text:
Through every season, we are here to serve your every reason for heating and air conditioning needs. Our primary goal at T.A. Kaiser Inc. is to make you feel as comfortable as possible in your home or business so you can focus on what is important to you.
At T.A.Kaiser, we want you to feel assured that you have chosen the most efficient and effective heating and cooling units on the market. Being a family owned business since 1987, we know what it takes to provide you with the best service and product in heating and air conditioning so that you will never have to look again.Our goal is to not just serve you, but be your trusted source for all your heating and cooling needs.
|Business Operation Hours
9:00 AM to 5:00 PM
9:00 AM to
9:00 AM to 5:00 PM
9:00 AM to 5:00 PM
9:00 AM to 5:00 PM
9:00 AM to 5:00 PM
9:00 AM to 5:00 PM
Other Services : Heating Installation, Preventative Maintenance, AC InstallationService Type : Yes, this business serves customers at their Location
HVAC Services
The highly trained technicians at T.A. Kaiser are ready to handle any your of your home comfort needs. Our technicians are the best in skill, attitude and workmanship, with experience in all areas of residential hvac. They will care for your home and complete the job with speed and precision.
We deliver on our satisfaction guarantee through constantly training our technicians on the latest service technology available. We teach all of our technicians to provide reasonable solutions to our customers heating and air conditioning problems. Also, prior to employment, all employees are required to go through a third party background check and pre-employment drug test.
T.A. Kaiser is a full service residential heating and air conditioning company. Whether your needs are for a new system or just maintenance on an existing one, give us a calland let us help make your home more comfortable! You can also Set up an appointment online for assistance with your residential hvac needs.
Languages SpokenEnglish
Payment OptionsVisaMaster CardCashAmerican ExpressCheck
Year Established1987


text:
Forums > General Industry > Model Scam" Here is my story"
Posts: 14
Corpus Christi, Texas, US
first of all, im a Bodybuilder competitor/personal trainer for 3 year . i'm also male stripper every sat night. Being run way model alway be my dream but im no tall enuf. Living in little Apt with a chihuahua dog name snowy. On jan 4 2013 i looking around craglist , i saw this guy have his own website casting model. i email him, we talking by email. he will fly me out to do photo shoot in Sounth American. I trust him. so feb 5 i recive a check "6000 dollars". i have to cash out and send to these adress. 2 hours after i sent the money. i notice the check is bounce and my friend telll me that i got a scam. so i call wester union to cancle everything. before the real money got into thier hand. it was "Chaos". Just be safe out there. God bless!!
Mar 23 13 01:53 am Link
Posts: 14671
Los Angeles, California, US
This is a classic scam. Please read up on things you can do to avoid this sort of thing in the future! But no one ever sends money before you do work in this industry. Heck, sometimes I have to wait 3 months to be paid for agency work!
Check out this link:
http://www.modelmayhem.com/education/us … the-basics
I'm glad you got the Western Unions cancelled in time! You are luckier than many.
Mar 23 13 02:00 am Link
Posts: 1654
West Hollywood, California, US
K I C K H A M wrote:
Mar 23 13 02:02 am Link
Posts: 10604
Los Angeles, California, US
Anh hoang wrote:
Why would you accept FUNDS..before the job was finished? Who does this? At Mr. Olympia Contest..Does Joe Weider hand out money...before the Bodybuilding Contest? NO!!!
Posts: 10604
Los Angeles, California, US
Anh hoang wrote:
....and Paul...Living in LA...I have been to plenty of Runway Shows...and not once...have I ever seen a 5'7 Model..Male or Female!!! lol
Posts: 14
Corpus Christi, Texas, US
K I C K H A M wrote:
Mar 23 13 07:07 pm Link
Posts: 14671
Los Angeles, California, US
DOUGLASFOTOS wrote:
I know two 5'7 models that are in almost every LAFW. But you don't KNOW they are 5'7. Oh, and they are chicks, too. So, that makes a difference.
Mar 24 13 08:16 am Link

subset: commoncrawl


text:
Bio-medical Research
Unlocking the Mechanism of tRNA Translocation Through the Ribosome Using Large-Scale Molecular Simulation
Karissa Sanbonmatsu, NMC Affiliated Research Scientist, LANL Staff Scientist
The ribosome is one of the largest and highly conserved molecular machines, whose function is essential to all life. Its operational principles, however, remain largely a mystery. Among the myriad of steps catalyzed by the ribosome during protein synthesis, translocation is considered the 'holy grail' by many. Translocation is an intrinsic property of the ribosome, involving complex conformational changes of the two ribosome subunits that move the mRNA and tRNA substrates by precisely three nucleotides in a unidirectional fashion. Recently, studies demonstrated that the majority of translocation occurs during head swivel (intra-subunit rotation within the small subunit). Understanding the connection between head swivel and mRNA/tRNA movement will unlock a key operational principle of translocation. While there is growing consensus that head swivel is crucial to translocation, a direct relationship between these conformational rearrangements and the translocation reaction coordinate is lacking. We will use closely integrated molecular simulation and single-molecule FRET imaging studies to bridge this gap. Development of Attenuated HCMV/HIV Vectors for an HIV/AIDS Vaccine
Discriminative Biomarker Assays for Detection of Shiga toxin-producing Escherichia coli
A Rapid Nucleic Acid-based Assay for the Detection and Characterization of E. Coli.
Shiga toxin-producing E. coli (STEC) are a serious threat to our food supply and public health, causing more than 265,000 infections each year. NMC participates with UNM on a large USDA grant that seeks to understand how E. coli pathogens travel throughout the beef production process and how outbreaks occur. The program seeks to find ways to prevent illness and improve the safety of our nation's food supply. The NMC/UNM team is applying LANL developed technology to develop and implement rapid detection technologies for pre-harvest, post-harvest and consumer environments. The team has developed a rapid nucleic acid-based assay for the detection and characterization of STEC in the beef chain. This assay is the multiplex oligonucleotide ligation-PCR (MOL-PCR) and unlike other ligation-based assays that require multiple steps, MOL-PCR consists of a single tube reaction.
