text:
A personal letter from Bob to you,
as a co-traveler down the road to
Financial Freedom God's Way
Greetings in Christ,
The Lord is calling Christian Financial Ministries (CFM) to prayerfully build and offer you several approaches to Financial Freedom God's Way:
- Financial Freedom God's Way Online Academy
- Crisis Financial Coaching
- E-Coaching
- Face Book
- Blog
Let me share with you just a bit more background
Since 1987 God has given Christian Financial Ministries and me the great pleasure and privilege to work with such godly men and organizations as Larry Burkett and Christian Financial Concepts, Howard Dayton and Crown Ministries/later Crown Financial Ministries, Dave Ramsey and his organization, as well as with a number of other godly folks and organizations. During all of this time the Lord was leading me to read, watch, listen, learn, apply, and pass along Bible-based spiritual and financial principles and applications.
Now the Lord is leading CFM to share every part of this ministry with you and others for FREE! Yes, you may receive any of our ministry services for absolutely no charge.!
Yes, we are a 501-c-3 non-profit ministry and we keep our overhead as low as possible. For example
- CFM is debt free–we budget on a cash basis
- In 2012 we sold our office building–no more building overhead
- Right now each of us at CFM work from home offices
- Our Ministry Leadership Team members serve on a volunteer basis. NOTE: the CFM Board of Directors has authorized me a salary and a housing allowance however my wife and I have chosen to defer those payments since 2013.
Ministry prayer and financial support
God is calling us to allow Him to support this ministry in one very traditional way and one very nontraditional and singularly unique way as well.
In the past we were very traditional.
- CFM charged variable fees (based on income) for all of our ministry programs, services, and materials, and
- Along the way we did have a few who regularly prayed for God's work through this ministry
Going forward you will find us to be BOTH singularly unique AND traditional.
- If you are an Academy Phase 1, 2, 3 or 4 Academy Team Member you are here "for such a time as this" because others have prayed for you to be here and have paid the full cost for you to be here. Therefore, for you to be part of the Academy we ask you to both PRAY IT FORWARD and DONATE IT FORWARD to help make it possible for others to follow you in the Academy.
- First, please "Pray It Forward" to help bring others
- Prayerfully commit to allow the Lord to work Bible-based spiritual and financial principles and applications into your personal life and finances.
- Prayerfully commit to pass along those same Bible-based spiritual and financial principles and applications to others as the Lord brings those "others" to you.
- Pray for your fellow Academy Team Members as part of your Academy Prayer Team (more information inside the Academy).
- Pray that the Lord will bring to your heart specific individuals who He is calling to follow you down the Road to Financial Freedom God's Way, then refer those folks to the Academy.
- Pray for Christian Financial Ministries as the Lord directs.
- Second, please "Donate It Forward" to help pay for others to come.
- How often? You will have access to Module 1 of the Phase I Seminar without being asked to "Donate It Forward". Then, if you believe the Lord would have you continue into Module 2 of the Seminar, we will ask you to please make a "Donate It Forward" contribution to the Ministry Scholarship Fund–Module 2 will only open up for you if you prayerfully choose to "Donate It Forward" at the end of Module 1. You will have the same choice to Donate It Forward and continue at the end of each Module throughout the Seminar and throughout the rest of the Academy Phases.
- How much? No donation is too small or too large. We just ask that you donate something as the Lord and you decide best.
- How? All you need to do is to click the green "Donate Now" buttons on the "Review and Look Forward" pages inside the Academy OR click the "Funded By Families Like Yours" at the top of this and other of our website pages. When you click on either location, you will go to a secure PayPal page where you may choose to donate using PayPal or your credit or debit card. You will be asked to signify at the bottom of your Academy Application that you prayerfully agree to commit to and support this Pray It Forward and Donate It Forward program.
- First, please "Pray It Forward" to help bring others
- If you are benefiting from E-Coaching, E-Books, Crisis Coaching, or "Ask Bob"
- First, please "Pray it Forward"
- Prayerfully commit to allow the Lord to work all He is giving you into your personal life and finances.
- Prayerfully commit to pass along to others what the Lord is giving you.
- Pray that the Lord will bring to your heart specific individuals who He knows will benefit from these ministry services–then refer those folks to CFM.
- Pray for Christian Financial Ministries.
- Second, please "Donate It Forward".
- How often? You will have a chance to donate to the Scholarship Fund any time you choose.
- How much? No donation is too small or too large. Please prayerfully donate what the Lord and you decide is best
- How? Please click on the "Funded By Families Like Yours" button at the top of every Ministry Web Page (upper right on this page). This will take you to a secure PayPal page where you may choose to donate using PayPal or your credit or debit card.
- First, please "Pray it Forward"
Please NOTE: The Lord may have equipped you to provide some of the skill sets we need to build and administer Christian Financial Ministries and the Financial Freedom God's Way Online Academy. Right now we need some help to edit the content of our Website and help administer our new Social Media program. If the Lord has given you either or both of these skill sets, and you would like to talk about about Donating these skill sets Forward, please let Bob know by email (firstname.lastname@example.com).
Please NOTE: According to a '"worldly perspective", the perceived value of what you receive can be greatly influenced by the financial cost or the fee you pay. However the true value of what God will give you and others through you is based on an eternal perspective, and you know from God's Word that things eternal can't be sold or bought. Therefore, I will never ask you to donate a specific dollar amount. However, there is a significant dollar value associated with our ministry services. For example, here are the fees charged for some similar Dave Ramsey programs and services:
- Financial Peace University: $100 per family
- Coach Training Program: $2,000
- In-house financial coaching: $100-$175 per hour
In Summary
You will only be able to receive the benefit of all that the Lord has to offer you through this Ministry and Academy because someone (possibly several "someones") Prayed It Forward and Donated It Forward. I ask that you do the same for those who will follow you. In addition (and this is critically important) you are welcome to be part of the Academy Seminar and Workshop (Academy Phases 1 and 2) if the Lord equips you and leads you to give a lot or a little. NOTE: Naturally all donations are fully tax-deductible.
Some folks say this approach to ministry won't pay for our continuing development and operating expenses much less make it possible for our Coaches to earn an income. I disagree! Why do I disagree? Because I didn't just dream all this up. I know this support approach is counter-intuitive to normal ministry operations and "best practices". However, I sense this is exactly the approach the Lord would have you and me follow. Therefore, I continue to trust and believe that what seems impossible with man is possible with God.
Rev. Bob Louder, MBA, PhD
Christian Financial Ministries


text:
New Delhi, Sep 18: Defence Minister Manohar Parrikar and Army chief General Dalbir Singh will on Sunday visit the army camp in Jammu and Kashmir where an audacious militant attack left 17 soldiers dead.
The early morning attack took place in Uri town.
Defence sources said that six soldiers were also injured in the attack who were airlifted to an army hospital in Srinagar.[Terror strikes at J&K: Rajnath cancels Russia visit]
"Terrorists attacked the rear base camp of an infantry battalion and not the headquarters of 12 Brigade in Uri today (Sunday) morning which is posted on the Line of Control (LoC)," the sources added.

subset: commoncrawl


text:
Mad Snail
[ Filters ]
|Cult of the Sacred Runes
Action Adventure Fantasy Harem Martial Arts Mature Xuanhuan
The formless ancient world was infested with despicable demons. They hunted and feasted on mankind, keeping the human civilization constantly in a weak and suppressed state.... more>>
Releases: 234
|Nine Heavenly Star Art
Action Adventure Comedy Drama Ecchi Fantasy Harem Martial Arts Xuanhuan
This is a world where humans, demons and mysterious beasts coexist. Humans cultivate mysterious qi. Demons cultivate corporeal bodies. Mysterious beasts cultivate soul perception. A... more>>
Releases: 6
|Rebirth of the Thief Who Roamed The World
Action Adventure Fantasy School Life Slice of Life
The world's largest VRMMO, Conviction, was almost like a second world for humanity. It had integrated itself into the real world's economy, with both corporations and individuals seeking their fortunes through the game. ... more>>
Releases: 287
|Star Martial God Technique
Action Fantasy Harem Martial Arts Xuanhuan
In the whole world there lays twelve paths to climb the Tower of God, and in legends these twelve pathways leads toward legendary road of immortality. However these paths in the Tower of God, are far too long, without end.... more>>
Releases: 40
|Tales of Demons and Gods
Action Adventure Comedy Fantasy Harem Martial Arts Shounen Xuanhuan
Killed by a Sage Emperor and reborn as his 13 year old self, Nie Li was given a second chance at life. A second chance to change everything and save his loved ones and his beloved city. He shall once again battle with the Sage Emperor to avenge his death and those of his beloved . With the vast knowledge of hundred years of life he accumulated in his previous life, wielding the strongest demon spirits, he shall reach the pinnacle of Martial Arts.... more>>
Releases: 463


text:
Lava Mobile Service Center in Aurangabad
Lava Mobile Service Center in Aurangabad, Maharashtra – Address, Contact Number are listed below. If you have any problem with your Lava mobile, kindly contact the service centers officials. There is 1 authorized mobile service centers in Aurangabad
List of Lava Mobile Service Centers in Aurangabad:
Lava mobile service center in Aurangabad
Name: M.M. Services ( Service Centre )
Address: Clony Chauraha , Sennari Raod , Ma Sharda Complex
City: Aurangabad
State: Maharashtra
Contact Person: Vishal
Contact Number 1: 90980304
Contact Number 2: 7560246644
Working Hours: 10:00 AM to 6:00 PM
LAVA Service Center timing : 10 AM to 6 PM on all working days. Collection Point only receives your LAVA handset/device and takes it to the nearest Authorized Service Partner for repair. All Servicing/Repair of your LAVA handset/device is done by LAVA Authorized Service Partner. All cost estimates (if any) will also be provided by Authorized Service Partner.


