text:
Let's celebrate the Oscars by pairing food and film. This year, I am only making one dish for one movie. I think Hidden Figures will be the movie that wins! It's such an inspiring and true story of Katherine Johnson, a brilliant NASA … Continue Reading
Tasting New Orleans: Dooky Chase's
"Food can make you happy. You can solve all kinds of problems over a plate of food. And that is a wonderful … [read more]
Tasting New Orleans: Shaya
Mardi Gras is almost here. Let's continue tasting New Orleans! What a delightful honor to … [read more]
Tasting New Orleans: Breakfast at Brennan's
New Orleans is where it all begins.... Many of you know that Kitchen Chat is a way to honor my late … [read more]
About Kitchen Chat:
Welcome to my kitchen – a gathering place to sample and share. Thank you for joining me on my culinary journey. It's a special way to honor my late father, who was an incredible gourmet home chef. My biggest regret in life was not going into the kitchen with him and learning how to cook. He passed away 25 years ago. Midlife, I am discovering my dad's joy of cooking along with great tips and trends from chefs and foodies. Please sign up for my newsletter and subscribe to my weekly podcast. Let's chat together in my kitchen. What are your favorite recipes? What are your questions for guests on the show? Please click on the Contact button and connect with me. I would love to hear from you.
As the Brazilians say, "Aproveite!" which means Enjoy! I, however, like to translate "Aproveite" to mean Savor the day!
Cook with Margaret
Culinary Travel
Tasting the World (Part Two): New York Times Travel Show
The Taste of the World Stage at The New York Times Travel Show featured great chefs such as Art Smith, Roy Yamaguchi, and Bill Yosses. My friend Doug Duda from IACP was the culinary director and host. Chef Bill Yosses, former White House pastry … Continue Reading
Chefs and Cookbooks
Tasting New Orleans: Shaya
Mardi Gras is almost here. Let's continue tasting New Orleans! What a delightful honor to interview the highly acclaimed Chef Alon Shaya at his modern Israeli restaurant, Shaya, which was awarded the 2016 James Beard Best New … Continue Reading


text:
TRENTON, N.J. (CBSNewYork/AP) — A Monmouth County, N.J.-based public adjusting company has admitted to submitting fraudulent insurance claims, in an attempt to bilk an area insurer out of more than $15,000.
Long Branch, N.J.-based Northeast Claims Group recently pleaded guilty to second-degree insurance fraud.
The firm also agreed to be dissolved and have its state public adjuster license revoked.
A public adjuster is an intermediary between an insured and their insurance company. The adjuster is hired by an insured after they suffer losses.
The company was also fined $7,500 criminal fine and its owner — Joseph Shammah, 40, of West Long Branch agreed to have his public adjuster's license revoked.
Shammah was allowed to enter a pretrial program, meaning the criminal charge personally against him would be dismissed if he successfully completes the program.
You may also be interested in these stories:
- Reward Offered For Information As Investigation Into Linden Suspicious Package Continues
- 1 Killed, 1 Injured In Small Plane Crash Near Tweed New Haven Airport
- Pit Bull Rescued From Abandoned Crack House Adopted By Lower East Side Firefighters
- Multi-Vehicle Crash, Debris Spill Snarls Traffic On Route 17 In Paramus
(TM and © Copyright 2013 CBS Radio Inc. and its relevant subsidiaries. CBS RADIO and EYE Logo TM and Copyright 2013 CBS Broadcasting Inc. Used under license. All Rights Reserved. This material may not be published, broadcast, rewritten, or redistributed. The Associated Press contributed to this report.)

subset: commoncrawl


text:
|Visitors Now:
|Total Visits:
|Total Stories:
|Story Views
|Last Hour:
|Last 24 Hours:
It's both a Friday news dump and an October surprise! FBI Director James Comey sent a letter to several members of the House today letting them know that the agency was reopening its investigation into Hillary Clinton's use of a private server to store emails related to her work as Secretary of State.
When Comey ended his investigation—over whether Clinton and her staff mishandled classified information—he determined that she had been being extremely careless with data but decided not to recommend charges, and the Justice Department agreed.
Today the situation has changed, but it's way too soon and Comey's being way too close-lipped to determine what is happening. The letter reads:
In connection with an unrelated case, the FBI has learned of the existence of emails that may be pertinent to the investigation. I am writing to inform you that the investigative team briefed me on this yesterday, and I agreed that the FBI should take appropriate investigative steps designed to allow the investigators to review these emails to determine whether they contain classified information, as well as to assess their importance to our investigation.
Comey concludes he doesn't know yet whether any of this information is significant and doesn't know how long it will take to find out. Check out the letter for yourself after the jump.
Jacob Sullum has noted that how Clinton has responded to this scandal has reinforced her reputation for dishonesty. And check out ReasonTV's hilarious-yet-horrifying supertake of misleading statements by Clinton:


text:
So, I am off to St. Louis----- I do have a food plan. Please critique or comment. I need chowhound experiences.
Saturday lunch---- Crown Candy Kitchen
Snack--- Drewes Frozen Yogurt
Saturday dinner---- Tony's
Sunday--I am at an event most of the day
I need a Sunday night idea.......
What am I missing? What is a must? THANKS!
Eat Rite looks so cute--I wish I could fit it in
Updated 1 year ago | 0
Updated 1 year ago | 29
Updated 1 month ago | 15
Updated 26 days ago | 254
Updated 24 days ago | 17


text:
Feher M.,Campbell University |
Williams C.I.,Chemical Computing Group
Journal of Chemical Information and Modeling | Year: 2012
This work examines the effect of small input perturbations on binding energies computed from differences between energy minimized structures, such as the Prime MM-GBSA and MOE MM-GB/VI methods. The applied perturbations include translations of the cognate ligand in the binding site by a maximum of 0.1 Å along each coordinate or the permutation of the order of atoms of the cognate ligand without any changes to the atom coordinates. These seemingly inconsequential input changes can lead to as much as 17 kcal/mol differences in the computed binding energy. The calculated binding energies cluster around discrete values, which correspond to specific ligand poses. It appears that the largest variations are observed for target-ligand systems in which there is a possibility for multiple poses with strong hydrogen bonds. The barriers between different poses can appear fractal-like, making it difficult to predict which solution will be produced from a given input. Including protein flexibility in MM-GBSA calculations further increases numerical instability, and the protein strain terms seem to be the major factor contributing to this sensitivity. In such calculations it appears unwise to extend the flexible region beyond 6 Å. © 2012 American Chemical Society. Source
Evaluating the Impact of Uncertainties in Clearance and Exposure When Prioritizing Chemicals Screened in High-Throughput Assays
Leonard J.A.,Oak Ridge Institute for Science and Education |
Sobel Leonard A.,Duke University |
Chang D.T.,Chemical Computing Group |
Edwards S.,U.S. Environmental Protection Agency |
And 6 more authors.
Environmental Science and Technology | Year: 2016
The toxicity-testing paradigm has evolved to include high-throughput (HT) methods for addressing the increasing need to screen hundreds to thousands of chemicals rapidly. Approaches that involve in vitro screening assays, in silico predictions of exposure concentrations, and pharmacokinetic (PK) characteristics provide the foundation for HT risk prioritization. Underlying uncertainties in predicted exposure concentrations or PK behaviors can significantly influence the prioritization of chemicals, though the impact of such influences is unclear. In the current study, a framework was developed to incorporate absorbed doses, PK properties, and in vitro dose-response data into a PK/pharmacodynamic (PD) model to allow for placement of chemicals into discrete priority bins. Literature-reported or predicted values for clearance rates and absorbed doses were used in the PK/PD model to evaluate the impact of their uncertainties on chemical prioritization. Scenarios using predicted absorbed doses resulted in a larger number of bin misassignments than those scenarios using predicted clearance rates, when comparing to bin placement using literature-reported values. Sensitivity of parameters on the model output of toxicological activity was examined across possible ranges for those parameters to provide insight into how uncertainty in their predicted values might impact uncertainty in activity. © 2016 American Chemical Society. Source
Feher M.,Campbell University |
Williams C.I.,Chemical Computing Group
Journal of Chemical Information and Modeling | Year: 2012
This work examines the sensitivity of docking programs to tiny changes in ligand input files. The results show that nearly identical ligand input structures can produce dramatically different top-scoring docked poses. Even changing the atom order in a ligand input file can produce significantly different poses and scores. In well-behaved cases the docking variations are small and follow a normal distribution around a central pose and score, but in many cases the variations are large and reflect wildly different top scores and binding modes. The docking variations are characterized by statistical methods, and the sensitivity of high-throughput and more precise docking methods are compared. The results demonstrate that part of docking variation is due to numerical sensitivity and potentially chaotic effects in current docking algorithms and not solely due to incomplete ligand conformation and pose searching. These results have major implications for the way docking is currently used for pose prediction, ranking, and virtual screening. © 2012 American Chemical Society. Source
Experimental versus predicted affinities for ligand binding to estrogen receptor: Iterative selection and rescoring of docked poses systematically improves the correlation
Wright J.S.,Carleton University |
Anderson J.M.,Carleton University |
Shadnia H.,Chemical Computing Group |
Durst T.,University of Ottawa |
Katzenellenbogen J.A.,Urbana University
Journal of Computer-Aided Molecular Design | Year: 2013
The computational determination of binding modes for a ligand into a protein receptor is much more successful than the prediction of relative binding affinities (RBAs) for a set of ligands. Here we consider the binding of a set of 26 synthetic A-CD ligands into the estrogen receptor ERα. We show that the MOE default scoring function (London dG) used to rank the docked poses leads to a negligible correlation with experimental RBAs. However, switching to an energy-based scoring function, using a multiple linear regression to fit experimental RBAs, selecting top-ranked poses and then iteratively repeating this process leads to exponential convergence in 4-7 iterations and a very strong correlation. The method is robust, as shown by various validation tests. This approach may be of general use in improving the quality of predicted binding affinities. © 2013 Springer Science+Business Media Dordrecht. Source
Developing a Physiologically-Based Pharmacokinetic Model Knowledgebase in Support of Provisional Model Construction
Lu J.,Oak Ridge Institute for Science and Education |
Goldsmith M.-R.,U.S. Environmental Protection Agency |
Goldsmith M.-R.,Chemical Computing Group |
Grulke C.M.,U.S. Environmental Protection Agency |
And 15 more authors.
PLoS Computational Biology | Year: 2016
Developing physiologically-based pharmacokinetic (PBPK) models for chemicals can be resource-intensive, as neither chemical-specific parameters nor in vivo pharmacokinetic data are easily available for model construction. Previously developed, well-parameterized, and thoroughly-vetted models can be a great resource for the construction of models pertaining to new chemicals. A PBPK knowledgebase was compiled and developed from existing PBPK-related articles and used to develop new models. From 2,039 PBPK-related articles published between 1977 and 2013, 307 unique chemicals were identified for use as the basis of our knowledgebase. Keywords related to species, gender, developmental stages, and organs were analyzed from the articles within the PBPK knowledgebase. A correlation matrix of the 307 chemicals in the PBPK knowledgebase was calculated based on pharmacokinetic-relevant molecular descriptors. Chemicals in the PBPK knowledgebase were ranked based on their correlation toward ethylbenzene and gefitinib. Next, multiple chemicals were selected to represent exact matches, close analogues, or non-analogues of the target case study chemicals. Parameters, equations, or experimental data relevant to existing models for these chemicals and their analogues were used to construct new models, and model predictions were compared to observed values. This compiled knowledgebase provides a chemical structure-based approach for identifying PBPK models relevant to other chemical entities. Using suitable correlation metrics, we demonstrated that models of chemical analogues in the PBPK knowledgebase can guide the construction of PBPK models for other chemicals. © 2016 Public Library of Science. All Rights reserved. Source


