Super Bowl rings (former San Francisco and Dallas linebacker Chuck Haley is the other). He's also the only four-time Super Bowl MVP and owns 25 individual playoff records, including most career home victories (15), most touchdown passes (69) and most passing yards (9,094).
Oh yeah ... Brady is also married to a supermodel (Giselle Bundchen), is on a first-name basis with President Donald Trump, has raised tens of millions of dollars for charity and is a coveted endorser for some of the world's most prestigious brands.
And if celebritynetworth.com is accurate, he's socked away a cool $180 million.
But perhaps most impressive is that after 15 seasons in the spotlight, Brady remains the same affable, unfailingly polite boy-next-door he was when the Patriots selected him in the sixth round - 199th overall - in the 2000 NFL Draft.
So how does he stay so motivated ... and unaffected?
"I'm sorry I don't have a more complicated answer for you, but I'd have to say it just comes down to my love of the game," Brady said at Beijing's Shi Cha Hai training center on Sunday afternoon, where he was promoting Under Armour's latest line of apparel.
"To be honest, it doesn't take much externally to motivate me. Everyone finds different ways of preparing themselves, mentally as well as physically ... and I'm very happy with the ways I've found. Football was my first love, and I've been very fortunate to have that feeling stay with me through all these years. In fact, it's grown stronger as I get older.
"Every practice, every game and every season presents new opportunities to challenge myself to be better, to not accept anything less than my best. That's all the motivation I need."
Like a high-school kid on his first field trip, Brady was bubbling over with excitement about visiting Asia for the first time.
"I've been fascinated with China my whole life, so to finally have the opportunity to come here and experience the people and the culture first hand is really incredible," he said.
"We arrived on Saturday night and we've already had a tour of the Great Wall and taken in some other sights. I'm really happy that my son made the trip, too. We tossed a football on the Great Wall, which made for a very special Father's Day moment that we'll cherish."
Brady, who will also visit Shanghai and Tokyo on this trip, said he hopes to have an opportunity to learn more about another subject close to his heart: Traditional Chinese Medicine.
"One of my best friends at college majored in TCM, and it has been part of my training and general health philosophy for years," he said.
"A lot of the value of TCM is preventing injuries before they happen by stressing hydration, nutrition and recognizing that what you put into your body is what you get out of it. It's a whole-body approach to better health, and I think it's wonderful."
Earlier this year, NFL commissioner Roger Goodell said the league is looking at staging a regular-season game in China by the end of the decade - and Brady would love to be calling signals in it.
"I think I can play at least another three or four years, and my dream is to one day play a game here in China. It's just a great spectacle, and knowing how passionate Chinese people are about sports, I think they would really enjoy it," he said.
Until that happens, Brady is happy to do all he can to promote the game in the world's most populous nation - including taking part in an energetic exercise session with aspiring Chinese players on Sunday.
"This country has a long history of producing great athletes, and with the dedication and work ethic these kids have, I don't think it will be too long before American football really takes off in China," he said.
murraygreig@chinadaily.com.cn
New England Patriots quarterback Tom Brady is flanked by a pair of trainers while learning wushu techniques during an impromptu workout at Beijing's Shi Cha Hai training center on Sunday.Wei Xiaohao / China Daily
(China Daily 06/20/2017 page24)
New bullet trains to depart on Monday
Top 11 fastest growing economies listed
The world in photos: June 19-25
Twin panda cubs celebrate upcoming first birthday in Macao
Rescue work underway after SW China's devastated landslide
Ivanka to visit China


Every morning the cat sits at the bottom of the steps and patiently stares at this spot on the wall, waiting for the shadows to show up. With any luck there is enough sun to give her some interesting stuff to jump at.
I'm enjoying Shadow Shot Sunday so much (over at Hey Harriet) that I'm playing along this week!
My sister and I both have birthdays within a week of each other in April, so we tend to celebrate together. This year, our parents came in and our other sister came down with her two girls for a family get together.
At one point, during all the very noisy bustle, it was announced that I had a birthday surprise, and had to cover my eyes for a few minutes while they brought it in.
Now, I've been campaigning hard for a new puppy, but want to wait until after summer vacations are over. When they were all talking about how heavy my present was and did they need a dolly to get it in, etc, and then there were lots of shhhh-s-ing noises, I was pretty convinced that, OH NO, they had gotten the dog early.
My daughter had come up from Florida for a birthday surprise.
It was a great weekend, even if it was over with too soon, with everybody going home on Sunday.
This evening, cleaning house one minute, then out in the yard cutting flowers the next. Thinking back, I honestly don't remember going from point A to point B ... I mean, really! I can't think of what made me put down whatever I was cleaning and wander out into the yard. I must have accidentally looked out the window.
Brrrr! Flowers and snow... a pretty combination early this morning.
What a beautiful weekend! Friday evening I got to visit with my parents who were in town for the evening, which started the weekend out right. Saturday morning my friend Ruth brought some flowers from her yard to our painting group, so I just had to do a quick sketch of them. I wish you all could have smelled those lilacs -- heaven. Later, after a diet-busting lunch with Doug and a nice long talk with my daughter on the phone, I got to meet up with Diahn to go see some great art and then hang out at the coffee shop (busting the diet even further) until nearly dark. Simply gorgeous.
And the gorgeousness continued today with lunch at Cosmo's and a new little piece of furniture and getting the wireless printer all set up, then opening the windows and turning on fans while doing a total rearrange of the guest room closets, my study closets and making a good dent toward cleaning and rearranging my painting space. All being done as I get ready for a piano to be delivered into the same area where I paint and (YAY!) company coming in next weekend. I worked like a dog but had a blast doing it, with the fresh air and the music blaring and knowing that the end result would be fantastic.


