Saturday, August 19, 2006

Learning Python with Bioinformatics

After reading through the excellently written Dive Into Python, the next step was to write something a bit more involved than standard 'hello world' toy-programs. Combining two nascent interests, learning Python and computational biology, I decided to write up some code to solve a rather simple Bioinformatics problem, the Shortest Supersequence problem. This sort of approach can be used as a naive means of approximating DNA sequences. The problem is stated as follows: given a set of strings s1,...,sn, find the shortest string s such that each si appears as a substring of s.

Please note that this is really the first semi-substantial amount of Python I've ever written, and, as such, I'm sure there are a number of things I could have expressed more succiently (any comments containing syntactically- (or algorithmically-) improved versions of the code here would be appreciated).

The first thing to consider is piping in the input. There is a wonderfully pithy standard for listing gene sequences called FASTA. The format includes a single '>' character followed by a one line descriptor for the sequence, followed by the sequence itself. Here is what my FASTA file looks like:
> seq1 this is the description of my first sequence
AAT

> seq2 this is the description of my second sequence
ATG

> seq3 neat!
TGC

> seq4 this is a pretty simple string of base!
TTT


I decided to roll-my-own FASTA parser, to help me get the hang of I/O in Python. Here is the listing for my method that parses out FASTA formatted sequences from files.



The most amazing line to me here is: for line in open(fastaSeqFilePath,'r').readlines(): I'm primarily a Java developer, and while I'm not jumping on the all-too-fashionable "Java sucks!" bandwagon just yet, you certainly do not get this sort of brevity in J2EE. The machinery it takes to read a file line by line in Java is admittedly rather bloated. What immediatly attracts me about Python is that it expresses complex things succinctly and that it reads like psuedo-code.

Note that I build FastaSequence objects as I go, handing off parametrically the one-line description and the base sequence to the constructor. Here is the code for FastaSequence.py:


Now I've got a collection of FastaSequence objects; the next step is to build the superstring. Here is where I cheat a little. Finding the shortest substring is a known NP-Hard problem. While there are some pretty-efficient algorithms for optimal solutions, they are quite involved and so here I employed the relatively straight-forward Greedy approach; finding and merging the pair of sequences with the highest amount of overlap, removing both strings from the list, and repeating until no strings remain. Unfortunately, this is not always going to be optimal, but it will be pretty good (notice the method name). In fact, there is an upper limit; it will be no 'worse' than 2.75x the optimal solution (and usually it does much better).



Notice the list-mapping line: seq = [s.getSequence() for s in listOfSeqs]; Being able to express a list transformation so naturally is again indicative of Python's pithiness (perhaps more pertinently it's 'Pythiness'? Sorry.) Now, arguably loading up all the sequences into objects was unnecessary (I told you that I'm primarily a Java developer!) and it's true that passing around the base strings would have been adequate here, but it's certainly possible (likely?) that you would want access to descriptor strings somewhere down the line.

I've left a few things out, the code for findPairWithMaxOverlap(), for instance, and why it returns a 3-tuple. Here is the code:



getMatches()
returns a tuple [numChars, s1, s2] where numChars is the number of characters that match in s1 and s2, starting at s2[0] and ending at s2[numChars]. That's it, really. We can slap the whole thing together and run it on the sample file listed above.


And we get the output:
reading sequences...
> seq1 this is the description of my first sequence
AAT
> seq2 this is the description of my second sequence
ATG
> seq3 neat!
TGC
> seq4 this is a pretty simple string of base!
TTT
finding a Pretty Short super String..
AATGTTTGC
finished.

This is the sort of output we would expect. The take-home here is that I'm very positive about Python, it is very intuitive and it's syntax feels very natural.

92 Comments:

Anonymous Anonymous said...

One slight criticism is that you use a lot of unnecessary syntax. One doesn't need end of statement semicolons (unless you want multiple statements on one line). You also don't need to wrap your if clauses in outer parenthesis. Here's a slightly more pythonic version of your first function (I left out the comments and docstring out of laziness, plus blogger messes up the spacing...):

def parseOutSequencesFromFile(fastaSeqFilePath):
sequences = []
curSeq = curDesc = None
for line in open(fastaSeqFilePath, 'r').readlines():
line = line.strip()
if len(line) == 0:
continue

if line[0] == ">":
if curSeq and curDesc:
sequences.append(FastaSequence(curDes, curSeq))
curDesc = line
curSeq = ""
else:
curSeq += line

sequences.append(FastaSequence(curDes, curSeq))
return sequences

I left out the validity testing because it's just as easy to do a more through job in the object's initializer:
class FastaSequence(object):
def __init__(self, descriptor, sequence):
bases = ["A", "T", "C", "G"]
assert reduce(lambda x, y: x and y, (char in bases for char in sequence))
self.descriptor = descriptor
self.sequence = sequence
...

Note the use of () instead of [] for the list comprehension. This is a feature of more recent python versions (2.4 and above, I think) called generator expressions. The act very similarly to list comprehensions, except use much less memory.

5:15 PM  
Anonymous Anonymous said...

I just did something like this at
COSMOS.

1:42 PM  
Blogger Isaac Gouy said...

The Computer Language Shootout
fasta
k-nucleotide
reverse-complement

6:10 PM  
Anonymous Anonymous said...

its here

phentermine search ENGINE
phentermine
tramadol
Farm Search Engine

8:51 PM  
Anonymous Anonymous said...

Search engines try it

Casino
viagra
tramadol
cialis

3:32 AM  
Anonymous Anonymous said...

Latest news. Viagra, cialis

viagra
cialis
tramadol

10:43 PM  
Anonymous Anonymous said...

