text:
AppCode Links
Tags3.3 3.3.2 3.3.3 3.4 5Shortcuts 2016.1 2016.1.2 2016.1.3 2016.2 2016.2.1 Bugfixes Champurrado CocoaPods code completion code generation code navigation Debug EAP EAP evaluators editor Espressino Espression examples Features guest post live template Marocchino materials meetup Pebble raffle RC Refactoring Release Roadmap swift Tascalate TDD team Tutorials UIKonf Webinar workshop Xcode Xcode7
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AppCode 3.1.3 update available
Hi everyone, AppCode 3.1.3 is now available for download. You'll also get an automatic notification about this update in the IDE. This update is mostly focused on Swift support, including some new features of Swift 1.2 introduced recently. Parameter Info … Continue reading
Posted in Uncategorized Tagged Champurrado, swift 15 Comments
Special thanks to the EAP builds evaluators
Hi, A week ago we've published AppCode 3.1 release, a new version of our iOS/OS X IDE with the Swift support on board. Now when you work with Swift in AppCode, you will be assisted with many essential tasks in … Continue reading
Posted in Uncategorized 7 Comments
Thanks & Gratitude from our team
Hello everyone, A week ago we've published AppCode 3.0 release. With all the new features and improved workflow with less switches we hope this version will make you more productive. And now it's time to thank those users who evaluated … Continue reading
Posted in Uncategorized 6 Comments
CocoaPods Bug Bash results
Hello, Here are some news about CocoaPods Bug Bash that has happened recently. It worked out amazingly with a nice statistics from the organizers. And the result is CocoaPods 0.31! We also would like to congratulate the winners! The AppCode licenses will … Continue reading
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Julian Król's Take on AppCode
This guest blog post is from Julian Król, Software Engineer at MiQUiDO, a JetBrains Training and Consulting Partner. He has been developing iOS apps for more than a year and a half at MiQUiDO and continues to find it very … Continue reading
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Lucky winners everywhere
October was full of different events which we attended and supported in different ways, and at some of those events there were lucky participants who got a chance to win a personal AppCode license. We would like to congratulate the … Continue reading
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Proudly Developed with AppCode
From time to time people ask us "who uses AppCode?" and we are curious about that too. This is exactly why we started a thread on our discussion forum encouraging you to tell us who you are and what awesome … Continue reading
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Get Involved in AppCode Community!
Do you know what we at JetBrains consider one of the key aspects of creating a great product? Building and maintaining a strong and committed community. You may have guessed that quite lots of people use AppCode on a daily … Continue reading
Posted in Uncategorized 2 Comments


text:
Australian state colours
From Wikipedia, the free encyclopedia
In Australia, state colours are frequently part of a state or territory's set of state symbols.
Some states have formally adopted a set of colours as their official "state colours" while others have de facto state colours that have become well-known through popular use. State colours often appear on a variety of different media, from the state's flag to the colours used in sports. In particular the Sheffield Shield team caps popularised the usage of single colours to represent each state.
|Primary colours
|Secondary colours
|Further information
|Blue and gold[1]
|see ACT Brumbies
|New South Wales
|Sky blue[2]
|see New South Wales rugby league team
|Northern Territory
|Black, white and ochre[3]
|see Northern Territory Football Club
|see Queensland rugby league team
|South Australia
|Blue and gold
|see South Australia Croweaters
|Bottle green[6][7]
|Maroon and yellow
|see Tasmanian Tigers
|Navy blue[8]
|White and Silver
|see Victoria Big V
|Western Australia
|Gold and black
|see Western Australia Sandgropers
See also[edit]
- National colours of Australia
- List of Australian states and territories symbols
- State of Origin
- Interstate matches in Australian rules football
- ^ "ACT Flags and Emblems". ACT Government Chief Minister's Department.
- ^ Symbols and emblems of NSW
- ^ "Northern Territory Flag". Department of the Chief Minister.
- ^ "State colour". Department of the Premier and Cabinet.
- ^ "The State Coat of Arms of South Australia".
- ^ Department of Premier and Cabinet - TASMANIA : Sporting colours
- ^ "Tasmanian Sporting Colours - Story of their Origin". The Mercury. 13 February 1925.
- ^ "Ensigns of Public Authority". Department of Premier and Cabinet.

subset: commoncrawl


text:
Thank you for your interest in Tiny Miracles Doula Services! My name is Jamie Canning. I am a fully certified doula who also has obtained a Masters in Counseling. I was trained and certified through First Breath Midwifery Care for my doula certification and graduated with a masters degree in counseling from the Montana State University-Billings. I also trained and certified through StillBirthday University for birth and berevement doula. I am a mother to two beautiful children and am lucky enough to be able to stay at home with them.
It is my goal to professionally assist expectant mothers, fathers and families through the birth process(s) from the birth plan, through labor and delivery, and the postpartum period. I will be your professional liaison between you and your healthcare providers to ensure your birth plan is honored and performed as completely and as safely as possible. I will be your emotional and professional support to whatever extent is desired. I will provide extended care, training and counseling in the recovery room or even at home if desired. A doula's services are many and cannot be summed up easily, please call with any questions, all consultations are free.
Serving the Billings, Mt area and Yellowstone County
Tiny Miracles Doula Services
Jamie Canning
P.O. Box 51061
Billings, MT 59105
Jamie 406 670-7433
Email: email@example.com
Or use my contact form.
Serving Yellowstone County, Billings area
" I KNOW for certain that if I had not had her as my doula, my experience would not have been what I wanted it to be."
What people are saying about their experience
Special Pricing for Military Families, teens, and adoption!
Payment options available!
Now accepting credit cards and payments by PayPal!!
Birth Pools now available for rent!


text:
3 posts tagged with language by colinmarshall.
Displaying 1 through 3 of 3.
All-Korean podcasts for Korean-language learning?
Which entertaining Korean-language podcasts intended for native speakers can best help a Korean-learner improve his listening skills? [more inside]
Why did so many (specifically) 80s computer games offer Afrikaans as a language option?
Why was Afrikaans so widely available as a language option in computer games of the late 80s and (maybe) the early 90s? [more inside]
Sources for short prose for public editing practice?
Where might I find real-life examples of 300-700-word pieces of prose that are neither perfect nor unsalvageable — "problematic," let's say — on which I can practice the craft of editing? [more inside]
Page: 1


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Search: magnoliaSearch profiles »
MagnoliaMatch % Unknown Join to find out!
Kissingan essayby GeorgiaMagnolia ©Satisfaction is all about just enough and never too much and sometimes leaving with just a little bit of want left, that thought that 'next time I want to try.
Cluracan13Match % Unknown Join to find out!
Alright fine, how about....I always thought it'd be cool to speak Yiddish. I've always wanted cowboy boots and a big belt buckle. The boots would be a bit worn in and dirty looking. I'm not saying I'd go dancing in them, but there are snakes out there in the hills. Sometimes a man...
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) Oddly enough, most of the time I was there I lived off of Magnolia Ave.
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The Jason Test
This test is basically just random questions I came up with based on environmental stimuli, and might possibly measure compatability.
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text: Posted April 9, 2014 2007 North Carolina 4A State Champion 2008 Class A South Atlantic League Champion 2009 Class A+ California League Champion 2010 MLB World Series Champion 2012 MLB World Series Champion Championship Statistics- 5 starts, 5-0 W/L, 36 IP, 3 ER, 6 BB, 44 K But hey, Kershaw gets all the media attention & media awards!!! Go Dodgers!!! Dude you are retarded. Stop sniffing Madison's jockstrap you fugging groupie. Share this post Link to post Share on other sites


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Volume 88 Issue 20 | pp. 25-26
Issue Date: May 17, 2010
Issue Date: May 17, 2010
Tech Assessment Makes A Comeback
Government, independent networks aim to enhance public understanding of science, technology impacts
Department: Government & Policy | Collection: Economy
Keywords: Technology assessment
Keywords: Technology assessment
After more than a decade of neglect, efforts to ramp up technology assessment—the practice of looking at the broad social implications of new science and technology—are under way in the U.S. Discussions are even starting to take place between the government and independent institutions on ways to integrate traditional models that incorporate expert technical opinions with emerging methods that consider the views of laypeople.
Technology assessment has been lagging in the U.S. since the Republican-led . . .
You Do Not Have Access to C&EN Protected Content.
- Chemical & Engineering News
- Copyright © American Chemical Society


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Fund Raising
Thanks for Your Generosity. Now Can You Give Again?
Going back, carefully, to core donors pays off for college fund raisers
As colleges grapple with declining endowments and increased financial needs, a growing number are asking their most generous supporters to dig even deeper.
To cover immediate shortfalls in fund raising and the growing demand for student aid...
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- Report: #472146
Report - Rebuttal - Arbitrate
Complaint Review: Debt Elimination Services
Debt Elimination Services29 Soundview Rd. Guilford, Connecticut U.S.A.
Debt Elimination Services will do NOTHING for you - except take your money! Guilford Connecticut
*REBUTTAL Owner of company: Angry Attorney Gets Fired and Spews at the Mouth
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They pass out 'legal advice' as if they are attorneys - they are not. They give you false, misleading advice, and lie.
Do yourself a favor - if you are in financial trouble, contact a Consumer Advocay Attorney (can be found at naca.net and several other places) - pay them for help - at least they can try to do something.
I'll be contacting the Better Business Bureau, Consumer Advocate of the State (they are not registered with them!) and Attorney General about this company.