text:
PerfectMatch.com seems to be one of the best dating sites around on the net today. In Perfect Match dating review you'll find detailed information about the sites structure and its best offers. Check out our online dating review or even review dating sites by yourself!
Vote to see results
Different Dates for Different States? What Are the Dating Trends in Your City?
The Toronto-based service analyzed data based on its 1.5 million users and discovered that dating trends vary from place to place, with certain regions showing a strong bent toward very specific interests. For example, based on the findings, Oklahoma ...
People Magazine
Bella Hadid Opens Up About Her Love Life: 'I'm Dating Abel. I Don't See Him as the Weeknd'
People Magazine
Bella Hadid has made it clear that she's dating Abel Tesfaye and not his superstar alter ego, The Weeknd. The 19-year-old model opened up about her love life in Glamour magazine's September cover story. While it's easy to love a multi-talented ...
Bella Hadid on dating The Weeknd: He's just Abel to meUPI.com
Bella Hadid talks dating, her sister and sudden fameNew York Daily News
Bella Hadid On What It's Really Like To Date The WeekndRefinery29
all 30 news articles »
Woman claims Chicago dating service didn't deliver, seeks $3800 refund
... Lunch Chicago declined an on-camera interview, but sent a one-page statement saying employees listen to customer concerns and they cannot guarantee chemistry, a long term relationship or marriage, and matchmaking differs from online dating in that ...
Online Dating: Boyfriend Flies to China, Waits Days at Airport
Alexander Pieter Cirk was due to arrive back home in Holland Tuesday after reportedly spending 10 days at a Chinese airport in a failed bid to meet a woman he'd fallen for on a dating app. Apparently frustrated by the time and distance separating the ...
and more »
The Cheat Sheet
Researchers have figured out the one thing not to do in your online dating profile
Washington Post
Online dating has made potential partners much more readily available than ever before -- and yet also, somehow, disposable. The other day I was sitting on a train with a friend as she flicked through profiles on Bumble, an online dating service in ...
4 Lies People Have on Their Online Dating ProfilesThe Cheat Sheet
all 2 news articles »
This interracial dating app is about more than love
After being selected for Y Combinator's 2014 class and working on creating an artificial intelligence-fueled chatbot, Vu Tran decided three months ago to scrap that project and create Color Dating, an app for interracial dating. It's been said that ...
Turns Out, Speed Dating May Actually Be a Great Way to Meet People in NYC
While online dating has totally shed the stigma that was long attached to it, speed dating is still largely seen as a last resort for desperate singles who have failed everywhere else in the dating pool... for clueless men and women who naively think ...
Asia Times
Lifestyle: Bae Suzy, Lee Min-ho are 'dating' but break-up rumours abound
Asia Times
After a long-long suspense over their relationship status that even triggered break-up rumours, star couple Lee Min-ho and Bae Suzy have reportedly broken the silence and confirmed that they are still dating each other. But still media speculations ...
Lee Min Ho and Suzy are Still Dating!China Christian Daily
all 13 news articles »
Fast Company
How Women Are Swiping The Weirdness Out Of Online Dating
Fast Company
As strange as her experience felt, Tweten realized after swapping stories with friends that it wasn't that unusual: For many women, oddly aggressive, sometimes threatening interactions with men are an all-too-familiar feature of Internet dating. The ...
People Magazine
Josh Groban and Kat Dennings Split After Nearly 2 Years of Dating: Report
People Magazine
Josh Groban and Kat Dennings Split After Nearly 2 Years of Dating: Report. 0SHARES. Reprint. subscribe now. Josh Groban and Kat Dennings Break Up After 2 Years of Dating. Kat Dennings and Josh Groban. Steve Granitz/WireImage. By Stephanie Petit.
Kat Dennings, Josh Groban Split After Nearly Two Years of DatingUs Weekly
Josh Groban & Kat Dennings Split After Two Years of DatingExtra (blog)
Josh Groban and Kat Dennings announce split after two years of datingDaily Mail
Just Jared -E! Online -UPI.com
all 30 news articles »


text:
Ricoh offers HZ15 digital camera, 5 HD Pentax lenses and auto flash units - all in September
Best DSLR cameras 2017: The best interchangeable lens cameras available to buy today
Best compact system cameras 2017: The best mirrorless interchangeable lens cameras...
Best compact cameras 2017: The best pocket cameras available to buy today
Panasonic Lumix GX800 review: An affable, affordable entry-level mirrorless...
Popular In Cameras On Pocket-lint
- Panasonic Lumix GX800 review: An affable, affordable entry-level mirrorless camera
- Canon EOS 77D preview: Lucky 7s for the '80D lite'?
- Canon EOS M6 preview: Canon's top-end mirrorless, minus the viewfinder
- Canon EOS 800D preview: Mid-range made accessible
- Canon EOS 77D and 800D expand mid-range EOS DSLR lineup
- Canon EOS M6 is Canon's new top viewfinder-free mirrorless camera
- Nikon scraps DL compact camera line promised last summer
- GoPro confirms Hero6 and other new cameras are coming this year
- Panasonic Lumix LX10 / LX15 review: The best high-end compact camera money can buy?
- Best compact system cameras 2017: The best mirrorless interchangeable lens cameras available to buy today
- Pokemon Go evolutions: Special items and how to evolve Polytoed, Slowking, Bellossom, Steelix and more
- Samsung Galaxy S8: Release date, rumours and everything you need to know
- This is what the Huawei P10 flagship phone will look like in full black
- Samsung Galaxy Tab S3 shows up wearing an iPad Pro-like keyboard
- Horizon Zero Dawn review: The best-looking game on PS4 by far
- This is what the LG G6 will (probably) look like
- New Samsung Galaxy S8 image leaks reveal phone, on-screen controls
- E3 2017: What to expect from the world's biggest games show
- Massive Sonos Brexit price hike coming: Buy your Playbar, Sub and speakers today!
- Best Android Wear 2.0 apps for your watch: Google Fit, Uber, and more