Zuma is committed to transparency regarding our charitable contributions by providing annual updates of our donations to the wonderful charities we support. We believe that Business can create positive social change through giving back, and that is our "reason for being" as a company.
Sometimes we struggle with our donation updates and marketing message because in a sense they lack true humility. They can focus too much attention promoting ourselves versus the charities and good work we aim to support. However, as an independently owned business in an extremely competitive market like office supplies, we feel it's necessary to communicate our mission to attract new loyal customers to spread the word about Zuma and help us grow.
The fact is if we don't grow, we don't give back. If we do not drive solid profits, our mission to help others is not accomplished. It is because of you, our loyal customers, that Zuma is able to exist and do our part to make the world a better place. So, please view our giving back results as your success!
Zuma has donated $24,281 to the 10 charities we support since our inception. Most new businesses take 3-5 years before they reach profitability due to the upfront investment, and Zuma is no exception. However we made the decision as an executive board to stay true to our mission and donate a minimum of $5000 per year, ($500 per charity) even before we were profitable. Although modest, fiscal 2012 was our first profitable year, and so our donations are now based on the preferences of our customers!
In relation to our true financial results, because of our commitment to give back from the start, we've donated many times over 100% of our profits. And the great news is, now that we are in the black, these donations will grow substantially as we gain momentum.
We appreciate the loyalty of our great customers, the relationships we've developed with the charities we support, and look forward to doing good work together for many years to come. Please tell a friend and other businesses about Zuma – you have no idea how much that helps our progress. Should you have any questions regarding our charitable giving, the voting preferences of our customers, or just want to talk about giving back, feel free to email me at gpierce@zumaoffice.com. Thank you for helping Zuma do good work!
This entry was posted in Charities and tagged Giving Back, office products, Office Supplies Online, Zuma, Zuma Charities. Bookmark the permalink.