Newtown, Connecticut
This report was posted on Ripoff Report on 07/22/2009 08:38 AM and is a permanent record located here: http://www.ripoffreport.com/r/Debt-Elimination-Services/Guilford-Connecticut-06437/Debt-Elimination-Services-will-do-NOTHING-for-you-except-take-your-money-Guilford-Conne-472146. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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#1 REBUTTAL Owner of company
Angry Attorney Gets Fired and Spews at the Mouth
AUTHOR: Businessman in CT - (United States of America)
SUBMITTED: Wednesday, September 19, 2012POSTED: Wednesday, September 19, 2012
Why would a disgruntled consumer refer anyone to legal network? This is the legal network I recommended you join. Just because we hired another law firm, so we could work with an attorney that doesn't lie and yell at my clients, you now want to badmouth me and my company. If your statements were true why did you work with me for six years?
The other reason we fired you is that our clients told us that you are trying to direct funds to you saying that they did not need to hire me. Did you really think I would continue to do business with an attorney that I could not trust? Did you forget you took an oath when you got your law degree.
We have integrity and good reputation. I am disappointed that you are so immature that you would stoop so low as to publish something like this, hiding behind your anonymity of being from Newtown, CT when in fact you are from Branford, CT. I can see from the judicial docket that you and your wife have not been paying your bills. I hope you get the professional help you need.
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With Instructables you can share what you make with the world, and tap into an ever-growing community of creative experts.
This is my first attempt at Instructables. I have been learning to build mountain dulcimers and stick dulcimers for a few months. This is not really a beginner ...
Some tools you may need: I used a band saw to cut the stick and fingerboard to size. Sanders for shaping and smoothing. Drill press for cutting holes ...
You can see in the photo the stick is bent. It enters and exits where it must to have the neck centered. I just cut small pieces of ...
I love how this worked out. Just a little whimsy!
Before anything is glued in place: measure, check and adjust everything several times. I placed dowels in the stick to support the fingerboard along the neck of the ...
I glued the lid to the box shut and dry fit the rest of the parts in place until the box had dried. Then I glued each end ...
Hope I remembered enough to be useful. The rest is up to you and your imagination.
Epic! Guitar Hero - Double Neck Guitar... Failby SCHLEPIC
How to Pick a Guitar by Eran-Li
How to Play Guitar for Beginnersby How to Play Guitar for Beginners
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on November 2, 2012 - 12:01 AM
Put in a quarter and 643-pound, 9-foot-tall bad guy Ralph will tear down an apartment building as its residents holler, but nimble-fingered game players can animate the sprightly Felix, who repairs the damage with his magic hammer. If Felix makes it to the top of the building, he gets a medal. Ralph gets tossed over the side into a mud puddle.
It's been going on like this for 30 years in the arcade game Fix-It Felix.
No wonder Ralph wants a break.
Ralph is describing his unhappiness at a Bad-Anon meeting, a support group for video game villains whose credo is "I'm bad and that's good; I will never be good, and that's not bad."
That's just not working for Ralph. He wants friends, he wants pie, he wants to be the guy who gets the medal sometimes. We've met him in other movies � think of the green ogre Shrek and Beauty's Beast, although he reminded me most of Andre the Giant's wonderful, big-hearted character Fezzig in "The Princess Bride."
It's that way all through "Wreck-It Ralph" � a fun and brilliantly designed derivative treat that we've seen in other shapes for years. Ralph (loudly voiced by John C. Reilly) is on a quest for respect and affection, so he travels among video games old and new to win his prize. Adults will have the most fun recalling their Pac-Man days, trying to speak Q*bert and spotting Montana Smith from Temple Run in the crowd of avatars at Game Central Station.
To get validation in his own game, Ralph heads out to win a medal to show he can be a good guy, too. His visit to the dark armored war game Hero's Duty snaps him into the aggressive reality of today's gamers � "When did video games get so violent and scary?!" he yells as he's pummeled by vicious mechanical bugs and shots fly around him. That's also where we pick up tough-as-nails and curvaceous as doughnuts Sergeant Calhoun (looking and sounding like Jane Lynch), who pursues Ralph to the Sugar Rush game to keep the giant bugs from taking it over.
When Ralph crashes into Sugar Rush, a pervasively pink Candyland car race game, he hooks up with another outcast, the glitchy little Vanellope (Sarah Silverman), a pixielike � and pixilating � nasty-mouthed minx who wants to win the big race so she, too, can be validated.
From here, the movie is a pileup of odd rules that amount to nothing ("if you die outside your game you don't regenerate" is repeated, but is never a threat); other bizarre characters, including King Candy, a mashup of the Mad Hatter and Willy Wonka wearing the face of Ed Wynn; and the most bizarre use ever of Mentos and Diet Coke.
The real drama, and danger, comes from outside the games, in the arcade where they are housed. Without Ralph, Fix-It Felix is declared � gasp!� "out of order." If a game can't be fixed � especially a little-played, decades-old "classic" � it is unplugged and its characters become homeless. We see some of them panhandling in Game Central Station.
Will Ralph get his medal? Will Venellope win her race? Will Sergeant Calhoun overcome being programmed with "the most tragic back-story ever?" Is Fix-It Felix destined to spend the rest of his days in a closet next to old Space Invaders consoles?
After a lot of very silly name-calling and childish fun with the word "duty" (or "doody") it all gets resolved, one way or the other. If you are worried about how, just remember the first name you see in this movie � Disney � and put your fears to rest.
email: email@example.com
2 1/2 stars (out of 4)
Starring: Voices of John C. Reilly, Sarah Silverman, Jane Lynch, Jack McBrayer
Director: Rich Moore
Running time: 83 minutes
Rating: PG-13 for mild violence and rude doody humor.The Lowdown: After 20 years as a video game bad guy, Ralph heads to other games to try to change his life in this Disney animated adventure.


text:
Live Prices Not Available
To show you the most accurate prices, TTPM only displays our pricing tool on products which we've reviewed in the past three years.
With the Fashion Marks Luscious Handbag Designer, kids can color and design their own handbag. The kit comes with one handbag that has outlined images on all sides. Kids use the four Fashion Marks markers to color in the designs any way they want. Then they can add sparkle to the color by putting on the Shimmer FX overlay.
The Fashion Marks Luscious Handbag Designer offers kids creative play. Kids will like the cool designs on the bag and being able to decorate it however they want. Plus, the handbag is functional, so kids can create a useful, personalized accessory.
The Fashion Marks Luscious Handbag Designer kit is for ages 7 and up.
The marker colors do bleed, so be careful not to get the ink on your clothes.
Candy Stripes Highlighters
from Mustard
Monster Art Center
from Alex
Harry Potter House Elves
from Incredibuilds
Harry Potter Golden Snitch
from Incredibuilds
Lil Lockitz Memory Studio
from Alex
SkyPaper High Velocity PPL-1 Paper Plane Launcher
from The Bridge Direct
Stick Lits Light-Up Stickers
from Funsparks
3Doodler Start Essentials Pen Set
from WobbleWorks
Bunchems Alive
from Spin Master
Calafant Pony Farm
from The Haywire Group


text:
- Report: #168741
Report - Rebuttal - Arbitrate
Complaint Review: Newport Marketing Group
Newport Marketing GroupNewport IM Corp., Box 1259 Seattle, Washington U.S.A.
Newport Marketing Group ripoff, spam, email fraud Seattle Washington
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I traced the spams using our domain and found my way to Robert Soloway, a.k.a. Newport Marketing Group.
I called a number I found on the site and left a message saying that I'd like to place an order. At the end of the day, around 6:00 eastern, I received a phone call from a caller I.D. bearing Robert Soloway's #. It was the same # I had called earlier in the day, 206 223 1270. The person I spoke with claimed to be just an "order fulfillment house." I told him, "here's my order; stop emailing me spam from my own domain name." As one might imagine, I got nowhere and I subsequently found this site, where I am leaving this information. There seems to be no shortage of this guy's spam cruising around online.
ny, New York
This report was posted on Ripoff Report on 12/20/2005 06:00 PM and is a permanent record located here: http://www.ripoffreport.com/r/Newport-Marketing-Group/Seattle-Washington-98111/Newport-Marketing-Group-ripoff-spam-email-fraud-Seattle-Washington-168741. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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text:
Starbucks Drinks
Treat Receipt is Back
For a limited time, get any cold grande beverage for $2.50 when you bring in your same day morning receipt after 2 pm. Try the new Iced Coconut Milk Mocha Macchiato. A hint of white chocolate mocha, Sumatran coconut milk, rich espresso shots, caramel and mocha drizzle create a perfectly-balanced, creamy flavor that'll keep you cool this season.
At participating stores. No cash redemption value. Void where prohibited. Cannot be combined with other discounts. Not valid if reproduced or transferred
Starbucks Refreshers™ Beverages
Starbucks Refreshers™ Beverages
Evolution Fresh™
Discover Evolution Fresh Cold-Pressed Juice.
Iced Coffee
Iced Coffee
Iced Tea
Iced Tea
Bottled Drinks
Now you can take Starbucks with you wherever you go.
Freshly Brewed Coffee
Artfully roasted and brewed fresh all day long, every day.
Chocolate Beverages
Luxurious hot chocolate for the sophisticated palate.
Starbucks Espresso Beverages
Lattes, cappuccinos, macchiatos, mochas and much more.
Frappuccino® Blended Beverages
The icy-cold original we made famous.
Kids' Drinks & Others
Some fun, flavorful alternatives for anyone to enjoy.
Nourishing blends made with real fruit and other good stuff.
Fizzio™ Handcrafted Sodas
Handcrafted sodas with no high-fructose corn syrup, artificial flavors, or preservatives.
Featured Offerings
Vanilla Sweet Cream Cold Brew
Just before serving, our slow-steeped custom blend Nariño 70 Cold Brew is topped with a delicate float of house-made vanilla sweet cream that cascades throughout the cup.
Starbucks Doubleshot® On Ice Beverage
Made with the rich, full-bodied espresso you love that's been chilled and mellowed with a touch of milk and lightly sweetened, this drink is delightfully convenient.