text:
Year Established:2007
Business Type:Manufacturer, Trading Company
Management Certifiation:
Operational Address:
Yongguang Industrial Zone, Luoge, Nanzhuang Town, Chancheng District, Foshan, Guangdong, China (Mainland)
Place of Origin: Guangdong, China (Mainland)
Brand Name: Falaiya
Model Number: DB1530BE
Material: Glass+stone+metal+shell
Material: glass,stone,stainless steel,shell and mesh.
Generally Size: 290x300mm,11 sheets/box
Color: Colorful (any color is available)
Finishing: transparent glass with color.
Usage: Indoor for the wall of Hall, Pub, Hotel, Night Club, KTV and Restaurant,balcony,or swimming pool.
Foshan Falaiya Decoration Material Factory is an effective enterprise specializing in glass Mosaic. We have TEN years' experience in decoration market. With four factories in Foshan, China, we can produce all kinds of mosaic, including glass mosaic, stone mosaic, ceramic mosaic and metal mosaic. You are welcome to know more about our culture and get in touch with us in advance. Falaiya has strong production strength. There are 1,023 staff members in our factories, and 20 different series of mosaic on show in our exhibition hall. Thanks to our strength and various products, we become a largest mosaic supplier in China. We focus on OEM service. Our main markets are the USA, Brazil, France and Italy.


text:
BBC loses Women's British Open to Sky SportsFebruary 12, 2015 Golf News
Ladies' British Major goes the same way as the Open
Just over a week since it was announced that the BBC is to lose the live rights to the Open Championship it has been revealed that the Women's British Open will follow the same path to Sky Sports.
The Ladies Golf Union and IMG who look after the championship have secured a five-year deal with the satellite broadcaster which will begin in 2017 meaning that all four days will be shown on Sky.
The statement read: "The Ladies Golf Union and IMG can confirm a new five-year broadcast agreement for the Ricoh Women's British Open with the UK's two largest sports broadcasters, Sky and the BBC, running from 2017-2021, in an arrangement similar to that announced last week by the R&A for the Open Championship."
Similar but also different in that the BBC has been left with just a one-hour highlights package for the WBO as opposed to two for the men's Open.
So, as from 2017 the only live golf that the BBC will televise will be the final two rounds of the Masters.
The BBC will be at the live controls for this year's Women's British Open at Turnberry before bringing the curtain down on a long-standing relationship at Woburn.
The brave new dawn gets underway at Kingsbarns in 2017, the first time the course has staged a Major.
For more golf news, discussion and videos follow @NCGmagazine on Twitter, like us on Facebook, subscribe to our YouTube video channel, and sign up to our newsletter


text:
Rugged, shock-proof polymer replacement for factory forend upgrades your Remington, or Mossberg pump shotgun with a heavily textured tactical forend for home defense or competition. Deep grooves give a secure purchase for fast, positive racking and enhanced recoil control. Installation requires some disassembly of shotgun, including barrel, forend, and forend nut.
SPECS: Injection-molded polymer, matte black. 6" (15.2cm) long, 17⁄8" (4.8cm) dia. 5.5 oz. (155g) wt. Remington - Fits Remington 870 and Winchester 1200/1300 - 12 gauge. Mossberg - fits Mossberg 500, 590, and 835 - 12 gauge. Includes wrench for installation.
Mfr Part: SFM1201B
Color: Matte Black
Gauge: 12 Gauge
Material: Injection-molded polymer
Mfr Part: MSF1201B
Would you like to be alerted when the item becomes available again?
Please enter your email address and quantity below andwe will send you an alert when the item becomes available.
Please note that alerted inventory is on a first come, first serve basis.
Thank you for contacting our Chat Team! Please visit us again.
If you would like a copy of the chat transcript please click "Send Transcript" below.
Thank you for contacting Brownells. You've reached us after business hours. Live Chat is available 7:30 am to Midnight (U.S. CST) Monday - Friday and 8:00 am to 5:00 pm (U.S. CST) Saturday - Sunday. You may also contact us via email at email@example.com.


text:
Kuhi Comfort Blog
Kuhi Comfort Travel Pillow FAQs
Can I wash my Kuhi Comfort Travel Pillow?
Absolutely! We understand that items often get dirty when traveling.
Both the Kuhi Comfort travel pillow and the carry case can be machine washed and gently tumbled dry.
Yes, the entire pillow can go in the washing machine!
Take the pillow out of the case, zip the case closed and toss them both in the washing machine with like colors (no bleach) on a gentle cycle.
Then you can toss them both in the dryer. And they will be fresh and clean for your next adventure.
Will any of the sold out models be available in the future?
We are doing our best to keep up with the demand for the best travel neck pillow on the market.
We understand than some models have been out of stock for some time.
We are working on having our complete line available.
For information regarding product availability, please sign up for our "Good News Newsletter" at: http://www.kuhicomfort.com/pages/subscribe-to-our-newsletter
Do you have a satisfaction guarantee?
Yes, We offer a 100% satisfaction guarantee!
If for any reason your Kuhi Comfort Travel Pillow does not meet your expectations you can return it for a full product refund.
For return/refund information please contact us at firstname.lastname@example.com.
What is the weight of Kuhi Comfort Travel Pillow?
The Kuhi Comfort Travel Pillow is a lean 10 ozs.
What is the size of the Kuhi Comfort Travel Pillow?
When it is in its carry case, the Kuhi Comfort Travel Pillow measures 6 inches x 8 inches.
Fully extended, the Kuhi Comfort Travel Pillow is 14 inches x 6 inches x 3.5 inches
Is the Kuhi Comfort Travel Pillow adjustable?
Yes, The Kuhi Comfort Travel Pillow is probably the most versatile travel pillow on the market.
The unique Kuhi Comfort shape is designed to be flipped and twisted and turned so you can find your perfect fit.
Try it with the handles front, then flip it 180 degrees so the handles are out to the sides, then for an even more custom fit you can twist it so one handle is front and one handle is out to the side.
Kuhi Comfort also makes a great back support, arm rest and even works as a foot rest.
There are more ways to adjust your Kuhi Comfort than you can imagine, which is another reason it's the best travel neck pillow you'll find. Shop our selection to find the perfect one for you!
Is the Kuhi Comfort Travel Pillow available in Canada?
Yes! Kuhi Comfort is available in Canada at
TravelSmarts Luggage and Accessories.
Online: http://www.travelsmartsproducts.ca/store/
Or at their store:
Metropolis at Metrotown
225 - 4820 Kingsway
Burnaby, BC
What is the difference between the Original and the Cuddle Fur?
Both the Original and the Cuddle Fur have the patented Kuhi Comfort design to bring you the best travel neck pillow available.
Both are sewn with a plush microfiber velvet fabric and stuffed with a hypoallergenic polyester loose fiberfill.
The only difference is that the Cuddle Fur cushions have faux sheepskin panels. Both are very soft.
It is a matter of what you like to feel against your skin. If you like a smoother feel, go with the Original. If you like a more cuddly, think Teddy Bear, feel then go with the Cuddle Fur.


text:
For your next construction project try using 20 ft. x 100 ft. Flame Retardant Plastic Sheeting. This polyethylene film can be used in a variety of applications including winterizing windows and as a cover for outdoor protection. This heavy-duty film can cover up to 2,000 sq. ft.
- Made of polyethylene
- 4 mil thickness
- Flame/fire-retardant
- Covers up to 2,000 sq. ft.
- Heavy-duty coverall
- Can be used for winterizing windows, as a paint drop cloth and as a cover for outdoor protection
- Get the right coverage - click here for Home Depot's buying guide
- Product packaging may vary by location