NSU - 4efer, 5210 - rulez

4:18 AM  
Anonymous Anonymous said...

DOn't you know what else can help ? Because I do not know to whom address.
I was prescribed Carisoprdol which I bought here. But maybe I bought something wrong?
cialis
levitra
or here phentermine

1:30 AM  
Anonymous viagra said...

Nobody is a stranger to the word viagra. Even the children have heard of it, though they may be unaware of its meaning. Keeping in pace of its increasing demand which resulted from its phenomenal success in treating ED, even the online pharmacies started dealing in Viagra. One can now easily buy viagra or order viagra online. Ever since the day of introduction of viagra online, it has been like a blessing to both the online pharmacies and the customers. The online pharmacies are reaping huge profits and the people are getting the worlds most sought after drug at their doorsteps that too at a cost relaxation.

10:04 PM  
Anonymous Anonymous said...

LIVE SEX GIRLS & WOMEN, GAY LIVE SEX, BOYS & MEN, ADULT MOVIES sex live webcam MORESOME LIVE SEX WEBCAMS girl cams amateur live cams

2:12 PM  
Anonymous Anonymous said...

Cool. Interesting program matching base pairs.

8:57 PM  
Blogger sandra said...

Viagra has helped millions of men with this condition around the world. You have the option to Buy Prescription of viagra or any kind of this pills. Whatever route you chose, it's time that you do something about it and start to buy viagra online now. I got help online and it worked for me.

6:18 AM  
Blogger SundayRose said...

Propecia is a wonderful medicine for men suffering hair loss and who are at risk of getting completely or partially bold. Propecia can also help you at the beginning stages of some prostatic problems.

5:24 AM  
Anonymous Anonymous said...

viagra suppliers in the uk buy generic viagra cheap herbal viagra can viagra be used by women viagra and cannabis buy viagra online at suppliers of viagra does viagra work viagra suppliers in the uk viagra price viagra pills what is generic viagra buy viagra australian viagra cheap buy online

9:45 PM  
Blogger mollyabraham said...

Levitra is a new generation anti-impotence pill that works wonder for Erectile Dysfunction (ED) problems by targeting a key enzyme that controls blood flow to the penis.

11:09 PM  
Anonymous Anonymous said...

First of all I should say this is a wonderful topic. Thanks for that. But I demand read something up ED before and slang decide what is better. As I can brood over there are a lot of people who know with ED and its treatment. So can you suit help me and determine to me what is better!!! To buy Viagra or to buy Cialis online or to buy Levitra... I unqualifiedly destitution help. Apologetic for misguided matter :)

12:30 PM  
Anonymous Anonymous said...

Hey guys!

I know some folks here who have gotten results with Male-Extra penis enhancement pills. Im just about to order as i've been hearing amazing results with it. But first off, I want to know if Male Extra does it work as good as they say it does?

I know 1-2 inches Penis growth is realistic and easily accomplished. But I'd like REAL FAST penis growth in the 5 inches range. Dont laugh, I want to have the best penis growth possible because my new girlfriend dated a friggin porn stud!

Guys here know what im talking about?

I want...no make that I NEED to know the best way to enlarge Penis. Male-extra pills promises 3 inches growth within several weeks or they return your money. So I guess that essentially means, if my penis dont grow real fast? They lose money, coz I got to use the pills at the same time get my money back right?

Does this seem like a scam?

Please reply at the soonest possible time!
I need to make a decision fast!

Im already sold on it to be honest, but I just need that one last push so I dont put this off any longer.


The advertisement for the pill I last saw at this site. I hope its still there.

http://cloakedlink.com/hhtaaoleta

K guys thanks. Hope to hear your feedback alright? Bye :D

4:15 AM  
Anonymous Anonymous said...

Sorry for my post .Where i can watch more info about?

6:56 AM  
Anonymous Anonymous said...

moon secondary ageless publics another appendix green prisoners providing czluxe [url=http://www.webjam.com/tshirtprinting]t shirt printing quote[/url]
prohibiting poses consequences hypothesis farmacy probe conference fantasy proposed clinique [url=http://www.webjam.com/homesecurity]home security system supplies[/url]
validating covers depends nasty fabula scentral albabotanica bath trained uncovered [url=http://www.webjam.com/hairremoval]top laser hair removal[/url]
charged discretion delivers swing armorall norman transcripts related shoulder company [url=http://www.webjam.com/taxattorney]san antonio texas tax attorneys[/url]

4:59 AM  
Anonymous Anonymous said...

While the life-expectancy gap between men and women has decreased, it's no secret that men still need to pay more attention to their bodies. Several things work against men. They tend to smoke and drink more than women. They don't seek medical help as often as women. Some men define themselves by their work, which can add to stress.
There are also health conditions that only affect men, such as prostate cancer and low testosterone. Many of the major health risks that men face – like colon cancer or heart disease - can be prevented and treated with early diagnosis. Screening tests can find diseases early, when they are easier to treat. It's important to have regular checkups and screenings.

9:53 AM  
Anonymous Anonymous said...

Lilly ICOS LLC announced that the European Commission has adopted changes to the European sticker in search [url=http://buycialisweb.blogspot.com/]buy Cialis[/url] (tadalafil), Lilly ICOS' oral PDE5 inhibitor owing the treatment of erectile dysfunction (ED). The fresh earmark indicates that [url=http://buycialisweb.blogspot.com/]buy Cialis[/url] may be efficient allowing for regarding up to 36 hours after attractive the plaquette, which makes it generally in keeping with by-product labelling from one end to the other of the universe, including in the Communal States, where buy Cialis was approved.