Teavana® Shaken Berry Sangria Herbal Tea
Iced tea is re-imagined thanks to inspiration from a classic summer cocktail. Freshly brewed Teavana® Iced Passion Tango™ Tea and apple juice are hand shaken with a medley of juicy berry flavors then accented with real blackberries and orange slices. The delicious result? A finish that's both sweet and tart.


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Evolution of Retinoid and Steroid Signaling: Vertebrate Diversification from an Amphioxus Perspective
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Although the physiological relevance of retinoids and steroids in vertebrates is very well established, the origin and evolution of the genetic machineries implicated in their metabolic pathways is still very poorly understood. We investigated the evolution of these genetic networks by conducting an exhaustive survey of components of the retinoid and steroid pathways in the genome of the invertebrate chordate amphioxus (Branchiostoma floridae). Due to its phylogenetic position at the base of chordates, amphioxus is a very useful model to identify and study chordate versus vertebrate innovations, both on a morphological and a genomic level. We have characterized more than 220 amphioxus genes evolutionarily related to vertebrate components of the retinoid and steroid pathways and found that, globally, amphioxus has orthologs of most of the vertebrate components of these two pathways, with some very important exceptions. For example, we failed to identify a vertebrate-like machinery for retinoid storage, transport, and delivery in amphioxus and were also unable to characterize components of the adrenal steroid pathway in this invertebrate chordate. The absence of these genes from the amphioxus genome suggests that both an elaboration and a refinement of the retinoid and steroid pathways took place at the base of the vertebrate lineage. In stark contrast, we also identified massive amplifications in some amphioxus gene families, most extensively in the short-chain dehydrogenase/reductase superfamily, which, based on phylogenetic and genomic linkage analyses, were likely the result of duplications specific to the amphioxus lineage. In sum, this detailed characterization of genes implicated in retinoid and steroid signaling in amphioxus allows us not only to reconstruct an outline of these pathways in the ancestral chordate but also to discuss functional innovations in retinoid homeostasis and steroid-dependent regulation in both cephalochordate and vertebrate evolution.
Branchiostoma floridae; cephalochordates; gene duplication; metabolic networks; nuclear receptor signaling cascades
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The globin gene family of the cephalochordate amphioxus: implications for chordate globin evolution
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Vinogradov, Serge N
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BMC Evolutionary Biology
The lancelet amphioxus (Cephalochordata) is a close relative of vertebrates and thus may enhance our understanding of vertebrate gene and genome evolution. In this context, the globins are one of the best studied models for gene family evolution. Previous biochemical studies have demonstrated the presence of an intracellular globin in notochord tissue and myotome of amphioxus, but the corresponding gene has not yet been identified. Genomic resources of Branchiostoma floridae now facilitate the identification, experimental confirmation and molecular evolutionary analysis of its globin gene repertoire.
We show that B. floridae harbors at least fifteen paralogous globin genes, all of which reveal evidence of gene expression. The protein sequences of twelve globins display the conserved characteristics of a functional globin fold. In phylogenetic analyses, the amphioxus globin BflGb4 forms a common clade with vertebrate neuroglobins, indicating the presence of this nerve globin in cephalochordates. Orthology is corroborated by conserved syntenic linkage of BflGb4 and flanking genes. The kinetics of ligand binding of recombinantly expressed BflGb4 reveals that this globin is hexacoordinated with a high oxygen association rate, thus strongly resembling vertebrate neuroglobin. In addition, possible amphioxus orthologs of the vertebrate globin X lineage and of the myoglobin/cytoglobin/hemoglobin lineage can be identified, including one gene as a candidate for being expressed in notochord tissue. Genomic analyses identify conserved synteny between amphioxus globin-containing regions and the vertebrate β-globin locus, possibly arguing against a late transpositional origin of the β-globin cluster in vertebrates. Some amphioxus globin gene structures exhibit minisatellite-like tandem duplications of intron-exon boundaries ("mirages"), which may serve to explain the creation of novel intron positions within the globin genes.
The identification of putative orthologs of vertebrate globin variants in the B. floridae genome underlines the importance of cephalochordates for elucidating vertebrate genome evolution. The present study facilitates detailed functional studies of the amphioxus globins in order to trace conserved properties and specific adaptations of respiratory proteins at the base of chordate evolution.
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BMC Neuroscience
Amphioxus, representing the most basal group of living chordates, is the best available proxy for the last invertebrate ancestor of the chordates. Although the central nervous system (CNS) of amphioxus comprises only about 20,000 neurons (as compared to billions in vertebrates), the developmental genetics and neuroanatomy of amphioxus are strikingly vertebrate-like. In the present study, we mapped the distribution of amphioxus CNS cells producing distinctive neurochemicals. To this end, we cloned genes encoding biosynthetic enzymes and/or transporters of the most common neurotransmitters and assayed their developmental expression in the embryo and early larva.
By single and double in situ hybridization experiments, we identified glutamatergic, GABAergic/glycinergic, serotonergic and cholinergic neurons in developing amphioxus. In addition to characterizing the distribution of excitatory and inhibitory neurons in the developing amphioxus CNS, we observed that cholinergic and GABAergic/glycinergic neurons are segmentally arranged in the hindbrain, whereas serotonergic, glutamatergic and dopaminergic neurons are restricted to specific regions of the cerebral vesicle and the hindbrain. We were further able to identify discrete groups of GABAergic and glutamatergic interneurons and cholinergic motoneurons at the level of the primary motor center (PMC), the major integrative center of sensory and motor stimuli of the amphioxus nerve cord.
In this study, we assessed neuronal differentiation in the developing amphioxus nervous system and compiled the first neurochemical map of the amphioxus CNS. This map is a first step towards a full characterization of the neurotransmitter signature of previously described nerve cell types in the amphioxus CNS, such as motoneurons and interneurons.
Branchiostoma; Cephalochordate; Chordate evolution; Lancelet; Neural patterning; Neurotransmitter
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Broken colinearity of the amphioxus Hox cluster
Pascual-Anaya, Juan
Adachi, Noritaka
Álvarez, Susana
Kuratani, Shigeru
D'Aniello, Salvatore
Garcia-Fernàndez, Jordi
In most eumetazoans studied so far, Hox genes determine the identity of structures along the main body axis. They are usually linked in genomic clusters and, in the case of the vertebrate embryo, are expressed with spatial and temporal colinearity. Outside vertebrates, temporal colinearity has been reported in the cephalochordate amphioxus (the least derived living relative of the chordate ancestor) but only for anterior and central genes, namely Hox1 to Hox4 and Hox6. However, most of the Hox gene expression patterns in amphioxus have not been reported. To gain global insights into the evolution of Hox clusters in chordates, we investigated a more extended expression profile of amphioxus Hox genes.
Here we report an extended expression profile of the European amphioxus Branchiostoma lanceolatum Hox genes and describe that all Hox genes, except Hox13, are expressed during development. Interestingly, we report the breaking of both spatial and temporal colinearity for at least Hox6 and Hox14, which thus have escaped from the classical Hox code concept. We show a previously unidentified Hox6 expression pattern and a faint expression for posterior Hox genes in structures such as the posterior mesoderm, notochord, and hindgut. Unexpectedly, we found that amphioxus Hox14 had the most divergent expression pattern. This gene is expressed in the anterior cerebral vesicle and pharyngeal endoderm. Amphioxus Hox14 expression represents the first report of Hox gene expression in the most anterior part of the central nervous system. Nevertheless, despite these divergent expression patterns, amphioxus Hox6 and Hox14 seem to be still regulated by retinoic acid.
Escape from colinearity by Hox genes is not unusual in either vertebrates or amphioxus and we suggest that those genes escaping from it are probably associated with the patterning of lineage-specific morphological traits, requiring the loss of those developmental constraints that kept them colinear.
Hox gene regulation; Hox cluster evolution; Amphioxus; Hox colinearity; Retinoic acid
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Genome-wide survey and expression analysis of the bHLH-PAS genes in the amphioxus Branchiostoma floridae reveal both conserved and diverged expression patterns between cephalochordates and vertebrates
Li, Kun-Lung
Lu, Tsai-Ming
Yu, Jr-Kai
The bHLH-PAS transcription factors are found in both protostomes and deuterostomes. They are involved in many developmental and physiological processes, including regional differentiation of the central nervous system, tube-formation, hypoxia signaling, aromatic hydrocarbon sensing, and circadian rhythm regulation. To understand the evolution of these genes in chordates, we analyzed the bHLH-PAS genes of the basal chordate amphioxus (Branchiostoma floridae).
From the amphioxus draft genome database, we identified ten bHLH-PAS genes, nine of which could be assigned to known orthologous families. The tenth bHLH-PAS gene could not be assigned confidently to any known bHLH family; however, phylogenetic analysis clustered this gene with arthropod Met family genes and two spiralian bHLH-PAS-containing sequences, suggesting that they may share the same ancestry. We examined temporal and spatial expression patterns of these bHLH-PAS genes in developing amphioxus embryos. We found that BfArnt, BfNcoa, BfSim, and BfHifα were expressed in the central nervous system in patterns similar to those of their vertebrate homologs, suggesting that their functions may be conserved. By contrast, the amphioxus BfAhr and BfNpas4 had expression patterns distinct from those in vertebrates. These results imply that there were changes in gene regulation after the divergence of cephalochordates and vertebrates.
We have identified ten bHLH-PAS genes from the amphioxus genome and determined the embryonic expression profiles for these genes. In addition to the nine currently recognized bHLH-PAS families, our survey suggests that the BfbHLHPAS-orphan gene along with arthropod Met genes and the newly identified spiralian bHLH-PAS-containing sequences represent an ancient group of genes that were lost in the vertebrate lineage. In a comparison with the expression patterns of the vertebrate bHLH-PAS paralogs, which are the result of whole-genome duplication, we found that although several members seem to retain conserved expression patterns during chordate evolution, many duplicated paralogs may have undergone subfunctionalization and neofunctionalization in the vertebrate lineage. In addition, our survey of amphioxus bHLH-PAS gene models from genome browser with experimentally verified cDNA sequences calls into question the accuracy of the current in silico gene annotation of the B. floridae genome.