text:
Transcriptional analysis of highly syntenic regions between Medicago truncatula and Glycine max using tiling microarrays
- Lei Li†1, 8,
- Hang He†1, 2, 3,
- Juan Zhang4,
- Xiangfeng Wang1, 2, 3,
- Sulan Bai5,
- Viktor Stolc6,
- Waraporn Tongprasit6,
- Nevin D Young7,
- Oliver Yu4 and
- Xing-Wang Deng1Email author
DOI: 10.1186/gb-2008-9-3-r57
© Li et al.; licensee BioMed Central Ltd. 2008
Received: 11 October 2007
Accepted: 19 March 2008
Published: 19 March 2008
Legumes are the third largest family of flowering plants and are unique among crop species in their ability to fix atmospheric nitrogen. As a result of recent genome sequencing efforts, legumes are now one of a few plant families with extensive genomic and transcriptomic data available in multiple species. The unprecedented complexity and impending completeness of these data create opportunities for new approaches to discovery.
We report here a transcriptional analysis in six different organ types of syntenic regions totaling approximately 1 Mb between the legume plants barrel medic (Medicago truncatula) and soybean (Glycine max) using oligonucleotide tiling microarrays. This analysis detected transcription of over 80% of the predicted genes in both species. We also identified 499 and 660 transcriptionally active regions from barrel medic and soybean, respectively, over half of which locate outside of the predicted exons. We used the tiling array data to detect differential gene expression in the six examined organ types and found several genes that are preferentially expressed in the nodule. Further investigation revealed that some collinear genes exhibit different expression patterns between the two species.
These results demonstrate the utility of genome tiling microarrays in generating transcriptomic data to complement computational annotation of the newly available legume genome sequences. The tiling microarray data was further used to quantify gene expression levels in multiple organ types of two related legume species. Further development of this method should provide a new approach to comparative genomics aimed at elucidating genome organization and transcriptional regulation.
The rapidly increasing number of genome and transcript sequences in recent years is having two marked, complementary effects on the relatively new discipline of plant genomics and transcriptomics. The newly available sequences need to be fully annotated to identify all the functional and structural elements. Because genome annotation is a reiterative process that is heavily dependent on large-scale, high-throughput experimental data, each additional genome sequence comes as a new challenge. On the other hand, the availability of multiple genomic and transcriptomic datasets fosters comparative analyses that improve structural annotation of the genomes and generate new insight into the function and evolution of protein-coding and non-coding regions of the genomes.
One approach to systematically characterize genome transcription is to use high feature-density tiling microarrays on which a given genome sequence is represented [1, 2]. Genome tiling arrays have been used in a number of model species for which the full genome sequence is available [3–8]. Results from these studies have shown that for well-documented transcripts, such as those of polyadenylated RNAs from annotated genes, hybridization signals from tiling arrays identify the transcriptional start and stop sites, the locations of introns, and the events of alternative splicing [3–8]. Tiling arrays therefore provide a valuable means for confirming the large number of predicted genes that otherwise lack supportive experimental evidence. However, tiling array signals also reveal a large number of putative novel transcripts for which no conventional explanations are yet available.
With respect to plants, the Arabidopsis thaliana genome was the first to be probed by tiling microarrays [5]. Tiling array analysis of the more complex rice genome has been carried out as well [8–10]. The rice tiling array data were used to detect transcription of the majority of the annotated genes. For example, of the 43,914 non-transposable element protein-coding genes from the improved indica whole genome shotgun sequence [11], transcription of 35,970 (81.9%) was detected [8]. On the other hand, comprehensive identification of transcriptionally active regions (TARs) from tiling array profiles revealed significant transcriptional activities outside of the annotated exons [8–10]. Subsequent analyses indicate that about 80% of the non-exonic TARs can be assigned to various putatively functional or structural elements of the rice genome, ranging from splice variants, uncharacterized portions of incompletely annotated genes, antisense transcripts, duplicated gene fragments, to potential non-coding RNAs [10].
In addition to detecting transcriptome components, genome tiling arrays in theory can be used to directly quantify the expression levels of individual transcription units. As an alternative approach to the surrogate expression arrays, tiling arrays offer two potential advantages. First, in tiling arrays each transcription unit is interrogated by hundreds of probes according to the actual genomic sequence. This strategy eliminates the need to arbitrarily select a small number of supposedly gene-specific probes and thus alleviates probe bias and improves cross-platform comparability in microarray experiments. Second, measurement of gene expression using tiling arrays allows averaging of the results from multiple probes per gene, which can reduce inconsistent probe behavior and thus provide improved statistical confidence.
Using DNA microarrays to study gene expression in closely related species has become an important approach to identify the genetic basis for phenotypic variation and to trace evolution of gene regulation [12–17]. However, expression levels as well as sequences may differ between species, creating additional technical challenges for inter-species comparisons. Current approaches to control for the effect of sequence divergence are either to mask probes with sequence mismatches [17, 18] or to use probes derived from the various species of interest to cancel out the sequence mismatch effect [19, 20]. Both approaches, however, rely on a few empirically or computationally selected probes for each gene of interest. Consequently, the effectiveness and accuracy of these approaches is still a matter of debate [18]. In related species for which genome sequences have all been determined, genomic tiling arrays could provide an alternative approach to inter-species comparison of gene expression. Again, the inclusion of multiple probes per transcription unit in tiling arrays could potentially improve the accuracy and fairness of the estimation of gene expression levels in each species, which in turn could improve cross-species comparison of the expression patterns of orthologous genes.
As the third largest family of flowering plants, legumes (Fabaceae) are unique among crop species in their ability to fix atmospheric nitrogen through symbiotic relationships with rhizobia bacteria [21]. Extensive expressed sequence tags have been collected for a number of legume species, including soybean (Glycine max), lotus (Lotus japonicus), common bean (Phaseolus vulgaris), and barrel medic (Medicago truncatula) [22, 23]. Genomes of barrel medic, soybean, and lotus are being sequenced because all are models for studying nitrogen fixation and symbiosis, tractable to genetic manipulation, and exhibit diploid genetics and modest genome sizes. Both barrel medic and lotus have a diploid genome of approximately 475 Mb while soybean has a diploidized tetraploid genome estimated at 950 Mb [24, 25]. Recently, preliminary genome assembly and annotation of barrel medic (Mt2.0) and soybean (Glyma0) became publicly available [26, 27]. As a result, legumes are now one of a few plant families in which extensive genome sequences in multiple species are available.
Comparisons of genome sequences have revealed various degrees of synteny (conservation of gene content and order) among species related at different taxonomic levels. For legume plants, early work based on DNA markers demonstrated substantial genome conservation among some Phasoloid species, including mungbean (Vigna radiata) and cowpea (V. unguiculata) [28], and between Vigna and the common bean [29]. Genome-wide gene-based analysis among legumes using a large set of cross-species genetic markers produced chromosome alignments from five species of the Papilionoid subfamily, including barrel medic and soybean [30]. More recently, direct synteny comparison of the finished and anchored genome sequences from barrel medic and lotus was made. Results from this study indicated that three-quarters of the genome of each species may reside in conserved syntenic segments in the genome of the other [25], which share at least ten large-scale synteny blocks that frequently extend the length of whole chromosome arms [26].
Two soybean regions comprising approximately 0.5 Mb each surrounding the soybean cyst nematode resistance loci, rhg1 and Rhg4, were extensively characterized [31]. Using these sequences, Mudge et al. [32] identified the syntenic regions from barrel medic. They found that many predicted genes in the syntenic regions were conserved and collinear between the two species. Here, we used tiling microarray analysis to verify the predicted genes, to identify additional transcripts, and to compare transcription patterns in six different organ types in each species. Our results provide transcriptional support to over 80% of the predicted genes and identified 499 and 660 TARs from barrel medic and soybean, respectively. The gene expression patterns in the six organ types of some collinear genes showed significant differences between the two species despite synteny at the DNA level, demonstrating the usefulness of genomic tiling analysis in comparative genomics.
Genes in the syntenic regions between barrel medic and soybean
In a previous study, two regions in the soybean genome comprising approximately 0.5 Mb each surrounding the soybean cyst nematode resistance loci, rhg1 and Rhg4, were used to identify syntenic regions in the Medicago genome [32]. Because there was a 2 cM gap in the first region, these sequences were referred to as synteny blocks 1a, 1b, and 2 [32]. The syntenic regions in barrel medic also totaled about 1 Mb, though they were scattered into smaller contigs. For example, synteny block 1b in barrel medic contained two additional gaps [32]. In barrel medic, there were two segmental duplications (block 2i and 2ii) that were both syntenic to soybean synteny block 2 [32].
Genes were predicted in the 1 Mb barrel medic and soybean sequence contigs using FGENESH [33]. Both the dicot plants (Arabidopsis) and the Medicago (legume plant) matrixes were used and their outputs compared [33]. Using the legume matrix, 229 and 217 genes were predicted for the barrel medic and soybean sequences, respectively (Additional data file 1). These represent significantly more but shorter genes (exons) compared with the Arabidopsis matrix outputs. However, the legume matrix prediction also resulted in more base-pairs in the exons (increases of 10.3% and 8.2% for barrel medic and soybean, respectively; Additional data file 1). These results clearly demonstrate that gene prediction output is sensitive to the training matrix and highlight the importance of experimental means in verifying and improving computational gene prediction. For simplicity, we selected the gene prediction from the legume matrix for further analysis.
Tiling microarray detection of predicted genes
Identification and characterization of TARs
The non-exonic TARs were further analyzed in terms of their physical location relative to the predicted genes. In this analysis, genome regions were divided into eight different configurations against the predicted exons (Figure 4b). Interestingly, in almost all antisense configurations, there were more TARs in soybean than in barrel medic (Figure 4b), suggesting that antisense transcription is more prevalent in soybean than in barrel medic. This analysis also revealed a surprisingly large number of intergenic TARs (36 in barrel medic and 45 in soybean) located in close proximity on the antisense strand 5' to the start of a predicted gene (Figure 4b). Because the predicted genes do not include untranslated regions, it is conceivable that transcripts derived from these TARs and the corresponding genes are arranged in a divergent antisense orientation and could potentially form duplex transcript pairs.
Differential gene expression in the syntenic regions
In soybean, all but one of the detected nodule-enhanced genes are known genes (Additional data file 3). In contrast, only three of the 11 nodule-enhanced genes detected in barrel medic match with a known gene while the other eight genes have no assigned functions (Additional data file 2). When the nodule-enhanced genes detected in barrel medic and soybean were compared for synteny, six of the ten soybean genes were found to have a collinear counterpart in barrel medic, although transcription of the collinear genes in barrel medic was not nodule-enhanced (Additional data file 3). Consequently, there was only one gene encoding a TGACG-binding transcription factor that is collinear as well as specifically expressed in the nodule in both species.
Transcriptional pattern of collinear genes in the syntenic regions
The barrel medic and soybean sequences interrogated by the tiling microarray are highly syntenic. In the previous report, a total of 68 pairs of genes were found to be collinear with both the gene order and orientation conserved between barrel medic and soybean homologs [32]. In the current study, we were able to identify 78 collinear gene pairs based on the gene prediction output from the legume matrix.
The rapidly accumulating amount of genome and transcriptome data in recent years is having profound effects on biological research. Elucidating all the functional and structural elements of the genome sequences and how they are organized and regulated, and how they evolved has thus become the focus of the next phase of genome projects. In these regards, genome tiling microarray analysis is emerging as a new powerful approach, which involves the development of tiling arrays containing progressive oligonucleotide tiles that represent a target genome. Recent advances in microarray technologies allow oligonucleotide arrays to be made with several hundred thousand to several million discrete features per array, which permits tiling complex genomes with a manageable number of arrays [1, 2]. This in turn has resulted in transcriptomic tiling data for a large number of model species [1–10].
Application of tiling array analysis in genomics studies has significantly broadened our understanding of the genetic information encoded in the genome sequences. When probed against various RNA samples, tiling array hybridization patterns identify transcript ends and intron locations [3–8]. Tiling array analysis thus provides a valuable means for verifying genome annotation, which is a challenge that must be met for each new genome sequence. In the current study, we generated tiling array data for a 1 Mb region syntenic between barrel medic and soybean in six different organ types (Figure 1). Analysis of the tiling array data detected 193 out of 229 (84%) and 176 out of 217 (81%) predicted genes in barrel medic and soybean, respectively (Figure 2), similar to results reported from tiling array analysis of the rice genome [8, 9]. Because genome annotation is a highly reiterative process that improves with the parallel refinement of gene-finding programs and the availability of experimental evidence, we anticipate further application of tiling array analysis to facilitate annotation of the fast emerging legume genome sequences [25, 30, 39].
Another use of the tiling array data is to identify transcription units in addition to the predicted genes [1, 2]. Previous tiling analyses indeed documented large numbers of putative novel transcripts in virtually all the genomes examined [3–10]. For example, detailed characterization of the non-exonic TARs identified in the japonica rice genome showed that they could be assigned to various putatively functional or structural elements of the genome, ranging from splice variants, uncharacterized portions of incompletely annotated genes, antisense transcripts, duplicated gene fragments, to potential non-coding RNAs [10]. In carrying out tiling array analysis of the legume sequences, we identified 499 and 660 unique TARs in barrel medic and soybean, respectively (Figure 3). Aligning against the predicted genes, 311 (62%) barrel medic and 355 (54%) soybean TARs were found to locate outside of or antisense to the predicted exons (Figure 4). Interestingly, in a promoter trapping study in lotus in which a promoter-less GUS reporter system was used, GUS activation, often tissue-specific, was found beyond the predicted genic regions [40]. Together, these observations indicate that novel transcripts missed by gene annotation account for a significant portion of the transcriptome in legume plants.
As a novel use of tiling array data for transcriptomic profiling, we used a median polishing-based procedure [10, 36] to determine the relative transcription levels and differential expression of the predicted genes. Because there are multiple probes involved in tiling a given gene, the median polishing-based procedure will have the corollary benefit of improved statistical confidence. Based on this method, approximately 30% of genes were found to be differentially expressed among the six examined organ types (Figure 5). The nodule-enhanced expression pattern of six selected soybean genes was subsequently verified by RT-PCR analysis (Figure 6). Collectively, these results indicate that genomic tiling array analysis can be extended to quantitatively examine the transcription levels of individual genes. This may prove particularly useful for quantifying transcription levels of members of paralogous gene families, which are notoriously hard to discriminate in conventional expression arrays that employ relatively fewer probes per gene.
Interestingly, 11 and 10 genes were identified as preferentially expressed in the nodule in barrel medic and soybean, respectively. These genes exhibited little overlap between the two species (Additional data files 2 and 3). Barrel medic and soybean diverged from a common ancestor approximately 50 million years ago, and represent two distinct groups of nodulating plants [41]. Barrel medic forms indeterminate nodules, which maintain an active meristem inside nodule primordia during the early stages of nodule development; while soybean forms determinate nodules that, after initial cell divisions, grow by cell expansions. These morphological differences may thus affect the architecture and gene expression in the nodules [42].
The availability of multiple genomic and transcriptomic datasets fosters comparative analyses that improve structural annotation and generate new insight into the function and evolution of coding and non-coding regions of the genomes [43]. A major principle of comparative genomics is that the functional DNA sequences in related species conserved from the last common ancestor are preserved in contemporary genome sequences, which encode the proteins and RNAs and the regulatory sequences controling genes with similar expression patterns [43]. Alignment of primary DNA sequences is the core process in most comparative analyses. The resulting information on sequence similarity among genomes is a major resource for infering gene functions, identifying other candidate funcational elements, and finding conserved genes missed from annotation in one genome or another.
Transcriptomic data from multiple species are also being extensively used in comparative analysis. For example, direct comparison of multiple transcript datasets using genome annotation tools has been shown as an effective way to uncover 'unannotated' genes. In rice, 255 new candidate genes were identified by cross-species spliced alignment of expressed sequence tags and cDNA to the genome sequence [44]. In this regard, the rich transcriptional activity documented from genomic tiling analysis constitutes an excellent complement to other tag-based transcriptome data. In the present tiling analysis of syntenic regions between two legume species, we identified over 300 unique TARs in both barrel medic and soybean in addition to the predicted exons. Transcripts tagged by these TARs should be useful for further comparison aimed at improving genome annotation and elucidating the transcriptome.