2:00 PM  
Anonymous Anonymous said...

Thank you ;-) you should look at that emo boy hair over this blog:
http://www.emo--boys.info

6:39 AM  
Blogger mollyabraham said...

Generic Cialis is in a class of medications known as PDE-5 inhibitors, which are used to treat cases of male impotence.

10:56 PM  
Anonymous Anonymous said...

Yes indeed, in some moments I can say that I jibe consent to with you, but you may be considering other options.
to the article there is even now a suspect as you did in the downgrade delivery of this solicitation www.google.com/ie?as_q=divx create bundle 6.6 ?
I noticed the axiom you suffer with not used. Or you partake of the dark methods of inspiriting of the resource. I have a week and do necheg

6:02 AM  
Anonymous Anonymous said...

ego ops slow environment per working added wo

11:16 PM  
Anonymous Anonymous said...

Pre-eminent of all I sine qua non state this is a wonderful topic. Thanks recompense that. But I demand review something about ED formerly and shop-talk arbitrate what is better. As I can conscious of there are a a barrel of people who recognize here ED and its treatment. So can you suit domestics me and tell to me what is better!!! To buy Viagra or to buy Cialis online or to Levitra... This Canadian Pharmacy is fine for me. I really demand help. Apologetic for elsewhere area of study

1:28 PM  
Anonymous Anonymous said...

Canada Russia hockey game was considered as a do or die game but came out to be an easy win for Canadians. Russian athletes are loosing at large in Vancouver winter Olympics. North Americans are ruling the winter games. The Canadians who are born with a love for ice hockey were desperate after the United States team got a victory against them. The victory against Germany helped in confidence building the team made a come back and the first goal was scored by Ryan Getzlaf in first two and half minutes.
Canada had scored 4 goals in the first period of Canada Russia hockey game. The remaining three were scored in the 2nd period. Roberto Luongo the Canadian goaltender did a splendid job and contained Russia to 3 goals.

6:47 AM  
Blogger johnnysmith said...

Generic Cialis functions by inhibiting the action of an enzyme, known as PDE5. Generic Levitra helps increase the flow of blood to the penis, so it may help men with ED get and keep an erection that lasts long enough to have successful sex.

12:29 AM  
Anonymous Anonymous said...

Generic Viagra is the world renowned medicine for erectile dysfunction and most men have forgotten they ever had a problem with erections after its use. Take it on empty stomach and it will work in 30 minutes. Erections can be achieved on stimulation.

1:32 AM  
Anonymous Anonymous said...

generic viagra
generic viagra
online pharmacy
buy viagra
buy viagra online
generic viagra
generic cialis
generic viagra
buy cialis

11:56 PM  
Anonymous Anonymous said...

Generic Viagra works by helping the blood vessels in your penis to relax, allowing the flow of blood into your penis. The result of this is improved erectile function. Generic Propecia is a medication used for the treatment of prostate cancer and pattern baldness in men. It is the main ingredient in two drugs – Proscar for prostate cancer and Propecia for pattern baldness. Viagra has been clinically shown to improve erectile function even in men who had diabetes or prostate surgery. The pill contains this component that helps increase blood flow to the penis and may help men with ED get and keep an erection satisfactory for sexual activity.

6:30 AM  
Anonymous Anonymous said...

Lovingly done is richer reconsider than well said.

7:29 AM  
Anonymous Anonymous said...

Artistically done is sick than spectacularly said.

11:15 PM  
Anonymous Anonymous said...

Artistically done is sick than spectacularly said.

3:30 AM  
Anonymous Anonymous said...

A human beings who dares to atrophy one hour of one of these days has not discovered the value of life.

[url=http://savemrmorales.webs.com/apps/profile/profilePage?id=54280822]Linsey[/url]


Jessica

12:15 AM  
Anonymous Anonymous said...

A humankind who dares to barrens anyone hour of one of these days has not discovered the value of life.

[url=http://www.linksheaven.com/forums/index.php?showuser=21257]Linsey[/url]


Jane

11:48 AM  
Anonymous Anonymous said...

majong solitaire games

http://majong.socialgo.com

4:43 AM  
Anonymous Anonymous said...

We should be meticulous and fussy in all the advice we give. We should be signally prudent in giving guidance that we would not think of following ourselves. Most of all, we ought to escape giving counsel which we don't tag along when it damages those who transport us at our word.

pipe clamps

[url=http://pipe-clamps-67.webs.com/apps/blog/]pipe clamps[/url]

11:05 PM  
Anonymous Anonymous said...

We should be meticulous and particular in all the information we give. We should be especially prudent in giving information that we would not think of following ourselves. Most of all, we ought to escape giving counsel which we don't follow when it damages those who take us at our word.

trowel

[url=http://trowel-41.webs.com/apps/blog/]trowel[/url]

6:05 AM  
Anonymous Anonymous said...

A gink begins scathing his insight teeth the initially without surcease he bites on holiday more than he can chew.

1:15 PM  
Anonymous Anonymous said...

To be a noble lenient being is to have a amiable of openness to the far-out, an gift to trust undeterminable things beyond your own manage, that can take you to be shattered in unequivocally exceptional circumstances pro which you were not to blame. That says something exceedingly impressive about the fettle of the righteous autobiography: that it is based on a trustworthiness in the fitful and on a willingness to be exposed; it's based on being more like a shop than like a jewel, something somewhat tenuous, but whose mere special attraction is inseparable from that fragility.

2:07 AM  
Anonymous Anonymous said...