Amphioxus; bHLH-PAS transcription factors; Branchiostoma floridae; Embryonic development; Molecular phylogeny
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Evolution of developmental roles of Pax2/5/8 paralogs after independent duplication in urochordate and vertebrate lineages
Bassham, Susan
Cañestro, Cristian
Postlethwait, John H
BMC Biology
Gene duplication provides opportunities for lineage diversification and evolution of developmental novelties. Duplicated genes generally either disappear by accumulation of mutations (nonfunctionalization), or are preserved either by the origin of positively selected functions in one or both duplicates (neofunctionalization), or by the partitioning of original gene subfunctions between the duplicates (subfunctionalization). The Pax2/5/8 family of important developmental regulators has undergone parallel expansion among chordate groups. After the divergence of urochordate and vertebrate lineages, two rounds of independent gene duplications resulted in the Pax2, Pax5, and Pax8 genes of most vertebrates (the sister group of the urochordates), and an additional duplication provided the pax2a and pax2b duplicates in teleost fish. Separate from the vertebrate genome expansions, a duplication also created two Pax2/5/8 genes in the common ancestor of ascidian and larvacean urochordates.
To better understand mechanisms underlying the evolution of duplicated genes, we investigated, in the larvacean urochordate Oikopleura dioica, the embryonic gene expression patterns of Pax2/5/8 paralogs. We compared the larvacean and ascidian expression patterns to infer modular subfunctions present in the single pre-duplication Pax2/5/8 gene of stem urochordates, and we compared vertebrate and urochordate expression to infer the suite of Pax2/5/8 gene subfunctions in the common ancestor of olfactores (vertebrates + urochordates). Expression pattern differences of larvacean and ascidian Pax2/5/8 orthologs in the endostyle, pharynx and hindgut suggest that some ancestral gene functions have been partitioned differently to the duplicates in the two urochordate lineages. Novel expression in the larvacean heart may have resulted from the neofunctionalization of a Pax2/5/8 gene in the urochordates. Expression of larvacean Pax2/5/8 in the endostyle, in sites of epithelial remodeling, and in sensory tissues evokes like functions of Pax2, Pax5 and Pax8 in vertebrate embryos, and may indicate ancient origins for these functions in the chordate common ancestor.
Comparative analysis of expression patterns of chordate Pax2/5/8 duplicates, rooted on the single-copy Pax2/5/8 gene of amphioxus, whose lineage diverged basally among chordates, provides new insights into the evolution and development of the heart, thyroid, pharynx, stomodeum and placodes in chordates; supports the controversial conclusion that the atrial siphon of ascidians and the otic placode in vertebrates are homologous; and backs the notion that Pax2/5/8 functioned in ancestral chordates to engineer epithelial fusions and perforations, including gill slit openings.
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Neofunctionalization in Vertebrates: The Example of Retinoic Acid Receptors
Escriva, Hector
Bertrand, Stéphanie
Germain, Pierre
Robinson-Rechavi, Marc
Umbhauer, Muriel
Cartry, Jérôme
Duffraisse, Marilyne
Holland, Linda
Gronemeyer, Hinrich
Laudet, Vincent
PLoS Genetics
Understanding the role of gene duplications in establishing vertebrate innovations is one of the main challenges of Evo-Devo (evolution of development) studies. Data on evolutionary changes in gene expression (i.e., evolution of transcription factor-cis-regulatory elements relationships) tell only part of the story; protein function, best studied by biochemical and functional assays, can also change. In this study, we have investigated how gene duplication has affected both the expression and the ligand-binding specificity of retinoic acid receptors (RARs), which play a major role in chordate embryonic development. Mammals have three paralogous RAR genes—RARα, β, and γ—which resulted from genome duplications at the origin of vertebrates. By using pharmacological ligands selective for specific paralogues, we have studied the ligand-binding capacities of RARs from diverse chordates species. We have found that RARβ-like binding selectivity is a synapomorphy of all chordate RARs, including a reconstructed synthetic RAR representing the receptor present in the ancestor of chordates. Moreover, comparison of expression patterns of the cephalochordate amphioxus and the vertebrates suggests that, of all the RARs, RARβ expression has remained most similar to that of the ancestral RAR. On the basis of these results together, we suggest that while RARβ kept the ancestral RAR role, RARα and RARγ diverged both in ligand-binding capacity and in expression patterns. We thus suggest that neofunctionalization occurred at both the expression and the functional levels to shape RAR roles during development in vertebrates.
In eukaryotic organisms, each gene is a stretch of DNA composed of control regions that bind transcription factors and coding regions that transcribe the mRNA that is later translated into proteins. At the molecular level, changes in control regions can affect the time and place at which a protein is synthesized, whereas changes in the coding region can alter the protein's function. Retinoic acid receptors (RARs) are chordate-specific transcription factors which, upon binding the natural morphogen retinoic acid, bind to and activate transcription from target genes. Here, the authors show how the ligand specificity of RARs has changed during vertebrate evolution in parallel with changes in expression. Through functional characterization of the RARs from several vertebrates, the chordate amphioxus, and the reconstructed ancestral RAR sequence, the authors show that of the three vertebrate RARs, RARβ has retained the ancestral characteristics in terms of both function and expression, while RARα and γ have evolved by acquiring new functions, both new binding specificity and new expression patterns. Thus both types of evolution have been important in the diversification of vertebrate RARs.
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Amphioxus (Branchiostoma floridae) has orthologs of vertebrate odorant receptors
Churcher, Allison M
Taylor, John S
BMC Evolutionary Biology
A common feature of chemosensory systems is the involvement of G protein-coupled receptors (GPCRs) in the detection of environmental stimuli. Several lineages of GPCRs are involved in vertebrate olfaction, including trace amine-associated receptors, type 1 and 2 vomeronasal receptors and odorant receptors (ORs). Gene duplication and gene loss in different vertebrate lineages have lead to an enormous amount of variation in OR gene repertoire among species; some fish have fewer than 100 OR genes, while some mammals possess more than 1000. Fascinating features of the vertebrate olfactory system include allelic exclusion, where each olfactory neuron expresses only a single OR gene, and axonal guidance where neurons expressing the same receptor project axons to common glomerulae. By identifying homologous ORs in vertebrate and in non-vertebrate chordates, we hope to expose ancestral features of the chordate olfactory system that will help us to better understand the evolution of the receptors themselves and of the cellular components of the olfactory system.
We have identified 50 full-length and 11 partial ORs in Branchiostoma floridae. No ORs were identified in Ciona intestinalis. Phylogenetic analysis places the B. floridae OR genes in a monophyletic clade with the vertebrate ORs. The majority of OR genes in amphioxus are intronless and many are also tandemly arrayed in the genome. By exposing conserved amino acid motifs and testing the ability of those motifs to discriminate between ORs and non-OR GPCRs, we identified three OR-specific amino acid motifs common in cephalochordate, fish and mammalian and ORs.
Here, we show that amphioxus has orthologs of vertebrate ORs. This conclusion demonstrates that the receptors, and perhaps other components of vertebrate olfaction, evolved at least 550 million years ago. We have also identified highly conserved amino acid motifs that may be important for maintaining receptor conformation or regulating receptor activity. We anticipate that the identification of vertebrate OR orthologs in amphioxus will lead to an improved understanding of OR gene family evolution, OR gene function, and the mechanisms that control cell-specific expression, axonal guidance, signal transduction and signal integration.
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Ion Channel Clustering at the Axon Initial Segment and Node of Ranvier Evolved Sequentially in Early Chordates
Hill, Alexis S.
Nishino, Atsuo
Nakajo, Koichi
Zhang, Giuxin
Fineman, Jaime R.
Selzer, Michael E.
Okamura, Yasushi
Cooper, Edward C.
PLoS Genetics
In many mammalian neurons, dense clusters of ion channels at the axonal initial segment and nodes of Ranvier underlie action potential generation and rapid conduction. Axonal clustering of mammalian voltage-gated sodium and KCNQ (Kv7) potassium channels is based on linkage to the actin–spectrin cytoskeleton, which is mediated by the adaptor protein ankyrin-G. We identified key steps in the evolution of this axonal channel clustering. The anchor motif for sodium channel clustering evolved early in the chordate lineage before the divergence of the wormlike cephalochordate, amphioxus. Axons of the lamprey, a very primitive vertebrate, exhibited some invertebrate features (lack of myelin, use of giant diameter to hasten conduction), but possessed narrow initial segments bearing sodium channel clusters like in more recently evolved vertebrates. The KCNQ potassium channel anchor motif evolved after the divergence of lampreys from other vertebrates, in a common ancestor of shark and humans. Thus, clustering of voltage-gated sodium channels was a pivotal early innovation of the chordates. Sodium channel clusters at the axon initial segment serving the generation of action potentials evolved long before the node of Ranvier. KCNQ channels acquired anchors allowing their integration into pre-existing sodium channel complexes at about the same time that ancient vertebrates acquired myelin, saltatory conduction, and hinged jaws. The early chordate refinements in action potential mechanisms we have elucidated appear essential to the complex neural signaling, active behavior, and evolutionary success of vertebrates.