Furthermore, comparison of transcription levels in six different organ types revealed that a large portion of the collinear genes between barrel medic and soybean exhibit different expression patterns (Figure 7). It should be noted that there is a segmental duplication of synteny block 2 (block 2i and 2ii) in barrel medic [32]. The process of subfunctionalization following gene duplication, where degenerative mutations in both genes result in the partitioning of ancestral functions or expression patterns in the duplicated genes, could, therefore, contribute to the observed expression divergence among the examined collinear genes between barrel medic and soybean. Further analysis of the cis-regulatory regions of the syntenic genes should help to identify the key regulatory sequence divergence that accounts for the differences in related legume species and add to our general knowledge of plant genome evolution and regulation.
We report here a transcriptional analysis using high-resolution tiling microarrays of syntenic regions totaling 1 Mb between the legume plants barrel medic and soybean in six different organ types. This analysis generated transcriptomic data that is useful for three purposes. First, we detected transcription of over 80% of the predicted genes in the interrogated genome regions in both legume species. As genome annotation is a reiterative process that is heavily dependent on experimental data, genomic tiling analysis is thus one valid option to meet the challenge of analyzing large-scale transcriptomic datasets for newly sequenced legume genomes. Second, we identified 499 and 660 TARs from barrel medic and soybean, respectively, over half of which are outside of the predicted exons. Further functional characterization of these candidate transcripts should be useful to better our understanding of the complexity and dynamics of the transcriptome of legume plants. Third, we used the tiling array data to detect differential gene expression and to compare transcription patterns of collinear genes. This novel approach was validated by the high confirmation rate by RT-PCR analysis of genes that are preferentially expressed in the nodule. Further investigation revealed that some collinear genes exhibited drastically different transcription patterns between the two species. Collectively, these results demonstrate that genomic tiling analysis is an effective approach to simultaneously complement computational annotation of newly available genome sequences and to facilitate comparative genomics aimed at elucidating genome organization and transcriptional regulation in closely related species.
Materials and methods
Plant materials and treatments
Barrel medic (Medicago truncatula cv. Jemalong A17) seed was treated with concentrated H2SO4 for 10 minutess, rinsed with water and then allowed to germinate on moist filter paper at room temperature for a week. Seedlings 1-2 cm in length were planted in soil and maintained in the greenhouse with nitrogen-free plant nutrient solution as previously described [45]. Soybean (Glycine max cv. William 82) seed was directly sown in soil and maintained in the greenhouse with nitrogen-free plant nutrient solution as described by Subramanian et al. [46].
The Rhizobium bacterium Sinorhizobium meliloti 1021 and Bradyrhizobium japonicum USDA110 was used to inoculate barrel medic and soybean plants, respectively. The bacteria were grown in a yeast extract-mannitol medium for three days at 28°C as previously described [47]. The bacterial cells were then suspended in nitrogen-free nutrient solution to an OD600 of 0.08 and used to water four-week-old plants. This flood-inoculation step was repeated after two weeks. The nodules were collected three weeks after the second treatment. Each nodule was separated from the roots with sharp tweezers and placed on dry ice immediately. The stem, root, and leaf organs were harvested from four-week-old plants that were maintained with nitrogen containing plant nutrient solution. The same plants were maintained until maturity for collection of the flower and seed organs.
Sequence selection and gene prediction
Soybean sequences from four bacterial artificial chromosomes (BACs) were obtained from GenBank (accession numbers: AX196294.1, AX196295.1, AX196297.1, and AX197417.1). The BACs AX196294 and AX196295, and AX196297 and AX197417 form two contigs. There is a physical gap (represented by 100 Ns) between AX196294 and AX196295, and an approximately 50 Kb overlap between AX196297 and AX197417. Thus, the two contigs represent a total of 977 Kb of non-redundant sequences. Putative homologs to these soybean sequences in barrel medic were identified from sequenced barrel medic BACs as previously reported [32]. A total of 12 BACs (accession numbers: AC141115.22, AC149303.10, CR378662.1, CR378661.1, AC142498.20, AC146585.18, AY224188.1, AC146706.8, AY224189.1, AC146705.11, AC144644.3, and AC146683.9) were identified. The sequences were aligned and merged in regions of sequence overlap on the basis of ≥99% identity [32]. The merged barrel medic sequences form 7 contigs and total 1,060 Kb. Genes were predicted from soybean and barrel medic sequence contigs using FGENESH [33]. Both the dicot plants (Arabidopsis) and the Medicago (legume plant) matrix was used and their output compared.
Tiling microarray design, production, and hybridization
Both the barrel medic and soybean tiling arrays were produced on the Maskless Array Synthesizer platform as previously described [6, 8, 34]. Briefly, tiling paths consisting of 36-mer oligonucleotides offset by five nucleotides were designed to represent both DNA strands of the selected barrel medic and soybean genome sequence. Probes were synthesized at a feature-density of 390,000 probes per array in a 'chessboard' design [34]. Microarray production and storage were carried out previously described [6, 8, 34].
Total RNA and mRNA were sequentially isolated using the RNeasy Plant Mini kit (Qiagen, Valencia, CA, USA) and the Oligotex mRNA kit (Qiagen), respectively, according to the manufacturers' recommendations. mRNA from different organ types was reverse transcribed using a mixture of oligo(dT)18 and random nonamer primers [6, 34], during which amino-allyl-modified dUTP (aa-dUTP) was incorporated. The aa-dUTP decorated cDNA was fluorescent labeled by conjugating the monofunctional Cy3 dye (GE Healthcare, Piscataway, NJ, USA) to the amino-allyl functional groups in the cDNA. We used 2 μg dye-labeled targets for hybridization as previously described [6, 8, 34]. Tiling microarray design and experimental data are available in the NCBI Gene Expression Omnibus under the SuperSeries GSE10151, which is composed of subset series GSE10055 and GSE10056 for the barrel medic and soybean arrays, respectively.
Tiling array analysis of gene expression
Finally, genes with a p-value smaller than 0.05 were considered as detected.
Determination of differential gene expression was carried out based on the Tukey's median polish procedure, which fits an additive linear model [10, 36]. The expression level of a given gene in the six organ types, M, is estimated by the equation:
O ij = M + a i + v j + e j
where O ij is the observed intensity of the ith probe in the jth organ type, a i is the probe affinity effect of the ith probe, v j is the organ type variation of the jth organ type, and e j is the experimental error. A series of iterations of residue subtraction was performed using Tukey's median polishing until the matrix reaches a stable status, and the organ type deviation v was determined for each organ type. To obtain the false negative error rate, a permutation test was applied. The intensities of every probe were randomly shuffled among the organ types and median polishing repeated 1,000 times. A significant level at p < 0.001 was selected for organ-specific expression calling.
Identification of TARs
To identify TARs in each organ type, the Log2-transformed, quantile-normalized data that were anchored to the chromosome coordinates were scanned from the 5' end of each DNA strand one probe at a time. TARs were identified as regions with length >70 nucleotides (spanning roughly 14 probes) with no 2 consecutive probes having an intensity below the cutoff of 11.0.
Identification of collinear genes
To identify collinear genes, repetitive sequences were masked and tandemly duplicated genes were counted as one. The BLAT [48] program was then used to search the protein sequences of the predicted genes between barrel medic and soybean. The gene pairs with more than 10 amino acids mapped and more than 40% identity over the entire mapped sequences were identified as potential collinear genes, which were further manually inspected for gene order and orientation.
Additional data files
The following additional data are available. Additional data file 1 is a table showing a summary of predicted genes in the 1 Mb syntenic regions between Medicago truncatula and Glycine max. Additional data file 2 is a table listing the Medicago truncatula genes preferentially expressed in the nodule. Additional data file 3 is a table listing Glycine max genes preferentially expressed in the nodule.
bacterial artificial chromosome
transcriptionally active region.
We thank Xuanli Yao at Yale University for assistance in plant sample collection, and Terry Graham at Ohio State University for the Sinorhizobium strain. This work was supported by a grant from the NSF Plant Genome Program (DBI-0421675) to X-WD.
Authors' Affiliations
- Mockler TC, Chan S, Sundaresan A, Chen H, Jacobsen SE, Ecker JR: Applications of DNA tiling arrays for whole-genome analysis. Genomics. 2005, 85: 1-15. 10.1016/j.ygeno.2004.10.005.PubMedView ArticleGoogle Scholar
- Johnson JM, Edwards S, Shoemaker D, Schadt EE: Dark matter in the genome: evidence of widespread transcription detected by microarray tiling experiments. Trends Genet. 2005, 21: 93-102. 10.1016/j.tig.2004.12.009.PubMedView ArticleGoogle Scholar
- Selinger DW, Cheung KJ, Mei R, Johansson EM, Richmond CS, Blattner FR, Lockhart DJ, Church GM: RNA expression analysis using a 30 base pair resolution Escherichia coli genome array. Nat Biotechnol. 2000, 18: 1262-1268. 10.1038/82367.PubMedView ArticleGoogle Scholar
- Kapranov P, Cawley SE, Drenkow J, Bekiranov S, Strausberg RL, Fodor SP, Gingeras TR: Large-scale transcriptional activity in chromosomes 21 and 22. Science. 2002, 296: 916-919. 10.1126/science.1068597.PubMedView ArticleGoogle Scholar
- Yamada K, Lim J, Dale JM, Chen H, Shinn P, Palm CJ, Southwick AM, Wu HC, Kim C, Nguyen M, Pham P, Cheuk R, Karlin-Newmann G, Liu SX, Lam B, Sakano H, Wu T, Yu G, Miranda M, Quach HL, Tripp M, Chang CH, Lee JM, Toriumi M, Chan MM, Tang CC, Onodera CS, Deng JM, Akiyama K, Ansari Y, et al: Empirical analysis of transcriptional activity in the Arabidopsis genome. Science. 2003, 302: 842-846. 10.1126/science.1088305.PubMedView ArticleGoogle Scholar
- Bertone P, Stolc V, Royce TE, Rozowsky JS, Urban AE, Zhu X, Rinn JL, Tongprasit W, Samanta M, Weissman S, Gerstein M, Snyder M: Global identification of human transcribed sequences with genome tiling arrays. Science. 2004, 306: 2242-2246. 10.1126/science.1103388.PubMedView ArticleGoogle Scholar
- Cheng J, Kapranov P, Drenkow J, Dike S, Brubaker S, Patel S, Long J, Stern D, Tammana H, Helt G, Sementchenko V, Piccolboni A, Bekiranov S, Bailey DK, Ganesh M, Ghosh S, Bell I, Gerhard DS, Gingeras TR: Transcriptional maps of 10 human chromosomes at 5-nucleotide resolution. Science. 2005, 308: 1149-1154. 10.1126/science.1108625.PubMedView ArticleGoogle Scholar
- Li L, Wang X, Stolc V, Li X, Zhang D, Su N, Tongprasit W, Li S, Cheng Z, Wang J, Deng XW: Genome-wide transcription analyses in rice using tiling microarrays. Nat Genet. 2006, 38: 124-129. 10.1038/ng1704.PubMedView ArticleGoogle Scholar
- Li L, Wang X, Xia M, Stolc V, Su N, Peng Z, Tongprasit W, Li S, Wang J, Wang X, Deng XW: Tiling microarray analysis of rice chromosome 10 to identify the transcriptome and relate its expression to chromosomal architecture. Genome Biol. 2005, 6: R52-10.1186/gb-2005-6-6-r52.PubMedPubMed CentralView ArticleGoogle Scholar
- Li L, Wang X, Sasidharan R, Stolc V, Deng W, He H, Korbel J, Chen X, Tongprasit W, Ronald P, Chen R, Gerstein M, Deng XW: Global identification and characterization of transcriptionally active regions in the rice genome. PLoS ONE. 2007, 2: e294-10.1371/journal.pone.0000294.PubMedPubMed CentralView ArticleGoogle Scholar
- Yu J, Wang J, Lin W, Li S, Li H, Zhou J, Ni P, Dong W, Hu S, Zeng C, Zhang J, Zhang Y, Li R, Xu Z, Li S, Li X, Zheng H, Cong L, Lin L, Yin J, Geng J, Li G, Shi J, Liu J, Lv H, Li J, Wang J, Deng Y, Ran L, Shi X, et al: The genomes of Oryza sativa: a history of duplications. PLoS Biol. 2005, 3: e38-10.1371/journal.pbio.0030038.PubMedPubMed CentralView ArticleGoogle Scholar
- Enard W, Khaitovich P, Klose J, Zöllner S, Heissig F, Giavalisco P, Nieselt-Struwe K, Muchmore E, Varki A, Ravid R, Doxiadis GM, Bontrop RE, Pääbo S: Intra- and interspecific variation in primate gene expression patterns. Science. 2002, 296: 340-343. 10.1126/science.1068996.PubMedView ArticleGoogle Scholar
- Ranz JM, Castillo-Davis CI, Meiklejohn CD, Hartl DL: Sex-dependent gene expression and evolution of the Drosophila transcriptome. Science. 2003, 300: 1742-1745. 10.1126/science.1085881.PubMedView ArticleGoogle Scholar
- Nuzhdin SV, Wayne ML, Harmon KL, McIntyre LM: Common pattern of evolution of gene expression level and protein sequence in Drosophila. Mol Biol Evol. 2004, 21: 1308-1317. 10.1093/molbev/msh128.PubMedView ArticleGoogle Scholar
- Saetre P, Lindberg J, Leonard JA, Olsson K, Pettersson U, Ellegren H, Bergstrom TF, Vila C, Jazin E: From wild wolf to domestic dog: gene expression changes in the brain. Brain Res Mol Brain Res. 2004, 126: 198-206. 10.1016/j.molbrainres.2004.05.003.PubMedView ArticleGoogle Scholar
- Gilad Y, Oshlack A, Smyth GK, Speed TP, White KP: Expression profiling in primates reveals a rapid evolution of human transcription factors. Nature. 2006, 440: 242-245. 10.1038/nature04559.PubMedView ArticleGoogle Scholar
- Khaitovich P, Hellmann I, Enard W, Nowick K, Leinweber M, Franz H, Weiss G, Lachmann M, Pääbo S: Parallel patterns of evolution in the genomes and transcriptomes of humans and chimpanzees. Science. 2005, 309: 1850-1854. 10.1126/science.1108296.PubMedView ArticleGoogle Scholar
- Kirst M, Caldo R, Casati P, Tanimoto G, Walbot V, Wise RP, Buckler ES: Genetic diversity contribution to errors in short oligonucleotide microarray analysis. Plant Biotechnol J. 2006, 4: 489-498.PubMedGoogle Scholar
- Gilad Y, Rifkin SA, Bertone P, Gerstein M, White KP: Multi-species microarrays reveal the effect of sequence divergence on gene expression profiles. Genome Res. 2005, 15: 674-680. 10.1101/gr.3335705.PubMedPubMed CentralView ArticleGoogle Scholar
- Oshlack A, Chabot AE, Smyth GK, Gilad Y: Using DNA microarrays to study gene expression in closely related species. Bioinformatics. 2007, 23: 1235-1242. 10.1093/bioinformatics/btm111.PubMedView ArticleGoogle Scholar
- Doyle JJ, Luckow MA: The rest of the iceberg. Legume diversity and evolution in a phylogenetic context. Plant Physiol. 2003, 131: 900-910. 10.1104/pp.102.018150.PubMedPubMed CentralView ArticleGoogle Scholar
- Cheung F, Haas BJ, Goldberg SM, May GD, Xiao Y, Town CD: Sequencing Medicago truncatula expressed sequenced tags using 454 Life Sciences technology. BMC Genomics. 2006, 7: 272-10.1186/1471-2164-7-272.PubMedPubMed CentralView ArticleGoogle Scholar
- The Gene Index Project. [http://compbio.dfci.harvard.edu/tgi/plant.html]
- Young ND, Cannon SB, Sato S, Kim D, Cook DR, Town CD, Roe BA, Tabata S: Sequencing the genespaces of Medicago truncatula and Lotus japonicus. Plant Physiol. 2005, 137: 1174-1181. 10.1104/pp.104.057034.PubMedPubMed CentralView ArticleGoogle Scholar
- Cannon SB, Sterck L, Rombauts S, Sato S, Cheung F, Gouzy J, Wang X, Mudge J, Vasdewani J, Schiex T, Spannagl M, Monaghan E, Nicholson C, Humphray SJ, Schoof H, Mayer KF, Rogers J, Quétier F, Oldroyd GE, Debellé F, Cook DR, Retzel EF, Roe BA, Town CD, Tabata S, Van de Peer Y, Young ND: Legume genome evolution viewed through the Medicago truncatula and Lotus japonicus genomes. Proc Natl Acad Sci USA. 2006, 103: 14959-14964. 10.1073/pnas.0603228103.PubMedPubMed CentralView ArticleGoogle Scholar
- Medicago truncatula Sequencing Resources. [http://www.medicago.org/genome/downloads/Mt2/]
- The Phytozome Project. [http://www.phytozome.net/soybean]
- Menancio-Hautea D, Fatokun CA, Kumar L, Danesh D, Young ND: Comparative genome analysis of mungbean (Vigna radiata L. Wilczek) and cowpea (V. unguiculata L. Walpers) using RFLP mapping data. Theor Appl Genet. 1993, 86: 797-810. 10.1007/BF00212605.PubMedView ArticleGoogle Scholar
- Boutin SR, Young ND, Olson TC, Yu Z-H, Vallejos CE, Shoemaker RC: Genome conservation among three legume genera detected with DNA markers. Genome. 1995, 38: 928-937.PubMedView ArticleGoogle Scholar
- Choi HK, Mun JH, Kim DJ, Zhu H, Baek JM, Mudge J, Roe B, Ellis N, Doyle J, Kiss GB, Young ND, Cook DR: Estimating genome conservation between crop and model legume species. Proc Natl Acad Sci USA. 2004, 101: 15289-15294. 10.1073/pnas.0402251101.PubMedPubMed CentralView ArticleGoogle Scholar
- Concibido VC, Diers BW, Arelli PR: A decade of QTL mapping for cyst nematode resistance in soybean. Crop Science. 2004, 44: 1121-1131.View ArticleGoogle Scholar
- Mudge J, Cannon SB, Kalo P, Oldroyd GE, Roe BA, Town CD, Young ND: Highly syntenic regions in the genomes of soybean, Medicago truncatula, and Arabidopsis thaliana. BMC Plant Biol. 2005, 5: 15-10.1186/1471-2229-5-15.PubMedPubMed CentralView ArticleGoogle Scholar
- FGENESH Gene Prediction Program. [http://www.softberry.com]
- Stolc V, Li L, Wang X, Li X, Su N, Tongprasit W, Han B, Xue Y, Li J, Snyder M, Gerstein M, Wang J, Deng XW: A pilot study of transcription unit analysis in rice using oligonucleotide tiling-path microarray. Plant Mol Biol. 2005, 59: 137-149. 10.1007/s11103-005-6164-5.PubMedView ArticleGoogle Scholar
- Royce TE, Rozowsky JS, Bertone P, Samanta M, Stolc V, Weissman S, Snyder M, Gerstein M: Issues in the analysis of oligonucleotide tiling microarrays for transcript mapping. Trends Genet. 2005, 21: 466-475. 10.1016/j.tig.2005.06.007.PubMedPubMed CentralView ArticleGoogle Scholar
- Wang X, He H, Li L, Chen R, Deng XW, Li S: NMPP: a user-customized NimbleGen microarray data processing pipeline. Bioinformatics. 2006, 22: 2955-2957. 10.1093/bioinformatics/btl525.PubMedView ArticleGoogle Scholar
- Flemetakis E, Agalou A, Kavroulakis N, Dimou M, Martsikovskaya A, Slater A, Spaink HP, Roussis A, Katinakis P: Lotus japonicus gene Ljsbp is highly conserved among plants and animals and encodes a homologue to the mammalian selenium-binding proteins. Mol Plant Microbe Interact. 2002, 15: 313-322. 10.1094/MPMI.2002.15.4.313.PubMedView ArticleGoogle Scholar
- Karaman MW, Houck ML, Chemnick LG, Nagpal S, Chawannakul D, Sudano D, Pike BL, Ho VV, Ryder OA, Hacia JG: Comparative analysis of gene-expression patterns in human and African great ape cultured fibroblasts. Genome Res. 2003, 13: 1619-1630. 10.1101/gr.1289803.PubMedPubMed CentralView ArticleGoogle Scholar
- Chen X, Laudeman TW, Rushton PJ, Spraggins TA, Timko MP: CGKB: an annotation knowledge base for cowpea (Vigna unguiculata L.) methylation filtered genomic genespace sequences. BMC Bioinformatics. 2007, 8: 129-10.1186/1471-2105-8-129.PubMedPubMed CentralView ArticleGoogle Scholar
- Buzas DM, Lohar D, Sato S, Nakamura Y, Tabata S, Vickers CE, Stiller J, Gresshoff PM: Promoter trapping in Lotus japonicus reveals novel root and nodule GUS expression domains. Plant Cell Physiol. 2005, 46: 1202-1212. 10.1093/pcp/pci129.PubMedView ArticleGoogle Scholar
- Shoemaker RC, Schlueter J, Doyle JJ: Paleopolyploidy and gene duplication in soybean and other legumes. Curr Opin Plant Biol. 2006, 9: 104-109. 10.1016/j.pbi.2006.01.007.PubMedView ArticleGoogle Scholar
- Subramanian S, Stacey G, Yu O: Distinct, crucial roles of flavonoids during legume nodulation. Trends Plant Sci. 2007, 12: 282-285. 10.1016/j.tplants.2007.06.006.PubMedView ArticleGoogle Scholar
- Hardison RC: Comparative genomics. PLoS Biol. 2003, 1: e58-10.1371/journal.pbio.0000058.PubMedPubMed CentralView ArticleGoogle Scholar
- Zhu W, Buell CR: Improvement of whole-genome annotation of cereals through comparative analyses. Genome Res. 2007, 17: 299-310. 10.1101/gr.5881807.PubMedPubMed CentralView ArticleGoogle Scholar
- Lullien V, Barker DG, de Lajudie P, Huguet T: Plant gene expression in effective and ineffective root nodules of alfalfa (Medicago sativa). Plant Mol Biol. 1987, 9: 469-478. 10.1007/BF00015878.PubMedView ArticleGoogle Scholar
- Subramanian S, Stacey G, Yu O: Endogenous isoflavones are essential for the establishment of symbiosis between soybean and Bradyrhizobium japonicum. Plant J. 2006, 48: 261-273. 10.1111/j.1365-313X.2006.02874.x.PubMedView ArticleGoogle Scholar
- Vincent JM: A manual for the practical study of root nodule bacteria. International Biological Program Handbook. 1970, Oxford: Blackwell Science, 1-13.Google Scholar
- Kent WJ: BLAT—the BLAST-like alignment tool. Genome Res. 2002, 12: 656-664. 10.1101/gr.229202. Article published online before March 2002.PubMedPubMed CentralView ArticleGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