text:
9 Oct 2009: Palmieri N, Schlötterer C (2009) Correction: Mapping Accuracy of Short Reads from Massively Parallel Sequencing and the Implications for Quantitative Expression Profiling. PLOS ONE 4(10): 10.1371/annotation/8d1d51b9-81be-4351-ba45-27e3afdea13a. https://doi.org/10.1371/annotation/8d1d51b9-81be-4351-ba45-27e3afdea13a View correction
Massively parallel sequencing offers an enormous potential for expression profiling, in particular for interspecific comparisons. Currently, different platforms for massively parallel sequencing are available, which differ in read length and sequencing costs. The 454-technology offers the highest read length. The other sequencing technologies are more cost effective, on the expense of shorter reads. Reliable expression profiling by massively parallel sequencing depends crucially on the accuracy to which the reads could be mapped to the corresponding genes.
Methodology/Principal Findings
We performed an in silico analysis to evaluate whether incorrect mapping of the sequence reads results in a biased expression pattern. A comparison of six available mapping software tools indicated a considerable heterogeneity in mapping speed and accuracy. Independently of the software used to map the reads, we found that for compact genomes both short (35 bp, 50 bp) and long sequence reads (100 bp) result in an almost unbiased expression pattern. In contrast, for species with a larger genome containing more gene families and repetitive DNA, shorter reads (35–50 bp) produced a considerable bias in gene expression. In humans, about 10% of the genes had fewer than 50% of the sequence reads correctly mapped. Sequence polymorphism up to 9% had almost no effect on the mapping accuracy of 100 bp reads. For 35 bp reads up to 3% sequence divergence did not affect the mapping accuracy strongly. The effect of indels on the mapping efficiency strongly depends on the mapping software.
In complex genomes, expression profiling by massively parallel sequencing could introduce a considerable bias due to incorrectly mapped sequence reads if the read length is short. Nevertheless, this bias could be accounted for if the genomic sequence is known. Furthermore, sequence polymorphisms and indels also affect the mapping accuracy and may cause a biased gene expression measurement. The choice of the mapping software is highly critical and the reliability depends on the presence/absence of indels and the divergence between reads and the reference genome. Overall, we found SSAHA2 and CLC to produce the most reliable mapping results.
Citation: Palmieri N, Schlötterer C (2009) Mapping Accuracy of Short Reads from Massively Parallel Sequencing and the Implications for Quantitative Expression Profiling. PLoS ONE4(7): e6323. https://doi.org/10.1371/journal.pone.0006323
Editor: Rodolfo Aramayo, Texas A&M University, United States of America
Received: April 7, 2009; Accepted: June 8, 2009; Published: July 28, 2009
Copyright: © 2009 Palmieri, Schlötterer. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work has been supported by grants of the Fonds zur Förderung der wissenschaftlichen Forschung (FWF) to C. Schlötterer (P19467-B11, P17373-B14). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Technological advances have revolutionized the analysis of the transcriptome, the set of genes expressed in a given tissue. Currently, a broad range of techniques is widely used for expression profiling, but each available technique has its specific limitations.
The first widely used microarrays were based on cDNA sequences [1]. PCR amplified cDNA fragments are spotted at a high density (10–50 spots per mm2) onto a microscope slide and probed against a labelled target. This technique offers the advantage that it is rather insensitive to mismatches between the probe and the cDNA sequence. The drawback of these probes is that they are very sensitive to cross-hybridization; hence it is impossible to contrast the expression pattern of genes with a similar sequence (e.g.: members of a multigene family). Microarrays with multiple short oligos per gene are used for most commercial gene expression platforms (e.g. Affymetrix). While longer oligos offer the advantage of more reliable hybridization, they are more prone to cross-hybridization than platforms using shorter ones.
Despite considerable effort to develop oligos with similar hybridization properties [2], the hybridization behaviour is very complex. The same target RNA molecule may hybridize with different efficiency resulting in different hybridization signals [3]. Hence, due to the sequence specific hybridization behaviour, the above-mentioned techniques are not well suited for measuring absolute expression levels and they are mainly used to compare the expression pattern of different samples relative to each other. As only relative gene expression levels can be inferred reliably, studies requiring measures of absolute gene expression are best advised to rely on different methods for expression profiling.
Expressed sequence tag (EST) sequencing is a very powerful, but also expensive method to measure gene expression. Building a cDNA library and sequencing a large number of clones provides a good overview on the absolute expression level, as the frequency of a given transcript is proportional to its expression level. Hence, it is possible to compare the expression of different genes without having to rely on the assumption of homogeneous hybridization behaviour. The high costs of EST sequencing however, make an extension to many species and tissues impractical. An alternative to EST sequencing is the Serial Analysis of Gene Expression (SAGE) technique [4]. Rather than sequencing individual cDNAs, for SAGE short 14 bp cDNA tags are concatenated and jointly sequenced. This allows an in depth analysis of gene expression at significantly lower, albeit still considerable costs.
The greatest challenge for all these methods is the comparison of gene expression across species. Despite that cDNA arrays potentially offer the advantage of only minor effects of mismatches, some studies of closely related species showed that cross-species expression profiling using one common cDNA array is complicated and may yield spurious results [5]. Furthermore, the well-known heterogeneity in evolution rates among genes further complicates the comparison of expression divergence between two species across genes. Recently, a modification of the cDNA arrays has been proposed, which requires a cDNA probe for each gene in every species included in the study [5], [6], which makes the generation of the arrays extremely labour intensive. Oligo arrays have also been used for cross-species expression profiling by restricting the analysis to those probes that were perfectly conserved between the species analyzed [7]. For obvious reasons, this approach is limited to closely related species for which a fully sequenced genome is available. In principle SAGE could be applied to any species, but the identification of the gene corresponding to a given SAGE tag does not only require a fully sequenced genome, but also a reliable gene annotation including a characterized 3′ UTR, as a large proportion of the SAGE tags is located in the 3′UTR [8]. Even the modifications of the traditional SAGE method, which result in longer tags [9] are not sufficient for poorly annotated genomes. Similar to SAGE, massively parallel signature sequencing approaches (MPSS), which result in 16–20 bases [10], suffer from the problem to assign the short sequences unambiguously to incompletely annotated genomes. EST sequencing is probably the most general method, but a broad application is prohibited by the high costs.
The increased read length of the second-generation sequencing technologies could potentially overcome most of the disadvantages mentioned above. Depending on the platform used, the read length of a single sequence is at least 35 bp, which provides a substantial improvement in the ability to accurately identify the gene corresponding to the short sequence read. Hence, by sequencing random pieces of cDNA molecules, massively parallel sequencing potentially provides an enormous potential to quantify gene expression.
Recently, Torres et al. [11] showed that the sequencing of 3′ ends of randomly sheared cDNA molecules provides an excellent tool for quantifying gene expression. Using the 454 sequencing technology they unambiguously mapped 97% of the sequence reads and showed that the obtained expression profiles were highly reproducible. Nevertheless, compared to the competing massively parallel sequencing technologies (Illumina, SOLiD), the cost per base with the 454 sequencing technology is about one order of magnitude higher. This raises the question if the shorter reads obtained with the more affordable sequencing technologies are sufficient for gene expression studies – i.e.: could they be mapped to the corresponding genes with a similar efficiency as longer sequence reads?
In this report we compare the mapping efficiency of reads of different length. We show that for smaller genomes, such as yeast and Drosophila, even the shortest 35 bp reads perform well and do not result in a major bias. For larger genomes, such as humans, a considerable bias could be observed for 35 bp reads.
We were interested to study how the accuracy of mapping short reads is affected by size of the reads, complexity of the reference genome, and the mapping algorithm used. As quantitative transcript profiling by massively parallel sequencing is potentially affected by the accuracy of the mapping of short reads, we performed an in silico analysis to evaluate this.
We generated in silico reads from Saccharomyces cerevisiae (yeast), Drosophila melanogaster (fly), Arabidoposis thaliana, and Homo sapiens (human) transcripts. Some genomic regions are occasionally used to encode for multiple transcripts, by either overlapping or nested genes. As this phenomenon would prevent the unambiguous mapping of reads to one gene, we only used genes that were encoded by a different genomic stretch of DNA.
Two methods for quantitative expression profiling by massively parallel sequencing have been described: whole transcript sequencing or sequencing of randomly broken 3′ fragments. We focus on the latter method, as it does not require an adjustment of the transcript length. Nevertheless, we obtained qualitatively similar results for both methods (Figure 1).
Reads are mapped against the genome of the corresponding species.
For every gene we selected the longest transcript, which was randomly sheared. After capturing the 3′ ends, we obtained the short reads from the 5′ end of the captured fragment. We generated sequence reads from four species and mapped them either to the transcriptome or genome using six different programs: BLAT [12], SSAHA2 [13], Bowtie [14], SeqMap [15], MAQ [16] and CLC NGS Cell [www.clcbio.com]. Then we classified the reads in two categories:
- Mapped: reads that mapped to a unique position in the genome (see Material and Methods)
- Correct: reads that mapped to the correct position.
Overall, we found that the mapping accuracy was high. Even the short 35 bp reads were mapped with a high precision. BLAT, SSAHA, SeqMap and CLC mapped almost all reads correctly. Only BLAT had a notable fraction of almost 7% of the 35 bp reads, which were incorrectly mapped against the human genome (Figure 2A). In the case of an ambiguous match Bowtie and MAQ randomly assign reads to one of the targets, which results in a higher proportion of incorrectly mapped reads (Figures 2C, 2E). Hence, neither Bowtie nor MAQ should be used for mapping reads to quantify gene expression.
In each simulation the labels for each bar indicate in the order: organism (y = S. cerevisiae, d = D. melanogaster, a = A. thaliana, h = H. sapiens), mapping against the transcriptome (t) or the genome (g), read length (35 bp, 50 bp, 100 bp).
While for species with a low complexity almost no difference in mapping accuracy could be noted between short 35 bp and long 100 bp reads, for species with a more complex genome the mapping improved with longer reads. In humans and Arabidopsis both the mapping accuracy as well as the number of reads mapped increased with read length. Furthermore, we found that mapping against the transcriptome was more effective and reliable than mapping against the genome.
Nevertheless, we found some notable exceptions to these general trends. First, in yeast MAQ and CLC mapped a larger proportion of the reads to the genome than to the transcriptome and we could not identify the reason for this counter-intuitive result (Figures 2E, 2F). Second, with Bowtie and SeqMap an increase in read length resulted in a decreased mapping efficiency to the genomic reference (Figures 2C, 2D). This observation is the result of a larger proportion of exon-intron junctions included in the longer sequence reads. As Bowtie and SeqMap do not map reads with more than three mismatches between the read and the reference sequence, reads spanning exon-intron boundaries have frequently more than three mismatches and are therefore not mapped. We validated this hypothesis by mapping reads that did not span two exons against the genome of D. melanogaster (see Material and Methods) and we observed an increase of mapping accuracy as the read length increases (Table S2).
For expression profiling the incomplete mapping of reads only becomes a problem if genes are differentially affected. If all genes have approximately the same number of reads that cannot be mapped, no bias would be introduced. We evaluated this by using BLAT, as this mapping tool was found to be sensitive to read length and genome complexity and should thus show the most pronounced effect. Figure 3 shows that for the majority of genes the inferred expression level does not deviate from the expectations. Nevertheless, a non-negligible number of genes deviated to a variable degree from the expectations. In D. melanogaster 97% of the genes deviate less than 10% from the true expression level for 100 bp reads and 91% for 35 bp reads. For humans only 85% (100 bp reads) and 65% (35 bp reads) of the genes deviate less than 10%. The differences are even more pronounced by considering the genes with more than 50% difference in expression, thus we systematically studied this effect for different species and mapping tools (Table 1). For yeast and D. melanogaster no major difference was detected among the read lengths tested. For humans and Arabidopsis, the bad performance of Bowtie and SeqMap for long reads is very prominent, but for CLC, SSAHA2 and BLAT the proportion of genes with more than 50% deviation from the expected expression intensity was substantially decreased (Table 1).
Interestingly, irrespective of the read length, we found a small proportion of genes (1.3% in Drosophila) which did not recover a single mapped read. We reasoned, that this observation could be attributed to recent gene duplications, which resulted in almost identical gene copies preventing unambiguous mapping. To test this hypothesis, we aligned the full transcripts of those genes for which we did not map a single read against the genome of the corresponding species. Consistent with the gene duplication hypothesis, we found two equally good hits for all the genes tested. Most importantly, our analysis indicated that even reads longer than 100 bp would not have improved the mapping accuracy for these duplicated genes.
So far, we only considered sequence reads generated from published genomes, thus no mutations were expected, which could further complicate the mapping of the reads. Nevertheless, many expression analyses focus on individuals with unknown genotypes. Hence, we evaluated the influence of base substitutions and insertions on the mapping accuracy. For simplicity, this analysis was conducted only in D. melanogaster. We mutated 3, 6 and 9% of the bases in a read and mapped the mutated reads to the original genome/transcriptome (Figure 4). The mapping of 100 bp reads was almost not affected by mutations. Even with 9% mutations, only a very limited effect on the mapping accuracy was noted. We point out that for 100 bp reads with 9% of mutation the comparison is possible only within the programs BLAT, SSAHA2 and CLC, because the mapping was not feasible with other programs due to high divergence of the reads. For 50 bp reads up to 6% of mutation did not have an effect on the mapping accuracy for all programs, except for Bowtie, which could not deal well with the divergent reads. The most pronounced effects were seen for 35 bp reads. While 3% mutations did not affect the mapping, 6% and 9% mutations had a substantial effect. Interestingly, we noted a marked difference in the performance among the programs used. BLAT could only map about 50% of the reads and SSAHA2 mapped more than 20% of the reads incorrectly. The best results were obtained by MAQ and SeqMap, which still mapped a very high proportion (>90%) of the reads correctly.
We also tested the effect of insertions by mapping D. melanogaster reads containing a single insertion of variable size (3, 6, 9% of the read length) in a random position. This analysis could be only conducted with BLAT, SSAHA2 and CLC (Figure 5), because the other programs cannot perform gapped alignments. While Bowtie also allows insertions, the high demand on CPU time prevented us from a systematic evaluation of this software. Already the insertion of a single base dramatically lowered the mapping efficiency of BLAT (from 97% to 67%). SSAHA2 and CLC were almost not affected by the presence of insertions, irrespective of their length. Nevertheless, SSAHA2 performed slightly better than CLC, which showed a small proportion (9.8% for a 9% insertion in 100 bp reads) of incorrectly mapped reads.
We were interested to test if some genes are more prone to suffer from the lower mapping accuracy of mutated reads than others, as this would result in a further bias in gene expression. Panel A in Figure 6 shows the variance in the observed eobs by two independent "sequencing runs", for which we generated two independent sets of fragments and mapped them to the genome with BLAT, as this mapping algorithm was found to be highly sensitive to mutations. The remaining panels show the comparison of eobs for no mutations to reads with 3%, 6% and 9% mutated bases respectively. Interestingly, eobs for reads with 3% and 6% mutations were highly correlated with eobs from reads without mutations. Only reads with 9% mutations showed a pronounced reduction in the correlation coefficient. Thus, up to 6% sequence divergence no pronounced gene specific effects could be detected, suggesting that we found no evidence for heterogeneity in the effect of mutations on the mapping efficiency.
A) Two independent runs of 35 bp reads without mutations, B) 1 mutation vs. no mutation, C) 2 mutations vs. no mutation, D) 3 mutations vs. no mutation. r indicates the correlation coefficient between the observed expression levels.
As our data set only contained non-overlapping genes, we repeated the mapping with CLC, using for each organism a cDNA database in which overlapping genes are not removed, in order to quantify the impact of overlapping genes. As expected we obtained a decrease in mapping efficiency (Figure 7). This is caused by more ambiguities in the mapping, as there are more genes that share partially the sequence. The impact was marginal for Drosophila, Arabidopsis and yeast, but for human there is a decrease of more than 2% in the genome mapping analysis).
Reads are generated from a transcript database in which the longest isoform is chosen, but overlapping transcripts are not removed.
Given that expression analyses produce a large number of reads, we evaluated not only the mapping accuracy, but also in the mapping speed of the programs used. For each program we determined the required CPU time on a 2×2.8 GHz Quad-Core Intel Xeon Mac Pro computer for 100,000 reads (100 bp), mapped against the three different species. The fastest mapping software is Bowtie, followed by CLC and SSAHA2. BLAT and SeqMap are the slowest ones (Table 2). Additionally, we report in table S3 a summary of all the relevant features of the mapping tools used.
Massively parallel sequencing offers an enormous potential for gene expression profiling. With the number of sequence reads being proportional to the transcript abundance, it is possible to make reliable absolute quantification of transcripts. Hence, the comparison of gene expression between species should be possible by massively parallel sequencing. Currently, researchers are facing the decision between more economic short reads and longer reads. The goal of this study was to compare both approaches with respect to their potential for accurate expression profiling. Apart from technological problems [11], the major challenge is the accurate mapping of the sequence reads to the corresponding gene.
Consistent with previous results [11], we showed that 100 bp reads produce highly accurate mapping results, almost independent of the mapping software used. Even mutations and indels did not show a major effect on the mapping accuracy. If adequate mapping software is used, 50 bp reads produce already highly reliable results for species with a small genome. For humans, however, sequence reads of 35 bp and 50 bp do not yield trustworthy results. Even without mutations, a considerable fraction of the reads were not mapped and more than 10% of the genes differed more than 50% for the true expression level.
The influence of sequence divergence on mapping accuracy is conceptually more difficult to handle. As genes differ in their sequence conservation, this will generate heterogeneity in mapping accuracy among genes that is difficult to predict. Hence, we restricted our analysis to a more general analysis of the impact of mutations and insertions on the mapping accuracy. To do so, we mutagenized each sequence read independently from the others. For 100 bp reads, sequence divergence had almost no effect on the mapping accuracy, even if up to 9% of the bases in the read were mutated. Also a single mutation in 35 bp reads or 3 mutations in 50 bp reads had almost no effect on the mapping accuracy. This suggests that even for polymorphic species, such as D. melanogaster, which has about 3% silent site polymorphism [17], short 35 bp reads will be sufficient to cover intraspecies variation without the loss of mapping accuracy. Interspecific comparisons are more problematic when the genome of one of the species is not known. The silent sequence divergence of the two closely related species D. melanogaster and D. simulans is slightly above 13% [17], suggesting that even the availability of a reference sequence from a close relative does not guarantee a reliable gene expression measurement with short reads. If the right software tool is used, insertions were not found to affect the mapping quality. Nevertheless, clustering of substitutions and indels [18] will probably further reduce the mapping efficiency of a large number of reads.
Based on these results, we conclude that gene expression studies in yeast and Drosophila could be reliably performed with 50 bp reads, even when some sequence divergence between the reference genome and the transcriptome is expected. For humans, and to some extent for Arabidopsis as well, 100 bp reads are preferable.
One important result of our study is the highly heterogeneous performance of the different mapping software tools. While Bowtie was undoubtedly the fastest mapping program, it performed very poorly with mutations and could not deal with insertions. Like Bowtie, SeqMap cannot deal with indels and the limitation to three mismatches strongly affects the mapping of 100 bp reads to genomic DNA. MAQ cannot map reads with indels. One further limitation of Bowtie and MAQ is the random assignment of equally well mapped reads to the corresponding genes. Unless, both genes are expressed at a similar level, this procedure will cause wrong expression estimates and should thus avoided. BLAT is a more general program, which could deal with insertions, but in the presence of mutations the mapping accuracy was only reasonably good for 100 bp reads. CLC and SSAHA2 were found the two programs that produced the most accurate results for most parameters tested. The relatively high rate of incorrectly mapped 35 bp and 50 bp reads with 9% mutations detected for SSAHA2, renders CLC the most versatile and accurate program in our comparison. Nevertheless, we would like to point out that mapping performance of all programs is severely affected by the choice of mapping parameters. As we tested a broad range of species and read lengths, we typically used only the recommended parameter settings. We cannot rule out that some optimization of the mapping parameters would have altered our conclusions about the mapping performance of the software tested.
Irrespective of the software used, we found that a given fraction of genes recovered fewer reads than expected. This under-representation of mapped reads is caused either by gene duplication or repetitive sequences in the transcript. While longer reads reduce this problem to some extent, for genes that were duplicated very recently even reads longer than 100 bp will not improve the situation. We propose a simple correction, which should account for the gene specific recovery of sequence reads. The expected eobs could be determined in a similar procedure as reported here. If the observed counts are weighted by eobs it is possible to account for the bias.
The results of our study are more general than just expression profiling by massively parallel sequencing. Recently, it has been suggested that massively parallel sequencing could also be used to estimate allele frequencies from DNA pools [19]. The accurate genome wide estimate of allele frequencies depends also on a highly accurate mapping of sequence reads. Our results show that the mapping accuracy will be highly dependent on the choice of the software, but also read length and genome complexity will have an effect. While paired end reads, substantially improve the mapping accuracy, our results also suggest that for species with a compact genome, such as D. melanogaster, a very high mapping accuracy could be obtained with 50 bp reads.
It is important to keep in mind that all our simulations used tags generated from the sequence to which the reads were mapped. Experimental data, however, may also contain reads for which no reference sequence is known. This could be either due to an incomplete genomic sequence or the presence of other organisms, such as Wolbachia and Viruses. In such cases fewer reads will be mapped and more reads may be incorrectly mapped.
Finally, we would like to point out that our analyses did not account for sequencing errors, as we do not think that they have a severe effect on expression profiling, as the position of the reads is governed by random breakage of the cDNA. Thus, even preferential mutations at the end of a read will not result in a systematic bias. Nevertheless, other biases caused by the sequencing process, such as a GC-content dependent production of reads [20], could also have a substantial effect on the estimated gene expression levels and these are not considered here.
Materials and Methods
Sequences of transcripts and chromosomes were obtained for S. cerevisiae and H. sapiens from the Ensembl database (www.ensembl.org), A. thaliana from the Arabidopsis Information Resource (http://www.arabidopsis.org/) and D. melanogaster from FlyBase release 5.4 (http://flybase.bio.indiana.edu/).
From the transcripts files we selected the longest isoform for all the protein coding transcripts (mRNA). To avoid the complication of overlapping genes, we removed all the genes that overlap at least with another gene. The number of transcripts before and after filtering is shown in Table S1.
Generation of sequence reads
We mimicked the generation of sequence reads from cDNA sequences as described in Torres et al. [11]. In brief, Torres et al. [11] randomly sheared the double stranded cDNA molecules, recovered the 3′ fragments and sequenced them from their 5′ end. They observed a pronounced bias against fragments shorter than 80 bp and longer than 300 bp. Thus, we used the experimentally derived distribution of 3′ fragment size distribution to generate 200 sequence reads from each transcript. We generated reads with three size classes, 35, 50 and 100 bp.
We also used a uniform distribution between 38 bp and 1000 bp to measure gene expression and obtained almost identical results (not shown). Hence, the results based on the experimental distribution of 3′ fragment sequences obtained by 454 sequencing could be generalized to any transcription analysis.
Sequence divergence
We studied the effect of sequence divergence by randomly mutating 3%, 6%, and 9% of the bases in a read following a Jukes-Cantor model of sequence evolution [21]. It should be noted that each read differed, thus our analysis does not reflect the effect of specific mutations in the genome. Rather we were interested in the general effect of mutations in the sequences. At the same way we evaluated effect of insertions introducing a single insertion in random position of each read, which size is 3%, 6% and 9% of the read length.
Mapping of the reads
The in silico generated reads were mapped either to the transcriptome or the complete genome using different programs: BLAT [12] (parameters –oneOff 1), SSAHA2 [13] (parameters –skip 2, -diff 0, -kmer 13, -solexa for 35 bp and 50 bp reads; -454 for 100 bp reads), Bowtie [14] (parameters –k 1, -n 3, -e 2000), SeqMap [15] (number of mismatches = 3), MAQ [16] (parameters –n 3, -e 2000) and the commercial tool CLC NGS Cell [www.clcbio.com] (algorithm: clc_ref_assembly_long, parameters: -r ignore). To keep the results most comparable as possible we set for each program only the essential parameters and assigned 3 mismatches to the programs that needed this parameter (Bowtie, SeqMap, MAQ), which is also the maximum number of mismatches allowed by these three programs.
Then we mapped the reads first against the transcriptome and then against the genome of the species considered. For each program, we used the coordinates of the best unique hit to determine if each read is mapped to the correct gene.
The proportion of correctly mapped reads eobs was determined for each gene and we produced, for each program, the percentage of genes that recover less than 50% of the expected expression (eexp = 200).
Exon analysis
This was done to confirm that Bowtie has difficulties mapping reads that span two exons. Rather than generating sequence reads from transcripts, we only used exons of genes with two exons. Hence, the reads did not contain exon junctions. We generated for each exon 100 reads of lengths 35, 50 and 100 bp. These reads were mapped with Bowtie against the transcriptome and the genome of D. melanogaster.
PERL scripts have been used for filtering the transcripts, generating the reads and counting the number of correctly mapped reads. They are available in the supplementary Material S1.
Supporting Information
We are thankful to the members of the Institute of Population Genetics for helpful discussion and comments on the manuscript. We especially acknowledge Ben Langmead for his insightful comments about the Bowtie software.
Author Contributions
Conceived and designed the experiments: NP CS. Performed the experiments: NP. Analyzed the data: NP. Contributed reagents/materials/analysis tools: NP CS. Wrote the paper: NP CS.
- 1. Schena M, Shalon D, Davis RW, Brown PO (1995) Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science 270: 467–470.M. SchenaD. ShalonRW DavisPO Brown1995Quantitative monitoring of gene expression patterns with a complementary DNA microarray.Science270467470
- 2. Rouillard JM, Herbert CJ, Zuker M (2002) OligoArray: genome-scale oligonucleotide design for microarrays. Bioinformatics 18: 486–487.JM RouillardCJ HerbertM. Zuker2002OligoArray: genome-scale oligonucleotide design for microarrays.Bioinformatics18486487
- 3. Draghici S, Khatri P, Eklund AC, Szallasi Z (2006) Reliability and reproducibility issues in DNA microarray measurements. Trends Genet 22: 101–109.S. DraghiciP. KhatriAC EklundZ. Szallasi2006Reliability and reproducibility issues in DNA microarray measurements.Trends Genet22101109
- 4. Velculescu VE, Zhang L, Vogelstein B, Kinzler KW (1995) Serial analysis of gene expression. Science 270: 484–487.VE VelculescuL. ZhangB. VogelsteinKW Kinzler1995Serial analysis of gene expression.Science270484487
- 5. Gilad Y, Rifkin SA, Bertone P, Gerstein M, White KP (2005) Multi-species microarrays reveal the effect of sequence divergence on gene expression profiles. Genome Res 15: 674–680.Y. GiladSA RifkinP. BertoneM. GersteinKP White2005Multi-species microarrays reveal the effect of sequence divergence on gene expression profiles.Genome Res15674680
- 6. Gilad Y, Oshlack A, Smyth GK, Speed TP, White KP (2006) Expression profiling in primates reveals a rapid evolution of human transcription factors. Nature 440: 242–245.Y. GiladA. OshlackGK SmythTP SpeedKP White2006Expression profiling in primates reveals a rapid evolution of human transcription factors.Nature440242245
- 7. Khaitovich P, Weiss G, Lachmann M, Hellmann I, Enard W, et al. (2004) A neutral model of transcriptome evolution. PLoS Biol 2: E132.P. KhaitovichG. WeissM. LachmannI. HellmannW. Enard2004A neutral model of transcriptome evolution.PLoS Biol2E132
- 8. Metta M, G R, G JM, Schlotterer C (2006) No Accelerated Rate of Protein Evolution in Male-Biased Drosophila pseudoobscura Genes. Genetics 174: 411–420.M. MettaR. GJM GC. Schlotterer2006No Accelerated Rate of Protein Evolution in Male-Biased Drosophila pseudoobscura Genes.Genetics174411420
- 9. Matsumura H, Reich S, Ito A, Saitoh H, Kamoun S, et al. (2003) Gene expression analysis of plant host-pathogen interactions by SuperSAGE. Proc Natl Acad Sci U S A 100: 15718–15723.H. MatsumuraS. ReichA. ItoH. SaitohS. Kamoun2003Gene expression analysis of plant host-pathogen interactions by SuperSAGE.Proc Natl Acad Sci U S A1001571815723
- 10. Brenner S, Johnson M, Bridgham J, Golda G, Lloyd DH, et al. (2000) Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays. Nat Biotechnol 18: 630–634.S. BrennerM. JohnsonJ. BridghamG. GoldaDH Lloyd2000Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays.Nat Biotechnol18630634
- 11. Torres TT, Metta M, Ottenwalder B, Schlotterer C (2008) Gene expression profiling by massively parallel sequencing. Genome Res 18: 172–177.TT TorresM. MettaB. OttenwalderC. Schlotterer2008Gene expression profiling by massively parallel sequencing.Genome Res18172177
- 12. Kent WJ (2002) BLAT–the BLAST-like alignment tool. Genome Res 12: 656–664.WJ Kent2002BLAT–the BLAST-like alignment tool.Genome Res12656664
- 13. Ning Z, Cox AJ, Mullikin JC (2001) SSAHA: a fast search method for large DNA databases. Genome Res 11: 1725–1729.Z. NingAJ CoxJC Mullikin2001SSAHA: a fast search method for large DNA databases.Genome Res1117251729
- 14. Langmead B, Trapnell C, Pop M, Salzburg SL (2009) Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biology. B. LangmeadC. TrapnellM. PopSL Salzburg2009Ultrafast and memory-efficient alignment of short DNA sequences to the human genome.Genome Biology
- 15. Jiang H, Wong WH (2008) SeqMap: mapping massive amount of oligonucleotides to the genome. Bioinformatics 24: 2395–2396.H. JiangWH Wong2008SeqMap: mapping massive amount of oligonucleotides to the genome.Bioinformatics2423952396
- 16. Li H, Ruan J, Durbin R (2008) Mapping short DNA sequencing reads and calling variants using mapping quality scores. Genome Res 18: 1851–1858.H. LiJ. RuanR. Durbin2008Mapping short DNA sequencing reads and calling variants using mapping quality scores.Genome Res1818511858
- 17. Andolfatto P (2005) Adaptive evolution of non-coding DNA in Drosophila. Nature 437: 1149–1152.P. Andolfatto2005Adaptive evolution of non-coding DNA in Drosophila.Nature43711491152
- 18. Averof M, Rokas A, Wolfe KH, Sharp PM (2000) Evidence for a high frequency of simultaneous double-nucleotide substitutions. Science 287: 1283–1286.M. AverofA. RokasKH WolfePM Sharp2000Evidence for a high frequency of simultaneous double-nucleotide substitutions.Science28712831286
- 19. Van Tassell CP, Smith TP, Matukumalli LK, Taylor JF, Schnabel RD, et al. (2008) SNP discovery and allele frequency estimation by deep sequencing of reduced representation libraries. Nat Methods 5: 247–252.CP Van TassellTP SmithLK MatukumalliJF TaylorRD Schnabel2008SNP discovery and allele frequency estimation by deep sequencing of reduced representation libraries.Nat Methods5247252
- 20. Dohm JC, Lottaz C, Borodina T, Himmelbauer H (2008) Substantial biases in ultra-short read data sets from high-throughput DNA sequencing. Nucleic Acids Res 36: e105.JC DohmC. LottazT. BorodinaH. Himmelbauer2008Substantial biases in ultra-short read data sets from high-throughput DNA sequencing.Nucleic Acids Res36e105
- 21. Jukes TH, Cantor CR (1969) Evolution of protein molecules. Mammalian Protein Metabolism 21–132.TH JukesCR Cantor1969Evolution of protein molecules.Mammalian Protein Metabolism21132