Author Summary
Because nervous systems generate behavior, innovations that confer new neuronal signaling functions are important potential factors in evolution. In mammals, clustering of ion channels on nerves is essential for electrical impulses used in rapid signaling. This channel clustering is generally absent in insects, worms, and other non-chordates. We traced the evolutionary emergence of mechanisms underlying channel clustering on nerves by analyzing the genomes of primitive chordates and studying the cellular distribution and functional properties of their channels. We found that sodium channel clustering evolved early in the chordate lineage, before the divergence of the earliest wormlike and planktonic groups (lancelets and sea squirts). Nerve fibers of the lamprey, a primitive fish, retained some invertebrate features but possessed dense sodium channel clusters like in more recently evolved vertebrates. A potassium channel clustering system evolved, after the divergence of lampreys, in a common ancestor of shark and humans. We conclude that the clustering of sodium channels on axons was the initial pivotal step in a chordate-specific series of evolutionary innovations, making nerve impulses more rapid and robust. The refinements in action potentials we have elucidated appear essential for the complex neural signaling and active behavior of vertebrates.
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Sequencing and Analysis of the Mediterranean Amphioxus (Branchiostoma lanceolatum) Transcriptome
Oulion, Silvan
Bertrand, Stephanie
Belgacem, Mohamed R.
Le Petillon, Yann
Escriva, Hector
PLoS ONE
The basally divergent phylogenetic position of amphioxus (Cephalochordata), as well as its conserved morphology, development and genetics, make it the best proxy for the chordate ancestor. Particularly, studies using the amphioxus model help our understanding of vertebrate evolution and development. Thus, interest for the amphioxus model led to the characterization of both the transcriptome and complete genome sequence of the American species, Branchiostoma floridae. However, recent technical improvements allowing induction of spawning in the laboratory during the breeding season on a daily basis with the Mediterranean species Branchiostoma lanceolatum have encouraged European Evo-Devo researchers to adopt this species as a model even though no genomic or transcriptomic data have been available. To fill this need we used the pyrosequencing method to characterize the B. lanceolatum transcriptome and then compared our results with the published transcriptome of B. floridae.
Starting with total RNA from nine different developmental stages of B. lanceolatum, a normalized cDNA library was constructed and sequenced on Roche GS FLX (Titanium mode). Around 1.4 million of reads were produced and assembled into 70,530 contigs (average length of 490 bp). Overall 37% of the assembled sequences were annotated by BlastX and their Gene Ontology terms were determined. These results were then compared to genomic and transcriptomic data of B. floridae to assess similarities and specificities of each species.
We obtained a high-quality amphioxus (B. lanceolatum) reference transcriptome using a high throughput sequencing approach. We found that 83% of the predicted genes in the B. floridae complete genome sequence are also found in the B. lanceolatum transcriptome, while only 41% were found in the B. floridae transcriptome obtained with traditional Sanger based sequencing. Therefore, given the high degree of sequence conservation between different amphioxus species, this set of ESTs may now be used as the reference transcriptome for the Branchiostoma genus.
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A dynamic history of gene duplications and losses characterizes the evolution of the SPARC family in eumetazoans
Bertrand, Stephanie
Fuentealba, Jaime
Aze, Antoine
Hudson, Clare
Yasuo, Hitoyoshi
Torrejon, Marcela
Escriva, Hector
Marcellini, Sylvain
Proceedings of the Royal Society B: Biological Sciences
The vertebrates share the ability to produce a skeleton made of mineralized extracellular matrix. However, our understanding of the molecular changes that accompanied their emergence remains scarce. Here, we describe the evolutionary history of the SPARC (secreted protein acidic and rich in cysteine) family, because its vertebrate orthologues are expressed in cartilage, bones and teeth where they have been proposed to bind calcium and act as extracellular collagen chaperones, and because further duplications of specific SPARC members produced the small calcium-binding phosphoproteins (SCPP) family that is crucial for skeletal mineralization to occur. Both phylogeny and synteny conservation analyses reveal that, in the eumetazoan ancestor, a unique ancestral gene duplicated to give rise to SPARC and SPARCB described here for the first time. Independent losses have eliminated one of the two paralogues in cnidarians, protostomes and tetrapods. Hence, only non-tetrapod deuterostomes have conserved both genes. Remarkably, SPARC and SPARCB paralogues are still linked in the amphioxus genome. To shed light on the evolution of the SPARC family members in chordates, we performed a comprehensive analysis of their embryonic expression patterns in amphioxus, tunicates, teleosts, amphibians and mammals. Our results show that in the chordate lineage SPARC and SPARCB family members were recurrently recruited in a variety of unrelated tissues expressing collagen genes. We propose that one of the earliest steps of skeletal evolution involved the co-expression of SPARC paralogues with collagenous proteins.
SPARC gene family; eumetazoans; ancient duplication; independent losses; vertebrate evolution
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Identification and characterization of novel amphioxus microRNAs by Solexa sequencing
Chen, Xi
Li, Qibin
Wang, Jin
Guo, Xing
Jiang, Xiangrui
Ren, Zhiji
Weng, Chunyue
Sun, Guoxun
Wang, Xiuqiang
Liu, Yaping
Ma, Lijia
Chen, Jun-Yuan
Wang, Jun
Zen, Ke
Zhang, Junfeng
Zhang, Chen-Yu
Genome Biology
An analysis of amphioxus miRNAs suggests an expansion of miRNAs played a key role in the evolution of chordates to vertebrates
microRNAs (miRNAs) are endogenous small non-coding RNAs that regulate gene expression at the post-transcriptional level. While the number of known human and murine miRNAs is continuously increasing, information regarding miRNAs from other species such as amphioxus remains limited.
We combined Solexa sequencing with computational techniques to identify novel miRNAs in the amphioxus species B. belcheri (Gray). This approach allowed us to identify 113 amphioxus miRNA genes. Among them, 55 were conserved across species and encoded 45 non-redundant mature miRNAs, whereas 58 were amphioxus-specific and encoded 53 mature miRNAs. Validation of our results with microarray and stem-loop quantitative RT-PCR revealed that Solexa sequencing is a powerful tool for miRNA discovery. Analyzing the evolutionary history of amphioxus miRNAs, we found that amphioxus possesses many miRNAs unique to chordates and vertebrates, and these may thus represent key steps in the evolutionary progression from cephalochordates to vertebrates. We also found that amphioxus is more similar to vertebrates than are tunicates with respect to their miRNA phylogenetic histories.
Taken together, our results indicate that Solexa sequencing allows the successful discovery of novel miRNAs from amphioxus with high accuracy and efficiency. More importantly, our study provides an opportunity to decipher how the elaboration of the miRNA repertoire that occurred during chordate evolution contributed to the evolution of the vertebrate body plan.
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No more than 14: the end of the amphioxus Hox cluster
Minguillón, Carolina
Gardenyes, Josep
Serra, Elisa
Castro, L. Filipe C.
Hill-Force, Alicia
Holland, Peter W.H.
Amemiya, Chris T.
Garcia-Fernàndez, Jordi
International Journal of Biological Sciences
The Hox gene cluster has been a key paradigm for a generation of developmental and evolutionary biologists. Since its discovery in the mid-1980's, the identification, genomic organization, expression, colinearity, and regulation of Hox genes have been immediate targets for study in any new model organism, and metazoan genome projects always refer to the structure of the particular Hox cluster(s). Since the early 1990's, it has been dogma that vertebrate Hox clusters are composed of thirteen paralogous groups. Nonetheless, we showed that in the otherwise prototypical cephalochordate amphioxus (Branchiostoma floridae), the Hox cluster contains a fourteenth Hox gene, and very recently, a 14th Hox paralogous group has been found in the coelacanth and the horn shark, suggesting that the amphioxus cluster was anticipating the finding of Hox 14 in some vertebrate lineages. In view of the pivotal place that amphioxus occupies in vertebrate evolution, we thought it of considerable interest to establish the limits of its Hox gene cluster, namely resolution of whether more Hox genes are present in the amphioxus cluster (e.g., Hox 15). Using two strategies, here we report the completion and characterization of the Hox gene content of the single amphioxus Hox cluster, which encompasses 650 kb from Hox1 to Evx. Our data have important implications for the primordial Hox gene cluster of chordates: the prototypical nature of the single amphioxus Hox cluster makes it unlikely that additional paralogous groups will be found in any chordate lineage. We suggest that 14 is the end.
gene clusters; Evx; gene duplication; vertebrate evolution
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The amphioxus genome provides unique insight into the evolution of immunity
Dishaw, Larry J.
Haire, Robert N.
Litman, Gary W.
Briefings in Functional Genomics
Immune systems evolve as essential strategies to maintain homeostasis with the environment, prevent microbial assault and recycle damaged host tissues. The immune system is composed of two components, innate and adaptive immunity. The former is common to all animals while the latter consists of a vertebrate-specific system that relies on somatically derived lymphocytes and is associated with near limitless genetic diversity as well as long-term memory. Deuterostome invertebrates provide a view of immune repertoires in phyla that immediately predate the origins of vertebrates. Genomic studies in amphioxus, a cephalochordate, have revealed homologs of genes encoding most innate immune receptors found in vertebrates; however, many of the gene families have undergone dramatic expansions, greatly increasing the innate immune repertoire. In addition, domain-swapping accounts for the innovation of new predicted pathways of receptor function. In both amphioxus and Ciona, a urochordate, the VCBPs (variable region containing chitin-binding proteins), which consist of immunoglobulin V (variable) and chitin binding domains, mediate recognition through the V domains. The V domains of VCBPs in amphioxus exhibit high levels of allelic complexity that presumably relate to functional specificity. Various features of the amphioxus immune repertoire reflect novel selective pressures, which likely have resulted in innovative strategies. Functional genomic studies underscore the value of amphioxus as a model for studying innate immunity and may help reveal how unique relationships between innate immune receptors and both pathogens and symbionts factored in the evolution of adaptive immune systems.
innate immunity; Toll-like receptors; expanded immune repertoire; allelic complexity; gut immunity
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An Otx/Nodal Regulatory Signature for Posterior Neural Development in Ascidians
Roure, Agnès
Lemaire, Patrick
Darras, Sébastien
PLoS Genetics
In chordates, neural induction is the first step of a complex developmental process through which ectodermal cells acquire a neural identity. In ascidians, FGF-mediated neural induction occurs at the 32-cell stage in two blastomere pairs, precursors respectively of anterior and posterior neural tissue. We combined molecular embryology and cis-regulatory analysis to unveil in the ascidian Ciona intestinalis the remarkably simple proximal genetic network that controls posterior neural fate acquisition downstream of FGF. We report that the combined action of two direct FGF targets, the TGFβ factor Nodal, acting via Smad- and Fox-binding sites, and the transcription factor Otx suffices to trigger ascidian posterior neural tissue formation. Moreover, we found that this strategy is conserved in the distantly related ascidian Phallusia mammillata, in spite of extreme sequence divergence in the cis-regulatory sequences involved. Our results thus highlight that the modes of gene regulatory network evolution differ with the evolutionary scale considered. Within ascidians, developmental regulatory networks are remarkably robust to genome sequence divergence. Between ascidians and vertebrates, major fate determinants, such as Otx and Nodal, can be co-opted into different networks. Comparative developmental studies in ascidians with divergent genomes will thus uncover shared ascidian strategies, and contribute to a better understanding of the diversity of developmental strategies within chordates.