text:
Traditional lockbox processing is being replaced with comprehensive Integrated Receivables solutions. Cabela's recognized this trend and has completely transformed their remittance processing operations.
Panelists will include:
Ben Martens - Accounting Operations Manager
In this webinar, we will define Integrated Receivables and discuss how Cabela's made the decision to move to an integrated platform. Primary objectives are:


text:
City: A Story of Roman Planning and Construction
Houghton Mifflin Harcourt, 1974 - Juvenile Nonfiction - 112 pages
Superb design, magnificent illustrations, and clearly presented information distinguish all of David Macaulay's books. Whether chronicling the monumental achievements of past civilizations or satirizing modern architecture, he is concerned above all in how constructions are made and what their effects are on people and their lives.
What people are saying - Write a review
LibraryThing ReviewUser Review - nathanmannn - LibraryThing
In the book "City" by David Macaulay, the reader is absorbed into the world of the Romans in the imaginary city of Verbonia. The reader is allowed an intimate view of how they planned and built their ... Read full review
LibraryThing ReviewUser Review - Stbalbach - LibraryThing
City is David Macaulay's second book. Unlike most of Macaulay's other books in this series, rather than a single building, he draws an entire city. It is interesting and I learned a lot, the Romans ... Read full review
Other editions - View all
Common terms and phrases
apartment buildings arch was constructed arena atrium Augustus bakery brick brick-faced concrete bridge built caldarium called cardo carts castrum cavea cement central market Ceres chorobate city wall CLOACA cofferdam compluvium connected courtyard CRENELATIONS cross decumanus ditch Emperor exterior walls farmland finished floor gate grain GROIN VAULT groma ground heated hypocaust impluvium insulae laborers land layers of concrete limestone low sections Lucius main streets marble Marcus Licinius master plan maximum population military camp mortar passages peristyle piers pipes planners Planning and Construction POMERIUM portcullis pozzolana process was repeated PROFILE MAP protected public baths public fountains Quintus rectangular religious center rented right angles river road Roman city Rome roof rooms row of curbstones seating area senator sewers shaft shops sidewalk slaves snack bars soldiers space stone and mortar stood stucco surveyors temples tepidarium tiles toilets tunnel vault valarium Verbonia Verbonia's population vertical weighted strings wooden arch