text:
The thrilling conclusion to the breathtaking saga of power, blood, birthright, and enchantment. The barbarian King Loethar may have gained his throne through brutality and bloodshed, but he has ruled Penraven with unexpected wisdom. Now his innumerable past sins threaten his reign as Valisars who escaped slaughter prepare to rise up. Loyalties shift and new alliances are formed as the truth of history begins to emerge. King-in-exile Leonel hungers for revenge against the man he considers the usurper of his throne. Yet Leo is unaware of another who wields the enchantment he covets the once-charming "halfwit" brother Piven, now a powerful youth whose exceptional cunning is matched only by his ruthless desire for the throne. But the efforts of all three to hold or gain control of Penraven may well be in vain, for the true inheritor of the Valisar Legacy is being called home . . . to claim her crown.
We've sent an email with your order details. Order ID #:
Listen on the Audible app or these devices.Launch Audible App
-or-Download Free Audible App
To access this title, visit your library in the app or on the desktop website.
Listen on the Audible app or these devices.Launch Audible App
-or-Download Free Audible App
Audible does not currently have the rights to sell this title in your country.
Please consider another book.Continue Browsing
Please add a payment method to continue.Add a Payment Method
Please edit your payment details or add a new card.
If the problem persists, contact customer service at (888) 283-5051Review My Payment Method
If the problem persists, contact customer service at (888) 283-5051
Please add your billing address to continue.Add My Billing Address
Please select a default credit card to continue.Select Credit Card
It looks like you've already used the specified payment method for a free trial.
Please contact customer service if you think we have made a mistake. (888) 283-5051Start Your $14.95 Membership
If the problem persists, contact customer service at (888) 283-5051
If the problem persists, contact customer service at (888) 283-5051
I'd probably put it at number two - close behind Theft of Swords books...
Kilt Farris... and Gavriel.
I laughed and cried. There were so many twists in this series - I would recommend you listen to all three books. Each one throws in shocking twists... laughter, tears... all things I love in a book series. You begin liking a character, and then they change on you - and you begin to hate them. My emotions went crazy in this series... which made it that much better in my opinion.
Fiona McIntosh is probably my favorite author because she's so consistent. Unfortunately this is the only series of hers that is in audio format. I'm hoping they add more of her books soon. Things happen in her books that will shock you, piss you off and then make you happy. Her endings always leave you ready to read something else of hers. Happy with the ending in this series. For those who haven't tried her work - I HIGHLY recommend you do so.
- Katy