text:
Tag / Psychological Suspense
I Let You Go by Clare Mackintosh
Adult Fiction, Artists, Auto Accidents, Coastal Towns, Domestic Abuse, Family Drama, Grief, Hiding, Hit & Run, Investigation, Loss, Loss of Child, Mystery, Police Procedural, Psychological Suspense, Psychological Thriller, Remote Locations, Romance, Secrets, Suspense, Thriller, Tragedy
I Let You Go will be published by Penguin Group on May 3, 2016.
Oh my goodness – this book is a roller-coaster ride! This book has it all – part police procedural, mystery, love story, and family drama. There are two story lines, both starting out with a tragic hit & run accident, killing a young boy named Jacob. Jenna Gray leaves her grief from losing her son and a fear-filled life for a remote seaside cottage on the Welsh coast. Meanwhile, seasoned police inspector, Ray, teams up with younger, beautiful inspector trainee, Kate, in an attempt to find the hit & run driver and bring some closure to Jacob's mother. Ray's home life is just about as stressful as his work life, due to the tension with his wife, a former police officer, and problems at school with his teenage son. Kate is young, energetic, easily accessible, and a great sounding board for Ray. Will he slip up and jeopardize his future, both at work and at home?
When Part 1 ended with a major twist, I had to go back and reread the first two chapters just to figure out what was going on. It really threw me for a loop, and after that I couldn't put the book down. Part 2 picks up quickly, with jaw-dropping scenes and unimaginable, terrifying moments, as Jenna's story is revealed piece by heartbreaking piece. The story line truly proves that things are not always what they seem.
This book made me cry for Jenna, and also gave me chills of terror. The writing is excellent, as even the ending leaves readers wondering if what people believed happened actually did transpire. Upon finishing, I was left with an unsettling, creepy feeling that can only be quelled with a sequel. I do hope Mackintosh writes many more novels. This is her first, and it's an amazing accomplishment. Ray's character reminds me a bit of Michael Connolly's famous character, Harry Bosch. Fans of the Bosch series and of psychological thrillers such as The Girl on the Train, The Luckiest Girl Alive, and The Good Girl will enjoy this book. I highly recommend it.
Wilde Lake by Laura Lippman
Adult Fiction, Coming of Age, Family Drama, Family Secrets, Lawyers, Mystery, Psychological Suspense, rape, Small Town Life, Suspense
Wilde Lake will be published May 3, 2016 by Harper Collins.
Wilde Lake is an atmospheric, family-centered, legal suspense novel which will keep readers interested with a quick paced plot, as layers of buried secrets are uncovered one at a time. Each family member: Lu, AJ, and their father, is hiding secrets of their own, and they all come to light, some very tragically, in the final chapters.
The story line might sound familiar to an old classic, To Kill A Mockingbird. This was by design, according to the author. She wanted to portray the idea that a crime, namely rape, committed in 1980 is looked at differently at the time, both legally and socially, than it would be in 2015.
Setting plays a large role in the story, beginning with the way the Brant family ended up in Wilde Lake, back when it was a newly developed area, offering such promise for success. Lu's mother was talked into moving there against her wishes. Interestingly, the Brant family has become very important to the area, and are well-known for having two state attorneys in the family, Lu's father, and Lu herself.
The dynamic between Lu and her older brother, AJ, adds a richness to the story. From as far back as Lu can remember, she has been in AJ's shadow, trying to measure herself to him, while others are drawn to him and his talents like a magnet. Their relationship is strained, and ultimately broken when Lu finds out the truth about the night when teenage AJ saved his best friend Davey's life and was deemed a hero. Is he really the hero that everyone believes he is? Is Lu forgetting something important about those events; or was she not privy to what really happened during her childhood?
The story switches back and forth between present day Lu at age 45, all the way back to when she was only 6 years old. Lu's mother passed away when she was 1 week old, so she was mainly raised by Teensy, a nanny hired by their family. Her father was a prominent lawyer and political figure who spent more time in the courtroom than he did at home. As snippets of Lu's childhood and her upbringing are revealed, readers may better understand Lu's demeanor and outlook on life. In the present, Lu, the first ever female state's attorney in the state of Maryland, is prosecuting a case against a young man who murdered a woman who comes home to her apartment to find him, a stranger, there. Their town doesn't see many murder trials, so Lu wants to do her best to seek the truth and prove herself worthy as the newly appointed state's attorney. The more involved she becomes in the investigation, the closer she gets to uncovering some long buried family secrets.
I enjoyed this novel, and would recommend it to any fans of suspense and/or legal suspense.
Missing Pieces by Heather Gudenkauf
Adult Fiction, Cold Cases, Family Life, Family Secrets, Farm Life, Murder, Mystery, Psychological Suspense, Suspense, Thriller
Married for many years and with twins in their first year of college, Sarah and Jack Quinlan live a pretty normal life. Sarah feels like she knows her husband pretty well. However, when Jack's aunt Julia is in a tragic accident, Jack and Sarah make their first trip back to Jack's hometown of Penny Gate to be there for his family. Julia was like a mother to Jack and his young er sister, Amy, after Jack's parents were killed when he was a teenager. Sarah was always under the impression that Jack's parents were killed in an auto accident. Once she begins talking to Jack's family members and others in the tight-knit community of Penny Gate, she realizes that Jack has been hiding the truth about his parents fate, as well as many other things from her throughout their whole marriage.
After Jack's sister, Amy, is suspected of hurting Julia, Sarah begins to wonder if Jack is not the true mastermind behind it all. It doesn't help his case when Sarah finds out that his cousin Hal's wife, Celia, was Jack's serious girlfriend in high school. Not one to back down easy, Sarah decides that she will not leave Penny Gate until she discovers the truth about Jack's parents, Julia's "accident," and other strange things she has witnessed in the short time since she arrived. The closer she gets to the truth, the closer she gets to someone playing a very dangerous game with her life.
This is a fast paced read, and the suspense builds all the way to the end. Gudenkauf is great at leaving little clues throughout the story, without giving away the outcome too early. I enjoyed the book, and I would recommend it to fans of Mary Kubica, Gillian Flynn, and Chevy Stevens.
Fool Me Once by Harlan Coben
Adult Fiction, Corruption, Family Secrets, Harlan Coben, Military, Murder, Psychological Suspense, PTSD, Surveillance, Suspense, Thriller, Trust, Wealthy Families