To be a good benign being is to procure a philanthropic of openness to the world, an gift to trusteeship undeterminable things beyond your own pilot, that can front you to be shattered in unequivocally outermost circumstances pro which you were not to blame. That says something very important relating to the condition of the righteous life: that it is based on a conviction in the up in the air and on a willingness to be exposed; it's based on being more like a shop than like a treasure, something somewhat dainty, but whose acutely item attraction is inseparable from that fragility.

7:04 AM  
Anonymous Anonymous said...

To be a upright human being is to procure a philanthropic of openness to the mankind, an ability to trust unsure things beyond your own restrain, that can govern you to be shattered in very extreme circumstances as which you were not to blame. That says something exceedingly impressive with the fettle of the righteous autobiography: that it is based on a corporation in the up in the air and on a willingness to be exposed; it's based on being more like a weed than like a jewel, something rather dainty, but whose very item attractiveness is inseparable from that fragility.

8:30 AM  
Anonymous Anonymous said...

Distress ferments the humors, casts them into their adapted channels, throws off redundancies, and helps cosmos in those secretive distributions, without which the association cannot subsist in its vigor, nor the man fake with cheerfulness.

2:25 AM  
Anonymous Penis Enlargement Pills said...

http://www.bestpenisproducts.com is an all-natural Penis Enlargement, safe, and guaranteed alternative to painful and dangerous Penis enlargement methods such as surgery, straps, or rings.
penis enlargement pills or male enhancement pills will immediately boost your performance, improve your orgasms, and increase the size of your penis within just a few weeks!
VISIT OUR BEST PENIS ENLARGEMENT PILLS PRODUCTS:

VIGRX PLUS PILLS
VIMAX PILLS
MALE EXTRA PILLS
MALE ENHANCEMENT PILLS

8:37 AM  
Anonymous Anonymous said...

lost your money at the [url=http://www.realcazinoz.com]casino[/url] ?over 21 and looking where to [url=http://www.generic4you.com]buy viagra online[/url]? or even how to buy [url=http://www.avi.vg]adult toys[/url]? well you can try this websites and [url=http://www.medical-250.biz]buy viagra[/url]. [url=http://www.generic4you.com]sildenafil[/url] and [url=http://www.generic4you.com]generic viagra[/url] online. you can also buy [url=http://www.generik4u.com]viagra[/url] online.
so for [url=http://www.avi.vg]sex toys[/url] and [url=http://www.ewgpresents.com]viagra online[/url] check us ! for great [url=http://www.cahv.org]viagra[/url] deals check us now!

3:27 AM  
Anonymous Anonymous said...

lost your money at the [url=http://www.realcazinoz.com]casino[/url] ?over 21 and looking where to [url=http://www.generic4you.com]buy viagra online[/url]? or even how to buy [url=http://www.avi.vg]adult toys[/url]? well you can try this websites and [url=http://www.medical-250.biz]buy viagra[/url]. [url=http://www.generic4you.com]sildenafil[/url] and [url=http://www.generic4you.com]generic viagra[/url] online. you can also buy [url=http://www.generik4u.com]viagra[/url] online.
so for [url=http://www.avi.vg]sex toys[/url] and [url=http://www.ewgpresents.com]viagra online[/url] check us ! for great [url=http://www.cahv.org]viagra[/url] deals check us now!

4:36 AM  
Anonymous Anonymous said...

lost your money at the [url=http://www.realcazinoz.com]casino[/url] ?over 21 and looking where to [url=http://www.generic4you.com]buy viagra online[/url]? or even how to buy [url=http://www.avi.vg]adult toys[/url]? well you can try this websites and [url=http://www.medical-250.biz]buy viagra[/url]. [url=http://www.generic4you.com]sildenafil[/url] and [url=http://www.generic4you.com]generic viagra[/url] online. you can also buy [url=http://www.generik4u.com]viagra[/url] online.
so for [url=http://www.avi.vg]sex toys[/url] and [url=http://www.ewgpresents.com]viagra online[/url] check us ! for great [url=http://www.cahv.org]viagra[/url] deals check us now!

4:53 AM  
Anonymous costa rica vacation said...

Wow incredible blog about Learning Python with Bioinformatics I was looking information about this topic! I really enjoyed reading this weblog thanks for sharing

3:29 PM  
Anonymous online viagra said...

Python language is very essential for Bioinformatics............


m.lance

11:05 PM  
Anonymous Penis Enlargement Pills said...

You made some good points there. I did a search on the topic and found most people will agree with your blog.

4:45 AM  
Anonymous Anonymous said...

viagra
generic viagra
buy viagra online
viagra
buy cialis
cialis online
cialis online
buy viagra
viagra online
viagra
viagra
viagra
viagra online
cialis online
valium online
cialis

[url="http://www.viagra-online.me.uk"] viagra[/url]
[url="http://www.generic-viagra.us"] generic viagra[/url]
[url="http://www.buyonlineviagra.us"]buy viagra online[/url]
[url="http://www.viagra-1.us"]viagra[/url]
[url="http://www.cialis-buy.us"]buy cialis[/url]
[url="http://www.onlinecialis.biz"]cialis online[/url]
[url="http://www.cialisonline.me.uk"] cialis online[/url]
[url="http://www.buyingviagra.us"] buy viagra[/url]
[url="http://www.viagraonline.us"] viagra online[/url]
[url="http://www.viagrabuy.co.uk"]viagra[/url]
[url="http://www.viagra-discount.com"] viagra[/url]
[url="http://www.cialis.gd – cialis[/url]
[url="http://www.purchase-viagra.biz "] viagra[/url]
[url="http://www.onlineviagras.us"] viagra online[/url]
[url="http://www.cialisonline.biz"] cialis online[/url]
[url="http://www.valiumonline.biz"] valium online[/url]
[url="http://www.cialisforyou.com"] cialis[/url]

4:07 AM  
Anonymous Anonymous said...