text:
Renal CPC. February 2009 'A 17 year old girl with chest pain and haemoptysis….'. Patient history. 17 y old girl admitted to hospital with a history of chest pain and haemoptysis Previous history of mild asthma Developed bi-pedal oedema 4 months previously
Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.
February 2009
'A 17 year old girl with chest pain and haemoptysis….'
Haemaglobin – normal
HcT –normal
WBCs – normal
Diff – normal
Platelets 497,000 [increased]
MCV – normal
PTT 70.3 [on heparin]
PT 1.1 [normal]
D-dimers –5,656 [increased]
Glucose – normal
Na+ – normal
K+ – normal
Cl- – normal
BUN – normal
Bilirubin –normal
total – normal
albumin – low
Ca++ – normal
CK – normal
CK-MB – normal
Alk phos – normal
AST – normal
ALT – normal
C-reactive protein –22.6 [increased]
Cardiac troponin – elevated
Amylase – normal
NT-pro-BNP 2571 [increased]
Lipoprotein [increased]
Lipase – normal
Activated protein C screen – normal
Functional protein S – normal
Anticardiolipin IgG phospholipid units [GPL] – normal
Anticardiolipin IgM phospholipid units [MPL] – normal
Antithrombin III – normal
Lupus anticoagulant – negative
PT gene G201210A – not detected
Antinuclear antibody 1:40 speckled
Anti-double stranded DNA Ab titre – negative
Anti-Ro [SS-A] antibody – negative
Anti-smooth muscle antibody – negative
Goodpasture's antigen – negative
pH 7.51
pCO2 increased
paO2 normal
Urine total protein [mg/24 hrs] – 2690 (increased)
Urine creatinine [g/24 hrs] 0.25
Small area of hypointense
thrombus extending from the
right renal vein into the
inferior vena cava
Proteinuria> 3.5 g of urinary protein:hypoalbuminaemia <3g/dl: oedema, hyperlipidaemia and lipiduria
in this case
B – membranous GN
Two general hypotheses have been proposed to explain the genetic association with disease.
a | The clearance hypothesis states that failure to clear apoptotic bodies — a principal source of SLE antigens — leads to inappropriate activation of mature, self-reactive B and T cells. Accordingly, apoptotic bodies are usually sequestered from the adaptive immune system by efficient recognition, binding and uptake by components of the innate immune system. So, SLE antigens are effectively 'hidden' from self-reactive lymphocytes. Defects in clearance of apoptotic bodies, such as those that occur in the absence of C1q or C4, would lead to the exposure of self-antigens to autoreactive lymphocytes and the development of SLE.
b | The tolerance hypothesis states that innate immunity is important in the negative selection of self-reactive lymphocytes, especially B cells that are specific for SLE antigens. The innate immune pathway functions to recognize, degrade and bind SLE antigens to stromal cells present in the bone marrow and spleen. Immature B cells that encounter concentrations of cognate antigen above a certain threshold are negatively selected; that is, they undergo receptor editing, clonal deletion or anergy. C1q and C4 function to enhance 'presentation' of SLE antigens to specific immature B cells in the bone marrow. Alternatively, coupling of C4b to self-antigen could result in co-receptor stimulation of immature B cells displaying the CR1–CD19–CD81 co-receptor. The co-receptor is expressed at the surface of immature B cells during transitional stages in the spleen, in which co-ligation with the B-cell receptor could result in enhanced negative selection and 'escape' of self-reactive B cells into the peripheral mature compartment, where they encounter SLE antigens and become activated.
Sandborg C et al. (2008) Therapy Insight: cardiovascular disease in pediatric /early adult systemic lupus erythematosus
Nat Clin Pract Rheumatol doi:10.1038/ncprheum0789