Fool Me Once will be released on March 22, 2016. This is a stand alone suspense novel from Harlan Coben, best known for writing the fast-paced, popular Myron Bolitar series. Coben is a master at writing hard-to-put-down novels, and his latest is no different.
Maya Stern, a former special operations pilot, has just buried her husband after he was brutally murdered in what appears to be a robbery gone wrong. Maya is no stranger to loss, having just buried her sister 4 months prior to her husband's death. A concerned friend, though Maya doesn't truly trust anyone, has brought her a nanny cam to help her feel at ease while at work away from her 2 year old daughter. One day while viewing the footage, she catches a glimpse of the unbelievable – her husband playing with their daughter. She was there when he was shot and she knows he's gone; so, how can this be? Is she suffering from PTSD, hallucinating, and imagining things, as her in-laws and friends start to suspect? Or, is Joe still alive? She starts to question the nanny, and realizes that someone is covering up the truth. She is determined to uncover the truth about Joe, and also his younger brother who died in a boating "accident" while the two were in college, despite being warned against such activity by those closest to her. Maya will not give up until she assures that the future is safe for her daughter.
This book will have you on the edge of your reading chair, biting your nails to the quick. It's trademark Coben, with a major plot twist which throws the reader for a loop, as he or she tries to figure out the truth. This is a quick, suspenseful story with an unexpected ending. Readers will not be disappointed.
Don't You Cry by Mary Kubica
Adult Fiction, Family Drama, Mystery, Psychological Suspense, Roommates, Suspense
Don't You Cry will be published and released on May 17, 2016. This book will surely be a hit summer read!
I've read and thoroughly enjoyed all of Mary Kubica's fast-paced suspense novels. She has a knack for hooking the reader into the story and then throwing a major plot twist in the mix, causing the reader to race to the end of the book with abandon. That is exactly how I read this book, all in one day because I couldn't put it down. It's like watching a scary movie; when you have to find out what will happen to the characters in the end.
Don't You Cry is the story of roommates Quinn Gallo and Esther Vaughn. While Quinn spends free time socializing, bar hopping like most single women her age, Esther is more content to stay at home. Esther is quiet, always willing to help others, and even sings in the local church choir. Quinn comes home late one evening, thinking Esther is already asleep, but in the morning she finds that Esther is gone, her window open and her cell phone left behind. Quinn is worried, and as she begins to search for clues to where Esther could be, she starts to find out some very puzzling things about her roommate. Namely, a disturbing letter addressed to "My Dearest," large ATM withdrawal receipts, and the truth surrounding the fate of Esther's previous roommate. Though Quinn was sure that she knew Esther and they were pretty close, she begins to wonder if she ever really knew her at all.
Also intertwined within the story is that of Alex Gallo, a recent high school graduate who turns down a full ride scholarship to take care of his alcoholic father, the only family he has left. Any money he works busing tables at the local diner goes to paying the bills and feeding his father's alcoholism. Alex just happens to be in the right place at the right time and catch a glimpse of an intriguing, mysterious girl. He becomes obsessed with getting to know her, helping her, and finding out why she turned up in his town. She refuses to tell him her name, so he refers to her as Pearl, because of the pearls on a bracelet around her wrist. Readers will really feel sorry for Alex, who is a genuine nice guy, the type that people like his father and Pearl will walk right over, when given the chance.
This story twists and turns like a roller-coaster, keeping the reader on the edge of their seat and racing to the finish. I loved the story, and the ending was a little bit crazy, but in a good way. Fans of The Girl on the Train, Gone Girl, and The Luckiest Girl Alive will be sure to enjoy Don't Your Cry, as well as Kubica's other two previously published, psychological suspense novels.
The Girl Without a Name by Sandra Block
1960s, Adult Fiction, Doctors, Missing Persons, Psychiatric Hospital, Psychological Suspense
This book will be published September 8, 2015 by Grand Central Publishing.
The Girl Without a Name is a psychological suspense novel that I really enjoyed. Dr. Zoe Goldman is a psychiatrist completing her residency in the children's psych ward. Certain events in Zoe's past are still coming back to haunt her, and she actually sees a psychiatrist herself, which is rather ironic. Zoe's real mother and foster mother have both passed away and her only remaining family is her brother, Scotty, and dog, Arthur. She also has a boyfriend, Mike, who is an ER doctor. When a new patient arrives, Zoe begins to doubt some of the treatment courses the attending physician, Dr. Tad Berringer, prescribes. The patient, referred to as "Jane," because her identity is unknown, is a young African American girl in a catatonic state who doesn't remember where she came from, or how she ended up in the hospital. She has a peculiar scar on her ankle, and is mysteriously being given medications even though no once claims to have prescribed them for her. Zoe begins working alongside a seasoned police detective with whom she has come into contact in her past. She starts to uncover clues about "Jane," and puts her own career in jeopardy to ensure that "Jane" is safe. It doesn't help that Dr. Berringer is a married, heartthrob who begins to take more interest in Zoe after he informs her that he is getting a divorce. Will Zoe fall for his charm? How can she know who to trust with the truth about the patient. The plot is fast paced, and I read it very quickly.
Pretty Girls by Karin Slaughter
Adult Fiction, Karin Slaughter, Kidnapping, Psychological Suspense, Thriller
This book is set to be published on September 29, 2015.
I raced through this book so quickly because the story line was fast-paced even though it jumps back and forth between past and present tense. It is definitely a nail biting, up-all-night-after-reading kind of scary suspense novel. Karin Slaughter has an uncanny knack of giving the reader the chills while also making he or she want to keep reading for more chills. There were a couple of plot twists in the book that were pretty shocking, but well worth it. The violence and description of such is pretty gory, so I wouldn't advise reading if you do not handle that kind of thing very well. I won't give many details as to the plot or synopsis of the book, because it becomes apparent rather quickly in the first few chapters. The craziest thing about this story is that one of the sisters was married to a man for almost 20 years and knew absolutely nothing about his actual lifestyle and killer instincts. Very creepy. I've read all of Karin's book so far, and this is one of my favorites, despite that fact that I was up thinking about it far after I had finished the last page. Trust me; this is way better than a scary movie or a creepy episode of Dateline!