Development of Novel Vector / Protein HIV-1 Vaccines
This program develops novel vector/protein HIV-1 vaccines by combining the vaccine platform technologies of Beth Israel Deaconess Medical Center (BIDMC) and GlaxoSmithKline Biologicals (GSK). The combination of BIDMC and GSK vectors, proteins, adjuvants, and expertise will result in a new academic-industry partnership that will rapidly translate innovative HIV-1 vaccine product concepts into clinical trials. The goal of this program is to define an optimized vector/protein HIV-1 Env vaccine with immediate clinical development potential. The project hypothesizes that priming with a novel adenovirus (Ad) vector and boosting with a stable Env trimer optimized for immunologic coverage of global virus diversity and formulated with the GSK AS01B Adjuvant System will induce potent and protective Env-specific antibody responses. The Ad vector priming will prove superior to poxvirus vector priming for a subsequent Env protein boost. This research will optimize each component of this vaccine regimen systematically in a logical, focused product development program involving a series of nonhuman primate challenge studies.
A High Throughput Pipeline to Select Renewable Recombinant Polyclonal Antibodies
This joint LANL/NMC research program develops high throughput affinity reagent selection and screening capabilities. In this project, the team is creating new methods to select and screen for antibodies. The research creates new antibody libraries, develops new selection methods using flow cytometry and explores novel ways to use these antibodies.
From the End to the Beginning: Termination and Ribosome Recycling in Eukaryotic Translation
Protein synthesis research is important to understanding the origin of many human diseases and devising treatments for them. This project examines a poorly understood step in protein synthesis: the structure of intermediate states in termination and eukaryote ribosome recycling. The research will model the transition between these intermediate states.


text:
Donate TODAY by clicking HERE!
Why Give?
Consider the numbers: There are more than 160,000 stray dogs and cats in San Antonio. The Animal Defense League saved more than 5,000 homeless pets in 2015, and performed more than 6,500 spay/neuter surgeries as part of the ongoing effort to solve this critical problem. Donating makes you part of the solution as we strive to save more lives going forward. As San Antonio's oldest no-kill shelter we continue to lead the city towards its live-release goals, but we need your financial support to make it happen.
As a 501(c)(3) nonprofit organization, ADL relies on the generosity of its supporters to operate and provide food, shelter, and medical care to the hundreds of homeless pets in our care. Your donation has a specific and immediate positive impact on the lives of these animals. Please consider making a donation today!
All donations are tax deductible and we have made available our 501c3 Letter and ADL W-9 form for download. Please be sure to also view our 2013 Audited Financial Statements. To request a copy of ADL's 2013 990, please send your request to Janice Darling, ADL Executive Director.
The Animal Defense League of Texas will never share or sell a donor's personal information with anyone else, nor send donor mailings on behalf of other organizations.
Ways You Can Support with a Donation:
How can I donate?
- MAIL your donation to 11300 Nacogdoches Rd., San Antonio, TX 78217.
- CALL 210-655-1481, ext. 117 to make a credit card donation by phone.
- FAX a completed donation form to the attention of Samantha Lee at 210-655-4489.
- Donate securely ONLINE using GiveDirect.
- VISIT the Adoption Center or ADL Hospital to make your donation in person.
- DROP OFF pet related items that help us care for our resident animals at our "Donation Station" located in the adoption center parking lot. Drop off daily from 11am-7pm. We accept the following items: newspaper, cat & dog toys, towels, cleaning supplies, pet supplies, etc.
- Shop from our Amazon Wishlist.
What kind of donations can I make?
- General Donations : A general donation will go into a general fund to provide daily care of our resident animals as well as ongoing operations.
- Tributes : Consider donating to ADL as a gift in memory or in honor of a special person or even a pet. A notification of your gift can also be sent to a recipient of your choice. Click here for a printable Tribute Donation Form.
- Sponsorship : Consider sponsoring a dog or cat of your choice! Our sponsorship program is a way for you to make a connection with a specific homeless pet in need and provide and advocate for it, even if you don't have the ability to adopt at this time. Click here for more information on our sponsorship program!
- Bequests : Bequests are excellent sources of support that guarantee ADL's ability to continue to rescue abused and abandoned dogs and cats in the future. A bequest can consist of money, securities,works of art and/or real estate. Please consider including ADL in your will. We also encourage you to speak with your financial advisor. For more information on making a bequest to the Animal Defense League, please contact Anne Meyer at 210-655-1481, ext. 111 or by email at firstname.lastname@example.com.
If you have any questions regarding making a donation to the Animal Defense League of Texas, the Development staff will be happy to assist you by phone at 210-655-1481, ext. 117 or by email at firstname.lastname@example.com.


text:
I've never read A Monster Calls, but if the traditional ratio of quality from books to movies is any indication, the film has inspired me to pick up the children's book. For those who never read the book, I won't spoil it. Having come fresh eyed from the film, I felt the journey was easily the most important element the story had to offer. There's a reason why it unfolds the way that it does. The visual style is also a very important element of the storytelling process. It may be easy for some to dismiss it as a failed attempt to integrate CGI into the live action, but those not too pretentious to see the film for the visual choices it makes to serve the story will find enjoyment. Every actor on screen gives the best performance possible, playing their characters as the subtle, complex people the story requires, recognizing that no one is purely any one thing but rather a sum of good and bad qualities, attempting to navigate a complicated world the best they can. To be sure, their performances are subtle, sometimes requiring a very keen eye to see the distinctly internal struggles that the film chooses not to hit the audience over the head with. Those facets of their performances, left unnoticed, will make or break anyone's opinion of the quality of acting, or even the effectiveness of the story. It's a bold move to leave that much of the success of the film in the hands of the audience to see what they aren't being directly told, and I can't say for sure if the movie will suffer for it, but I appreciate that they're willing to trust us in a way most filmmakers refuse to. They left many layers to this film, many of which will go undiscovered if it isn't properly scrutinized, but those types of choices are the kinds that put authors into the halls of literary canon, and in those ways I think they succeeded most of all.