text:
Returning here (past mc's and pg ment)
View Single Post
September 11th, 2013, 06:00 PM
Just keep breathing.
Join Date: Sep 2012
Posts: 1,734
Hey everyone,
I'm Mel, 34, from Chicago. I've been on and off this board (though active on other JM forums) for the last year. I've had 3 miscarriages in the last year, and I am newly pregnant again. I'm hopeful- or as hopeful as I can really be after 3 losses. I'm currently 4w4d.
I had extensive RPL testing, with no real results. I am hypothyroid, but have been managing that for years just fine. I also have single mutation MTHFR a1298c- it's a gene mutation related to blood clotting disorders, but my specific variant doesn't cause problems. I had chromosomes tested on 2 of the lost pregnancies- they were normal. Third loss was a blighted ovum.
I'm currently seeing a great RE, and will get weekly appointments and ultrasounds. My hcg levels yesterday (at 17dpo) were 2914! A great, encouraging number. My first ultrasound was today! Not much to see, being so early. Just a sac- but good to know it was in the right place, and appropriately sized for where I should be.
I'm looking forward to being here and connecting with you all!
View Public Profile
Find all posts by MelChicago


text:
Can't We Talk About Something More Pleasant?: A Memoir
This is a best-selling book a few days, a lot of people who want to read it, because it is very good books. You can download ebook, i provide downloads as a pdf, kindle, word, txt, ppt, rar and zip. A book is very important in life. Life without book makes many people cannot know about any attractive things in the world. Hightechnology in this modern era makes many people can easy to get many books that they want and love in online book. The online book has same function looks like offline book. One of the best books is the book entitled Can't We Talk about Something More Pleasant?: A Memoir that gives the readers good inspiration.
Can't We Talk about Something More Pleasant?: A Memoir
Books Spesification :
Title : Can't We Talk about Something More Pleasant?: A Memoir
Books Rank : 8588
Author : Roz Chast
Publisher : Bloomsbury USA
Pages : 228
Type : Paperback
Language : English
Label : Bloomsbury USA
Creator :
Categories: Graphic Design
Type : Paperback Paperback at Amazon
Release : 2016-09-13
Price : $19.00 on amazon store
This book is also rich of knowledge. It makes thereaders are interesting to read this online book. When you open Can't We Talk about Something More Pleasant?: A Memoir you willfind the attractive content. This book is delivered in simple word. It makes the readers are easy to know the meaning of the content of this online book Can't We Talk about Something More Pleasant?: A Memoir When you read Can't We Talk about Something More Pleasant?: A Memoir you will find attractive content that cannot be found in other online books. The content of this book is reallyawesome. This book gives the real examples about the real life in the world.
A short description :
#1 New York Times Bestseller
2014 National Book Award Finalist
Winner of the inaugural 2014 Kirkus Prize in nonfiction
Winner of the National Book Critics Circle Award
Winner of the 2014 Books for a Better Life Award
Winner of the 2015 Reuben Award from National Cartoonists Society
In her first memoir, New Yorker cartoonist Roz Chast brings her signature wit to the topic of aging parents. Spanning the last several years of their lives and told through four-color cartoons, family photos, and documents, and a narrative as rife with laughs as it is with tears, Chast's memoir is both comfort and comic relief for anyone experiencing the life-altering loss of elderly parents.
While the particulars are Chast-ian in their idiosyncrasies–an anxious father who had relied heavily on his wife for stability as he slipped into dementia and a former assistant principal mother whose overbearing personality had sidelined Roz for decades–the themes are universal: adult children accepting a parental role; aging and unstable parents leaving a family home for an institution; dealing with uncomfortable physical intimacies; and hiring strangers to provide the most personal care.
An amazing portrait of two lives at their end and an only child coping as best she can, Can't We Talk about Something More Pleasant shows the full range of Roz Chast's talent as cartoonist and storyteller.
Please Register & Join Membership Get Unlimited Books


text:
For all press enquiries please contact: David Deaves T. 07793 319116
For general enquiries please contact Premier Cheese at: firstname.lastname@example.com
Cheese Makers Who WIsh To Exhibit
The Cheese Makers Market is aimed at cheese makers throughout the United Kingdom and Europe who are passionate about their products. It's also open to producers of cheese related products.
The Cheese Makers Market is held at The Market Place, Old Beaconsfield, Buckinghamshire HP9 2JW, and offers a fantastic opportunity to showcase cheeses rarely seen by the public. Meet and chat to the public, professional chefs and sell your products to enthusiastic cheese lovers.
As with all public markets, producers must be able to document the production and traceability of their products in response to questions either from the organisers or visitors to the market.
The Cheese Makers Market is a one day event and available only to cheese makers who have booked prior to the event. The event will include a professional on site management team, set up and infrastructure, and cold storage will be available for both traders and visitors should they want to purchase and keep products cold during their time on the market.
Price: £50 + £10 VAT (Total: £60)