Author Summary
The Chordate phylum groups vertebrates, tunicates (including ascidians) and cephalochordates (amphioxus). These animals share a typical body plan characterized by the presence during embryonic life of a notochord and a dorsal neural tube. Ascidians, however, took a significantly different evolutionary path from other chordates resulting in divergent morphological, embryological and genomic features. Their development is fast and stereotyped with very few cells and ascidian genomes have undergone compaction and extensive rearrangements when compared to vertebrates, but also between ascidian species. This raises the question of whether developmental mechanisms controlling typical chordate structure formation are conserved between ascidians and vertebrates. Here, we have studied the set of ascidian genes which control the formation of the posterior part of the nervous system. We uncovered original usages of the signaling molecule Nodal and the transcription factor Otx. For example, Otx, which is a specific determinant of anterior identity in most metazoans, has been co-opted for the formation of the ascidian posterior nervous system. These two factors define a regulatory signature found in enhancers of posterior neural genes in two genomically divergent ascidian species.
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FGFRL1 is a neglected putative actor of the FGF signalling pathway present in all major metazoan phyla
Bertrand, Stephanie
Somorjai, Ildiko
Garcia-Fernandez, Jordi
Lamonerie, Thomas
Escriva, Hector
BMC Evolutionary Biology
Fibroblast Growth Factors (FGF) and their receptors are well known for having major implications in cell signalling controlling embryonic development. Recently, a gene coding for a protein closely related to FGFRs (Fibroblast Growth Factor Receptors) called FGFR5 or FGFR-like 1 (FGFRL1), has been described in vertebrates. An orthologous gene was also found in the cephalochordate amphioxus, but no orthologous genes were found by the authors in other non-vertebrate species, even if a FGFRL1 gene was identified in the sea urchin genome, as well as a closely related gene, named nou-darake, in the planarian Dugesia japonica. These intriguing data of a deuterostome-specific gene that might be implicated in FGF signalling prompted us to search for putative FGFRL1 orthologues in the completely sequenced genomes of metazoans.
We found FGFRL1 genes in the cnidarian Nematostella vectensis as well as in many bilaterian species. Our analysis also shows that FGFRL1 orthologous genes are linked in the genome with other members of the FGF signalling pathway from cnidarians to bilaterians (distance < 10 Mb). To better understand the implication of FGFRL1 genes in chordate embryonic development, we have analyzed expression patterns of the amphioxus and the mouse genes by whole mount in situ hybridization. We show that some homologous expression territories can be defined, and we propose that FGFRL1 and FGF8/17/18 were already co-expressed in the pharyngeal endoderm in the ancestor of chordates.
Our work sheds light on the existence of a putative FGF signalling pathway actor present in the ancestor of probably all metazoans, the function of which has received little attention until now.
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Genomic organization and gene expression of the multiple globins in Atlantic cod: conservation of globin-flanking genes in chordates infers the origin of the vertebrate globin clusters
Wetten, Ola F
Nederbragt, Alexander J
Wilson, Robert C
Jakobsen, Kjetill S
Edvardsen, Rolf B
Andersen, Øivind
BMC Evolutionary Biology
The vertebrate globin genes encoding the α- and β-subunits of the tetrameric hemoglobins are clustered at two unlinked loci. The highly conserved linear order of the genes flanking the hemoglobins provides a strong anchor for inferring common ancestry of the globin clusters. In fish, the number of α-β-linked globin genes varies considerably between different sublineages and seems to be related to prevailing physico-chemical conditions. Draft sequences of the Atlantic cod genome enabled us to determine the genomic organization of the globin repertoire in this marine species that copes with fluctuating environments of the temperate and Arctic regions.
The Atlantic cod genome was shown to contain 14 globin genes, including nine hemoglobin genes organized in two unlinked clusters designated β5-α1-β1-α4 and β3-β4-α2-α3-β2. The diverged cod hemoglobin genes displayed different expression levels in adult fish, and tetrameric hemoglobins with or without a Root effect were predicted. The novel finding of maternally inherited hemoglobin mRNAs is consistent with a potential role played by fish hemoglobins in the non-specific immune response. In silico analysis of the six teleost genomes available showed that the two α-β globin clusters are flanked by paralogs of five duplicated genes, in agreement with the proposed teleost-specific duplication of the ancestral vertebrate globin cluster. Screening the genome of extant urochordate and cephalochordate species for conserved globin-flanking genes revealed linkage of RHBDF1, MPG and ARHGAP17 to globin genes in the tunicate Ciona intestinalis, while these genes together with LCMT are closely positioned in amphioxus (Branchiostoma floridae), but seem to be unlinked to the multiple globin genes identified in this species.
The plasticity of Atlantic cod to variable environmental conditions probably involves the expression of multiple globins with potentially different properties. The interspecific difference in number of fish hemoglobin genes contrasts with the highly conserved synteny of the flanking genes. The proximity of globin-flanking genes in the tunicate and amphioxus genomes resembles the RHBDF1-MPG-α-globin-ARHGAP17-LCMT linked genes in man and chicken. We hypothesize that the fusion of the three chordate linkage groups 3, 15 and 17 more than 800 MYA led to the ancestral vertebrate globin cluster during a geological period of increased atmospheric oxygen content.
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Roles of retinoic acid and Tbx1/10 in pharyngeal segmentation: amphioxus and the ancestral chordate condition
Koop, Demian
Chen, Jie
Theodosiou, Maria
Carvalho, João E
Alvarez, Susana
de Lera, Angel R
Holland, Linda Z
Schubert, Michael
Although chordates descend from a segmented ancestor, the evolution of head segmentation has been very controversial for over 150 years. Chordates generally possess a segmented pharynx, but even though anatomical evidence and gene expression analyses suggest homologies between the pharyngeal apparatus of invertebrate chordates, such as the cephalochordate amphioxus, and vertebrates, these homologies remain contested. We, therefore, decided to study the evolution of the chordate head by examining the molecular mechanisms underlying pharyngeal morphogenesis in amphioxus, an animal lacking definitive neural crest.
Focusing on the role of retinoic acid (RA) in post-gastrulation pharyngeal morphogenesis, we found that during gastrulation, RA signaling in the endoderm is required for defining pharyngeal and non-pharyngeal domains and that this process involves active degradation of RA anteriorly in the embryo. Subsequent extension of the pharyngeal territory depends on the creation of a low RA environment and is coupled to body elongation. RA further functions in pharyngeal segmentation in a regulatory network involving the mutual inhibition of RA- and Tbx1/10-dependent signaling.
These results indicate that the involvement of RA signaling and its interactions with Tbx1/10 in head segmentation preceded the evolution of neural crest and were thus likely present in the ancestral chordate. Furthermore, developmental comparisons between different deuterostome models suggest that the genetic mechanisms for pharyngeal segmentation are evolutionary ancient and very likely predate the origin of chordates.
Electronic supplementary material
The online version of this article (doi:10.1186/2041-9139-5-36) contains supplementary material, which is available to authorized users.
Cephalochordate; Cyp26 function; evolution of developmental mechanisms; evolution of the vertebrate head; functional knockdown; pharmacological treatments; pharyngeal patterning; retinoic acid signaling; Tbx1/10
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Characterization, developmental expression and evolutionary features of the huntingtin gene in the amphioxus Branchiostoma floridae
Candiani, Simona
Pestarino, Mario
Cattaneo, Elena
Tartari, Marzia
BMC Developmental Biology
Huntington's disease is an inherited neurodegenerative disorder that is caused by the expansion of an N-terminal polyQ stretch in the huntingtin protein. In order to investigate the hypothesis that huntingtin was already involved in development of the nervous system in the last common ancestor of chordates, we isolated and characterised the huntingtin homologue from the amphioxus Branchiostoma floridae. In the present paper the amphioxus general term must be referred to Branchiostoma floridae.
In this report, we show that the exon-intron organization of the amphioxus huntingtin gene is highly conserved with that of other vertebrates species. The AmphiHtt protein has two glutamine residues in the position of the typical vertebrate polyQ tract. Sequence conservation is greater along the entire length of the protein than in a previously identified Ciona huntingtin. The first three N-terminal HEAT repeats are highly conserved in vertebrates and amphioxus, although exon rearrangement has occurred in this region. AmphiHtt expression is detectable by in situ hybridization starting from the early neurula stage, where it is found in cells of the neural plate. At later stages, it is retained in the neural compartment but also it appears in limited and well-defined groups of non-neural cells. At subsequent larval stages, AmphiHtt expression is detected in the neural tube, with the strongest signal being present in the most anterior part.
The cloning of amphioxus huntingtin allows to infer that the polyQ in huntingtin was already present 540 million years ago and provides a further element for the study of huntingtin function and its evolution along the deuterostome branch.