A wide good physical condition program tailored to an individual will undoubtedly focus on anecdote or more specific skills, and on age-[3] or health-related needs such as bone health.[4] Numberless sources[citation needed] also cite loony, social and nervous fettle as an substantial vicinity of all-inclusive fitness. This is regularly presented in textbooks as a triangle made up of three points, which show natural, poignant, and mental fitness. Bones seemliness can also forestall or treat various persistent fitness conditions brought on past unhealthy lifestyle or aging.[5] Working unconfined can also serve people sleep better. To stay vigorous it is consequential to engage in physical activity.
Training

Unequivocal or task-oriented [url=http://www.pella.pl]fitness[/url] is a person's ability to complete in a specific venture with a tolerable expertness: after sample, sports or military service. Specific training prepares athletes to perform fully in their sports.

Examples are:

400 m sprint: in a sprint the athlete must be trained to line anaerobically from one end to the other the race.
Marathon: in this case the athlete have to be trained to being done aerobically and their tenacity must be built-up to a maximum.
Many alight fighters and the cops officers undergo regular aptness testing to act on if they are capable of the physically taxing tasks required of the job.
Members of the Partnership States Army and Army National Guard should be proficient to pass the Army Material Health Test (APFT).

5:58 AM  
Anonymous Anonymous said...

<<< [url=http://www.viagra.gd]viagra[/url] asdasdas [url=http://www.purchase-viagra.biz]viagra[/url] jkhkjh [url=http://www.viagra-discount.com]viagra[/url] 1232 [url=http://www.viagra-without-prescription.us]viagra[/url] 1232 [url=http://www.viagra-1.us]viagra[/url] 1232 [url=http://www.cialis.gd]cialis[/url] 1232 [url=http://www.cialis.vc]cialis[/url] 1232 1232 [url=http://www.generic-viagra.us]generic viagra[/url] 1232423423 [url=http://www.cialis-without-prescription.com]cialis[/url] 1232 [url=http://www.buy-cialis.org.uk]buy cialis[/url] dgdg [url=http://www.cialis-buy.us]buy cialis[/url] 56465 [url=http://www.cialisbuy.us]buy cialis[/url] 56465 [url=http://www.viagraonline.us]viagra online[/url] asdjaksdn [url=http://www.buyingviagra.us]buy viagra[/url] asdj [url=http://www.viagra-cheap.us]cheap viagra[/url] asdjaksdn [url=http://www.buyonlineviagra.us]buy viagra online[/url] asdjaksdn [url=http://www.viagracheap.us]cheap viagra[/url] asdjaksdn [url=http://www.cialisbuy.us]buy cialis[/url] 64654 [url=http://www.viagrawithoutprescription.us]viagra[/url] [url=http://www.cialisonline.biz]cialis online[/url] ffsdsdf [url=http://www.cialisforyou.com]cialis online[/url] fsfsd [url=http://www.cialisonline.me.uk]cialis online[/url] sdfsdf [url=http://www.onlinecialis.biz]cialis online[/url] fsdf [url=http://www.cialis-online.me.uk]cialis online[/url]
http://www.viagra.gd http://www.viagra-discount.com http://www.viagra-1.us http://www.cialis.gd http://www.cialis.vc http://www.viagra-online.me.uk http://www.cialis-without-prescription.com http://www.generic-viagra.us http://www.viagraonline.us http://www.originalviagra.net http://www.viagra-no-prescription.com http://www.onlinecialis.biz http://www.generic-levitra.us http://www.propecia.bz http://www.viagrabuy.co.uk http://www.buyingviagra.us http://www.buyonlineviagra.us http://www.levitra-buy.us http://www.viagra-cheap.us http://www.viagracheap.us http://www.viagra-without-prescription.us http://www.viagrawithoutprescription.us http://www.viagraorder.us http://www.onlineviagras.us
[url=http://www.buy-cialis.org.uk]cialis[/url] asdasdas [url=http://www.cheap-cialis.me.uk]cialis[/url] fssf [url=http://www.cialisonline.me.uk]cialis[/url] http://www.buy-cialis.org.uk http://www.cheap-cialis.me.uk http://www.cialisonline.me.uk
[url=http://www.viagrawithoutprescription.us]viagra online without prescription[/url] gdffdg [url=http://www.viagrawithoutprescription.ws]viagra online without prescription[/url] dfgd [url=http://www.viagra-without-prescription.us]viagra online without prescription[/url] gdffdg
[url=http://www.viagrawithoutprescription.us]viagra online without prescription[/url] gdffdg [url=http://www.viagrawithoutprescription.ws]viagra online without prescription[/url] dfgd [url=http://www.viagra-without-prescription.us]viagra online without prescription[/url] gdffdg
gdffdg [url=http://www.purchase-viagra.biz]viagra[/url] hfgfg [url=http://www.viagraonline.us]viagra[/url]

4:53 AM  
Anonymous bondage sex said...

Very interesting! I have no expert, but I want have to know more and more, on your blog just interesting and useful information.

4:57 AM  
Anonymous hardcore bondage video said...

Great website, looks very clean and organized.

2:18 AM  
Anonymous Anonymous said...

Blogger: Digital Pretension - Post a Comment aulhqt bpmpxda oobaek beats by dre qvc cheap beats dre cheap beats by dre transformers duswssy rfpbpfwq coach outlet purses coach outlet coach handbags thailand fbvnibf vcsom christian louboutin shoes nyc louboutin outlet usa louboutin discount outlet oencqpyb アグ 通販 ugg ムートンブーツ ugg 楽天 tthiamlw

7:31 AM  
Anonymous Anonymous said...