text:
Q: GoogleTrans Python not translating I am using GoogleTrans for my project. Actually I have a text in Marathi which I got from "speech to text" but when I am converting that text into English it is not translating properLY.
But when I am using Google Translate from website, it is translating properly with 100% accuracy ?
Image
Here is the piece of Code :
import speech_recognition as sr
from cltk.corpus.utils.importer import CorpusImporter
from googletrans import Translator
import goslate
import googletrans

print("Hello World")

filename = r"C:\Users\TOSHIBA\Documents\Mini Project-2 (Trimester 4)\Project 1- (Document Summary)\Dataset\Voice\Gaurav.wav"
r = sr.Recognizer()

with sr.AudioFile(filename) as source:
    audio_data = r.record(source)

    text = r.recognize_google(audio_data,language='mr')
    print("Text\n",text)

    print(type(text))

    translator = Translator()
    converted_text = translator.translate(text)
    print("Converted text\n",converted_text)

    with open("D:/output.txt",'w',encoding="utf-8") as f:
        f.write(text)
        f.write(str(converted_text))

Please suggest, what should I do?

A: https://pypi.org/project/googletrans is an unofficial package (not created by Google). Also in the disclaimer, it has the following "...this API does not guarantee that the library would work properly at all times...". If you want stability, use official Google API https://cloud.google.com/translate/docs
Anyway below is a test to show the translator work:
from googletrans import Translator

translator = Translator()
results =translator.translate('हॅलो वर्ल्ड')
print(results.text)

Output:
Hello World

I'm not sure what the issue you're experiencing. However I would suggest refactoring your code into methods and then you can test in isolation.
The code below has not been tested, but should help debug:
import speech_recognition as sr
#from cltk.corpus.utils.importer import CorpusImporter
from googletrans import Translator
#import goslate
#import googletrans

def provideAudioToText(filename):
    r = sr.Recognizer()
    with sr.AudioFile(filename) as source:
    audio_data = r.record(source)
    return r.recognize_google(audio_data,language='mr')

def translate(text):
    translator = Translator()
    results = translator.translate(text)
    return results.text

def Save(filename, original_text, converted_text):
    with open(filename,'w',encoding="utf-8") as f:
        f.write(original_text)
        f.write(str(converted_text))

original_text = provideAudioToText(r"C:\Users\TOSHIBA\Documents\Mini Project-2 (Trimester 4)\Project 1- (Document Summary)\Dataset\Voice\Gaurav.wav")
print("Text\n", original_text)
print(type(original_text))
converted_text = translate(original_text)
print("Converted text\n",converted_text)
Save("D:/output.txt", original_text, converted_text)

A: Upgrade your googletrans module as below.
pip install googletrans==4.0.0rc1

I see the default installation googletrans uses the 3.x version and it has the problems that you mentioned above.
See the conversation on Github page