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Evolutionary changes in the notochord genetic toolkit: a comparative analysis of notochord genes in the ascidian Ciona and the larvacean Oikopleura
Kugler, Jamie E
Kerner, Pierre
Bouquet, Jean-Marie
Jiang, Di
Di Gregorio, Anna
BMC Evolutionary Biology
The notochord is a defining feature of the chordate clade, and invertebrate chordates, such as tunicates, are uniquely suited for studies of this structure. Here we used a well-characterized set of 50 notochord genes known to be targets of the notochord-specific Brachyury transcription factor in one tunicate, Ciona intestinalis (Class Ascidiacea), to begin determining whether the same genetic toolkit is employed to build the notochord in another tunicate, Oikopleura dioica (Class Larvacea). We identified Oikopleura orthologs of the Ciona notochord genes, as well as lineage-specific duplicates for which we determined the phylogenetic relationships with related genes from other chordates, and we analyzed their expression patterns in Oikopleura embryos.
Of the 50 Ciona notochord genes that were used as a reference, only 26 had clearly identifiable orthologs in Oikopleura. Two of these conserved genes appeared to have undergone Oikopleura- and/or tunicate-specific duplications, and one was present in three copies in Oikopleura, thus bringing the number of genes to test to 30. We were able to clone and test 28 of these genes. Thirteen of the 28 Oikopleura orthologs of Ciona notochord genes showed clear expression in all or in part of the Oikopleura notochord, seven were diffusely expressed throughout the tail, six were expressed in tissues other than the notochord, while two probes did not provide a detectable signal at any of the stages analyzed. One of the notochord genes identified, Oikopleura netrin, was found to be unevenly expressed in notochord cells, in a pattern reminiscent of that previously observed for one of the Oikopleura Hox genes.
A surprisingly high number of Ciona notochord genes do not have apparent counterparts in Oikopleura, and only a fraction of the evolutionarily conserved genes show clear notochord expression. This suggests that Ciona and Oikopleura, despite the morphological similarities of their notochords, have developed rather divergent sets of notochord genes after their split from a common tunicate ancestor. This study demonstrates that comparisons between divergent tunicates can lead to insights into the basic complement of genes sufficient for notochord development, and elucidate the constraints that control its composition.
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Decelerated genome evolution in modern vertebrates revealed by analysis of multiple lancelet genomes
Huang, Shengfeng
Chen, Zelin
Yan, Xinyu
Yu, Ting
Huang, Guangrui
Yan, Qingyu
Pontarotti, Pierre Antoine
Zhao, Hongchen
Li, Jie
Yang, Ping
Wang, Ruihua
Li, Rui
Tao, Xin
Deng, Ting
Wang, Yiquan
Li, Guang
Zhang, Qiujin
Zhou, Sisi
You, Leiming
Yuan, Shaochun
Fu, Yonggui
Wu, Fenfang
Dong, Meiling
Chen, Shangwu
Xu, Anlong
Nature Communications
Vertebrates diverged from other chordates ~500 Myr ago and experienced successful innovations and adaptations, but the genomic basis underlying vertebrate origins are not fully understood. Here we suggest, through comparison with multiple lancelet (amphioxus) genomes, that ancient vertebrates experienced high rates of protein evolution, genome rearrangement and domain shuffling and that these rates greatly slowed down after the divergence of jawed and jawless vertebrates. Compared with lancelets, modern vertebrates retain, at least relatively, less protein diversity, fewer nucleotide polymorphisms, domain combinations and conserved non-coding elements (CNE). Modern vertebrates also lost substantial transposable element (TE) diversity, whereas lancelets preserve high TE diversity that includes even the long-sought RAG transposon. Lancelets also exhibit rapid gene turnover, pervasive transcription, fastest exon shuffling in metazoans and substantial TE methylation not observed in other invertebrates. These new lancelet genome sequences provide new insights into the chordate ancestral state and the vertebrate evolution.
The lancelet, or amphioxus, is an extant basal chordate that diverged from other chordate lineages about 550 million years ago. Here the authors sequence and assemble the diploid genome of a male adult of the Chinese lancelet, B. belcheri, and highlight genomic features that may have played an important role in the origin and evolution of vertebrates.
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Evolutionary Origin of GnIH and NPFF in Chordates: Insights from Novel Amphioxus RFamide Peptides
Osugi, Tomohiro
Okamura, Tomoki
Son, You Lee
Ohkubo, Makoto
Ubuka, Takayoshi
Henmi, Yasuhisa
Tsutsui, Kazuyoshi
PLoS ONE
Gonadotropin-inhibitory hormone (GnIH) is a newly identified hypothalamic neuropeptide that inhibits pituitary hormone secretion in vertebrates. GnIH has an LPXRFamide (X = L or Q) motif at the C-terminal in representative species of gnathostomes. On the other hand, neuropeptide FF (NPFF), a neuropeptide characterized as a pain-modulatory neuropeptide, in vertebrates has a PQRFamide motif similar to the C-terminal of GnIH, suggesting that GnIH and NPFF have diverged from a common ancestor. Because GnIH and NPFF belong to the RFamide peptide family in vertebrates, protochordate RFamide peptides may provide important insights into the evolutionary origin of GnIH and NPFF. In this study, we identified a novel gene encoding RFamide peptides and two genes of their putative receptors in the amphioxus Branchiostoma japonicum. Molecular phylogenetic analysis and synteny analysis indicated that these genes are closely related to the genes of GnIH and NPFF and their receptors of vertebrates. We further identified mature RFamide peptides and their receptors in protochordates. The identified amphioxus RFamide peptides inhibited forskolin induced cAMP signaling in the COS-7 cells with one of the identified amphioxus RFamide peptide receptors expressed. These results indicate that the identified protochordate RFamide peptide gene is a common ancestral form of GnIH and NPFF genes, suggesting that the origin of GnIH and NPFF may date back to the time of the emergence of early chordates. GnIH gene and NPFF gene may have diverged by whole-genome duplication in the course of vertebrate evolution.
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Origins of anteroposterior patterning and Hox gene regulation during chordate evolution.
Schilling, T F
Knight, R D
Philosophical Transactions of the Royal Society of London. Series B
All chordates share a basic body plan and many common features of early development. Anteroposterior (AP) regions of the vertebrate neural tube are specified by a combinatorial pattern of Hox gene expression that is conserved in urochordates and cephalochordates. Another primitive feature of Hox gene regulation in all chordates is a sensitivity to retinoic acid during embryogenesis, and recent developmental genetic studies have demonstrated the essential role for retinoid signalling in vertebrates. Two AP regions develop within the chordate neural tube during gastrulation: an anterior 'forebrain-midbrain' region specified by Otx genes and a posterior 'hindbrain-spinal cord' region specified by Hox genes. A third, intermediate region corresponding to the midbrain or midbrain-hindbrain boundary develops at around the same time in vertebrates, and comparative data suggest that this was also present in the chordate ancestor. Within the anterior part of the Hox-expressing domain, however, vertebrates appear to have evolved unique roles for segmentation genes, such as Krox-20, in patterning the hindbrain. Genetic approaches in mammals and zebrafish, coupled with molecular phylogenetic studies in ascidians, amphioxus and lampreys, promise to reveal how the complex mechanisms that specify the vertebrate body plan may have arisen from a relatively simple set of ancestral developmental components.
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A SINE in the genome of the cephalochordate amphioxus is an Alu element
Holland, Linda Z.
International Journal of Biological Sciences
Transposable elements of about 300 bp, termed "short interspersed nucleotide elements or SINEs are common in eukaryotes. However, Alu elements, SINEs containing restriction sites for the AluI enzyme, have been known only from primates. Here I report the first SINE found in the genome of the cephalochordate, amphioxus. It is an Alu element of 375 bp that does not share substantial identity with any genomic sequences in vertebrates. It was identified because it was located in the FoxD regulatory region in a cosmid derived from one individual, but absent from the two FoxD alleles of BACs from a second individual. However, searches of sequences of BACs and genomic traces from this second individual gave an estimate of 50-100 copies in the amphioxus genome. The finding of an Alu element in amphioxus raises the question of whether Alu elements in amphioxus and primates arose by convergent evolution or by inheritance from a common ancestor. Genome-wide analyses of transposable elements in amphioxus and other chordates such as tunicates, agnathans and cartilaginous fishes could well provide the answer.
Branchiostoma; transposon; cis-regulation; SINE; Alu; chordate evolution
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Coon, Steven L.
Klein, David C.