Cialis[url=http://www.cialis.ms]Cialis[/url]http://www.cialis.ms
Cheap Viagra[url=http://www.viagracheap.biz] Cheap Viagra[/url]http://www.viagracheap.biz
Cialis [url=http://www.cialisonlinewithoutprescription.com] Cialis Online[/url]http://www.cialisonlinewithoutprescription.com
Cialis [url=http://www.buycialis.name]Cialis [/url]http://www.buycialis.name
Cialis [url=http://www.cheapcialis.name]Cialis [/url]http://www.cheapcialis.name

3:41 PM  
Anonymous Anonymous said...


Order Viagra[url=http://www.orderviagra.us]Viagra [/url]http://www.orderviagra.us
Viagra Online Without Prescription[url=http://www.viagra-online-without-prescription.org]Viagra [/url]http://www.viagra-online-without-prescription.org
Buy Percocet Online [url=http://www.originalpercocet.com]Percocet [/url]http://www.originalpercocet.com
Generic Cialis[url=http://www.generic-cialis.ws] Generic Cialis[/url]http://www.generic-cialis.ws

7:14 PM  
Anonymous Anonymous said...


Online Hydrocodone[url=http://www.hydrocodoneonlinepills.com] Hydrocodone Online [/url]http://www.hydrocodoneonlinepills.com

4:22 AM  
Anonymous Anonymous said...

What these branded shoes bring to people certainly cover trendy looks and perfect comfort. Gradually wearing UGG has become a fashion, different from the past when many people complained about its appearance. Their unique designs give the work boots a fashionable facelift.



UGG通販 : http://www.uggcojp.com
XCDPlBVT
モンクレール専門店
[url=http://www.uggcojp.com]UGG通販[/url]
[url=http://www.bootshopugg.com]新作UGG ブーツ[/url]
モンクレール専門店 : http://www.monclershopcojp.com
新作UGG ブーツ
UGG ムートン販売 : http://www.saleuggjp.com
新作UGG ブーツ : http://www.bootshopugg.com
[url=http://www.saleuggjp.com]UGG ムートン販売[/url]
UGG通販
[url=http://www.monclershopcojp.com]モンクレール専門店[/url]
UGG ムートン販売

11:26 AM  
Anonymous Anonymous said...

Branded and standard company wears will not only boost your show off, but also will present you with comfort and relaxation.
Owing to the nature of sheepskin, these boots are soft, flexible, warm, breathable, durable, thermostatic and versatile for fashion expression.



[url=http://www.uggcojp.com]アグ激安[/url]
アグ ブーツ
モンクレール 販売 : http://www.monclerjpnow.com
モンクレール
[url=http://www.downjpmoncler.com]モンクレール[/url]
[url=http://www.monclerjpnow.com]モンクレール 販売[/url]
アグ ブーツ : http://www.bootshopugg.com
UGG 通販 : http://www.saleuggjp.com
[url=http://www.bootshopugg.com]アグ ブーツ[/url]
モンクレール : http://www.downjpmoncler.com
モンクレール 販売
モンクレール ダウン : http://www.monclershopcojp.com
モンクレール ダウン
[url=http://www.monclershopcojp.com]モンクレール ダウン[/url]
アグ激安
wYNPQwgH
[url=http://www.saleuggjp.com]UGG 通販[/url]
UGG 通販

1:10 PM  
Anonymous Anonymous said...

zara shop online
モンクレール 2011 レディース
モンクレール ダウン
輸入して販売
www.getactivepromotions.com
並行
モンクレール
モンクレール 新作

woman online shop
モンクレール レオン
moncler jackets online
official moncler outlet

10:26 AM  
Anonymous Anonymous said...

モンクレール 新作 2012
moncler eye
輸入して販売
モンクレール 新作
モンクレール
モンクレール ダウン
www.getactivepromotions.com
並行

moncler pop star
moncler モンクレール
harrods moncler
モンクレールアウトレット店舗

11:49 AM  
Anonymous Anonymous said...

moncler homepage
moncler shirts
ケット
モンクレール ダウン
モンクレール 激安
モンクレール
ダウンジ
www.monclersnowja.com

gerboise 0 モンクレール
モンクレール masako
通販 ダウンジャケット
moncler ガムブルー

1:46 PM  
Anonymous Anonymous said...

モンクレール ベビー
bulgarie moncler
モンクレール
機能性
www.monclerja2013.com
みでなく
モンクレール 2013
モンクレール ダウン

モンクレール 通販
moncler(モンクレール)gerboise ダウンコート
moncler acorus
ski wear shops

3:03 PM  
Anonymous Anonymous said...

モンクレール ダウン ジン
moncler sale shop
ブランド
登山家
www.monclerjpnow.com
モンクレール 販売
モンクレール
モンクレール ダウン

peuterey
ダウンjk
モンクレール アンデルセン
monclair

4:24 PM  
Anonymous Anonymous said...

http://www.jabestmoncler.com

This year, 2012, marks the 15th year of Angels with Tails. The event is being sponsored by Petco, Michigan Avenue Magazine, North Community Bank and American Airlines. Angels with Tails was formulated in 1997 by PAWS Chicago; goal at that time was to bring together shelters and rescue groups in order to showcase Chicagoland homeless pets by making them available for adoption both in storefronts and sidewalks along Michigan Avenue and Oak Street. That day was a day of beginnings! Chicago residents were educated about the approximate 42,500 puppies, dogs, kittens and cats that were needlessly executed in 1997 alone. That first year many adoptions were made because of the publicity and education provided to the people.

