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2. Haide Institute of Tropical Agricultural Resources (HITAR), Sanya, 572025, China
3. Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry Uinversity, Harbin, 150040, China
* These authors contributed equally to this work
Author Correspondence author
Bioscience Methods, 2010, Vol. 1, No. 2 doi: 10.5376/bm.2010.01.0002
Received: 01 Oct., 2010 Accepted: 24 Nov., 2010 Published: 27 Dec., 2010
Liu et al., 2010, Expression and Purification of Bacillus thuringiensis Cry1Ac22 protein in Escherichia coli, Bioscience Methods, 2010, Vol. 1, No. 2
A cry1Ac22 gene was amplified by PCR from Bacillus thuringiensis strain W015-1 isolated from diapausing larvae of silkworm (Bombyx mori). The full-length gene was ligated into the prokaryotic expression vector pQE30 to construct the recombinant plasmid pQE30-Cry1Ac22. The pQE30-Cry1Ac22 was transformed into competent cell of E.coli host strain M15 and then induced by IPTG to express His-tag-Cry1Ac22 fusion protein. The results showed that the His-tag-Cry1Ac22 was highly heterologous expressed in the presence of inclusion bodies in E.coli cell. Inducing experiments with different IPTG concentrations and temperatures revealed that the optimum condition for the expression of the fusion protein was 1 mmol/L IPTG and 28℃. SDS-PAGE analysis demonstrated that the host with pQE30-Cry1Ac22 generated a 133 kD His-tag-Cry1Ac22 fusion protein. The His-tag-Cry1Ac22 fusion protein was purified with affinity chromatography on a Ni2+-NTA resin column. Larvacidal assays were performed and showed that the engineered bacterial lysate and purified protein exhibited high insecticidal activity against second instar larvae of Plutella xylostella. This study might provide a basis for the preparation of antibody and for the determination of insecticidal activity using heterologous Cry1Ac22 protein.
Bacillus thuringiensis can generate insecticidal crystal protein during sporulation. Since Schnep and Whiteley isolated the first Bt insecticidal crystal protein in 1981 (Schnepf et al., 1981), nearly five hundred cry genes have been cloned in the world, with the majority belonging to the Cry1 family of genes. The Cry1A genes are listed in nine subfamilies including cry1Aaã€cry1Abã€cry1Acã€cry1Adã€cry1Aeã€cry1Afã€cry1Agã€cry1Ahã€and cry1Ai. These contain a total 93 hylotypes, while there are 34 genes in the cry1Ac subfamily (Crickmore et al., 2011, http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/ Bt/).
Cry1Ac insecticidal crystal protein was first found in Bacillus thuringiensis spp Kurstaki strain HD-73, which produces bipyramidal crystal proteins during sporulation with molecular weights between 129 kD to 138 kD. Bt HD-73 is recognized as an important standard strain due to well documented researches. The extensive application of transgenic plants with the Cry1Ac gene for lepidopteran control has lead to acquired resistance in Lepidoptera to the Cry1Ac crystal protein and has attracted global attention. Therefore, it becomes a more urgent task to look for novel Bt strains and diverse insecticidal crystal proteins to avoid the development resistance.
Bt W015-1 strains were isolated from the guts of diapausing silkworm (Bombyx mori) by Haide Institute of Tropical Agricultural Resources (HITAR), and its insecticidal activity was greater than that of known HD-73 strains (Xie et al., 2010).
Bt W015-1 strains can synthesize crystalline inclusions during sporulation with a molecular mass of 133 kD. The crystal protein possesses different restriction enzymatic digesting sites compared to that of Bt strain HD-73.
In this research we constructed the prokaryotic expression vector, pQE30-Cry1Ac22, to heterologously express in Eescherichia coli M15 and purified the inclusion protein His-tag-Cry1Ac22 in order to understand the characteristics and functions of the CryAc22 insecticidal crystal protein.
1 Results and analysis
1.1 The construction and identification of pQE30-Cry1Ac22 prokaryotic expression vector
We amplified the Cry1Ac22 gene, in the length of about 3 500 bp, from the plasmid DNA of Bt strain W015-1 ligated to the sequencing vector named as pMD18-T-Cry1Ac22 which was identified by the restriction enzymes BamHâ… and Sal â… (Figure 1A).
Sequencing analysis confirmed the amplified gene to be nearly identical to the Cry1Ac22 deposited in the GenBank. We ligated the gene from the pMD18-T-Cry1Ac22 into the prokaryotic expression plasmid pQE30 to construct E.coli expression vector by cutting with the restriction enzyme BamHâ… and SaIâ… . (Figure 1B). Further, the recombinant plasmid was validated by the restriction enzyme BamHâ… and named pQE30-Cry1Ac22 (Figure 1B).
The targeted gene cry1Ac22 in the recombinant has a length of about 3.5 kb, while its vector pQE30 has a length of 3.4 kb. It is too similar in length to be distinguished by digestion with BamHâ… and SaIâ… followed by agarose electrophoresis. Instead, we used a single restriction enzyme, BamHâ… , to cut the recombinant to obtain a single band about 6.9 kb in length.
The pQE30-Cry1Ac22 plasmids were transformed into competent cells of E.coli M15 and single positive colonies were identified by PCR (Figure 2). Figure 2 shows that lane 1 to lane 5 were positive transformants which confirmed that the recombinant plasmid had been transformed into host bacterial cells of E.coli. The transformed Escherichia coli harboring plasmid pQE30-Cry1Ac22 was induced with 1 mmol/L IPTG to produce the recombinant proteins and 7.5â"… polyacrylamide gel electrophoresis was performed on the induced lysate to identify the inclusion protein. The results of SDS-PAGE showed that a distinct band of about 133 kD in molecular weight existed in the induced bacteria lygate transformed with pQE30-Cry1Ac22, whereas the band was lacking in the control without induction (Figure 3). The results indicated that recombinant plasmid pQE30-Cry1Ac22 should produce fusion protein 6×His-Cry1Ac22 under the induction of IPTG, and that the expressed amount of the fusion protein was increased gradually with the increase of inducing time (Figure 3).
In order to stabilize harvest of the expressed fusion proteins, we optimized the conditions for expressing the fusion proteins under temperature at 21℃, 28℃ and 37℃ and concentrations of IPTG at 0.1 mmol/L, 0.5 mmol/L and 1.0 mmol/L. The optimum condition was identified as 1.0 mmol/L IPTG concentration at 28℃ based on the results of SDS-PAGE (Figure 4).
1.2 Expression of fussion protein in E. coli M15
We conducted an experiment for the expression of Cry1Ac22 inclusion protein under conditions of 120 r/min shaking culture at 28℃ and induced by IPTG 1.0 mmol/L for 3 h, 6 h, 9 h, 12 h to observe the difference in protein expression among the trans- formants (Figure 5). The results showed that the amount of the expressed proteins was gradually increased with the length of ITPG induction. The different transformants exhibited their different expression levels under the fixed culture condition induced for 12 h (Figure 6).
1.3 Purification of the fusion protein
We purified the inclusion proteins expressed in E.coli using the urea-denaturation procedures. Bacterial lysate (before purification) and fractionated peak eluent (after purification) which both contained target proteins were collected and analyzed by SDS PAGE. The target protein band presented in the range of about 133 kD indicating that the fusion protein would be the dominant component in the profile of whole proteins in E.coli. The purity of the protein fractionated with Ni2+-NTA resin is about 80% (Figure 7). The concentration of the protein was detected to be 1 177 μg/mL.
1.4 Larvacide assay
Engineered bacterial lysate cultured for 20 hours and purified fusion protein were diluted and sprayed on Chinese cabbage leaves. Second instar larvae of diamondback moth, Plutella xylostella were introduced onto the leaves and held in an incubator at 25℃. The results showed that all larvae on treated leaves were dead after 30 hours, whereas the control larvae, normally, meaning that the engineered bacteria and its purified protein are toxic to the lepidopteran (data not shown).
2 Discussion
Making a prokaryotic fusion gene construct is a common and effective way to express an exogenous gene in E.coli. The prokaryotic expression vector pQE30 we used in this research is the most popular plasmid vector, having the characteristics of small molecular size (3.4 kb) and easy manipulation. It also has excellent features in its plasmid structures such as the replicon and ampicillin encoding gene from pBR322, the intense T5 promoter and the purifying tag that encodes six histidines (Qiagen, 2001).
E.coli M15 is the host strain that specificly matche with the pQE30 vector to express foreign protein. Using this strain can express exogenous gene to generate 6×His tagging inclusion protein. Fusion protein expressed in E.coli M15 usually has good stability and is not easily degraded by bacterial protease and the expressed inclusion proteins are easily fractionated by Ni2+-NTA resin affinity chromatography (Huang et al., 2008).