text:
Is Mediamindz incorrect or missing key business information? Update your Hotfrog listing today.
Contact details
Are these details correct?
Hotfrog INDIA provides information regarding Mediamindz in Ahmedabad Gujarat. Mediamindz is located at 1 Devpreet Complex 2nd Floor, Opposite New Iit Campus, Gandhinagar Highway, Ahmedabad Gujarat 380005 and provides Advertising And Marketing services. Contact them on 099746 89299 or by visiting them on their website http://www.mediamindz.in.
Is Mediamindz in 1 Devpreet Complex 2nd Floor, Opposite New Iit Campus, Gandhinagar Highway, Ahmedabad Gujarat 380005 your business? Claim your listing and attract more leads by adding more content, photos and other business details.
We have more Advertising And Marketing services in Ahmedabad Gujarat available on Hotfrog INDIA. You can update your search for Ahmedabad Advertising And Marketing by location, keyword or service options.
SWASTIK CHAR RASTA NAVRANGPURA Ahmedabad Gujarat 380009
NAVRANGPURA Ahmedabad Gujarat 380009
C G ROAD NAVRANGPURA Ahmedabad Gujarat 380009
NR JODHPUR CROS ROAD SATELLITE Ahmedabad Gujarat 380015
Small Business Hub
Sharing tips, ideas and advice to help grow your small business.
Your profile without ads
Have you considered upgrading to Hotfrog AdVantage? No competitor advertising means exclusive focus on your business. You take centre stage.
Create your free Hotfrog business profile
Follow Hotfrog on:
Share on:
Hotfrog Small Business Hub


text:
My Favorite Flirts is a feature that lets you reach your favorite Flirts easily from My Account or on 1-800-TO-FLIRT.Manage My Favorites
(18 Reviews)
You meet me and are so enchanted with my beauty, intelligence and charm, within a month you have sold your house and moved into my ranch.
Taking you over is just forcing you into your destiny. You never were any good at running your life as a man. I will be much better at running your life as a whore.
Let's face it. Men aren't anal. Men are gay and like taking it in the ass.Submitting to me means facing all your little flaws, and all your big flaws.
Welcome to your inevitable break down, your next fag jag, your next sexual obsession. Even the financial drain turns you on. . . .
I know your secrets are precious to you. But that's why I want them.
Your anus needs what I can do. You can't deny it any more. You cum like a bottom bitch when you have a rubber cock up your ass.
I understand you like no other Mistress can.
None of your struggles can save you from the excitement that already corrupts you.


text:
DesperatelySeekingSleep (17-04-2015)
Mokeybear! That pic gives things a whole new meaning! Barry has his back?!
harvs (17-04-2015),~Marigold~ (17-04-2015)
Ha ha I'm just too excited that @~Marigold~ is back.
This is seriously my face right now
ImageUploadedByThe Bub Hub1429277281.989562.jpg
BH-KatiesMum (18-04-2015),Chunkydunks (17-04-2015),DesperatelySeekingSleep (18-04-2015),Little Ted (17-04-2015),Mokeybear (17-04-2015),~Marigold~ (17-04-2015)
@deku yes, you should start a reveal yourself thread! It's been ages since Siberian did that!
Last edited by ~Marigold~; 17-04-2015 at 22:38.
BH-KatiesMum (18-04-2015),DesperatelySeekingSleep (18-04-2015),harvs (17-04-2015)
DesperatelySeekingSleep (18-04-2015),harvs (17-04-2015),~Marigold~ (17-04-2015)
Pregnant for the first-time?
Not sure where to start? We can help!
Our Insider Programs for pregnancy first-timers will lead you step-by-step through the 14 Pregnancy Must Dos!


text:
This Australiian link is pretty amazing - A great link for the ACGA website.
Best wishes,
Adam Honigman,
Clinton Community Garden
The American Community Gardening Association listserve is only one of ACGA's services to community gardeners. To learn more about the ACGA and to find out how to join, please go to http://www.communitygarden.org
To post an e-mail to the list: firstname.lastname@example.com
To subscribe, unsubscribe or change your subscription: https://secure.mallorn.com/mailman/listinfo/community_garden


text:
Arizona Diamondbacks' Bullpen Bungling Robs D-Backs of Victory Over Colorado Rockies
Arizona Diamondbacks fans, meet your newest scapegoats: Bobby Korecky, Doug Slaten, Juan Gutierrez, and Jon Rauch.
What's their crime? The quartet of relief pitchers collectively screwed the D-Backs out of a two-game win streak (not to mention their first series clinch of the season) by blowing a three-run lead and essentially gift-wrapping a 9-6 comeback victory for the Colorado Rockies last night at Chase Field. Korecky & Company qualified for the Byung-Hyun Kim Boner Award by allowing six runs over three innings late in the game, pooching a potential victory for the hometown heroes.
Every time the Rox scored early on, the Snake Squad answered back. After Colorado went 2-0 in the second when Brad Hawpe and Clint Barnes were plated by successive RBI singles (courtesy of Franklin Morales and Yorvit Torrealba, respectively), Arizona's Chris Young and Eric Byrnes retorted with back-to-back home runs over the center field wall off lefty Morales minutes later. ("Who says we can't hit against a left-handed pitcher?" asked KTAR colorman Tom Candiotti after the pair of dingers, referencing his team's problems against southpaws as of late.) The call-and-response run-making continued in the third when Todd Helton scored thanks to a swat single by Ian Stewart, only to have the D-Backs battle back by driving home Felipe Lopez and Augie Ojeda with RBIs. (Mark Reynolds also homered with Conor Jackson on second base in the fifth).
Then it all went to hell. The bullpen meltdown went down after D-Backs head coach Bob Melvin nixed starter Yusmeiro Petit (who racked up 90 pitches while struggling through five innings) in favor of Korecky in the sixth. The former Minnesota Twin proceeded to load the basses via an Omar Quintanilla double and walks of Torrealba and Dexter Fowler. But instead of fighting his way out of the inning, Ryan Spilborghs then grounded off Korecky's next pitch, scoring Torrealba. (It could've been much worse had Ojeda not whipped out a 6-4-3 double play). When Doug Slaten took the mound, the floodgates opened. A seemingly non-stop parade of singles up the middle ensued, causing Quintanilla, Hawpe, and Garrett Atkins to notch runs. Even Gutierrez couldn't stop the bleeding. And to think, a single out could have stopped it all from happening. But just in case the score wasn't lopsided enough, Rauch (who KTAR sports wag Dave Byrnes has dubbed "piñata," due to his loathsome habit of "always getting hit on") allowed Helton to homer in the eighth with Hawpe on second, making things 9-6.
Upcoming Events
Arizona Diamondbacks vs. Colorado Rockies
TicketsSat., Feb. 25, 1:10pm
Cincinnati Reds vs. San Francisco Giants
TicketsSun., Feb. 26, 1:05pm
Colorado Rockies vs. Arizona Diamondbacks
TicketsSun., Feb. 26, 1:10pm
Arizona Rattlers vs. Iowa Barnstormers
TicketsSun., Feb. 26, 3:00pm
When Rauch finally left, a porcine Colorado Rockies fan in the bleachers yelled out the following:
"What's the quickest way to get to Mobile?"
Although technically Rauch would likely be sent down to the Triple-A Reno Aces, the heckler's point is nonetheless astute.
It makes you wonder how embattled pitcher Dan "Hard Luck" Haren would've fared. We'll find out this afternoon during the rubber match (which starts at 12:40 p.m.) as the 0-3 righty (1.89 ERA) attempts to not only grab his first win of the season, but just maybe give the 'Backs that elusive series win they sorely need. He'll face off against Jorge De La Rosa (0-1, 4.50 ERA). TV: FSN AZ. Radio: KTAR-AM.
For tickets or more info, visit www.dbacks.com.
Get the ICYMI: Today's Top Stories Newsletter Our daily newsletter delivers quick clicks to keep you in the know
Catch up on the day's news and stay informed with our daily digest of the most popular news, music, food and arts stories in Phoenix, delivered to your inbox Monday through Friday.
In Case You Missed It
Upcoming Events
Scottsdale Arabian Horse Show
TicketsWed., Feb. 22, 8:00am
Arizona Diamondbacks v. GCU Antelopes
TicketsWed., Feb. 22, 1:10pm
Cleveland Indians vs. Cincinnati Reds
TicketsSat., Feb. 25, 1:05pm
Los Angeles Angels vs. Milwaukee Brewers
TicketsSat., Feb. 25, 1:10pm
Sponsor Content