I'm going to break one of my cardinal rules I set out in the beginning, but I feel strongly enough to make a recommendation. Watch this movie. Make time to watch this movie. If you'll only be able to watch two movies this year (who are we kidding? Star Wars: Episode VIII is coming out this year), A Monster Calls needs to be the other one.


text:
I've finally gained some ground this month! Not only did I launch a full on attack of the children's rooms, but I finally began weeding through our storage shed. Our storage shed is packed plum full of STUFF! In the past when I was done with something, or the kids outgrew clothes, I packed them away and stored them. Absolute foolishness on my end. There will be no more of that in the future. I'm so tired of storing stuff, and hanging onto things I don't need and/or want any longer.
Here's what left our home this month, either by way of trash or donation:
*Please forgive me, I thought I had snatched a picture of everything before I dropped it off, but I can't seem to find it now.
3 Awards w/frames
Box of Jr. High/HS Report Cards –And please tell me why I had these for so long! My 10 year HS Reunion is this year!
Alarm Clock
Horse Poster
2 Pillows
Corner Shelf
Apple Corner Shelf
1 Belt
2 Doilies
Fajita Cooker
2 Mattress Pad
2 Rugs
3 Stuffed Animals
2 Dale Earnhardt Bears
2 Glass Flowers
Dishwasher Basket–Not sure why I had this either!
3 Baskets
Broken Basket
Apple Salt Shaker
Apple Towel Holder
Glass Apple Bowl
Plastic Canvas Apple Basket
Wooden Apple Basket
7 Random Apple Decorations
Fake Flower Boquet
3 Blankets
Glass Casserole Dish
Plastic Shelf
Plastic Onion Cooker–I have no idea where this came from.
Wooden Patio Spinner Decoration
3 Flat Sheets
Tin Planter
Boy Shadowbox
174 Clothing items (!)
4 coats
5 pairs of shoes
12 books
Phonic Book/Letter Block Set
Letter Match Game
Kids Video Recoder
Mini Train Set
Travel Train Set
Car Carrier for Dolls
Barbie Hair Styling Head
Plastic Play Table
Kleenex Cover
Wool Blanket
Napkin Holder
Set of Drapes
Swiffer Sweeper
Toy Car
2 Dolls
2 Purses
2 Jewelry Cases
Broken Apple Plate
Wisteria Flower Swag
Barbie Tackle Box
Bag of Glow in the Dark Stars
2 Baby Gates
Metal Standing Shelves
Metal Corner Standing Shelves
Noah's Ark Decorations:
- Pillow
- Diaper Stacker
- Crib Sheet
- Picture in Frame
- Figurine
- Book Ends
- Clock
- Stool
- 2 Wall Hangings
Car Seat
Plastic Jungle Gym
Plastic Bag Holder
295 Items Total!! Add that to my previous total for a grand total of 369 items! I'm more than half way toward my goal–only 361 items left.
Be sure to check out everyone else's progress over at Amy's Finer Things.


text:
WIN! A DeLonghi Kettle and Toaster with Wayfair.co.uk
Top home ware website www.wayfair.co.uk is giving one At Home reader the chance to win a stylish Delonghi Kettle and Toaster Faceted Set in black, worth £159, to suit all decors in all homes.
These two high quality products boast a black polished, faceted finish with chrome details for sheer design luxury. The kettle has a 360° swivel base, an anti-scale filter and comes with a detachable base for cord-free convenience. The toaster has four-slice capability; reheat, defrost and cancel mechanisms and is non-slip. Not only are they chic looking kitchen must haves but their first rate quality means they're guaranteed to last for years in any home.
To find a zillion other great home ware products at competitive prices, head to www.wayfair.co.uk.
For your chance to win the Delonghi kettle and toaster just fill in the form below:
Terms & Conditions
- Closing date for this competition is 15th June 2013.
- This offer is only open to UK residents.
- For standard EMP Competition terms and conditions click here
Please note: When you enter our competitions you are also opting in to the At Home email newsletter (you can unsubscribe at any time) and email and mail communications from the competition prize provider. Your phone details should only be used for prize notification purposes by the competition prize providers as they are only given to them in that capacity.
Click here to find out what other competitions at home magazine is running
Keep up to date with our latest competitions and special offers by following us on Twitter and joining us on Facebook
**This competition has now closed
Related posts
Comments (2)
sharon wormald
Very funky!!


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Blatantly Political Request Also Marks Height of Desperation from Kelly Ayotte, Whose Own Travel Has Gotten Her in Hot Water
Concord, N.H. – New Hampshire Democratic Party Chair Ray Buckley issued the following statement on the Right to Know request filed by Jennifer Horn while traveling overseas on the dime of noted hate groups:
"Jennifer Horn issued what may be the most ironic Right to Know request of all time while traveling overseas on a trip funded by extremist hate groups that has come under fire from organizations across the political spectrum. The blatantly political request also marks the height of desperation from Kelly Ayotte, whose own travel has gotten her in hot water as she exploits 'loopholes' to hold expensive Utah ski weekends paid for by lobbyists and corporate interests."