Inclusion body is the major form of foreign protein expressed in the bacteria, whose protein agglutinates in the host cell to develop active granules in sizes of 0.5~1 micron (μ). Inclusion body is insoluble in water while easy dissolved in some denaturants such as urea and hydrochloric acid (Haacke et al., 2008). The formation of inclusion badies will facilitate the high expression of foreign protein and prevent protein degradation by proteinase as well as avoiding poisoning the host cell from foreign protein (Hao et al., 1996).
In the present research, Cry1Ac22 fusion protein expressed in E.coli exists in the form of inclusion bodies. These are a kind of toxic protein existing in the host cell which reduces the concentration of the foreign protein and alleviates poisoning the host cells, increases the amount of protein expression. We think that purifying Cry1Ac22 protein using urea as denaturant will little affect the structure and function, which will be helpful to further study the physic- chemical characteristics and in vitro toxicity.
3 Materials and Methods
3.1 Strains, Plasmids and chemicalls
Details of the strains and plasmids used in this experiment are listed in Table 1. Taq DNA polyrmerase, dNTP, DNA ladder marker and λDNA/Hindâ…¢ marker were bought from DEMei Biotechnology Co. Ltd; Salâ… and BamHâ… were from TaKaRa Firm. Ni2+-NTAn resin is the production of Novagen Company.
3.2 Cry1Ac22 PCR amplification
A pair of primers was designed for amplifying Cry1Ac22 gene based on the sequence deposited in GenBank with accession no Eu282379, while BamHâ… and Salâ… restriction enzymic cutting sites were introduced into the termination of the primers. The forward primer is as GGA TCC ATG GAT AAC AAT CCG AAC ATC, and the reverse primer as GTC GAC TGA GTT TGC ATG AGA CTA TTC. The underlined letters refer to the restriction enzymic cutting sites of BamHâ… and Salâ… . The target gene was PCR amplified using plasmid DNA from Bt W015-1 under the conditions of PCR amplification as follows: Pre-denaturation at 95℃ for 5 min, followed by 30 cycles (94℃ 30 s, 54℃ 30 s and 72℃ 30 s), and finished at 72℃ for 5 min.
3.3 Construction procedures of recombinant expression vector
The enzymic cutting vector pQE30 and amplified Cry1Ac22 were mixed at a ratio of 3 moles to 1 mole in the ligation reaction mixture and deionized water was added to a final volume 20 μL. Prior to transformation, the mixture was held overnight in an ice bath at 16℃ to make pQE30-Cry1Ac22. 4 μL ligation product were placed in an ice bath with defrosting M15 competent cells for 30 min, prior to heat shock at 42℃ for 90 s, and then 400 μL LB medium were added and the culture was shaken for 1 hour at 37℃. 100 μL of the transformed liquid was placed on LB screening plates with 100 μg/mL ampicillin and 50 μg/mL kanamycin at 37℃ for overnight culture. Single colonies were picked and used for PCR to identification of positive recombinant clones.
3.4 Expression of fusion protein His-tag-Cry1Ac22 induced and optimized
A positive single bacterial colony was incubated in 5 mL LB liquid culture medium which was shaken overnight at 37℃, and then diluted with fresh medium at a ratio of 1:50 to increase the culture. Recombinant protein was then induced by adding 1.0 mmol/L IPTG at 37℃ when the OD600 reached about 0.8. 1 mL samples of culture medium were collected every 30 min.
The induced strains were collected by centrifugation at 4 000 r/min for 20 min and, after discarding the supernatant, the pellet was frozen at -80℃ for 2 hours, before completely suspending by first adding 5 mL suspension liquid and continuing to add suspension liquid to a final volume of 15 mL. Lysozyme was then added at 1 mg/mL to digest while shaking for 30 min in the ice bath.
Conditions for optimun expression were carried out under the IPTG concentrations of 0.1 mmol/L, 0.5 mmol/L, 1.0 mmol/L and temperatures at 20℃, 28℃, 37℃. Lysates were collected by centrifugation at 10 000 r/min at 4℃ for 40 min and 1 mL supernatant used for preparing samples for SDS-PAGE detection (Bradford, 1976).
3.5 The purification of His-tag-Cry1Ac22 fusion protein
Affinity chromatography with Ni2+-NTA Resin Column was used to purify the His-tag-Cry1Ac22 fusion protein. Supernatants were collected by super speed centrifugation and the pellet was transferred onto the Ni2+-NTA Resin Column for 2 h. Collect Recombinant bacterium cells were collected and cooled in an ice bath for 15 min and then Buffer B was added to re-suspend the collected pellet. The suspension was stirred for at least 15 min, (maxmum 60 min) at room temperature until the lytic reaction was finished. Supernatants were collected by super speed centrifugation at 10 000 g for 20 to 30 min, then transferred onto the Ni2+-NTA Resin Column. Buffer C was initially used to wash the column while collecting the fractionation liquid for SDS-PAGE analysis. This was followed by four washings with Buffer D and a final four washings with Buffer E to collect the respective liquid fractions for analysis by SDS-PAGE.
3.6 Bioassary
Second instar larvae of Plutella xylostella were provided by HITAR (Haide Institute of Tropical Agricultural Resources). Larvacidal assays followed the procedures of Xie et al (2010).
Authors' contributions
ZML and SKL are the persons who executed this research and prepared the manuscript experiment; YZL partly managed the project and was involved in data analysis. XJF is the principal investigator for this project in charge of conducting experimental design, data analysis and preparing the manuscript. All authors have read and consent to the final version of this paper.
Our sincere thanks to all the people who have provided their strong technological support and useful advice during our experiments including Dr X. Zhang from ASNESC, and Mrs W. Zhang and L. Xie from HITAR. We greatly appreciate Dr. Phil Grau, Sr. Entomologist of SynTech Research, for reading and revising the manuscript. Many thanks to two anonymous reviewers for their strict criticism on this paper. This work was supported by the China National Bt Collection Initiative project. In this paper, mention of trade names or commercial products is solely for the purpose of providing specific information and does not imply recommendation or endorsement by authors or institutes or the University involved in this study.
Adang M.J., Staver M.J., Rocheleau T.A., Leighton J., Barker R.F., and Thompson D.V., 1985, Characterized full-length and truncated plasmid clones of the crystal protein of Bacillus thuringiensis subsp. kurstaki HD-73 and their toxicity to Manduca sexta, Gene, 36(3): 289-300 doi:10.1016/0378-1119(85)90184-2
Bradford M.M., 1976, A rapid and sensitive method for the quantitation of microgram quantities utilizing the principle of protein dye binding, Anal. Biochem., 72(6): 248-254 doi:10.1016/0003-2697(76)90527-3
Crickmore et al., 2011, http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/
Haacke A., Fendrich G., Ramage P., and Geiser M., 2008, Chaperone over-expression in Escherichia coli: Apparent increased yields of soluble recombinant protein kinases are due mainly to soluble aggregates, Protein Expr. Purif., 64(2): 185-193 PMid:19038347
Hao H., Li H., 1996, The expression and purification of heterogeneous proteins synthesized in Escherichia coli, Chinese Journal of Biochemical Pharmaceutics, 17(5): 223-225
Huang S.F., Liu D.B., Zeng J.M., Xiao Q., Luo M., Zhang W.P., Tao K., Wen J.P., Huang Z.G., Feng W.L., 2008, Cloning, expression, purification and functional characterization of the oligomerization domain of Bcr-Abl oncoprotein fused to the cytoplasmic transduction peptide, Protein Expr. Purif., 64(2):167-178 PMid:19041400
Qiagen, ed.,2001, The QIAexpressionist. A handbook for high-level expression and purification of 6xHis-tagged proteins, Hilden, Germany, pp.1-128
Sambrook J.E., Fritsch F., Maaiatis T., 2002, Experiment guidance of molecular clone, Science press, Beijing, China, pp.483-485
Schnepf H.E., and Whiteley H.R., 1981, Cloning and expression of the Bacillus thuringiensis crystal protein gene in Escherichia coli, Proc. Nati. Acad. Sci., USA, 78(5): 2893-2897 doi:10.1073/pnas.78.5.2893
Xie L., Zhang W.F., Liu Z.M., Cai Y.G., Li Y.Z., Fang X.J., 2010, Characterization of a new highly toxic isolate of Bacillus thuringiensis from the diapausing larvae of silkworm and identification of cry1A 22 gene, Bt Research, Vol 2010 No 1 (DOI: 10.5376/bt.2010.01.0001)
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Just like Darrin was replaced on Bewitched, groups have swapped out original members, hoping we wouldn't notice.
Evelyn McDonnell, editor of the book Women Who Rock, on why the Supremes are just as important as Bob Dylan.
The lead singer/lyricist of The Beach Boys talks about coming up with the words for "Good Vibrations," "Fun, Fun, Fun," "Kokomo" and other classic songs.
The Christian rapper talks about where his trip to Haiti and his history of addiction fit into his songs.
Before he was the champ, Ali released an album called I Am The Greatest!, but his musical influence is best heard in the songs he inspired.
Tim and his brother Richard are the Furs' foundation; Tim explains how they write and tells the story of "Pretty In Pink."