Molecular and cellular endocrinology
The melatonin rhythm-generating enzyme, arylalkylamine N-acetyltransferase (AANAT) is known to have recognizable ancient homologs in bacteria and fungi, but not in other eukaryotes. Analysis of new cDNA and genomic sequences has identified several additional homologs in other groupings. First, an AANAT homolog has been found in the genome of the cephalochordate amphioxus, representing the oldest homolog in chordates. Second, two AANAT homologs have been identified in unicellular green algae. The homologs in amphioxus, unicellular green algae, fungi and bacteria are similarly primitive in that they lack sequences found in vertebrate AANATs that are involved in regulation and that facilitate binding and catalysis. In addition, all these sequences are intronless. These features are consistent with horizontal transfer of the AANAT ancestor from bacteria to green algae, fungi and chordates. Lastly, a third AANAT gene has been found in teleost fish, suggesting that AANAT genes serve multiple functions in addition to melatonin synthesis.
arylalkylamine N-acetyltransferase; AANAT; evolution; melatonin
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Ouagadougou Hotels
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Ouagadougou Hotels
Venere.com is the perfect place to make all of your hotel bookings in Ouagadougou, Burkina Faso! Check out our 6 Ouagadougou hotels, including a wide selection of accommodations, ranging from the super luxurious 5-star to the budget-conscious 2- and 1-star. Get the best value for your money with Venere. No matter what kind of accommodations you are looking for - ski, family, spa, sports, business, wellness, city center, adventure, golf or boutique hotels - we have the perfect hotel for your next vacation or business trip. Not sure which type of Ouagadougou hotel you want? Browse the 6 hotels on our interactive map to see exactly where they are located in Burkina Faso. Carefully read the detailed hotel descriptions for even more information on the Ouagadougou area and the hotel you are interested in booking. Find out about the nearby tourist attractions, such as museums, beaches, shops, monuments, churches and exhibition centres. Learn all you need to know about each of the 6 accommodations offered at Venere. Even find hotels with onsite restaurants, pools, gyms, internet, concierge and business services. When traveling to Ouagadougou, Burkina Faso - for work or pleasure, a quick weekend or an extended stay - check out Venere.com first. Venere has something to fit any budget. Get amazing discounts on Ouagadougou hotels. We update our inventory daily and are always adding new deals and sales for our 6 accommodations. Treat your special someone to a romantic weekend or book a fun-filled vacation for the entire family. Start by searching our easy-to-navigate website. Simply type Ouagadougou into the search box, then put in your travel dates and the number of rooms you need. Then simply look through all of the available accommodations among the 6 that Venere has to offer. Browse through all of the Ouagadougou hotels in the list and choose the ones that look good. Our gallery of photos will let you see what the hotel looks like inside and out. Read hotel descriptions for an overview of the hotel you chose in Burkina Faso. Be sure to check for any available freebies, such as free Wi-Fi, breakfast and parking! Get all of the information about your room and location before making your final choice. Check out the hotel reviews from genuine Venere guests and Tripadvisor comments to see what others have to say about your Ouagadougou, Burkina Faso accommodations. Learn all you can about your hotel choice and them book the perfect vacation accommodations with Venere.
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Kevin Johnson
From BoxRec
(Redirected from Human:173416)
Name: Kevin Johnson
Alias: Kingpin
Born: 1979-09-07
Birthplace: Asbury Park, New Jersey, USA
Hometown: Atlanta, Georgia, USA
Stance: Orthodox
Height: 6′ 3″ / 191cm
Reach: 82″ / 208cm
Boxing Record: click
Amateur Career
Kevin Johnson had a brief 14-2 record as an amateur. He was a New Jersey Open Heavyweight Golden Gloves champion in 2002, but was defeated in the first round of the National Golden Gloves tournament.
Professional Career
Johnson holds wins over former WBA titleholder Bruce Seldon, Damian Wills, U.S. Olympian Devin Vargas, and Albert Sosnowski in the Prizefighter Tournament. Johnson lost in the tournament final to Tor Hamer. In 2009, Johnson went the distance with WBC Heavyweight Champion Vitali Klitschko (38-2).


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I'm new to both WordPress and Gravity Forms. I'm a salesforce administrator and helping a nonprofit get set up. They use WordPress - this plug-in looks like it would be a good option for them. I'm a bit confused though on licensing: Developer, Business, Personal
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Thanks so much,


text:
COMIN' UPSEPTEMBER 16, Friday, 7:00 - p.m. "Red House Live from Landmark Center" Butch joins Prudence Johnson and Dean Magraw at the F.K. Weyerhaeuser Auditorium in St. Paul's historic Landmark Center.
Event page
Hey, collectors...going...going...The very last of Butch's solo series on Daring Records. Order while they last!
It is a great honor to be the 2015 recipient of the A.P. Anderson Award from the Anderson Center at Tower View in Red Wing, Minnesota. "Each year the Anderson Center presents the A.P. Anderson Award for Outstanding Contributions to Literature and the Arts in Minnesota."
Born in a small Minnesota river town, Thompson played Christmas carols on his mother's upright piano by age three, and began formal lessons at six. He studied clarinet in high school, and as a teenager led his first professional jazz group, Shirt Thompson & His Sleeves.
Read more
Stan Hall on piano leads the Hall Brothers and guest musicians from New Orleans in the mid 1960s. My Blue Heaven. Stan Hall, piano; Russ Hall, trombone; Mike Polad, banjo; Kid Thomas Valentine, trumpet; Joe Butler, bass; Emanuel Paul, tenor sax; Sammy Penn, drums, and Butch on clarinet.
Been doing my radio show Jazz Originals for over 25 years now on @KBEMfm Here's a sample: https://t.co/cExPbDB0mf— Butch Thompson (@butchthompson) July 2, 2016
Butch hosts the Jazz Originals radio show on KBEM 88.5 FM in Minneapolis, Tuesdays at 7PM, and Sundays at 6PM (repeat). Latest shows on demand here.
Evolving Jazz Tradition
Butch brings his own voice to the classic jazz repertoire of blues, stomps, ragtime and pop songs. From Scott Joplin's Maple Leaf Rag to Duke Ellington's Mood Indigo, from Jelly Roll Morton's New Orleans jazz to the virtuosic stride piano of 1920s Harlem, Butch has played his personal style from Cairo to Kansas City.
Whether he's working solo, with his well-known trio, or other ensembles around home or in New Orleans, Butch delivers variety and the pure, contagious joy of music making.
Jelly Roll Morton said it best
Jazz was born in New Orleans, he said, from a gumbo of "everything from blues to opera. Jazz music is to be played sweet, soft, plenty rhythm. When you have your plenty rhythm with your plenty swing, it becomes beautiful."
One afternoon on a New Orleans porch. From left: Tom Ebbert, Brian O'Connell, Butch, Bill Huntington, Chuck Badie, Duke Heitger.


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Hotels in Lampeter, United Kingdom
Search & Compare Lampeter Hotels
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Hotels in Lampeter
- Most bookedTy Haf at Tyglyn Lodge
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Most recent review
"Learning day to day living Lifes mid South Wales lampeter felt just a big topsoftheworld apart feeling the comforts not to socialable but happy university Accomodations again apart from this I enjoyed my paid stay wonderful cheerful Welsh country for summers 2016 ok det"
A Traveller, May 2016, GB
4 people looked at this hotel in the last hour
Most recent review
"Lovely place, friendly and helpful staff, fantastic surroundings. I would definitely stay again"
A Traveller, Sep 2015, GB
Hotels within 10 miles of Lampeter
9 people looked at this hotel in the last hour
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"We had a lovely stay in the hotel on Saturday night, then had breakfast at 7.30 and caught the early tide. We sailed for a couple of hours and moored up before the harbour dried out. Most enjoyable time. Its our idea of a good weekend."
A Traveller, May 2016, GB
9 people looked at this hotel in the last hour
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"The owner and staff were quite pleasant, there was ample parking and the room was very cozy. Because our room was on the edge of the house, our wi-fi was a bit spotty, but overall this was an excellent experience. Aberaeron was a close drive and was a wonderful little town with a quiet beach and ..."
A Traveller, Jul 2016, US
- Aromatherapy Reflexology Centre - B&BNo Guest Reviews yet
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6 people looked at this hotel in the last hour
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"Lovely view of the bay from hotel window. From the hotel We went to new quay, which was about 12 miles away. Paid only £17 for a 1 1/2 hour boat trip dolphin watching. Loads of sightings of bottle nose dolphins. Amazing experience"
A Traveller, Aug 2016, GB
5 people looked at this hotel in the last hour
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"Friendly owner, very clean and well decorated room.excellent service. Would go again."
A Traveller, Aug 2016, GB
5 people looked at this hotel in the last hour
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"Very comfortable hotel in an ideal spot for walking into the very picturesque town of Aberaeron, Only disappointment with the hotel, for me personally, was that the bar did not serve real ale, but everything else was very good."
A Traveller, Jul 2015, GB
Best hotels in Lampeter
If you are looking for Lampeter hotels in a particular area, you can use the search feature at the top of the page to search by neighborhood, landmark, address or zip code, or you can search by using the built-in map feature. You can also browse hotels in Lampeter by type, amenities, and star-rating so that you can find the best hotel. Currently, top rated hotels in Lampeter are:
- Ty Haf at Tyglyn Lodge
- Falcondale Mansion Hotel
- University Of Wales Trinity St David - Lloyd Thomas Centre
- TyGlyn Restaurant & Conference Centre
- Merwerydd Guest Accommodation
The location of your Lampeter hotel is important. If you are traveling to Lampeter, &StateorCountry for business, you may want to consider getting a hotel close to the airport or near your meeting venue. By doing so, you can really cut down on transportation costs and time. You may also want to book a hotel that offers free wifi just in case you need to get some work done.
If you are traveling for vacation, figure out which attractions or landmarks you want to see and book a hotel that is within close proximity. Although hotels near attractions might cost a little more, you may find that spending those extra few dollars is worth the convenience. You can even find hotels that align with your interests. For example, if you are looking for a relaxing vacation, a spa hotel may be the perfect choice for you.
You'll also want to determine which type of accommodations will suit your needs before booking a hotel in Lampeter. If you're traveling with the kiddos, a family friendly hotel will offer larger rooms and more sleeping arrangements. For those traveling for an extended period of time, booking a hotel suite with a kitchenette is a smart move and will feel a little more like home. If you are budget conscience and just need a place to rest your head, a hostel or motel may be all you need.
If you are looking to score the best discounts and deals, use the filter to sort by price from low to high. You can book a hotel in Lampeter for as low as $49 per night. If you are looking to cut down on expenses, look for hotels that include free breakfast and parking. Some hotels even offer free shuttle service to and from nearby places. You can also sort by star rating, proximity, and guest rating.
Before you book, check out user reviews. Hotels.com has 35 reviews in Lampeter. We show reviews from Expedia and TripAdvisor so you know that you are reading genuine, reliable reviews. Be sure to download the Hotels.com app so you can book your reservations on the go, view amenities, and save the hotels you want to view and compare later.


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Thunder Beast Sound Suppressors - Built For Accuracy, By Shooters - Tactical, Competition, Sporting
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