Jennifer Horn Traveling Overseas with Hate Groups Known for "Incendiary and Bigoted" Language
New Hampshire Republican Party Chairman Jennifer Horn is touring Israel with this week with other members of the Republican National Committee courtesy of two evangelical groups that have come under fire for controversial language directed at gays, Mormons and Muslims. Sponsoring the trip, reportedly for about 60 RNC members, is the American Family Association and the American Renewal Project. Politico reported today that about a week ago, officials at the Anti-Defamation League privately reached out to the RNC saying it would be "inappropriate" for RNC members to travel to Israel with members of those groups because they are known for "incendiary and bigoted" language. [NH Journal, 2/3/15]
Criticism of Horn's Trip "Is Spreading Across the Political Spectrum"
"Now, criticism of the trip, which has been mounting for days among left-leaning groups and journalists, is spreading across the political spectrum. In interviews with POLITICO on Monday, evangelical leaders and right-leaning groups said the national party showed poor judgment at best in associating with "extremists," as evangelical Alan Noble described the groups. AFA has been labeled a "hate group" by the Southern Poverty Law Center — a description it has disputed." [Politico, 2/3/15]
Ayotte Under Fire for Exploiting "Loophole" that Allows Lobbyists and Corporate Interests to Pay for "Destination Fund-Raisers"
"New Hampshire Sen. Kelly Ayotte's campaign finance chairman and spokesman defended her fund-raising practices Monday after she was mentioned prominently in a weekend report on a "loophole" in a gift-ban law that allows lobbyists and corporate interests to indirectly pay for "destination fund-raisers," where they then mingle in exclusive vacation spots with members of Congress. Republican Ayotte, according to The New York Times, "kicked off the new year" at the Park City, Utah, ski resort, where she held an event for her campaign committee at the "multimillion-dollar home of Gordon Smith," a former senator from Oregon who is now a lobbyist and chief executive of the National Association of Broadcasters. The association, according to the report, "has a long list of high-stakes matters before the Senate Commerce Committee," on which Ayotte sits as a member." [Union Leader, 1/20/14]


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text: Hi all, OK, trying to fill my time this afternoon as the weathers **** & I'm nursing a slight hangover!! Before the new S3 arrives (looking like the 1st march now!), thought I'd check out some insurance quotes. Vital stats are: 30, 6yr's no claims protected, 3pts SP30 (******* speed cameras!), £250 voluntary excess, living in devon, std S3 (with extras obviously but no 30" alloys or blue under body neons!!). Std alarm, no tracker or anything. Best quote I've found so far was with Tesco for £448.35. Anyone in the same sort of bracket able to better that? And if so, please share the wealth!! Cheers Ivan


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RExPrimer: an integrated primer designing tool increases PCR effectiveness by avoiding 3' SNP-in-primer and mis-priming from structural variation
Eighth International Conference on Bioinformatics (InCoB2009): Computational Biology
Asia Pacific Bioinformatics Network (APBioNet) Eighth International Conference on Bioinformatics (InCoB2009)
Polymerase chain reaction (PCR) is very useful in many areas of molecular biology research. It is commonly observed that PCR success is critically dependent on design of an effective primer pair. Current tools for primer design do not adequately address the problem of PCR failure due to mis-priming on target-related sequences and structural variations in the genome.
We have developed an integrated graphical web-based application for primer design, called RExPrimer, which was written in Python language. The software uses Primer3 as the primer designing core algorithm. Locally stored sequence information and genomic variant information were hosted on MySQLv5.0 and were incorporated into RExPrimer.
RExPrimer provides many functionalities for improved PCR primer design. Several databases, namely annotated human SNP databases, insertion/deletion (indel) polymorphisms database, pseudogene database, and structural genomic variation databases were integrated into RExPrimer, enabling an effective without-leaving-the-website validation of the resulting primers. By incorporating these databases, the primers reported by RExPrimer avoid mis-priming to related sequences (e.g. pseudogene, segmental duplication) as well as possible PCR failure because of structural polymorphisms (SNP, indel, and copy number variation (CNV)). To prevent mismatching caused by unexpected SNPs in the designed primers, in particular the 3' end (SNP-in-Primer), several SNP databases covering the broad range of population-specific SNP information are utilized to report SNPs present in the primer sequences. Population-specific SNP information also helps customize primer design for a specific population. Furthermore, RExPrimer offers a graphical user-friendly interface through the use of scalable vector graphic image that intuitively presents resulting primers along with the corresponding gene structure. In this study, we demonstrated the program effectiveness in successfully generating primers for strong homologous sequences.
The improvements for primer design incorporated into RExPrimer were demonstrated to be effective in designing primers for challenging PCR experiments. Integration of SNP and structural variation databases allows for robust primer design for a variety of PCR applications, irrespective of the sequence complexity in the region of interest. This software is freely available at
Polymerase Chain Reaction (PCR) is a common laboratory technique in biological and medical sciences, with a wide range of applications such as DNA cloning, DNA resequencing for single nucleotide polymorphism (SNP) discovery and quantification of gene expression. In the design of any PCR experiment, the first step of designing oligonucleotide primer pairs is crucial for the success of the experiment. Selection of inappropriate primers can result in no amplification (PCR failure) or amplification of non-targeted regions (mis-priming). Therefore, the primer pair is tailored to be specific to the desired target sequence.
The design of target-specific primers for PCR experiments typically requires consideration of different types of genomic information besides the target DNA sequence, such as repetitive DNA elements, intron/exon boundaries, and SNPs, which must be retrieved from various databases. All information is then combined to construct a sequence template for a primer design program. To confirm their specificity, designed primers are usually aligned against the corresponding genome sequence using tools like BLAST
To resolve this situation, a number of automated primer designing tools have been developed based on Primer3
While these programs provide some solutions related to the aforementioned primer design process, they are not always able to effectively design primers for two main problems, namely 1) no amplification due to severe mismatching, or lack of target and 2) mis-priming (non-specific binding besides the target sequence). These two problems lead to increased PCR failure rate
To address these issues, we present a graphical web-based tool, named RExPrimer, which allows users to automatically design PCR primer pairs for amplifying human genomic sequence without leaving the website. RExPrimer uses Primer3 as the design core, since this open source software has been continuously adopted by research communities as the
Input processing
RExPrimer offers three modules of primer design: 1) for resequencing genomic DNA (promoter, exon/intron boundary, any genomic region), 2) for SNP genotyping (gene based and region based), and 3) for oligonucleotide checking. The following types of identifiers are supported as input: HUGO gene name, NCBI Gene ID, and chromosomal locations. Since there are also other interesting non-gene regions, such as regulatory regions and intergenic regions, our program also supports arbitrary genomic regions as input based on chromosome location. For the SNP genotyping module, the program allows one additional input format as a SNP ID (rs-number) or set of SNP IDs. Because this application supports batch design for SNP genotyping primers, it can enhance productivity of high-throughput SNP sequencing projects.
After the input is received, the local human genome database module is interrogated and the corresponding target sequence information, including the genomic sequence, annotations (e.g., promoters, introns, exons) and polymorphisms is subsequently identified and retrieved using query language. When a gene name is used as an input, more than one associated sequence, such as splicing variants may be found in the database. In this case, users have an option to choose the desired isoform. Then, the program supplies the option of selecting the gene regions to be amplified: whole gene (only exons or all gene regions), region of interest (specified by intron/exon number e.g. from intron/exon x to intron/exon y, list of non-contiguous intron/exon).