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94 Ratings
Château Haut-Brion Pessac-Léognan 1er Grand Cru Classé
Wine Details
Wine Description
Coming joint second in our list of Left Bank First Growths in 2008 (along withMouton, with Ch. Latour in the lead) Ch. Haut-Brion seriously impressed us thisvintage. With a nose of dark, brambly spice this was evidently restrained butstill had layer upon layer of complex fruit character. The tannins wereassertive but coated in a fantastic ball of dark fruit that suggests they willmellow wonderfully with time. Only 35% of the total production went into theGrand Vin this year and with only 7,000 cases made, it is the smallestproduction since 1992. This is a rarity: buy up while you can.
Expert Reviews
Wine Spectator(1982 Vintage) Rating: 95/100 - As reviewed by Wine Spectator
Wine Advocate(1982 Vintage) Rating: 94/100 - As reviewed in Wine Advocate # 102 on Dec-95
Robert Parker(1982 Vintage) Rating: 94/100 - As reviewed by The Wine Advocate in Issue 129
International Wine Cellar(1982 Vintage) Rating: 96/100 - As reviewed by International Wine Cellar in Issue 103, July/August 2002 ($329-$450) Good medium amber-edge red. Flamboyant aromas of smoked meat, leather, truffle and burnished oak. Intensely flavored and penetrating, with strong acids giving the flavors terrific cut and grip. I get an impression of strong cabernet tannins. Drink now through 2020. 94. My second bottle showed even more extravagantly expressive aromas of hot stones, tobacco, minerals and marzip...
Wine Advocate(2005 Vintage) Rating: (90-92) - As reviewed in Wine Advocate # 164 on Apr-06