text:
|Page (1) of 1 - 02/09/06
|email article
|print page
Tiger Technology Introduces New File Sequence Optimizer to Streamline DI WorkflowAnnounces release 2.1 of its flagship MetaSAN software (February 09, 2006)
Tiger Technology GmbH (www.tiger-technology.com), developer of high performance, affordable storage area networking (SAN) management software solutions, today announced that release 2.1 of its flagship MetaSAN software consolidates its technological leadership position by introducing an innovative File Sequence Optimizer and a high-performance Dynamic Disk Optimizer, in addition to offering support for Mac OS X Tiger 10.4.
File Sequence Optimizer - for Windows and Linux operating systems
3D animators, colorists, and special effects artists working on digital intermediates using image file sequences are confronted daily with significant performance problems as the large number of individual files they use quickly clutters their disks.
"A huge amount of time is wasted in post-production facilities worldwide de-fragmenting terabytes of storage weekly" says Alexander Lefterov, CTO and co-founder of Tiger Technology "our technology dramatically increases streaming and editing performances of large image file sequences by transparently storing multiple images inside a single packed file, thereby eliminating problems associated with disk fragmentation."
Any image file type can be packed using the Tiger Technology's File Sequence Optimizer (DPX, TGA, JPG, BMP, TIF, etc.). Files that match a given pattern (number of digits and file extension) are packed together. A typical pack holds 100 frames. Multiple packs are created as needed to accommodate very large image sequences. The operation remains fully transparent to the user.
Dynamic Disk Optimizer - for Windows operating systems
"Reading and writing performances are optimal when application's accesses and disk data chunks match in size and are fully aligned. Unfortunately, the formatting options available to the users under Windows are very limited." explains Lefterov, "We developed a RAID and Stripe Set Optimizer utility that lets users align disk chunk sizes when formatting their drives with the application's reading pattern. As a result, we have observed performance gain exceeding 50% on storage units used in facilities."
Mac OS X Tiger support
The legion of Final Cut Pro users that have been waiting for an alternative to Apple Xsan will be pleased to know that the MetaSAN port over to Mac OS X Tiger is now completed. "Between its Panther and Tiger releases, Apple has made many significant changes - mostly undocumented - to its file system. This gave us, and a number of other storage-related manufacturers a lot of grief," notes Lefterov. "But unless Apple goes through another round of significant changes, we should be able to catch up with their subsequent releases much faster, like we've always done!"
Report from the field
"MetaSAN is the ideal SAN management solution for Fibre Channel, iSCSI, and Gigabit Ethernet networks." highlights J�rg Hoffman of PSIBER GmbH "In the past 30 days, we have successfully completed two complex SAN integrations that involved a mix of Apple OS-X and Windows workstations. MetaSAN was chosen over Apples's Xsan because it not only allowed us to save money, but also fulfilled all the demands for performance, features, and reliability that demanding video professionals have come to expect. So far, we have tested MetaSAN with Huge MediaVault and Fibrenetix Storage systems, Qlogic SANBox switches, and Atto Celerity host bus adapters and have never experienced any compatibility issues."
About MetaSAN
MetaSAN is a comprehensive, time-proven, high-speed file sharing SAN management software solution that sets new standards for workgroup collaboration. MetaSAN enables multiple users to share access to common data files in workgroups where heavy bandwidth requirements are the norm. Targeted to the needs of digital content creation professionals involved with the creation of animation, digital intermediates, audio, graphics, satellite imagery, video clips, etc., as part of the production process, the software also meets the needs of a wide range of enterprise users in other markets including imaging, digital prepress and corporate/government.
MetaSAN is fully compatible with leading standard applications including Adobe Digital Video Collection and Apple Final Cut Pro. In addition, the Avid Unity MediaFile virtualization feature enables efficient collaboration among multiple users of Avid software. MetaSAN also supports leading vendors of SAN technology including ADTX, Atto, IBM, Cirprico/Huge Systems, Emulex, LSI, Qlogic and Seagate.
Tiger Technology MetaSAN coupled with MetaLAN provides the very best that SAN and NAS architectures have to offer. Tiger Technology turns a simple gateway server into an extremely cost effective, high performance, and scalable NAS-like cluster capable of advanced features such as cross-platform support, 24/7 failover, and load balancing while allowing the performance of multiple cost-effective servers to be aggregated together.
Pricing and Availability
MetaSAN and MetaLAN support Windows, Mac OS X, and Linux platforms. MetaSAN licenses are priced at $995.00 USD. MetaLAN licenses are priced at $295.00 USD. Tiger Technology software products are currently available from the company's OEM partners and its international reseller representatives. For sales inquiries please email Tiger Technology at [email protected]. Additional information on the Tiger Technology product line is available at www.tiger-technology.com.
About Tiger Technology
Founded in 2003, Tiger Technology GmbH, is a privately held company based in Zurich, Switzerland, delivering new technologies and OEM solutions in the field of storage area networking. The company's flagship product MetaSAN builds upon years of SAN-related expertise and development to bring digital content creation and enterprise professionals involved with handling large bandwidth files a unique, cross-platform, hardware-agnostic, high-speed file sharing SAN management solution that sets new standards for workgroup collaboration and reliability.
For further information on Tiger Technology please visit www.tiger-technology.com.
Page: 1
Related Keywords:Tiger Technology, File Sequence Optimizer, MetaSAN, SAN, Dynamic Disk Optimizer, digital intermediat, colorists, special effects, storage, MetaLAN, NAS,


text:
Print this Page | BACK
Wednesday, November 15, 2017 10:30am - 11:30am Location: The L.E. Shore Memorial Library Gallery - 173 Bruce St. S.
Phone: 519-599-3681 Email: email@example.com
©2017. All Rights Reserved.
All data is subject to errors, omissions or revisions & is not warranted.
Site Design & Development by Running Tide
Funded by a grant from the Trillium Foundation


text:
Dr. David Perry – At Davidson, we often tie the concepts of honor and trust so closely together that I fear we miss some important distinctions between the two, and obscure the extent to which our actual practices reflect trust and whether we are according students the right amount of trust.
We faculty at Davidson place a lot of trust in our students to uphold academic integrity, more so than at any other educational institution that I know. That's not only because you have individually pledged to be honest: you genuinely seem to take ownership of the Honor Code as a kind of sacred collective promise, not an alien thing imposed upon you. But as you know, in practice we don't completely trust you to do the right thing. One way to make that clear is to think what would be entailed if we did grant you unconditional, boundless trust.
Imagine, first, that we had a way to test incoming freshmen on their comprehension of what academic integrity entails, employing myriad examples of what counts as cheating and plagiarism. If some didn't pass that test initially, we might coach them intensively until they did pass. But let's say that before classes start, they all pass the test, and solemnly pledge to be scrupulously honest and uphold academic integrity throughout their sojourn at Davidson.
So based on our confidence in your understanding and commitment to academic integrity (i.e. we'd assume that no one will cheat, because everyone knows what cheating looks like, and everyone's pledged not to cheat), let's then imagine that we've decided to place complete trust in every student. What would that entail? 1) We could delete from the Honor Code the requirement to report cheating by one's peers and the penalty for not reporting. 2) We could dissolve the Honor Council, because we would never expect any Honor Code infractions fro them to investigate or try. 3) We could make all written tests take-home, and assume that no one will use any unauthorized materials during tests, collaborate with others in prohibited ways, or exceed specified time limits. We could also eliminate the restrictions on backpacking and electronic devices during self-schedules exams in Chambers (Why even require such exams to be held in online one building?). And 4) when reading your course papers, we wouldn't need to check whether you cited your sources properly or paraphrased thoroughly, because of course you would always give credit where credit is due.
In fact, if we really trusted you, we wouldn't need to require you to take any tests or submit any papers or other work products Instead, at the end of each course, we could ask you to tell us how well you've mastered the course material and what grade you deserved. Some students who may not be entirely confident about their mastery of the material or what grade they deserve might then ask us to test their comprehension and skills. But if any student said to us, "I'm confident that I thoroughly mastered the material in this course, and deserve an A," what reason would we have to doubt them, seeing as they satisfied our "rigorous" assessment of their comprehension of and commitment to integrity as incoming freshmen?
Now, I'm sure you'll agree that it would be a huge mistake to trust you to that degree. But why? 1) Because we know better than our students what it takes to exhibit mastery and excellence in our fields. That's parly what it means to have earned a Ph.D., to have proved that mastery to others who previously proven theirs. 2) Because we know that even generally consientious people don't always do the right thing (E.g., our students procrastinate, they panic, they get wasted the night before a test, or they don't check their eses of sources carefully enough). So we need to have ways to monitor conduct and performance according to high ethical standards, and punish violations of academic integrity.
Part of our solemn responsibility as faculty is to ensure fairness: 1) Fairness of students toward one another, that none takes unfair advantage of an opportunity to chear. 2) Fairness to other researchers and writers, giving them due credit for their ideas. And 3) fairness to ourselves, in students not decieving us to believe that they merit grades that they haven't earned.
A commitment to fairness is the main point of an honor node, not trust per se. Trust can be the happy consequence of a demonstrated commitment to honesty and fairness. But trust freely bestowed can also be forfeited. Our obligation to academic integrity justifies our having a degree of prudent mistrust of our students.
Given the nature of our honor system at Davidson, we can at least take some comfort in the findings of careful researchers like Dan Ariely ("The [Honest] Trust About Dishonest") that people are less likely to cheat: 1) When they've made a public commitment to practice ethical conduct (like the pledge-signing ceremony that Dean Shandley instituted years ago). 2) When they've also pledged to hold one another accountable for cheating. 3) When their peers openly value honesty. 4) When they regard honesty and fairness as integral to their personal identity. 5) When they're reminded of their obligations every time they take a test or submit a paper. 6) When their professors guard against cheating and plagarism, so that honest students don't feel like dishonest ones are free riders on their ethics.
On that last point, I worry that many of our faculty greatly underestimate the risks of cheating on take-home exams and plagiarism on papers by our students. We don't actually have a reliable estimate of the percentage of Davidson students who cheat, apparently because we're afraid to do that research, even though national surveys indicate that over 70% of college students overall admit to cheating, as do about 50% of students at schools with explicit honor codes. I'd wager that our percentage is lower than that, maybe even much lower, but I have no solid evidence to support that belief. No one knows, because we haven't asked. I think we should be surveying our senior students anonymously every year.
To be sure, some faculty report even minor errors in citation as full-blown plagiarism, which is unjust to their students. But other faculty don't place adequate checks on potential test-cheating opportunities, or use the best tools to identify plagiarism. I'm personally conflicted about the requirement that our students report others' cheating, in other words, that failing to report is as much an Honor Code violation as the cheating itself. On the other hand, the requirement makes me extremely confident that there will be no cheating during an in-class exam after I've distributed that test questions and left the room! That's a welcome change from the in-room proctoring that I felt compelled to do in other college settings.
In any case, I hope that these thoughts will stimulate further reflection and dialogue.
Dr. David Perry is a Professor of Applied Ethics and the Director of the Vann Center for Ethics at Davidson College. Contact him at email@example.com.
This article was originally published on February 26, 2016.