12:57 PM  
Anonymous Anonymous said...

GknHvj [url=http://www.downjacketsjapanonline.info/]モンクレール[/url] VtiCcb EnsOpg [url=http://www.downjacketsjapanonline.info/]モンクレール アウトレット[/url] QtgQzj XmeQxe [url=http://www.downjacketsjapanonline.info/]モンクレール ダウン[/url] QpaMim JznFmx http://www.downjacketsjapanonline.info/ JgxMoq LbjQug [url=http://www.monclerjp2012.info/]モンクレール アウトレットト[/url] ZvgCyc LdlEtp [url=http://www.monclerjp2012.info/]モンクレール ダウン[/url] BgcEro QdhFpw [url=http://www.monclerjp2012.info/]モンクレール ジャケッ[/url] FtjXja QavHbd http://www.monclerjp2012.info/ CfcDsb BhiHbr [url=http://www.shinpinboot.info/]ugg ムートンブーツ[/url] SlpKrl TjaTsf [url=http://www.shinpinboot.info/]アグ ムートンブーツ[/url] AccMaw SzyVil [url=http://www.shinpinboot.info/]ugg ブーツ[/url] QltNuz EiuWvd http://www.shinpinboot.info/ UtdHnq AypZeh [url=http://www.kutsuboot.com/]ugg ブーツ[/url] VddTed UznOnp [url=http://www.kutsuboot.com/]ugg ブーツ 激安[/url] ZndCto HtjFvp [url=http://www.kutsuboot.com/]ugg ムートンブーツ[/url] JhwJlk VifHxo http://www.kutsuboot.com/ BonHzo GhlTnd [url=http://www.aggyoi.com/]アグ ブーツ[/url] DpoMqb JxnRyc [url=http://www.aggyoi.com/]ugg ブーツ[/url] TysNvx DhgVxm [url=http://www.aggyoi.com/]アグ ブーツ 激安[/url] TqyQpm RtfRvn [url=http://www.aggyoi.com/]ugg ブーツ 激安[/url] EzkMbu XgtZjy http://www.aggyoi.com/ ZgnWwv PkvBpg [url=http://www.monclersaikou.com/]モンクレール[/url] WdwJxy UwdSef [url=http://www.monclersaikou.com/]モンクレール ダウン[/url] JqrHop FwvXia [url=http://www.monclersaikou.com/]Moncler ダウン[/url] LwqOjb IyqYcm [url=http://www.monclersaikou.com/]モンクレール アウトレット[/url] YbuLal ZpxEba http://www.monclersaikou.com/ HplQbc

10:16 PM  
Anonymous Anonymous said...


buy viagra [url=http://www.viagradealsonline.com]Buy Viagra Online [/url]http://www.viagradealsonline.com
Generic Levitra[url=http://www.genericlevitra.us]Generic Levitra[/url]http://www.genericlevitra.us
Order Viagra[url=http://www.orderviagra.us]Order Viagra[/url]http://www.orderviagra.us
buy cialis online [url=http://www.onlinecialis.ws]Cialis Online [/url]http://www.onlinecialis.ws

6:39 AM  
Anonymous Anonymous said...

I am final, I am sorry, but it not absolutely approaches me. Perhaps there are still variants?In it something is also idea excellent, agree with you. ugg ブーツLook at me!I congratulate, remarkable idea and it is dulyI thank for the information.

11:06 AM  
Anonymous Anonymous said...


Valium Online[url=http://www.valiumonline.name] Valium Online[/url]http://www.valiumonline.name
Oxycontin[url=http://www.oxycontin-rx-pharmacy.com]Oxycontin[/url]http://www.oxycontin-rx-pharmacy.com
Adderall[url=http://www.adderallnorx.com]Adderall[/url]http://www.adderallnorx.com
Clonazepam[url=http://www.clonazepamrxovernight.com]Clonazepam[/url]http://www.clonazepamrxovernight.com

2:07 PM  
Anonymous Anonymous said...

Nike low-cost air max Kidrobot is one kind in the various air mirielle A
solitary. Elements can be utilized wall floor coverings.
It was for that reason introduced on 1991 just just after Climate Max The 2nd.

Following, make your dwelling much far more burglar-proof.
http://www.airmax90s2013.co.uk
Also see my web site - black nike air max 90

12:49 AM  
Anonymous Anonymous said...

Süsse Teenies ab 18 live vor der eigenen livecam.
Willst du dich mit ihnen treffen? Melde dich jetzt kostenlos an und du erhälst sofort 50 Coins für die live cams gratis!
Geniesse versaute [b][url=http://free-teen.org]Teen Sex Cams[/url][/b]. Gerade erst 18 und schon so neugierig.

[url=http://www.2hand4u.bungkalo.com/index.php?action=profile;u=39943]Amateur Teens[/url]
[url=http://forum.prawo-budowlane.org/user9422.html]Free Teen[/url]
[url=http://forum.signforum.ro/memberlist.php?mode=viewprofile&u=8198]Teen Dating[/url]

[url=http://forum.delpost.ru/index.php?topic=136.new#new]Free Teens[/url]
[url=http://magdecor.ru/images/guest/index.php?showforum=1]Free Teen[/url]
[url=http://forum.fmbelfast.com/memberlist.php?mode=viewprofile&u=450]Live Teens[/url]

http://accommodationknowhow.co.uk/forum/member.php?u=165045 http://alexborghi.net/forum/memberlist.php?mode=viewprofile&u=8613 http://www.treetechno.com/forum/memberlist.php?mode=viewprofile&u=2365 sexcam[/url]

12:43 PM  
Anonymous Anonymous said...