Non-unique PCR primers are one of the main factors that lead to PCR failure. Multiple regions of homologous, or near-homologous sequence in the genome can compete with the target sequence for primer hybridization leading to PCR artifacts. The human genome contains many pseudogenes, which can have near-perfect homology to their functional counterparts. To avoid designing non-specific primers for genes with pseudogene counterparts, the input sequence's pre-defined pseudogene sequences are retrieved from the pseudogene database module. Multiple sequence alignment of these pseudogene sequences and target gene including alignment scores by the MUMmer program
Figure 1
Detection of
Detection of
After all of the target DNA sequence information is retrieved from the local database module, it is passed as input to Primer3 for the automatic generation of primer pairs. The RExPrimer user interface allows the users to specify parameters for Primer3 primer selection, such as product size, melting temperature (Tm), GC content. If desired, each designed primer could be screened against a repeat database to reduce nonspecific priming by choosing this available option. The amplicon size is user-defined according to the constraints of the experiment, e.g. accurate sequencing limits amplicons to a few hundred base pairs. If the input sequence is larger than a user-specified product size limit, the program automatically subdivides the template sequence into smaller segments with user-defined segment overlap size in order to ensure that the overlapping sequences can give high quality sequencing data. Primers are then designed separately for each overlapping fragment.
Sequence variation in the 3' end, particularly the last three bases can severely disrupt primer hybridization and thus decrease PCR efficiency to the point of PCR amplification failure
Figure 2
The SVG graphical output displaying resulting primer pairs with other genomic features
The SVG graphical output displaying resulting primer pairs with other genomic features. The designed primer pairs are represented by double-headed arrows along with gene structure (intron/exon) and other genomic features (pseudogenes, indels, CNVs, etc.). Pseudogenes and CNVs/indels are shown as multiple sequence alignments with the target gene. SNP-in-Primers are demonstrated with different colors according to the degree of destabilizing effects introduced by such SNPs. Green color presents primer without SNPs inside. Blue color represents primer with SNP in any position but not within 7 bp of the 3'end while red color specifies primer with SNP found within 7 bp of the 3'end. Each designed primer pair can be linked-out to a redesign module.
Output report
Results reported from RExPrimer include two parts: SVG graphical display and HTML primer information summary. The graphical representation displays resulting primer pairs with their respective locations in the target gene in the same window (Figure
Figure 3
HTML report of designed primers
HTML report of designed primers. The designed primer sequences are shown with assigned PCR conditions. Redesign module and uniqueness test by In-Silico PCR are provided as the links from this report.
When strong primer-destabilizing effects such as SNP-in-Primer are reported, users can immediately redesign primers without repeating the same procedure again. Optionally, the users can directly specify the approximate range of primer or target region, e.g., to avoid regions of high SNP density. The redesign of primer pairs can then be forced to a confined user-specified region.
Case study: CYP2D6
To validate whether RExPrimer is effective in designing primers in challenging PCR applications, RExPrimer was used to design primers for the Cytochrome P450 2D6 locus (
• Pseudogenes (
• CNVs from unequal crossing-over
• SNPs reported to influence the enzyme activity
Existing primer design tools could have failed to consider the effect from
In this study, primers were cautiously designed by considering every possible problem mentioned above using RExPrimer. The primers designed by RExPrimer for the
Table 1
The designed primers for
Sequence 5' → 3'
whole gene amplification
penta-plex PCR
penta-plex PCR
penta-plex PCR
one-copy internal standard in penta-plex PCR
two-copy internal standard in penta-plex PCR
Rows 2-6 represent semiquantitative analysis of CNV
Figure 4
PCR amplification of
PCR amplification of
Figure 5
Semiquantitative analysis of
Semiquantitative analysis of
In this study, we have developed a comprehensive tool for PCR primer design which covers a broad range of functionalities including resequencing target genes and SNP genotyping. The primer design pipeline is composed of three key components: 1) pseudogene detection, 2) primer design core, and 3) SNP-in-Primer/genomic variation notification. Additional file
Additional file 1
The feature comparison of existing primer designing software. The table displays the features provided in currently available primer designing software compared with those present in RExPrimer program. The cells with bold italic letters indicate unique features offered by RExPrimer.
Click here for file
RExPrimer utilizes the publicly available sequence information including the human genome and annotation database, SNP databases, and the pseudogene database. By incorporating these databases and the Primer3 program, reliable and accurate primer designs can be achieved. The current pseudogene database module comprises of approximately sixteen thousand pseudogenes
Most primer design tools verify the uniqueness of the PCR target sequence by using UCSC In-Silico PCR
RExPrimer appends several key features before and after the primer design process using Primer3, enabling the selection of unique primer sequences and reliably amplifiable targets, which other currently available software cannot match. However, with the caveat that the primers designed are unique and the targets are amplifiable (no CNV or SNP-in-Primer), no extra claims for the actual performance of the primers beyond what is predicted by Primer3 are made. Hence, we have not attempted to determine the success rate of RExPrimer for designing primers, since the cost of performing multiple PCR experiments is not justified. Finally, RExPrimer has a major strength on its graphical web interface, especially the gene structure visualization, which makes RExPrimer intuitive and user friendly.
Future development
Currently, RExPrimer can offer the oligonucleotide primer design for human genomic sequence in which SNP and pseudogene data are largely available. In the future, the program and locally built databases will be expanded to support primer generation for different organisms once their pseudogene and polymorphism data are readily available. To enhance the quality of primer design, the local SNP databases will be automatically updated to make use of the latest information, including as wide a range of population diversity as available.
RExPrimer is a one-stop tool for PCR primer design, which can support high-throughput resequencing and mutation screening research. The incorporation of large SNP and genomic variation databases make it possible to efficiently detect sequence variation in the designed primers that might cause PCR failure. The notification system of target pseudogenes before the primer design step helps users to select the appropriate target regions, which can significantly shorten the design process. RExPrimer is shown in this study to be indeed effective for designing primers for
RExPrimer is a web-based application that was written in Python language. The web interface was created by Python Webware framework
Figure 6
The organization of RExPrimer
The organization of RExPrimer. The workflow of RExPrimer following the steps described in methods section is displayed.