A: had same issue with 3.0.0, worked with latest version . 4.0.0rc1

redpajama_set_name: RedPajamaStackExchange


text:
info@cefne.com
WHAT DOES HE DO?
CEFNE
AUTHOR & LECTURER.
CONSULTANT & COACH
EDUARD BELTRAN
An expert in negotiation, Eduard received training in the Negotiation Program at Harvard University. He was awarded a Law degree from the University of Barcelona, a Political Science degree from the Institute of Political Studies in Paris and a master's degree in International Law (LLM) from New York University. He is also an alumnus of the prestigious Ecole Nationale d'Administration (ENA).
As a lawyer, he has worked on international investment projects and international conflict management. As head of the Department of International Cooperation at the Ministry of Justice for the French government he was responsible for relationship management with the European Union, international organizations, and foreign governments.
In the field of negotiation, he has trained over 5,000 professionals, been involved in negotiations in more than 25 countries in Europe, America, Africa, and Asia, and has advised over 100 companies and international groups as a senior consultant at the Centre Européen de la Négociation in Paris. Currently, he runs the Center for Negotiation (CEFNE) and collaborates as an expert and professor in Institut d'Etudes Politiques, Paris, Universidad Complutense, Madrid and Universitat Pompeu Fabra, Barcelona.
Director of CEFNE - Center For Negotiation
Professor of International Negotiations & Leadership (ENA France, Sciences Po Paris, Univ Complutense Madrid, Pompeu Fabra Barcelona...) Author of many books and articles, speaker and trainer
Principal Consultant of the European Negotiation Center (Paris)
Attorney at Law in France, Belgium and Spain
Head of the Department of International Cooperation of the Ministry of Justice (France)
Negotiation & Leadership certificate, PON Harvard University (USA) 2016
Master of Laws (LLM), New York University (USA) 2013
Ecole Nationale d'Administration (France) 2006
Institut d'Etudes Politiques de Paris (France) 2004
Bachelor of Law, University of Barcelona (Spain) 2002
HIS INSPIRATION
Aurélien Colson
Daniel Shapiro
David Lax
Deepak Malhotra
Diamond Stuart
François de Caillères
James K Sebenius
John C Maxwell
Howard Raiffa
Max Bazerman
Peter F Drucker
Richard Shell
Robert B Cialdini
Roger Fisher
Stephen R Covey
Tahl Raz
"A book is a garden, an orchard, a storehouse, a party, a company by the way, a counselor, a multitude of counselors."
Contact with his team
Some customers
TALENT PREMIUM GROUP AMERICA
224 Fifth Avenue, Suite E208
New York, NY 10.001, USA
TALENT PREMIUM GROUPE EUROPE
31-35 rue de la Fédération
2022 © eduardbeltran.com - All rights reserved.
That makes


We've all figured out long ago that we are what we eat.
However, now more than ever, it seems like the challenge is to confirm exactly what it is we're eating.
In theory, we're supposed to have a bit of help with that task. In the USA, for example, we'd turn to the Food & Drug Administration. They do what they can, but compared to the marketing mavens at Big Food, their message is quite often dizzied by all the spin.
Incidentally, one of the ingredients that got top billing on a label in this report is absolutely the worst stuff you can put into your body: high fructose corn syrup.
The FDA's authority over food labeling terms generally limits itself to nouns. It's referring to categories.
All marketers know the saying to sell the sizzle, not the steak. In other words, it's the adjectives that appeal to a consumer's emotions and biases.
Many do an effective job of couching what they're not in terms of what a customer is actually seeking.
Thus, you've got to be aware of what you're reading as opposed to what you think you're reading.
One of the most misunderstood term sets is non-GMO vis-a-vis GMO.
First of all, what is a genetically modified organism, really?
Take a carrot, for example.
They actually come in a multitude of colors. However, orange became the default option when the Dutch cultivated them to celebrate their fearless leader, William of Orange, back in the late 1600s, and its popularity stuck. They did it by selective breeding, aka genetic modification.
Yes, GMO's been around that long and longer. Much longer.
If you're dug in against the concept, though, and want to stay non-GMO, then stick to heirloom vegetables and heritage meat. Many of those will taste better, although you often pay more for the privilege.
Basically, this is a term reserved for foods that are minimally if ever exposed to pesticides and is free of antibiotics or hormones.
The key here is there are standards for organic food. As Dr Mike explained, terms such as natural and healthy are nice to know, but they don't have an objective meaning.
That makes it even more important for us to know what we're eating.
Next articleIs Your Social Media Message Really Connecting with Your Target Market?