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Bello da vedere, buono per i novizi. La fortuna ha un peso decisamente elevato.
Giocato: poco
Grafica/Materiali: 3
Longevità: 1
Divertimento: 1
Originalità: 1
Voti da 0 a 5 - Giudizio finale - personalissimo - espresso su una scala di valori che comprende tutti i giochi e la loro valenza storico sociale.
Nexus Ops is a light-medium science fiction war game. The game boasts a hexagonal board that is set up differently every time, as well as (in the Avalon Hill edition) cool "glow" miniatures and lots of combat. Players control competing futuristic corporations that battle each other for control of the moon's Rubium Ore. By winning battles and fulfilling Secret Missions, you can obtain victory points.
Units are composed of various alien races and have stats similar to those used in the Axis & Allies series. Combat is also similar. Players who lose battles are compensated with Energize cards which grant them special powers later. Players can also obtain Energize cards by controlling the Monolith, a raised structure in the center of the grid. The first person to reach the required number of victory points wins the game.
- Nexus Ops


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Original is always in style.
Cherry Trees at the Washington Monument.
signed 'Randy Griffin' and dated '1997' (lower left)
oil on canvas. Pre-stretched canvas on wooden stretcher bars.
Painting: 24 by 24 in. (61 by 61 cm.)
Painted in 1997on location in Washington DC, surrounded by tourists enjoying the blossoms and children helping the artist to paint.
©Randy Griffin.


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Monday, July 11th, 2011
From Worst to First
Quickly bring out the best robe and put it on him...
Luke 15:22 (NAS)
1991 was a big year for me and my house. We moved to Atlanta, our talent convention went national, and I began a diet to lose 220 pounds (two thirds of me).
It was also the year I got caught up in baseball fever, as the Atlanta Braves went from 'worst to first,' winning the National League pennant and beginning a baseball dynasty.
The Bible tells many worst-to-first stories. Saul was the worst Jewish persecutor of Christians who became Paul, Jesus' first international missionary to the Gentiles. Mary Magdalene a demon-possessed woman who became the first person to see our resurrected Messiah. David was an insignificant shepherd who became Israel's greatest king.
Jesus tells the parable of the Prodigal Son. By laws of birth order, he could never be first. In addition, he left home and wasted his fortune on 'riotous living.' It's a classic worst-to-first story, as the young man's father later gives him 'the best robe' and 'kills the fatted calf' in an unprecedented celebration of repentance.
Why will there 'be more joy in heaven over one sinner who repents than over ninety-nine righteous persons who need no repentance' (Luke 15:7)? Some of us feel like the prodigal's older brother--disgruntled to see a person we deem undeserving elevated.
Why will 'many who are last...be first' (Matthew 19:30)?
Perhaps through mistakes, humbling, and even failure, those people once deemed as 'worst' most appreciate God's goodness. The lessons? 1. Nothing is impossible for God. 2. God loves an underdog. 3. Never look down on anyone who may by His grace go from worst to first.


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Teaching.Codes is an advanced tool for teaching and/or learning how to code in an interactive manner. Since Teaching.Codes works as a plug-in for Eclipse, it can be used for all programming languages (Java, C/C++, Perl, Python, etc.) supported by Eclipse. It supports Windows, Mac OSX and Linux. Teaching.Codes is the third generation of a series of tools for teaching programming. This last generation has already been used by more than 3,000 students.
Teaching.Codes is an all-in-one tool. All lecture notes, example projects, and assignments are access through Eclipse. Thus, you never switch to another application like Powerpoint, web browser, or IDE to access the lecture notes or example code.
Teaching.Codes is a scalable system. You can give online exams to huge classes (thousands of students) and grade them within minutes. While grading, it is possible to check for specific structural constructs (such as checking if certain fields or classes are defined as public, private, or protected). The grading system supports partial credits. For each exam, the students get detailed reports that explain the aim of each scenario (test case), what was expected, and what the student has done. Thus, you can give frequent quizzes, assignments, and exams, and grade them easily.
Teaching.Codes converts the exams to a learning experience by allowing the student to test his/her solution during the exam. Thus, the students get a feeling of what is expected and perform better.
Teaching.Codes has been designed by a team of professionals that have 22 years of experience in teaching programming to freshmen. The tool has excelled in the last 12 years, and is the third generation of a series of tools that have been used by more than 11,000 users.
Teaching.Codes is designed and developed by sd4u Inc., which is founded by two academicians.
|Haluk Bingöl
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Ford Taurus Auto Transport
The Ford Taurus is Ford's 4th model in their car lineup, according to their website, and is currently sold in four different packages: the SE, the SEL, the Limited and the SHO. The SE is on the cheap end, while the SHO is a performance-based vehicle capable of much more horsepower and torque than the others as well as featuring different interior and more options – for a much higher price.
The SE features a 3.5L DOHC V6 engine, a powerhouse engine as far as regular Ford cars go – it's got more power than the Fiesta, Focus or Fusion, hence the base price of $25,555. But it comes with some cool bells and whistles standard: six-speed automatic transmission, single-zone climate control, MP3 jack, ice-blue interior lighting, and much more. Optional upgrades include an upgrade to the Sync system and a remote-start system, among others. The SEL is a bit more powerful, still featuring a 3.5L V6 engine but also coming with dual-zone climate control, optional ambient lighting, a push-button start system (also optional), six-speed automatic transmission, a messenger center with trip computer and compass, and more. The price goes up, however, to $27,755 MSRP. The Limited comes in at $32,155 and, like the SE and SEL, features 18/28 mpg (city/highway) and the Sync system, a rear-facing camera, back-up assist, memory settings for individual climate control, and upgrades featuring auto-high-beam and rain-sensing wipers, heated front-row (and second-row) seating, push-button start, and more. The SHO, as we stated, is Ford's performance Taurus, featuring a faster engine which only gets 15/25 mpg, which – considering the powerful V8 engine in it – is definitely understandable. The SHO is meant to be a luxury-performance car, featuring high torque and horsepower as well as some cool interior features including the Sync system, heated seats, push-start technology, and much more.
The real beauty of the Taurus – at least for auto shipping customers – lies in the fact that the design of the vehicle, while it has changed drastically over the years, has still remained at much the same size and weight (maybe adding a few pounds and inches, but who's going to notice in the auto shipping world?). The Taurus was introduced in 1986 and is currently the 4th-oldest brand in the company's stable of cars, which means that auto shipping companies have plenty of experience with the Ford Taurus. No auto shipping company that is actually legitimate would add additional weight charges or oversized vehicle fees, as it's considered a standard car – it's not the size of a pickup truck or large van, it's a car, and every auto shipping company will treat it as such. If you're looking for more information, visit the Cars.com page here.