1. Users provide input parameters required to design primer pairs such as gene name, SNP ID, genomic location.
2. The input parameters are used to query the targeted sequence from the local human genome sequence database, which was retrieved from NCBI and stored locally on our server.
3. The target sequence is checked using MUMmer
4. RExPrimer prepares the input file for Primer3 from the primer conditions and target sequence. If the target sequence is greater than the target size parameter set in Primer3, the system will construct a set of overlapping fragments that covers the entire target sequence.
5. Primer3 processes the input files prepared in step 4 and generates the primer pair results in Primer3 format.
6. The primer pair results are then crosschecked against different locally constructed SNP and genomic variation databases covering a wide range of human SNP variation, CNV, and indel polymorphisms.
7. Primer pairs are visualized on the locus-specific region using scalable vector graphics (SVG). The software also provides primer information in HTML format (see Figure
8. Each resulting primer pair can be validated for uniqueness using local In-Silico PCR (see Figure
RExPrimer stores public sequence information and genomic variant information in order to accelerate the speed of processing and allow the user to seamlessly unify the required information used in RExPrimer. These databases are hosted on MySQLv5.0, which offers several important services to RExPrimer. The human genome sequence database module was downloaded from NCBI build 36.3 to be used as template sequence as well as providing gene organization information. The SNP database module comprises of common and population-specific SNPs from various databases, namely NCBI dbSNP
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
JP wrote the manuscript and designed RExPrimer. CN developed RExPrimer and wrote the manuscript. AA wrote the manuscript, designed RExPrimer and
Other papers from the meeting have been published as part of
The authors would like to thank Dr. Chanin Limwongse for supporting with
This article has been published as part of
Basic local alignment search tool
BLAT--the BLAST-like alignment tool
Primer3 on the WWW for general users and for biologist programmers
SNPbox: web-based high-throughput primer design from gene to genome
ELXR: a resource for rapid exon-directed sequence analysis
ExPrimer: to design primers from exon--exon junctions
MutScreener: primer design tool for PCR-direct sequencing
EasyExonPrimer: automated primer design for exon sequences
PrimerZ: streamlined primer design for promoters, exons and human SNPs
UCSC In-Silico PCR
Predicting failure rate of PCR in large genomes
Global variation in copy number in the human genome
Detection of large-scale variation in the human genome
Versatile and open software for comparing large genomes
Minimal homology requirements for PCR primers
Amplification-refractory mutation system (ARMS) analysis of point mutations
The human debrisoquine 4-hydroxylase (CYP2D) locus: sequence and identification of the polymorphic CYP2D6 gene, a related gene, and a pseudogene
Denaturing HPLC coupled with multiplex PCR for rapid detection of large deletions in Duchenne muscular dystrophy carriers
Pseudogene.org: a comprehensive database and comparison platform for pseudogene annotation
Webware for Python
dbSNP: the NCBI database of genetic variation
A haplotype map of the human genome
JSNP: a database of common gene variations in the Japanese population
ThaiSNP database


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The 50 m (164 ft) luxury motor yacht LUMIERE comfortably accommodates 12 guests in 6 luxurious cabins and is operated by 11 experienced and dedicated crew.
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Jeff's Bronco Graveyard - BAK Tonneau Covers in Brighton, MI
Jeff's Bronco Graveyard
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Phone: 248-437-5060
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Matt Masotti offers an interesting article on Microsoft Dynamics CRM and how it makes enterprise architecture more important than ever. Masotti is a Senior Consultant, Business Analysis, CRM at Avanade. No longer is, "Microsoft Dynamics CRM implementation just about whether or not the software had the capabilities to support business requirements."
Enterprise architecture is sometimes viewed as an expensive and overly abstract discipline in the context of a Dynamics CRM implementation. I would argue that today, even smaller organizations benefit from an initial assessment of when CRM is and is not going to be the appropriate answer when considering it as part of an overall technology strategy.
Masotti concludes with:
Starting with a completed enterprise architecture assessment and a well-aligned CRM strategy is not new advice, in fact it has always been true and is a regular subject of discussion in the CRM world – especially when you consider that more than half of today's buyers will pay more for a better customer experience. Today, however, when CRM is likely to have the potential for a larger technology footprint in your organization, EA is more relevant than ever.
Check the video below from Microsoft Dynamics CRM to find out more. It offers an example of Microsoft Dynamics Intelligent Customer Engagement in use:
Read the article by Matt Masotti at MSDynamicsWorld.com.


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|About IPO
Lincoln's Changing Views on Slavery
Jamie Schaefer
Abraham Lincoln, the sixteenth president of the United States, entered office during a critical period in American history. During his presidency he dealt with the cohesiveness of the Union and slavery. Lincoln had formulated opinions of slavery from the time he was young; he grew up with slavery, and had the opportunity to determine whether slavery would be upheld or abolished while he was president. As a political leader, Lincoln's views about slavery changed as he had to worry about the Union and all of the people within it. No longer could he view slavery as he had in his earlier life.
Lincoln's views on slavery can be attributed to childhood and family experiences. There are two factors that are part of why Lincoln formulated anti-slavery thoughts and feelings. Lincoln and his family were very religious. They were of the Separate Baptist faith. Many of the Separate Baptists during this period opposed slavery. Ill feelings toward slavery also came from growing up in Kentucky in the early 1800s. During the early 1800s, Kentucky's land was not surveyed, but was settled at random. This caused a number of land disputes between the larger farmers, who typically utilized slave labor, and the smaller farmers who did not utilize slave labor. Lincoln's father, Thomas, and other small farmers, began to have problems with land disputes and claims that had to be settled in court. Due to a lack of money, many of the small farmers decided they could not afford the legal expenses and were forced to leave Kentucky. Thomas and his family moved to Indiana as a result of being put out of business by wealthier landowners.