text:
Guy Robertson
Clinic Schedule 2022
Cow Horse Clinics Schedule 2022
10 Day Starting Clinic
Terms & Conditions of All clinics
Starting & Breaking
Ground Work & Young Horse Training
Racehorse Retraining
Bespoke Horse Training
Horsemanship Weeks
Horsemanship School
GRH Online Training HUB
Horsemanship Demonstration March 2019
Trailer Loading newsletter
Guy has travelled extensively, and considers himself a student of horsemanship and great horseman. Guy has trained and started horses all over the world, including UK, Australia, USA, Poland and New Zealand.
victoria robertson
Victoria grew up around horses living on her family's equestrian centre and riding school in Bradford, West Yorkshire. It was this upbringing that sparked a passion for horses and she spent much of her formative years riding and showing, native ponies, at county shows up and down the country.
It was in 2010 Victoria first met Guy when she retained his help with her Welsh Section C stallion Rembrandt. Fast forward 10 years Rembrandt now belongs to Guy and Victoria's daughter Summer and is still very much part of the family.
In 2011 victoria joined the business and was instrumental in taking it forward with her hard work, photography skills and social media expertise. She still runs the office and takes care of the day to day running of the yard in addition training and working her own horses.
our extended team
All Creatures Animals Care
Karen is a regular face on the yard helping us with all aspects of animal care she is very knowledgable and has been with us now since December 2020.
Registered Veterinary Nurse
Qualified Companion Animal Behaviourist
Qualified Lecturer
Martin brookes - equine Dentist
Guy & Martin have been friends for over 25 years, they first met at a Polocrosse open day in 1996.
After Martin completed a degree in Mechanical Engineering at the University of Sheffield he attended the Academy of Equine Dentistry, USA in 2002. He certified in 2003 and has attended for at least 10 days each year to complete CPD.
Martin passed the DEFRA/RCVS approved exam in the UK in 2006 and represented the WWAED (UK) on the Veterinary Activities Committee for LANTRA in developing the National Occupation Standards for equine dentistry. He is currently a member of BAEDT.
Martin has ridden since 5 years old in a variety of disciplines, being a member of local riding clubs and Rockwood Harriers Pony Club. The highlight so far was representing the UK Polocrosse team in the 2007 World Cup in Australia, coming second. He currently plays polo at Leadenham Polo Club.
Charlotte Innes - the horse vet
Dr Charlotte Inness BVSC MRCVS
Charlotte qualified as a veterinary surgeon from the University of Liverpool then worked in Australia, Hong Kong, Staffordshire and finally Yorkshire as an Equine Veterinarian. During this time she has worked with all types of horses from racehorses, brood mares, foals, competition horses and companions. Always having an interest in lameness and poor performance, Chiropractic was appealing as another skill to aid lameness work up and diagnosis.
Having carried out further training in Germany at the International Academy of Veterinary Chiropractic she decided to incorporate chiropractic more prominently in her veterinary work.
The Horse Vet was established in 2015 to provide a service to treat poor performance in horses, focussing on the musculoskeletal system, using a combination of traditional veterinary medicine, equine chiropractic, equine acupuncture (traditional Chinese and Western approach) and state of the art gait analysis technology.
Outside of work Charlotte enjoys going to the races, playing polo, hunting, walking her energetic German Short Haired Pointer and relaxing when time allows.


Christmas Wishes, Mick Jagger and No Resting on Laurels!
As 2013 draws to a close there could not be a better time to "take stock"( if you'll pardon the pun), of the housing market in Coventry and it's surrounding areas. Before that, 2013 has been a landmark year for Elizabeth Davenport Estate Agents. Not only have we opened our first, prominent High Street Office and employed our first non director staff, we have been consistently ranked either 1st or 2nd place for the click through rates on our website and the speed of our sales! With over 85 Estate Agents in competition with us this is something we are obviously very proud of. 2014 will see no resting on our Laurels (Christmas Pun intended) and we are investing in dramatic new sign boards that really compliment the nature of Agency and most importantly will get your property noticed first! We plan to further recruit and are constantly improving our website. We have invested further in professional photographic equipment too which is evidenced within our brochures and proven by way of our number 1 "Click Through" ranking by Rightmove. Highlights of the year for us include the dramatic amount of viewings and sale of "Whitegates", a characterful and unique cottage residence in Keresley that had sat on the market gathering dust with other agents for over six months. We sold this within 4 weeks. "Armorial Road" is a similar success story with us having a number of offers and plentiful viewings when other agents had again failed. The sale of "Cannon Close" was another proud moment. A number of agents had valued the house and many unusual tales arose from these valuations too which quite honestly don't depict Estate Agents well at all. Myself and the owners got on really well and the offers we received came thick and fast. It was certainly lovely to deal with a seller with such moral virtues. We had offers from investors and owner occupiers but they wanted the house to sell to family who would cherish and develop it over the years and that's exactly what happened. The seller, as it transpired, was attracted to us by the quality of our photography which was certainly saying something as that was his profession! He kindly photographed George and I outside the house as a memento. For us it was great. For him probably a step down as he's previously photographed Prince Charles, Mick Jagger and a host of real talent!
So what can we expect of the housing market in 2014? Firstly throw away the crystal ball and think about your own needs first. Don't get excited or depressed by national statistics or the media's declaration of a new housing boom with property prices increasing 6% per annum. In the West Midlands property prices are still considerably lower than they were at peak and only London and the South East are showing above inflation linked asking prices. However the "West Midlands " statement is also not specific enough. In Coventry alone the growth in one area has been substantially greater than another with property prices literally at the end of one road remaining quite static whilst the other end see's growth. This is why the only real monitor of worth is to the buyer. We need to put ourselves in there shoes when valuing correctly. In 2013 despite record instructions our stock levels remain low. When we price and market a property it sells very quickly. So for every new instruction a sale also occurs. This is an unlikely business model that must be retained. Why would any agent want a stock of 100's of unsellable houses because they haven't addressed the real reasons as to why they are not selling? If we can keep marketing to the precedent we have set then we will keep selling. We just need more, because there are buyers waiting, as my earlier examples proved. The stock level will then always remain manageable allowing us the time to productively work for you in finding your perfect buyer!
So let's hope for New Year's Health and Happiness and continuing the success of 2013. Thank you to all of our Customers and Christmas best wishes to all.
We will be closing for Christmas at Midday on 23rd December 2013 and opening again between 10 and 4pm on December 28th, Saturday 29th and Monday 30th December. Normal office hours will resume on Thursday 2nd January 2014.
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text:
Его́ршино — узловая станция Свердловской железной дороги в городе Артёмовском Свердловской области, Россия.