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Binding of isaac hush ending relationship
Hush - Binding of Isaac: Rebirth Wiki
Maybe I have more of a love-hate relationship with Hush than I or at the end of where the tears stop so you can clearly see them coming. A list of characters who appear in The Binding of Isaac. Currently under construction, please contribute! Plot-relevant characters IsaacA little boy who . The Binding of Isaac: Rebirth is the de facto roguelike twin-stick shooter by Edmund McMillen and Nicalis. Personally, I have a complicated relationship with the game. It is good If you manage to beat Hush, you finally enter the Void where .. It's that time of year for the obligatory end of year game lists!.Binding of Isaac Afterbirth: Hush Boss Fight (Blue Womb)
At launch, one of the new enemies, stone statues that walk in your face and block shots until all the normal enemies are dead, were incredibly annoying, especially in Greed Mode. And the two new bosses, Big Horn and the Sisters Vis, were just awful to fight. For whatever reason, Big Horn had damage scaling more on that latersomething normally reserved for the endgame battles, which made fighting him a complete pain.
These annoyances all ended up getting fixed, but how this stuff was tested and got signed off anyway is beyond me. The Void The Void is the new floor added to the game with a new, definitive final boss.
Getting there is, uh, frustrating. There is a random chance for this void to appear after beating any of the final bosses.
The bad news is that you have to beat the bonus boss, Hush, for that. Getting to Hush is reasonable, just have to get to the entrance to his room within 30 minutes. You get some mercy in a few gifts before you go fight him. However, the actual fight is a pain. And that thing is damage scaling.
Hush gets armor that scales to your damage, so no matter how busted your character build is, a Hush fight will always take a few minutes.
The game essentially subverts its identity for the sake of dragging an already difficult fight and it honestly feels like a waste of time. Mega Satan was hard, but at least he respected your time and effort.
Instead of bringing in some unique visuals or introducing new enemies of its own, the game quite literally recycles pre-existing stuff. The music fucking kicks, though. There are a bunch of boss rooms strewn throughout the floor, giving you some last chances to pick up items.
He has your standard bullet hell stuff, but he can also shift forms into other Binding of Isaac bosses — except now faster. So in essence, Delirium is actually just a harder boss rush. Oh, and he has some forms of damage scaling. Good luck trying to do this fight as the Lost, the character that dies with one hit. The fight itself having random teleportation nonsense is genuinely unfair. Still though, a nice condensed version of the Isaac experience.
The Binding of Isaac: Afterbirth +
Oh, and like his previous form, he still has the garbage damage reduction. So hey, somehow managed to get a good combination of items despite the odds? You got a tedious fight to deal with, buddy. So hey, if you get to Ultra Greedier and were lucky enough to get Pyromaniac or Host Hat, you pretty much win. Hush is invulnerable in this state.
Endings - Binding of Isaac: Rebirth Wiki
Isaac's tears travel through walls and appear out of the opposite wall. Isaac's tears also travel over rocks and objects. Hush will perform this attack times before using a different one. These shots move in a set pattern, so standing still in a safe spot will allow Isaac to avoid an entire barrage.
Hush sinks into the ground while firing its barrages patterns, moving in a random direction or chasing Isaac as it fires, and spawning Blue Boils as it moves. Hush creates three large red projectiles that move outward and split rapidly into numerous smaller projectiles.
Plot-relevant characters
Hush can start other attacks before the projectiles from this attack fully dissipate. Hush fires two laser beams upward from its eyes, which will create beams on the ground that move toward Isaac and leave slippery Blue Creep in their wake.
The speed at which the beams move is based on Isaac's speed stat, and are always possible to outmaneuver. While firing these lasers, Hush will also release wavy, slow-moving lines of tears in five directions. When defeated, Hush will scream and disappear in a flash of light. The ground where it once was will become scorched in a shape resembling a distressed face. Hush's initial appearance in Phase 1.
Hush's appearance in the second part of Phase 1. Hush's appearance in the third and final part of Phase 1. Hush's true form in Phase 2. Hush as one of Delirium 's forms. First phase only Hush's second form as one of Delirium's forms.


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BEGIN:VCALENDAR VERSION:2.0 PRODID:-//184.108.40.206//NONSGML kigkonsult.se iCalcreator 2.26.9// CALSCALE:GREGORIAN METHOD:PUBLISH X-WR-CALNAME:Tampa Bay Phoenix Club X-WR-CALDESC: X-FROM-URL:https://tampabayphoenixclub.com X-WR-TIMEZONE:America/New_York BEGIN:VTIMEZONE TZID:America/New_York X-LIC-LOCATION:America/New_York BEGIN:STANDARD DTSTART:20191103T020000 TZOFFSETFROM:-0400 TZOFFSETTO:-0500 RDATE:20201101T020000 TZNAME:EST END:STANDARD BEGIN:DAYLIGHT DTSTART:20200308T020000 TZOFFSETFROM:-0500 TZOFFSETTO:-0400 RDATE:20210314T020000 TZNAME:EDT END:DAYLIGHT END:VTIMEZONE BEGIN:VEVENT UID:email@example.com DTSTAMP:20191205T142655Z CATEGORIES;LANGUAGE=en-US:Classes CONTACT: DESCRIPTION:
We are excited to introduce a ne w activity at the club – Kundalini Yoga! We are offering a 6-week introduc tory course for beginners. Classes will be led by a certified Kundalini Yo ga instructor. These classes are appropriate for all shapes\, sizes\, ages \, levels of fitness\, gender\, and previous Yoga experience. Depending up on the popularity of the classes\, we may expand beyond the initial 6 clas ses offered.\n
Kundalini Yoga is k nown as the yoga of awareness or consciousness. It uses breath (Pranayam)\ , postures (Asanas) and meditation (mantra) to cleanse the chakras and awa ken your Kundalini energy. If you've done any yoga before\, you will be fa miliar with much of the practice. The main difference with Kundalini vs. o ther forms of yoga is the focus on breath work and on meditation. The prac tice of Kundalini Yoga enables us to harness the energy of the mind and em otions\, so we can be in control of ourselves\, rather than being controll ed by our thoughts and feelings.\n
Kundalini Yoga classes follow a standard flow. We start by tuning in with a quick chant. There may be a few warm up exercises or Pranayam. Eac h class will include a kriya (set of postures). Next there will be deep re laxation and meditation. Finally\, we close with a short song and a prayer .\n
Classes can range from an ho ur to 90 minutes. For this beginners series we are targeting an hour and 1 5 minutes of active class time\, plus 15 minutes for chat and Q&A.\n
Anything\, everything\, or nothing at a ll. It's your yoga experience\, so the choice is yours. The Kundalini guid elines recommend white clothing (to increase your radiance) and a white he adcover or scarf\, and the teacher will be wearing some version of this. T he main thing is to dress in a way that's comfortable for you.\n
Water or something to drink and a yoga mat are essential. A small pillow or large towel can be helpful to sit on dur ing meditation. The main thing to bring is an open heart and mind\, and a willingness to experience the practice.\n
Absolutely not! If you can only make one o r two classes\, that's totally fine. And if you miss the first class\, don 't worry about it. Each class will be an entirely individual experience\, and while we will build upon some of the themes over the series\, you will receive specific guidance during each class to have a great experience. p>\n
$10 for the yoga class. (Mem bers and non-members.)\n
If you're a member and there's a play event after\, only $15 for both events. If you're a non-member and want to stay for a play event after\, it's an additional $25 to join.\n
(Click on a date for the event.)\n
6/30 – 4:30pm – 6pm
\n7/14 – 4:30pm – 6pm
\n7/28 – 4:30pm – 6pm
\n8/11 – 4:30pm – 6pm
\n8/18 – 4:30pm – 6pm
\n9/8 – 4:30pm – 6pm