Even though Lincoln did not approve of slavery throughout his life, he realized it would be improbable that blacks and whites could live with equality, since they had to deal with too many prejudices. During the Lincoln-Douglas debate at Charleston on September 18, 1858, Lincoln stated:
It was early in this debate that Lincoln was tested on questions dealing with the admission of additional slave states into the Union, the repeal of the Fugitive Slave Act (which required that all slaves who escaped to northern states must be returned), and the abolition of slavery in the District of Columbia. He answered that he was not in favor of repealing the Fugitive Slave Act, he was not pledged against the admission of another slave state into the Union, and he did not at that time favor abolition of slavery in the District of Columbia. However, toward the end of the debates, Lincoln started to convey his feelings regarding slavery. He stated that the inferior races were equal in their right to life, liberty, and the pursuit of happiness, but he also knew that it was impossible to produce a perfect social and political equality between black and white races.
Lincoln had a number of ideas on how to tackle slavery. He suggested setting up a black colony in Central America in 1863 so that blacks could seek rights and freedoms where prejudice did not exist. He also thought colonization was the best solution. Many people, including blacks, did not believe colonization was a viable solution and believed it would never be carried out.
The Emancipation Proclamation (1863) stated
Fugitive slaves often successfully escaped to free states, which was no guarantee of their permanent freedom.
that slaves in the seceded states were free. Lincoln knew that he could not declare total emancipation: hence, he decided this was the only legal action he could take under the Constitution. This proclamation was designed to maintain a balance in the nation so the border states, which had slavery, but were still part of the Union, would remain loyal. Lincoln knew that if he made a move toward immediate emancipation it may cause the border states to secede. Instead of ordering a dramatic form of emancipation in the border states Lincoln decided to urge the states' governments to accept a program of gradual emancipation. Because Lincoln was unsure of the legality of his proclamation and whether it would be sustained by the Supreme Court, he strongly urged Congress to adopt the Thirteenth Amendment abolishing slavery throughout the nation. Lincoln insisted that this was necessary for the restoration of the nation after the Civil War.
Lincoln was concerned about freeing the slaves, but he also stated, "My paramount objective in this struggle is to save the Union, and it is not either to save or destroy slavery. If I could save the Union without freeing any slaves, I would do it, if I could save it by freeing all the slaves I would do it." This shows that Lincoln's primary objective at that time was to save the Union, because he did not believe it should be split by an issue such as slavery. Even though he did not approve of slavery, it did not mean that he wanted to abolish slavery.
The issue of slavery was a complicated one that Lincoln had to deal with his entire life. From his childhood to his later years, his views on slavery were altered. Lincoln did not necessarily change his views on slavery, but rather his ideas and policies changed from compensation and gradual emancipation to immediate abolition during his political career.—[From Roger Bridges and Rodney Davis, eds., Illinois: Its History and Legacy; David Herbert Donald, Lincoln; John Hope Franklin, "The Emancipation Proclamation: An Act of Justice," Quarterly of the National Archives; Fred Kerner, Lincoln: A Treasury of Lincoln Quotations.]


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Nelson Mandela
January 15, 2011 by USA Post
Nelson Mandela, Last Sunday, in an opinion piece, Budalang'i MP Ababu Namwamba, gave interesting examples of successful leaders years.
The Parliamentary Committee on Justice and Legal Affairs quoted President Abraham Lincoln, President Kibaki, Abdoulaye Wade of Senegal, Jaramogi Oginga Odinga and Nelson Mandela"en years as leaders who contributed greatly to various political and historical achievements.
He argued, if a leader is good enough, it is quite old and the young generation or the change should not be a key factor in the search for leadership.
Let me start by saying all the great leaders start young. In Kenya, leaders like Kibaki, Raila Odinga, John Michuki, Kenneth Matiba and Tom Mboya all began public life while young. They made their greatest achievements in the days before.
Second, those who begin well does not always end well, as is the case with Robert Mugabe. I would like to see a generational change in the life and times of Madiba. "Nelson Mandela"On remembered in history as one of the great moral and political leaders of our time: an international hero whose life dedication to the fight against racial oppression in South Africa earned him the Nobel Peace and the Presidency of his country.
Mandela is on the high pedestal with Mahatma Gandhi, Martin Luther King Jr and Thomas Jefferson. It did not start his fight as an old man, but as a young lawyer. He challenged the oppression throughout his life and that Pinnacle can not be considered in isolation from its beginning.
In his advanced age, Mandela has served a term in Pretoria, his dream was not a power, but to ensure peace and a constitution for South Africa. At one point he was even a delegate to the presidency during a trip abroad with a political rival Mangosuthu Butelezi, chief minister of KwaZulu and President of Inkatha.
In Kenya, some leaders are still talking about the structure of power more than two years after the National Accord and Annan (Kofi) mission. We read in the press. It is more about power, more teeth, more than protocol, a greater sharing of power, several committees, commissions and more … Period. However, since his triumphant release in 1990 of more than a quarter century of imprisonment, Mandela has been at the center of history's most compelling and inspiring political. Unification and break whatever barriers the power play.
As chairman of the African National Congress and head of the movement in South Africa against apartheid, he was instrumental in moving the nation toward multiracial government and majority rule.
The "Long Walk to Freedom is his moving and exhilarating autobiography, a book destined to take its place among the fondest memories of the great figures of the world. Here for the first time, Nelson Rolihlahla Mandela tells the extraordinary story of his life – an epic struggle, setback, renewed hope, and ultimate triumph.
The adopted son of a Thembu chief, Mandela was raised in the traditional tribal culture of his ancestors, but at an early age learned the modern, inescapable reality of what would be called apartheid. He brings vividly to life the escalating political warfare in the 1950s between the ANC and the government, culminating in his struggle as a leader of the underground and notorious Rivonia Trial of 1964, when he was sentenced to life imprisonment. He recounts the surprisingly eventful 27 years in prison and the complex, delicate negotiations leading to his freedom and the beginning of the end of apartheid.