Название
Село Егоршино известно с 1665 года и сыграло важную роль в истории Урала, закрепившись в ряде топонимов и других названий — например, в его честь называется магистральная улица Егоршинский подход в Екатеринбурге. Город Артёмовский был образован в 1938 году путём слияния села Егоршино с посёлком имени Артёма. Название Егоршино при этом сохранилось за станцией, соответствующее пригородное железнодорожное направление из Екатеринбурга также называется Егоршинским.

Описание станции
Узел, расположенный на пересечении магистрали Екатеринбург — Устье-Аха (однопутная, на тепловозной тяге) и меридионального направления Каменск-Уральский — Богданович — Алапаевск - Нижний Тагил (южная часть (до станции Рефт) двухпутная, далее - однопутная; электрифицирована, постоянный ток, 3000v). Пригородное сообщение с Екатеринбургом, Нижним Тагилом, Алапаевском, Туринском (ст. Туринск-Уральский), Каменском-Уральским, Рефтинским, Буланашем и Богдановичем. Пассажирское сообщение представлено единственным пассажирским поездом местного сообщения № 610 Екатеринбург — Устье-Аха

Станция относится к Екатеринбургскому региону обслуживания Свердловской железной дороги. В пределах станции расположено локомотивное депо Егоршино.

Ссылки
 Страница станции на сайте РЖД
 Егоршино на Яндекс-Расписаниях

Железнодорожный транспорт Артёмовского
Станции Екатеринбургского региона Свердловской железной дороги

redpajama_set_name: RedPajamaWikipedia


text:
Australia captain Steve Smith named ICC Cricketer of the Year for 2015
The Aussie skipper scored a world leading 1,734 runs at an average of 82.57 with seven centuries over the voting period
On the rise: Smith has had a standout year (Image: Getty)
Australia captain Steve Smith was named ICC Cricketer of the Year for 2015 on Wednesday after scoring a world leading 1,734 runs at an average of 82.57 with seven centuries over the voting period.
The 26-year-old right-hander kept the Sir Garfield Sobers Trophy in Australia for the third straight year after pace bowler Mitchell Johnson and his predecessor as skipper, Michael Clarke, won in 2014 and 2013.
Smith also scored four centuries and 1,249 runs in 25 one-day internationals from Sept. 18, 2014 to Sept. 13, 2015, notching up the winning runs as Australia stormed to a fifth World Cup triumph.
"I'm obviously completely surprised by it, very humbled," said Smith, who was also part of the team that lost the Ashes to England during the voting period.
Losing battle: Smith during Australia's Ashes loss (Image: Reuters / Philip Brown)
"You don't think about these accolades, you just want to go out there and do the best you can for the team and try and win as many games as possible.
"My favourite moment would have to be winning the World Cup, playing against New Zealand at a packed house at the MCG. It was an amazing feeling to be able to hit those winning runs."
The Australian also scooped the ICC Test Cricketer of the Year award while South Africa batsman AB de Villiers was rated the best player in the one-day game, winning the award for the second successive year.
Magic man: De Villiers has shone for South Africa (Image: Matt King/Getty Images)
De Villiers, South Africa's captain in the ODI format, also won the award in 2010.
The 31-year-old's brilliant quickfire batting lit up the World Cup, not least his unbeaten 162 against West Indies in Sydney, and he racked up 1,265 runs in total at an average of nearly 80 an innings over the relevent period.
His team mate Faf du Plessis won the award for best Twenty20 performance of 2015 for his 56-ball innings of 119 against West Indies in January, the third highest score of all time in T20 internationals.
Australian paceman Josh Hazlewood was named best emerging player, while the Spirit of Cricket Award went to New Zealand captain Brendon McCullum, who made a virtue of his team's sportsmanship as he led them to the final of the World Cup.
Up and coming: Hazlewood has broken through (Image: Ryan Pierse/Getty)
McCullum said on Tuesday that he would be retiring from international cricket after the two-test series against Australia in New Zealand in February.
Australia captain Meg Lanning was the women's One Day International Cricketer of the Year, while her West Indies counterpart Stafanie Taylor won the women's Twenty20 award.
Khurram Khan of the United Arab Emirates was named Associate and Affiliate Cricketer of the Year, while England's Richard Kettleborough was named best umpire for a third year in a row.
mirrorsport
Josh Hazlewood
Brendon McCullum