text:
I receive this for being a carer of one of my other children. So the child who is in care does not have a disability but I still receive ccr as I am a carer to my other child who has a disability.
I have also received it while on Mat leave in the past too
You are eligible for rebate if you are a carer.
Maybe she's mixed it up with the child care benefit.
ChelleBH (03-07-2015)
I was made redundant last year in July so it's going to be interesting as I was looking for work then decided to start my own business so I think I still qualify. Normally it gets paid in a lump sum in August so I have my fingers crossed.
It would be very hard to police. Especially if you do a combination of part time study and casual work. Though my study is supposed to be for 20+ hours a week but its almost completely autonomous. How do you prove that? Lol
Just updating for those following. My CCR Came in today. It was automatic. I haven't even done tax returns yet. Very happy. I was looking for work and then set up a little business from home that's not yet earning an income. Very happy
BlondeinBrisvegas (04-08-2015),JustJaq (04-08-2015),WannaBMummy82 (04-08-2015)
That's good to know @lilypily
Pregnant for the first-time?
Not sure where to start? We can help!
Our Insider Programs for pregnancy first-timers will lead you step-by-step through the 14 Pregnancy Must Dos!


text:
Football's tide of history has turned and the former kings of the castle are left with the waves lapping over their feet. But that doesn't mean the Manchester United-inspired publishing flood has abated. The biggest literary sporting splash this year was another book of revelations from a son of Old Trafford.
Keane: The Autobiography (Michael Joseph, £17.99) concerns Roy of that ilk, possibly the hardest man in football, and certainly one of the most controversial. The latest controversy must surely be about why it was left off the shortlist for the William Hill Sports Book of the Year. Eamon Dunphy co-writes with a heroic dose of the hard-bitten realism that made his diary of a footballing journeyman, Only A Game?, such gripping stuff. These two bad boys are a match made in sports-page heaven, and one can only wonder what the William Hill judges were thinking of.
Another strong contender must have been the latest labour of tough love for Michael Crick, the investigative journalist and lifelong United fan. The Boss: The Many Sides of Alex Ferguson (Simon & Schuster, £17.99) takes the club's manager, another gritty controversialist, pins him to the dissecting table, and gives him a good going-over – even if most of his supposed sins were the kinds of things that most managers get up to, especially the most successful.
In the William Hill gap where Keane should have been was a relatively anodyne work by one of his former Ireland team-mates: someone who, having donated the million-pound proceeds of his testimonial to charity, could be said to come from the opposite end of celebrity's moral spectrum. Niall Quinn: Head First – The Autobiography (Headline, £17.99), co-written with Tom Humphries, is an affable account of a solid professional's career, laced with a pleasing lyricism and peppered with good lines. But his story of life with the national side and Keane's are like dispatches from parallel universes.
Quinn's was one of three autobiographies on the William Hill shortlist. The other two were self-written, one of them a book to give to anyone, not just sporty or nautical types. Taking on the World by the global circumnavigator Ellen MacArthur (Michael Joseph, £17.99) tells the story of an extraordinary woman, from the tear-jerking prologue recounting her triumphant entry into Les Sables d'Olonne last year at the end of the Vendée Globe Race which made her name, through the Swallows and Amazons tales of childhood adventures, to her awe-inspiring grown-up exploits on the high seas (sometimes very high indeed). Fired by her drive, warmth and passion, and beautifully illustrated by her line drawings, this book is a constant delight.
From the evidence of Opening Up: My Autobiography (Hodder & Stoughton, £17.99), Michael Atherton demonstrates that his post-cricket career as a journalist enjoys a sound footing. As considered and thoughtful as the man himself, it affords the reader a wealth of insights into the modern game.
The fourth William Hill book was A Season In Verona (Secker & Warburg, £16.99), by the Booker Prize nominee, Tim Parks. He chronicles over the course of a season his support for Hellas Verona, the Millwall of Italian football. Parks has all the insight of an accomplished novelist, and is good on the sport's hallowed, mystical place in the national culture. But he can never reconcile himself with the racism of the Verona hardcore.
However, the star and deserved winner of the William Hill show was Donald McRae's In Black & White: The Untold Story of Joe Louis and Jesse Owens (Scribner, £18.99). These two friends were sporting gods with the extreme misfortune to be black in 20th-century America, when the colour of your skin overshadowed trifling matters such as four Olympic gold medals or the heavyweight championship of the world. Owens managed to play the system and stayed afloat; Louis all but went under. McRae tells their stories brilliantly.
William Fotheringham must have been disappointed that Put Me Back on My Bike: In Search of Tom Simpson (Yellow Jersey, £15.99) did not make the shortlist. It is partly a straightforward but compelling biography of the charismatic rider who died during the 1967 Tour de France, partly an account of how the seasoned cycling journalist found it no easy matter to separate myth and reality. Each upholder of the Simpson flame had different memories, and Fotheringham does well to pick his way through them.
Most memories of Alex Higgins are the same, it seems – traumatic. His nickname was apt: at the snooker table, and especially away from it, he left a trail of devastation in his wake for 25 years. In The Hurricane: The Turbulent Life and Times of Alex Higgins (Atlantic, £16.99), Bill Borrows catalogues the multiple varieties of vileness perpetrated by a man who comes across as a supremely talented, but basically obnoxious creep. It's hugely entertaining, of course: a caustic antidote to Christmas, perhaps, and a dire warning about the evils of the demon drink.
- More about:
- Bestsellers (books)
- Biography
- Castles
- Literature
- Manchester United
- Nonfiction Literature
- Premier League
- Publishing
- Realism
- Sir Alex Ferguson


text:
Need a Titer Test in Tustin, CA? Simply order online 24/7 and get a fast, affordable Titer Test at a lab near you (Find More Locations):
What is a Titer Test?
A Titer Test is a blood test that checks your titer level for one or more diseases. People in Tustin, CA order a Titer Test for compliance with hospital, vendor credentialing, schools, and other organizations.
How much does a Titer Test cost?
Our Titer Test prices include the doctor's order that you need to get tested in Tustin, CA, all lab fees and taxes, and a pdf copy of your results. For current prices, please CLICK HERE.
What does a Titer Test report look like?
For a sample PDF report of one of our most popular Titer Test lab panels, please click below:
We are delighted to offer the Titer Test in Tustin, CA.
Titer Test in other cities near Tustin, CA:Aliso Viejo | Anaheim | Atwood | Costa Mesa | East Irvine | Fountain Valley | Fullerton | Garden Grove | Irvine | Newport Beach | Newport Coast | Orange | Placentia | Santa Ana | Villa Park


text:
Budget Beauty: Organic beauty buys under £25
September 21st 2016
November 6th 2013
Santa Montefiore decided to kick start her detox with all the facts thanks to The Organic Pharmacy - with impressive results
After a long summer eating my way through Italy's pasta and parmesan supplies, and knocking back glasses of white wine as if it were water, I felt heavy, sluggish and in dire need of getting back into shape. This September, I told myself, would be the month I set about losing the weight I had never quite lost since giving birth to my two children ten years ago. This time, I vowed, I'd really do it. (As opposed to the other countless times when I had nearly done it!)
I joined the Equinox gym in Kensington, which was a good start. But we all know it's about 80% diet and 20% exercise. However, cutting out the usual 'white things' didn't appeal. I wanted to know exactly which foods my body finds hard to break down and I wanted to be advised on supplements. I'm 43; I have to start taking care of myself.
I have always been a big fan of The Organic Pharmacy. I love their ethos and their products. Everything smells so good in there, I never come out empty handed. So, I booked an appointment for a detox with Vidhi, a qualified doctor and the most charming Indian lady who immediately made me feel that I was in the very best of hands.
Professional and friendly, she discussed my diet and lifestyle in great detail for about half an hour before strapping me gently on the wrists and ankles, connecting me to the Quantum QX machine, a genius device invented by a NASA scientist, which scans your body and all its organs, glands and lymphs in great detail. The results were fascinating. Not only did it reveal my vitamin, mineral, serotonin and hydration levels, but the foods my body is unable to process efficiently, which results in toxic waste piling onto my stomach and thighs as fat and cellulite.
Vidhi went through the results with me. I was very dehydrated, in spite of drinking masses of water. That was because of the low mineral levels, which meant I was not absorbing the water I drank. I was deficient in vitamin B, hence the weak, breaking nail and hair, and so toxic, she nearly jumped when she saw the result. No surprise then that wheat, dairy and white wine (my summer diet) came up as foods to avoid. My summer was literally sitting on my thighs and stomach!
I began a detox immediately, conscientiously, with no cheating. Three detox pills before bed and on waking, with plenty of water. Drops to support the kidneys as I expelled all the toxicity, and various other bottles of supplements. I avoided a small list of foods and all alcohol, and added kale and spinach smoothies with walnuts, almonds, Dr Udo's Oil and protein powder to my porridge breakfast. I bought spelt pasta, vegan pesto, spelt and nut bread and discovered that it's not so hard to avoid wheat and dairy after all. Fortunately, although I can't process white wine, red is fine – I looked forward to that post detox.
The first week I got a sty, a cold sore, and the cold that was coming when I went for my appointment developed into a stinker. But the following week I felt fantastic. My skin glowed, the whites of my eyes were brighter, my energy levels were off the scale and I felt generally lighter. By week three I was down a jeans size and had to go to Trilogy to replenish my supply of denim (no hardship there!).
I have finished the detox pills but continue taking the supplements and avoiding the list of foods, although I am drinking red wine in moderation. It's now week seven and I've dropped from 71 kilos to 65, which is what I was before I got pregnant and my ideal weight (I'm 5ft 11"). I went for a follow-up scan to find that my hydration levels are now 100%. My vitamin and mineral levels are strong and the detox has brought my toxicity down. It was really interesting to compare the results: the detox really worked.
I am so excited by the way I feel and that I have lost the weight I've spent the last ten years trying to shift, that I am persuading all my friends to go. It's not just about the weight, though, it's about being healthy and feeling full of energy and vitality, and looking your best. It works and it's not hard to do. It costs £150 for the first consultation (supplements on top of that) and another £70 for the follow-up scan.
Hence I have decided to share this gem of an experience with GTG readers. Which, in my opinion, is the best way to review something – when you are so happy with a result you can't keep it to yourself!
Find out more at www.theorganicpharmacy.com and book in with Vidhi on 0207 376 9200
Login to add a comment
September 21st 2016
October 17th 2014
20 hours ago
February 17th 2017
February 16th 2017
February 16th 2017
February 7th 2017
January 23rd 2017
January 17th 2017
January 16th 2017
January 13th 2017
Sana Jardin - Vice President EMEA
B Public Relations - Social Media Manager
Elemis International Marketing Manager - Spa and Salons
SEEN Group - Senior Beauty Publicist/Account Director
Noble Isle - Marketing Manager