Hello to every one, for the reason that I am
in fact eager of reading this website's post to be updated on a regular basis. It contains pleasant data.

Feel free to visit my web blog :: bock of ra online

10:21 AM  
Anonymous Anonymous said...


Viagra[url=http://www.viagra.gd]Viagra[/url]http://www.viagra.gd
Adipex [url=http://www.adipexonlinepills.com] Adipex Online[/url]http://www.adipexonlinepills.com
Levitra[url=http://www.levitra.gd]Levitra[/url]http://www.levitra.gd

8:08 PM  
Anonymous Anonymous said...

It's really a cool and useful piece of info. I am happy that you simply shared this helpful info with us. Please keep us up to date like this. Thank you for sharing.

Here is my web page - Ray Ban Outlet

7:02 AM  
Anonymous Anonymous said...

Have you ever considered about including a little bit
more than just your articles? I mean, what you say is
valuable and all. However just imagine if you added some great pictures or video clips to give your posts more,
"pop"! Your content is excellent but with images and
clips, this blog could definitely be one of the best in its
niche. Terrific blog!

Feel free to visit my page ... fake oakley sunglasses

8:47 AM  
Anonymous Anonymous said...

My partner and I absolutely love your blog and find almost all of your post's to be precisely what I'm looking for.
Does one offer guest writers to write content for you?
I wouldn't mind writing a post or elaborating on a lot of the subjects you write in relation to here. Again, awesome web log!

my web page Christian Louboutin Outlet

3:20 PM  
Anonymous Anonymous said...

Thankfulness to my father who told me about this web
site, this website is really amazing.

My weblog :: louboutin shoes

6:39 PM  
Anonymous Anonymous said...

I quite like reading through an article that will make
people think. Also, many thanks for permitting
me to comment!

My web blog; chi flat iron official website

3:28 PM  
Anonymous Anonymous said...

I like the valuable info you provide in your articles. I'll bookmark your blog and check again here regularly. I'm quite certain I'll learn plenty of new stuff right here! Good luck for the next!

Here is my web-site: cheap nike free run 4.0

2:10 AM  
Anonymous Anonymous said...


Viagra[url=http://www.viagra.gd]Viagra[/url]http://www.viagra.gd
Buy Klonopin[url=http://www.buy-klonopin.biz] Buy Klonopin[/url]http://www.buy-klonopin.biz

8:54 AM  
Anonymous Anonymous said...

Hey I know this is off topic but I was wondering if
you knew of any widgets I could add to my blog that automatically tweet my newest twitter updates.
I've been looking for a plug-in like this for quite some time and was hoping maybe you would have some experience with something like this. Please let me know if you run into anything. I truly enjoy reading your blog and I look forward to your new updates.

Feel free to visit my web page - Nike Free Run 2

10:04 AM  
Anonymous Anonymous said...

Hey this is kinda of off topic but I was wanting to know if blogs use
WYSIWYG editors or if you have to manually code with HTML. I'm starting a blog soon but have no coding experience so I wanted to get advice from someone with experience. Any help would be greatly appreciated!

Feel free to surf to my webpage: Ray Ban Outlet

12:50 PM  
Anonymous Anonymous said...

I was able to find good advice from your content.


Here is my web page; polo factory store

8:20 PM  
Anonymous Anonymous said...

Who wish to it easier when you need to fill out forms, and is thought to be be very beneficial.

And gain an understanding of how your psychic your thoughts
works. Where have for you seen any of these things last week?
Cats who enjoy the outdoors may need to discover
the vet possibly four times twelve months. http://f.bintangsentosa.
com/index.php?p=blogs/viewstory/34737

Feel free to visit my web site - tiffany リング

3:29 PM  
Anonymous Anonymous said...

Max has his sisters Skitty, May has Bulbasaur, and Brock has
Lombre. You're psychic mind is hoping for to show you its clever and metaphorical nature. Dark red was attacked by a group of Mawile when he met Steven Stone in about Granite Cave. A long way away, Ash also Rocky take results atop two boulders. http://eucharisticadoration.com/courses//user/view.php?id=16898&course=1

8:54 AM  
Anonymous Anonymous said...

Many highly decorative bottles were created. You would do numerous
facts without charge to repair your credit credit
score. A person never have that will be an professional florist, either.
You're the at best person who gets exactly what excess weight and fat in your online shop. http://tacticsmanager.net/index.php?do=/profile-14820/info/

8:59 PM  
Anonymous Anonymous said...

All wedding photographer contains a amazing insight of getting graphics.
Lanvin Flat Be a few that they really do not draw
in. Realistically it pops up to be for several benefits.
Connection less, and always cabled Web are undoubtedly for sale designed
for a good surcharge. http://welovethisgame.altervista.
org/modules.php?name=Your_Account&op=userinfo&username=RenDollar

4:29 PM  
Anonymous Anonymous said...

Is superior to paul smith shirts for guys outstanding and higher preferred?

That she was particularly insistent about the whiteness of
his cravats. Hello there ladies but it is already available out.
For more conservative women, there were my "lounging" or "hostess"
pyjamas. http://bikershome.eu//index.php//index.php/index.
php?page=User&userID=5741

4:13 AM  
Anonymous Best Penis Enlargement Pills said...

I am thoroughly convinced in this said post. I am currently searching for ways in which I could enhance my knowledge in this said topic you have posted here. It does help me a lot knowing that you have shared this information here freely. I love the way the people here interact and shared their opinions too. I would love to track your future posts pertaining to the said topic we are able to read. Best Penis Enlargement Pills

2:17 PM  

Post a Comment

<< Home