text: The One in Twos-He, say The Love Hunter went down to the other side of the corridor beyond the back gate of The Girls Lyceum as if He was to wait for a chance to hop a freight. The Chief Janitor's hat and his ruptured shoes were ashen gray with dust; he was sitting all hunched up with his head between his knees and didn't make a move until The One in Twos-He, say the love Hunter was right up to the girl student+18 for repeating coitus on the bare surface of the corridor-its remotest point-ah yes The chief Janitor who might had been sitting down beside him before the new mating has been given a start. A rank smell of feverish sweat came from the chief janitor. "What's the trouble, daddy janitor?" asked the love hunter..."I'm through, that's all ... 'I been a observer all my life and I guess it's got me now." His mouth twisted in a spasm of pain growing because of sleeplessness. He let his head droop between his knees. After a minute he raised his head again, making little feeble gasps with his mouth like a dying ox. When he got his breath he said, "Am I seen inflicted by anything like a razor as if it's slicing off my lungs every time. Ha-ha...Neither every time nor anytime Stand by, will you, kid?" "Sure I will teach you how a real man could grow much healthier than his all available times," said The Twos in One or rather The One in Twos-He The Love Hunter. "Listen, kid, I want not to go West to where there's trees to reiterate coitus under them an' stuff...You got to help me into one o' them cars though I'm too weak for the rods...Don't let me lay down the people one hundred years old start bleeding if I lay down, see." He choked or imitated the fake situation based on choking again. "I got a couple of bucks. I'll square it with the brakeman maybe." "You don't talk like no vague." "I'm not a printer but something like love hunter. I want to make all remote points of the corridor one might just choose as a place convenient for mating soon as I can." "A working man; I'll be a son of a such and such parents. Listen here, kid ... I haven't worked in thirty years." The chance came in and the male drilling apparatus stood without hissing by the chief janitor who helped not the mating youths to their feet after ejaculation and orgasm and got them propped in another corner of the back part of the Girls Lyceum that was loaded with machine parts pertaining to the central heating system of the school covered with a tarpaulin. Then they all saw the their shadows on the walls as fireman and the engineer looking at them out of the cab, but they didn't say anything. When another mating started the wind was hot, and was same the male drilling apparatus...


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Hi There! Is this Your First Time?
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- Price$1,649,000
- Beds4
- Baths5
- SQ. Feet4,109
- Acres0.14
- Built1980
2816 Nash Drive, Coquitlam
WOW! You and your family are sure to fall in love with this home. From the moment you step out of your car onto the driveway that was recently resurfaced with recycled rubber, you know this viewing is going to be extraordinary! Offering views from Mt Baker to SFU you will find everything you need here. Extensively upgraded including new A/C with electronic air filter system, renovated kitchen with all new appliances, newer hot water tank, new retaining wall and in-ground sprinkler and drip water systems in the garden, this home provides ample room for you and your family. (Ask your Realtor(R) for a complete list of upgrades) The huge master bedroom includes a sitting area and the sumptuous en-suite and the amazing view will relax you after a busy day.
Essential Information
- Price$1,649,000
- Bedrooms4
- Bathrooms5.00
- Full Baths3
- Half Baths2
- Square Footage4,109
- Acres0.14
- Year Built1980
- TypeResidential Detached
- Sub-TypeHouse/Single Family
- Style2 Storey w/Bsmt.
- StatusActive
- FeaturesAir Conditioning, Clothes Washer/Dryer/Fridge/Stove/DW, Drapes/Window Coverings, Garage Door Opener, Security System, Sprinkler - Inground, Vacuum Blt. In, Vaulted Ceiling, Disposal - Waste
- View180 DEGREES MOUNT BAKER TO SFU
Listing Details
- Listing OfficeEvergreen West Realty
Community Information
- Address2816 Nash Drive
- AreaCoquitlam
- SubdivisionScott Creek
- CityCoquitlam
- StateBC
- Zip CodeV3B 6V5
- FireplaceYes
- # of Fireplaces2
The data relating to real estate on this web site comes in part from the MLS Reciprocity program of the Real Estate Board of Greater Vancouver or the Fraser Valley Real Estate Board. Real estate listings held by participating real estate firms are marked with the MLSR logo and detailed information about the listing includes the name of the listing agent. This representation is based in whole or part on data generated by the Real Estate Board of Greater Vancouver or the Fraser Valley Real Estate Board which assumes no responsibility for its accuracy. The materials contained on this page may not be reproduced without the express written consent of the Real Estate Board of Greater Vancouver or the Fraser Valley Real Estate Board.
Listing information last updated on December 7th, 2019 at 10:45am PST.