text:
I can't believe I've been here so damn long, it feels like it was only yesterday I made an account. I remember when animeplanet.com was a porn site, and I found it all the more humorous since I forgot the hyphon. That was hard to explain at work.
I adore this site and the service it provides.
9,048 total episodes
Hi Mainks! Doing the anime list is actually pretty easy... though I see you figured it out, awesome! ^_^ Very soon you'll have the ability to do a mini review for anything on your list, too.
Also, ahem... I am one of those wild ultima snobs you may have heard about... grew up playing them on the Atari 1040ST (shadowlords ftw!) and proudly proclaim that 7/7.5 are the best PC rpgs ever made -- it's also why I don't play any RPGs past them (and they are what, 12 years old now?) because all RPGs nowadays hold your hand and suck compared to Ultimas of olde.
But it is my duty to inform you that you suck, because Ultima online is a bastard child that Richard Garriot created, has gameplay like 8 (UGH) and basically gives the good Ultimas a bad name
:( boooooo!!!
Vodka ninja, eh? Sounds... unstable. Lol. But awesome to see you introduced yourself in the forum. If you're like a friend of mine,a nd your list is quite substantial, then the lists will be a few days' job. But it's well worth it to have it all organised in some way. But don't forget to do recommendations for the stuff you've watched. PS. I am overjoyed to see Akira as your number one. It's one of my top favourites too (although not in my Top 5) and has a special place in my heart.
Lol. You won't believe how many ninjas there are on this site. Me, I'm gonna stick to samurai. We might not have cool costumes, but at least we don't have to sneak around in the shadows, which I assume gets well boring after a while. :P Anyway, glad to see someone start with their bio for once. Wonder what the rest of your Top 5 will look like... Oh, and welcome! Don't forget to join us in the forums afterwards.


text:
Contests for people in Guyana - 794
Note: The contests shown here are not filtered in accordance with your preferences. To see only contests that match your preferences, view the "Your new contests" page.
Sorted by date added Sort by closing date Sort by rating
SubPac S2 Seatback Physical Sound System
Expires: 25 February 2017
Razer Blackwidow Chroma V2
Expires: 10 March 2017
Jabra Sport Coach Wireless Headset
Expires: 25 February 2017
You can win a Kylie Mini kit by participating in this contest.
Expires: 21 February 2017
1 x iPhone 7 (Matt Black); 4 x $50Amazon Gift Cards; 2 x 18k Real Gold Chains
Expires: 5 March 2017
Wall Hanging/Tapestry designed in India
Expires: 13 March 2017
OPI's Fiji Spring/Summer Collection
Expires: 5 March 2017
Set of Estee Lauder Bronze Goddess 2017 Products
Expires: 5 March 2017
• Acqua di Gioia • Air di Gioia • Sun di Gioia • Armani Eau de Cèdre • Signature Armani pouch
Expires: 27 February 2017
$50 Amazon or eBAy Gift Card
Expires: 26 March 2017


text:
The Berkshire Hathaway-owned Class I freight railroad will pay $50,000 in civil penalties related to a March 2015 crude oil train derailment in Galena, Ill., according to a statement from Illinois Attorney General Lisa Madigan.
Additional shippers and logistics companies have expressed support on a call for the Federal Maritime Commission to provide guidelines on detention and demurrage charges.
Hyundai Heavy Industries (HHI) is putting its restructuring plans on a shareholders' meeting agenda for Feb. 27, but HHI workers plan to strike from Feb. 22-27 in attempts to block the meeting.
Registration takes less than 1 minute.
The new president immediately put the world on notice that his administration will combat unfair trade practices, which many interpret as a return to protectionism.
The National Transportation Safety Board on Dec. 13 will open the docket for its investigation into the sinking of the TOTE Maritime cargo ship last October to the public, including a detailed transcript of audio from the vessel's voyage data recorder.
Elaine Chao has served as Secretary of Labor under President George W. Bush and Deputy Secretary of Transportation under President George H.W. Bush.


text:
Tom Jones became one of the most popular vocalists to emerge from the British Invasion. Since the mid-'60s, Jones has sung nearly every form of popular music pop, rock, show tunes, country, dance, and techno, he's sung it all. His actual style a full-throated, robust baritone that had little regard for nuance and subtlety never changed, he just sang over different backing tracks.
""Rather than make a traditional covers record, I thought it would be much more fun to create a new type of project in which artists communicated with each other and swapped a song for a song, i.e. you do one of mine and I'll do one of yours, hence the title - Scratch My Back - And I'll Scratch Yours." Peter Gabriel
Taken from the 2002 album, 'Up'. The title track is backed with two non-LP tracks, 'Growing Up' (Trent Reznor Remix & Tom Lord Alge Radio Edit)…
Learn seven Tom Petty tracks, guitar lessons by Danny Gill. This superb 2 DVD set will teach you to play seven tracks from this award winning American singer-songwriter spanning his 30 year career.


text:
Wholesale Online Specialist - FlowerWorld, Derby
|Manufacturing - Flowerworld
|Job type:
|Full time
|Closing date:
We are recruiting a Wholesale Online Specialist to join our FlowerWorld operation in Derby. This is a new and truly exciting role which is pivotal to the success of FlowerWorld's planned growth.
Reporting to the Flower Manager- who manages the journey of flowers from grower to customer- you'll work cross functionally with Manufacturing Operations including Technical, Wholesale and Online.
You'll be an exceptional salesperson with a track record of using your negotiation skills to deliver great results. You'll increase our business-to-business accounts and online business-to-customer sales, whilst all the time delivering outstanding account management. This is a unique opportunity to join the Morrisons' family, yet have responsibility for developing an emerging brand.
This vacancy is offered as a permanent opportunity and we're looking for someone who is ambitious, customer focused and passionate about flowers!
About FlowerWorld:
FlowerWorld is unique to Morrisons manufacturing as it is our only non-food producing site.
With over 300 florists, each and every bouquet is treated with care and consideration before it leaves site ensuring the best possible quality to our customers. Not only that but we also use specialised techniques to first check how our flowers are going to develop through their life and then to keep them at their very best.
Here we make over 400,000 bouquets per week using both hand tying methods and machinery; during peak periods such as Valentines and Mothers Day this increases significantly.
Key accountabilities:
Knowledge and experience:
What's stopping you?
We're a supermarket with a difference, in that we make food as well as sell it. Take fresh beef for example, we buy cattle direct from the farmer, take it through our own preparation sites and deliver the meat to the store ourselves. We are the UK's second largest food manufacturer and no other British supermarket has greater traceability and direct control over its supply chain than we do.
We have 16 manufacturing sites including abattoirs, meat processing plants, produce packaging sites, a seafood site and a bakery. What's more, we're investing massively in our manufacturing capability - over £200m over the last three years alone, plus our talent agenda as we develop our people, creating exciting career prospects at all levels.
Our aim to offer more of what matters extends to our employees too. Join us and you can expect to enjoy great career prospects and a very competitive pay and benefits package. In addition to an attractive salary, we offer a discount card for you plus two more for friend or family member giving you 10% off your shopping in Morrisons. We also offer company share options, a highly competitive pension scheme, Life Assurance and more besides.
We are an equal opportunities employer and welcome applications from all sections of the community.
|Back to Search Results
|New Search


text:
Las Cruces is located in Dona Ana county and has a population of 74,267.
Physician Assistant schools in Las Cruces charged students an average tuition of $2,412.00 if they were New Mexico residents, and $8,844.00 if they were non-residents, during the 2009 - 2010 school year.
Books and supplies for physician assistant schools in Las Cruces cost an average of $1,032.00. The average cost for room and board for students living on campus at physician assistant schools in Las Cruces was $7,526.00. The average cost for students living at home was $3,878.00 for the 2009 - 2010 school year. There is one physician assistant college in Las Cruces for students to choose from. In the 2008 - 2009 school year, reportedly 34 students completed physician assistant programs in Las Cruces.
|Average Salary
|Number Employed in City
|Medical Assistants


text:
- 0 reviews
- 0 gaji
Key Role & Responsibilites
- To do Mechanical, Electrical and Plumbing design for building project
- S1 in Teknik Elektro, IPK min. 3
- Have good understanding in building code and regulation
- Min 3 year work experience with MEP Consultant
- Experience in MEP design for high rise building projects
- Passionate in MEP building design work
- Good command in English
- Opportunites to work with Top Indonesia developer
- Various types of projects
- Good renumeration and bonus
- Insurance: BPJS Kesehatan & Ketenagakerjaan
If you feel that you can meet the qualification and up for the challenge, please send your complete resume and current color photograph by clicking button bellow
Lowongan pekerjaan ini kemungkinan telah berakhir.
Deskripsi Perusahaan
PT Prima Detailindo (Detail Studio) is a professional Architecture & Engineering consultant with core team in Architecture and inhouse Structure & MEP engineer. Top Indonesia Developer are among our list of loyal clients.We have been doing projects as Local Architect for International Architecture firm as well as collaborate with Indonesian architecture firm. We are a fast growing company and now are seeking energetic and creative talent to further expanding our company.