In Kenya, it's time for youth. This is your moment Mandela. You must redefine the future of the nation. Stand of solidarity, equity and demand justice and the right to a dignified life. Even President Barack Obama, who is so popular worldwide, is still not a Mandela; King Jr. and Gandhi are of this caliber. Kenya needs a new leadership with the simplicity and sincerity of purpose.
The journey began in 2012 and youth cannot ignore its importance, and crying after the event. We must reject the politics of deception that America did in 2008. United Youth must know not even the most powerful army cannot defeat an idea whose time has come. Consider this: "This is not the size of the wave, but the motion of the ocean that moves the ship."
Source: http://www.standardmedia.co.ke/InsidePage.php?id=2000026789&cid=15
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|"Alex Karasulu (JIRA)" <j...@apache.org>
|[jira] Closed: (DIRSERVER-883) Improve the way Schema are loaded
|Sat, 18 Aug 2007 02:40:30 GMT
[ https://issues.apache.org/jira/browse/DIRSERVER-883?page=com.atlassian.jira.plugin.system.issuetabpanels:all-tabpanel ] Alex Karasulu closed DIRSERVER-883. ----------------------------------- Resolution: Invalid The present design was conceived to make sure (at all costs) to remove the authoritative copy problem of schema files. Your solution re-introduces that but in the form of an LDIF file. If users don't have a clue and they modify the LDIF file thinking they will modify the schema there then we have a problem. Furthermore if they modify the schema via the protocol then what is in the DIT is different from the LDIF file. Again we have the same authoritative copy problem. The present solution although a bit involved removes this issue entirely. > Improve the way Schema are loaded > --------------------------------- > > Key: DIRSERVER-883 > URL: https://issues.apache.org/jira/browse/DIRSERVER-883 > Project: Directory ApacheDS > Issue Type: Improvement > Affects Versions: 1.5.0 > Reporter: Emmanuel Lecharny > Fix For: 1.5.1 > > > As far as I understand the way schema are loaded into 1.5.0 version of the server, we can improve it to be more versatile. > We have 5 kind of elements which are part of the schema repository : > 1) mandatory elements, like ObjectClass, Top, Name, Desc, any objects without which the server can't load a schema. > 2) meta-schema elements, which are used to declare a new schema > 3) core schemas, like Core, System, ApacheDs, which are loaded first > 4) Extension schema, like Samba, Nis, etc. > 5) User defined schema > Right now, we have a bootstrap sequence which is quite complicated : > 1) the server is started, and declare the mandatory elements > 2) The metaSchema is loaded, with the core schema, from some generated java classes (the schema are only used when compiling the server, to generate those java classes) > 3) The extended classes are loaded from some generated partition (there is a file named DBFILES which contains the list of all the backend files, files which will be created when compiling the server from the generated java classes) > 4) the server is available. > IMO, we can do the bootsrap differently : > 1) server startup : we load the mandatory elements > 2-a) if this is the first time the server is started, load the Ldif files representing the MetaSchema, and all the other Ldif files, so that the cn=schema partition is fed with the base and extended schema > 2-b) if the schema is already loaded, do nothing except if we have one file which date is younger than the last modification date in the cn=schema partition. > The list of schema ldif files will be stored into the server.xml file, assuming that the order must be corect (we must load core.schema before any other shcema, etc.) > This process will be much simpler and easier to modify than the first one, and we will be able to remove some projects and plugins, like the apacheds-bootsrap-schema and apacheds-schema-extra projects. > Not sure I'm totally right... > Wdyt ? -- This message is automatically generated by JIRA. - You can reply to this email to add a comment to the issue online.
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Auto Repair Services
Some of the services performed at PILOT'S AUTO:
Axle / CV Joint Repair (FWD)
Brakes: Standard or ABS
Clutch Repair
Electric: Batteries - Sell and Install
Electric: Electrical and Electronic Systems
Electric: Starting and Charging
Engine: Diagnostics and Performance
Engine: Service/Repair - Major/Minor
Exhaust Systems
Heating and Air Conditioning
Preventive Maintenance
Quick Lube and Oil Change
Steering and Suspension
Wheels/Tires: Alignment
Wheels/Tires: Mounting and Balancing
Vehicle Inspection: Total
Towing/Roadside Assistance
Radiator Repair/Replacement
Axle / Driveshaft Repair (RWD, 4WD)
Four Wheel Drive Systems
Factory Recommended Inspections (15k,30k,60k,90k miles)
Preventive Maintenance
The best way to minimize more serious breakdowns and the higher costs associated with automotive repair is by doing routine maintenance.
Preventive vehicle maintenance is simply what the manufacturer of your vehicle suggests getting done to keep your vehicle going. Whether you have a car or truck we provide the maintenance services to keep it running well.
Oil Change & Lube Service
As our mission is to provide Chicago, IL and the surrounding areas with a place that takes care of all their vehicle needs, we provide oil changes for all cars and trucks. We additionally offer a complimentary vehicle inspection with your oil change as we care about your safety.
Brake Repair & Service
Brakes wear out over time requiring service. How long your brakes last is based on various things such as driving conditions, how you brake and other factors. For this reason, it is necessary to service your brakes regularly.
We are familiar with the driving conditions here in Chicago and know that your brakes need care. So we provide a complimentary inspection with every service to make sure that any problem is taken up before it becomes a major concern.
Transmission Service & Repair
The transmission may well be one of the most complicated and important components of your car or truck. That's why it's important to trust its maintenance and repair to a full-service shop. There are a number of problems that can be resolved simply while others may require a complete transmission overhaul. Our skilled mechanics have the training and expertise to properly diagnose your transmission problem and get you back on the road.


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Hotel has an indoor pool, health club, and lobby coffee bar round out the opportunities to unwind and enjoy at every turn. Total 160 guest rooms redefine contemporary luxury and elegance. Sophisticated travellers will appreciate comforts such as our signature bed and rainforest showers, paired with the high tech style of large flat screen TVs and compact virtual office hubs.
Located in the dynamic central business district, Le Méridien Taipei is a refined stimulating starting point for exploring this intriguing city. Discover nearby cultural attractions, the impressive Taipei 101 and extensive shopping opportunities.
A modern, siliver, high-rise building.
Large in size and bright with a comfortable seating area.