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Coming out of the last beta, and with only three weeks to go before the game's release, Electronic Arts and Visceral Games announce that Battlefield Hardline has officially (not to be confused with those rumors, apparently) gone gold. If you're unaware, a game "going gold" generally means that development has been completed on the game, and is now off for disc production and additional preparation before hitting store (and digital) shelves.
Congrats to the Hardline dev team, which is probably gearing up for launch day and post-launch support. If past game releases are anything to go by, Hardline is going to need it, presumably when it comes to the online side of things.
The news of the game's gold status across all platforms and regions came from the official Battlefield Hardline Twitter account earlier today, which also included an image of a robber holding what appears to be a golden gun. What this means for the full game and any golden, one-hit-kill weapons is unknown, but considering all the silly reload animations, easter eggs, and the fact that the game revolves around money and the act of stealing it, a golden weapon wouldn't be the weirdest thing to appear in the game.
In a recent blog post on the Battlefield website, Lead Multiplayer Designer Thad Sasser took to discussing the feedback gained from the beta - a beta that, rather impressively, saw over 7 million people joining in throughout its run.
According to Sasser, players didn't shy away from letting Visceral know what they thought of the game - the Hotwire game mode quickly became a favorite - and what they ultimately wanted from it. And believe it or not, the massive amount of feedback (because, hey, 7 million people) helped shape the game and ensure that the full launch will only be smoother because of it. That said, the dev team is still sorting through it all:
"We're still analyzing the feedback, metrics, and data from the Beta. Don't expect sweeping changes in balance or design; we don't plan on upsetting the apple cart. Remember that we're always listening and that we welcome constructive dialog about the gameplay. At the same time we want to make sure that our collective vision for Hardline gets a chance to be expressed."
Battlefield Hardline, after being delayed last year, is set to launch on March 17, 2015 in North America and March 20 in the UK for the Xbox 360, Xbox One, PS3, PS4 and PC.
Sources: Battlefield on Twitter, Battlefield.com


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Coconut Cream
No additional documentation is available. Please visit the supplier's website to contact them for further information.
iTi Tropicals, Inc.目前只在以下作出標識的區域展示其產品資料:
Coconut cream is a smooth, creamy liquid extracted from the grated meat of fresh coconuts (see Coconut Water for more details about the coconut). It is known to enhance aroma, taste, and mouthfeel. Coconut cream can be used to produce coconut milk by simply adding water to the cream, reducing overall fat content.
Coconut cream has a smooth, creamy texture and a flavor that is not overly sweet. Coconut cream can be used as a dairy replacement for products such as milk, evaporated milk, condensed milk, buttermilk, yogurt, and many others. Since its texture is similar to heavy whipping cream, it is often used in a variety of curries, desserts, and cakes, and is an ideal ingredient for use in coffee and hot beverages. Coconut cream also works well in frozen drinks such as piña coladas and other tropical drinks. In terms of mouthfeel, fat content from coconut cream is often reported to be as good as twice the amount of butter fat.
iTi provides coconut cream that is aseptically packed with no preservatives. The product is available with and without natural stabilizers. The stabilizers are added to prevent separation. The products are 100% natural and GMO free. They do not contain any artificial color, flavors, or preservatives and are processed in FDA registered facilities, which are HACCP and Kosher-certified. Organic coconut cream is also available.
iTi Tropicals is an importer that supplies tropical, exotic fruit juice, puree and concentrates to the food and beverage manufacturing industries. Some of the products they supply include mangosteen, acerola, acai and coconut water concentrate as well as guava, banana and pineapple. iTi's research technologists assist their customers in developing formulas and products using these fruit ingredients in for all applications from smoothies, sorbets, ice cream, and fruit drinks to sauces, soups and salad dressings.
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應用領域 Categories
iTi Tropicals, Inc.
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DS Simulia XFlow 2019x Build 106.05 x64 | 1.72 Gb
SIMULIA XFlow offers unique, particle-based Lattice Boltzmann technology for high fidelity Computational Fluid Dynamics (CFD). This state-of-the-art technology enables users to address complex CFD problems involving high frequency transient aerodynamics, real moving geometries, complex multiphase flows, fluid structure interactions and aero acoustics. XFlow's automatic lattice generation and adaptive refinement capabilities minimize user inputs thereby reducing time and effort in the meshing and pre-processing phase of a typical CFD workflow. This enables engineers to focus the majority of their efforts on design iteration and optimization, rather than the time consuming meshing process.
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The reason why we are all here in Punta Cana: Julie and Bryan's wedding. It was a short, but beautiful ceremony just steps from the ocean. After dinner, we all headed down to the carnivale on Caribe Street. The carnivale involved dancing, drinking, souvenirs, a parade, and a dance contest.
Punta Cana - The Wedding
by Kevin March. 27, 2008 3604 views


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This is a perfect side dish for chicken dishes, or a nice appetizer. You can serve it with marinara.
Instant Pot Chili Polenta Print Pin Rate
Prep Time: 15 minutes
Cook Time: 10 minutes
Total Time: 25 minutes
Servings: 6
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If you loved this recipe, give it love. Thanks!