text: I have a customer that wants me to level his yard but the darn thing is always soaked! You can step on it, with regular pressure, and it'll move around your foot, with grass. I was curious, do you folks think there may be a busted line under his yard? Can I just use a heavy sand mix to help with the drying?


text:
Take it to Toccoa
4 p.m. Sunday, Georgia Baptist Conference Center, 462 Sonrise Way, Toccoa
Athens isn't the only town in Northeast Georgia to hear top-notch holiday music. The Toccoa Symphony Orchestra will present its annual Christmas concert this Sunday. Conductor David Stufft will lead the orchestra through featured selections, which include the debut of a new piece that answers the age-old question: "Yes, Virginia, there is a Santa Claus." Jarred Hettrick will narrate the piece written by Georgia composer Chuck Sayre. Other holiday selections include "Christmas Rocks at the Pops," an updated medley of holiday favorites performed pops-style and "A Bach Christmas." The 100-member Toccoa Symphony Orchestra Choir will lend voices to the orchestra for "Let it Snow," the Hallelujah Chorus and more. $10. For more information and tickets, visit www.toccoasymphony.org.
That's pretty sick
9 p.m. Tuesday, Georgia Theatre, 215 N. Lumpkin St.
After a head-turning first album, Olubowale Victor Akintimehin — you probably know him as Wale — is back with his sophomore effort, "Ambition." The Washington, D.C., rapper and producer will show off the lion's share of the new record, which he calls a classic, Tuesday at the Georgia Theatre. To Wale, "Ambition" and hard work walk hand in hand. The 27-year-old isn't afraid to tell everyone how hard he worked on this November release under the Maybach and Warner Music Groups. How great and clutch is he? To take a line from "Barry Sanders," Wale is as sick as (Michael) Jordan before Game Six. That's as sick as you can get. Wale already has a BET Hip Hop Award and more than 100,000 album sales for "Ambition" alone under his belt. And he's just getting started. To hear some of Wale's music, visit www.ralphfolarin.com. $18. www.georgiatheatre.com.
Make your list
7 p.m. Wednesday, Avid Bookshop, 493 Prince Ave.
Do you have readers on your holiday present list? Are you not quite sure what books to get them? Avid Bookshop has your back this year. Random House Books representatives Sarah Nasif and Toni Hetzel will present mini book talks about the best bets for everyone on your holiday shopping list — that includes you, by the way. Avid Bookshop will have treats from a neighbor, Heirloom Cafe and Fresh Market, and will raffle off a gift basket and Avid gift certificate. Also, Avid will showcase Books for Keeps, a local nonprofit that gives books to those who need them. Ten percent of sales during the evening will go to the charity. Reservations are required and can be made by calling (706) 352-2060. www.avidbookshop.com.


text:
Colchester, Essex United Kingdom (PressExposure) December 04, 2008 -- Tradeplumbing , one of UKâs leading bathroom and kitchen plumbing retailers, has today launched the article section on their website, dedicated to customers and people wishing to improve the look and style of their homes. This section aims at becoming a reliable information source for both end-users and specialists in the plumbing industry.
âEvery person that buys a product for their home would like to have some additional information about their purchase or some practical advice on what to do in order to improve the look of their home. By developing an article section on our website, we aim to help even more of our customers to achieve their dream homeâ, says Peter Clayton, CEO at TradePlumbing.
The first article in the section is called How to hire a Plumber and is a must-read for people considering hiring plumber, giving them the necessary knowledge in order to get the most efficient professionals with the least outlay. The article will help people understand:
- What things they should think about before hiring a plumber - What qualifications a plumber needs to increase credibility - How to make the most of plumbing services in as short time as possible - The services for which a plumber actually charges.
The articles will be free to read on the companyâs official website, http://www.tradeplumbing.co.uk. For further articles, make sure you check the resources section regularly.
TradePlumbing is committed to providing quality knowledge and free access to basic resources & tools for people interested in improving their homes and for plumbing and heating professionals, while continuously updating its product portfolio with stylish, high quality products, perfectly fitted for every bathroom, kitchen or any other room in the house.


text:
Posted 28 Dec 2008
Link to this post
I have a RadGrid issue. My grid binds data via ObjectDatasource and my inserts, updates and deletes are happening in my codebehind. Editing and inserts are contained the the Grids FormTemplate. I use ItemDataBound event to preload two dropdownlists in the formtemplate. When a user clicks "Add a record" the formtempate displays on the first page first row, this is fine. When the user clicks on a record to edit, the formtemplate displays on the last page of the grid at the top. Example: pagesize is set to 25 and we have 70 records and you click on edit on the 4 record on the 1st page, the formtemplate will display on the 3rd page at the top. The grid works as far as insert, editing and deleting just can't figure out why the formtemplate doesn't display where the row was originally. Any suggestions would be great. I'm including code, I stripped out most everything that isn't "relevant", to keep it brief.
.aspx page
Posted 30 Dec 2008
Link to this post


text:
Found 1 products
Detroit Red Wings Retro Brand NHL Women's Nubby Scoop T-ShirtStyle - 20886837Detroit Red Wings Retro Brand NHL Women's Nubby Scoop T-ShirtClearance$36.00 $18.00You Save: $18.00
Added to Your Shopping Cart.
Detroit Red Wings Fan Gear
Shop Detroit Red Wings NHL gear from lids.com. Find officially licensed Red Wings NHL fan gear repping Texas hockey including Detroit Red Wings Hats, Red Wings NHL jerseys and Red Wings Shirts % more. Cheer on the Detroit Red Wings with official team gear for the biggest NHL fans from lids.com today!


text:
GraphicRiver Christmas Web Banners 6339823
Photoshop \ Web Element \ Banners & Ads
Editable \ Christmas \ Layers \ Groups \ Photoshop \ Details \ Deviantart
Web Element - Christmas Web Banners 6339823 by bluenila @ GraphicRiver
Photoshop \ Web Element \ Banners & Ads
- All elements are fully editable.
- All layers are organized in groups.
- 3 different sizes with 4 color schemes.
What you get
1. 2 full layered editable Photoshop psd file.
2. 2 EPS file.
3. Help file.
I have added 2 psd files which are well named and having editable layers, well organized and separated in groups.
Click on the color Name to identify the layer group for that particular color.
Customization .
Photoshop files are fully customizable.
All PSD layers are fully editable and specified into separate groups.
Sources and Credits.
FONT used are : ShowcardGothic, Vani
ShowcardGothic is available to download at http://fontzone.net/font-details/showcard-gothic
Vani is available to download at http://fontzone.net/font-details/vani
- Special thanks to http://camxso.deviantart.com , http://hurleyrocks.deviantart.com for Ornamental Christmas graphics.
Beautiful Christmas Newsletters
High Resolution: No; Layered: Yes; Graphics Files Included: Photoshop PSD, Transparent PNG, Layered PNG, JPG Image, Vector EPS; Minimum Adobe CS Version: CS3.
High Quality Leaves textures; Clean Business Card - 6 colors, 3 different style; Ecommerce/Web button.
Keywords: blue, christmas, discount, festive sale, Google Adwords Banners, grey, holiday, holiday sale, merry christmas, promotion, red, snow, Static Banners, vintage, web banners.


text:
One person is dead and another injured in a shooting at a grocery store parking lot in Jennings, Missouri, Friday night.
The shooting happened on the parking lot of the Aldi store on the 8500 block of Lucas And Hunt Road Friday night at around 7:50.
Police say when they arrived they found 27-year-old Jacobi Bolden shot to death and another victim, a 28-year-old man, with a gunshot wound to the torso.
The victim stated they were shot while sitting in a vehicle on the parking lot. He told police the suspects then stole their car and drove away from the scene. The vehicle was recovered in Riverview.
The incident is under investigation by the St. Louis County Police Department Bureau of Crimes Against Persons.


text:
Nicole T.
Fredericton, Canada, E3C ...
Joined on August, 2013
Gender: Female
Average Rating:
Weekdays in the evening
Weekends in the afternoon
Tues/Thurs in the afternoon
Weekends in the afternoon
Tues/Thurs in the afternoon
Lives Near:
Can Meet:
Up to 15 minutes away for no additional charge
EducationPursuing a Bachelor of Science in Chemical Engineering from University of New Brunswick.
Harbour View High School class of 2012.
ExperienceTook AP Physics and Chemistry in high school.
Achieved high honors upon graduation.
Certified advanced oral French proficiency.
Completed first year of four year degree program.
Enthusiastic, dedicated and loves working with people.
HobbiesSports including swimming, rugby, and field hockey.
Environmental Sustainability - everything from recycling to researching renewable energy sources.
International Development - returning August 27th from a four month volunteer placement in Zambia, Africa working in Business Development
Preferred Subjects
Nutrition, Health, Biology, Chemistry, Science, Geometry, Trigonometry, Physics, Beginner French, ADD, ADHD, Environmental Science
Inquire about this tutor
The tutor will respond if they are available/interested.