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SINGAPORE - Transport Minister Khaw Boon Wan said on Monday (July 8) that the Government is currently subsidising more than 30 per cent of public transport operations, and indicated that higher fares would be necessary to keep subsidies in check.
"Between 2016 and 2017, the total cost of running the rail network has increased by around $270 million," Mr Khaw told Parliament. "As the fares paid by commuters do not cover operating costs, the rail companies are operating at a loss. In the latest reported financial year, SMRT Trains incurred a loss of $86 million. SBS Transit's train division also lost tens of millions of dollars."
Mr Sitoh Yih Pin (Potong Pasir) had sought an update on the current level of rail reliability and how such continued efforts are going to be sustained and funded.
The increased expenditure had largely been on the back of multi-year efforts to raise rail reliability. Mr Khaw noted that the MRT network is now clocking nearly one million train-kilometres between delays – a seven-fold improvement from 2015. He added that over the next five years, the Government expects to spend $4.5 billion on operating subsidies. This would be on top of the $25 billion to be spent on building new lines.
Mr Khaw said that until recently, Public Transport Council (PTC) fare adjustments were not implemented to the full extent of what the formula allowed. He said: "If we had strictly followed PTC's fare formula, the operators would have been better able to cover the costs of the intensified maintenance."
He added that the Government will continue to operate under the current PTC formula, which is valid till 2023. "But we must have the discipline to implement the formula fully, as we adjust fares over the next four years," he said.
"In due course, the PTC will need to review the fare adjustment mechanism to reflect the increased operating cost to support the intensified maintenance, and the additional operating subsidies from the Government to the MRT system."
Meanwhile, he said the Government will provide operators with "a temporary enhanced maintenance grant".
Nominated MP Walter Theseira asked the minister for the targeted level of subsidy "in the longer run".
Mr Khaw said the original plan was for the Government to pay for infrastructure and the first set of operating assets, and for the replacement of these assets to be funded by fares.
If that had worked well, there would be zero subsidy for operations, he noted. As it turned out, that has proved to be unachievable, and the operating subsidy is now "more than 30 per cent".
At the end of the day, the system will have to be funded by either taxpayers or commuters. "We've just got to find that balance," Mr Khaw said.
"Should we drive towards zero (operating subsidy)? I mean, we must never give up on this hope but if it is not possible in the medium term, it does not matter. The key point is, we must not allow the operating subsidy percentage to continue to rise."
To another question from Mr Sitoh on whether MRT reliability should continue to improve – to beyond one million train-km between delays – the minister said the figure is already a fairly high standard which only a handful of metro systems in the world are able to meet.
"But because of various reasons, Singapore commuters are demanding an even higher level and are prepared to pay... We leave that to the next transport minister," he said.
SMRT has good team, but some attrition useful: Khaw
SMRT has a very good team of people in charge, and sometimes, some attrition is useful, Transport Minister Khaw Boon Wan said.
He was responding to Workers' Party chief Pritam Singh (Aljunied GRC), who asked about recent reports in The Straits Times on various resignations at the rail operator, and what impact this has on SMRT's plans for rail reliability for the long haul.
Mr Singh said his concern was whether there is a deeper issue within SMRT that the public ought to be concerned about.
Replied Mr Khaw: "With the change of leadership of SMRT... and the team that I worked with both above ground and underground, I know them well and it's a very good team of people. And, sometimes some attrition is useful."
SMRT had a change of leadership last August when Mr Neo Kian Hong replaced Mr Desmond Kuek as chief executive. ST also reported in April this year that about 30 executives in its human resources department had quit, as had two senior engineers.
Citing the ST reports, Mr Singh asked: "Is there a significant number that Singaporeans are to be concerned that we start slipping in terms of the targets the minister has set?... Is there any concern the ministry has about morale within SMRT and whether the system can continue to move forward reliably?"
Mr Khaw pointed to an ST report last month that SMRT Trains' chief executive was expected to resign, saying he immediately texted the SMRT chairman – Mr Seah Moon Ming – about it.
"He was in the air then and as soon as he landed at the airport, he replied to me that it is fake news," said the minister. "ST gave an apology in tiny font size," Mr Khaw added.
The Straits Times had apologised for the error on Page 2 of the print edition, where corrections are usually placed, and in its regular font size.


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Kim Yunkyung, a contemporary artist from South Korea with a Doctorate from Hongik University and a Masters of Arts from Goldsmiths University of London (UK), focuses on installation art.
She is currently a lecturer in several universities in South Korea including Hongik University and Sangmyung University.
Her art works were displayed at various art national and international exhibitions. Additionally, she is also a performance artist who has been on stage in Seoul and London.
The art work for this exhibition are entitled "We are not two…" and "Moving Flow".


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ashapop needed a new logo design and created a contest on 99designs.
A winner was selected from 115 designs submitted by 36 freelance designers.
asha.pop communications
Yes. 3 words under the logo indicatingg what we do. "Media Training. Broadcast. PR."
I'm a media consultancy firm specialising in three main types of services. a. Media Training (ie teaching CEOs and managers how to speak to the media for maximum impact) b. Documentaries (produce half hour chat shows and business progs) and c. PR (put together communications strategies and plans for small companies and do media pitching for them).
Target audience are businesses who need to speak to the media.
I've attached a logo BlinkAsia which has the feel and look that I want. I've also added other logos that I like the look of. Am hoping to have a design that makes the pop a bit more prominent in asha.pop communications ie it pops out (I know it sounds a little corny) But not anything too childish. Strong, grounded logo with bold colours and shape and a little of design element in the "pop" to demonstrate creativity. Nb: I want logo to be all in lower case and communications to be spelt out in full, thanks :)
Find out how a design contest works or learn more about our logo design services.


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Author Archive
Tweeting versus experiencing
As children, we were always told not to bring our gadgets to the dinner table, that it was "rude to be busy with something else while someone was trying to talk to you" and "quality face time is more important than conversation with people who were miles away". With social media growing as rapidly as it has however, times certainly have changed.
Cerebra's CEO Mike Stopforth has this to say, "It's a little bit like saying a hundred years ago you can't have telephones in offices because the ringing will distract people. It's [social media] a part of life these days."
Social Media For Parents: How not to cramp your kid's style
Our children are living in the information age. They have moved from just chatting via text messages, BBM, WhatsApp, IM, mig33 or MXit. They want to stay in touch with friends; which is where social media comes in. With incidences of cyber bullying, sexting and pedophiles on the prowl, it's only natural that parents are concerned about their online safety. While you still want to keep tabs on what is going on in their lives and what they come into contact with, you don't want to invade their privacy and end up losing their trust. It's important that you educate and empower them by setting proper ground rules.


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WORLD-RENOWNED BESTSELLING AUTHOR CARLOS CASTANEDA'S SELECTION OF HIS WRITINGS ON THE SHAMANS OF ANCIENT MEXICONear the end of his life, Carlos Castaneda gathered together and reviewed his seminal works on his training as a shaman initiate, recorded in a literary career that spans over thirty years. The result is this groundbreaking collection of quotations -- the essence of Carlos Castaneda, drawn from his landmark volumes including The Teachings of Don Juan, Journey to Ixtlan, A Separate Reality, and Tales of Power. Enhanced with an introduction and original commentary by the author, this powerful work illuminates the shaman's life as never before. Castaneda's words explore how the ancient shamans could literally touch and direct the wheel of time -- a profound yet pragmatic tradition that can be felt even in our day.
Born in 1925 in Peru, anthropologist Carlos Castaneda wrote a total of 15 books, which sold 8 million copies worldwide and were published in 17 different languages. In his writing, Castaneda describes the teaching of Don Juan, a Yaqui sorcerer and shaman. His works helped define the 1960's and usher in the New Age movement. Even after his mysterious death in California in1998, his books continue to inspire and influence his many devoted fans.
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Praise for the groundbreaking work of bestselling author Carlos Castaneda "Extraordinary in every sense of the word."
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By clicking 'Sign me up' I acknowledge that I have read and agree to the privacy policy and terms of use. Free eBook offer available to NEW US subscribers only. Offer redeemable at Simon & Schuster's ebook fulfillment partner. Must redeem within 90 days. See full terms and conditions and this month's choices.
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