text:
I worked out back too. Felt good to not have my livelyhood depend on it.
My back is fine but oh my hamstrings. The hammies need a work out then
maybe I can get out of the Flat Butt Society.
Bought a couple of Salvias today...Salvia nemorosas. I still have the
tropical thing going but want to lean more toward butterfly and
hummingbird plants. Speaking of which I hacked down my buddleia plant
which thought was supposed to get to 6'. I planted it WAY to close to
the magnolia grandiflora and it was over 12' tall. Down it came and I
left two feet of the plant to leaf back out and hopefully attract some
monarchs or pink ladies.
Theresa did you go down to Richmond to see Annie or did she move again?
----- Original Message ----
From: Theresa G. <firstname.lastname@example.com>
To: email@example.com
Sent: Saturday, March 29, 2008 5:39:10 PM
Subject: Re: [CHAT] oh my aching...
My aching back too. I re-doing the sink area of the master bathroom and
today i finally managed to get the old vanity out. Good lord that thing
was heavy. But at least it is gone, and most of the walls are painted.
The plan for tomorrow is to get the tile down on the floor, so that I
can return the tile saw first thing Monday morning.
Weather is beautiful here too, and I got some plants from Annie's
Annuals (nursery) on Friday. But they'll have to hang out and wait
until next weekend I think.
Kitty wrote:
> back.
> Glorious day! A bit chilly, but that makes it easier to work. I
> cleaned up most of the back today and removed the straw from around my
> overwintered pots. I've removed some of the top layer of leaves too,
> but not all. Some is frozen. It was quite a thrill to remove the the
> top and find bright green leaves on the Chimney Bellflowers and the
> Hydrangeas with good buds on them.
> I was out there this aftn until 7:30. I'm tired and achy from lifting
> and lugging a lot of heavy stuff. But things are looking good. I
> have iris and crocus blooming and so much coming up!
> Tomorrow we open the MG nursery. I hope it doesn't rain as we have
> lots to do.
> Kitty
> neIN, Zone 5
> ---------------------------------------------------------------------
> To sign-off this list, send email to email@example.com with the
> message text UNSUBSCRIBE GARDENCHAT
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text:
Written on: 27/12/2008 by SolHut (1 review written)
Good Points
No more worries on smearing my lipstick on things.
Bad Points
NO bad points.
General Comments
Thank you revelon overtime lipstick. I have been waiting for something like this to come out. I love it. And I will continue to buy it. Sincerely, Happy Customer.
As rated by our community of reviewers
" Blistering on outside and inside after week of use. " Read More
" Color stay lipstick " Read More
Written by missydenny22
" Best ever!!! " Read More
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" Became allergic after many years of use! " Read More

subset: commoncrawl


text:
More Videos...
White Zombie vs Tesla Roadster NEDRA PiR
White Zombie electric vehicle (EV) crazy launch Sept. 11, 2010
Whoever says electric vehicles are slow hasn't seen the White Zombie!
Running On Lithium: White Zombie vs. Mazda Turbo
This is the beginning of the docu "Running On Lithium" and it features John Wayland's Electric White Zombie vs. Mazda turbo Rotary Power. Clip from the documentary: Running On Lithium. Are backyard engineers pioneering the future of the electric car? As street-legal electric cars race against muscle cars, this documentary provides a thrilling insight into the future of renewable energy: Journeyman Pictures. Thanks for looking!
White Zombie - Fastest Electric Car
This clip is about the fastest street-legal electric car, and how it works.


text: Hoehle, Joerg-Cyril writes: > Wim Oudshoorn wrote: > > (asdf:operate 'asdf:load-op 'clsql-sqlite3) > > FFI::FOREIGN-CALL-OUT: no dynamic object named "sqlite3_open" in > library :DEFAULT >=20 > I haven't looked at current CLSQL. In version 3.5.4, I see that > sqlite-api supplies the :module "sqlite" argument, which is good. You > need to provide a mapping from that name to the shared object file > (assuming UNIX). >=20 > No, wait, you're using the CFFI backend, which does not know about > modules and libraries AFAIK. CFFI only looks up names in the current > process namespace AFAIK. >=20 > Maybe a simple (ffi::foreign-library "/usr/lib/sqlite.dylib") does the > trick for your current session?? > It should open the library and make all its names available (at least i= n > Linux). > You need to call that before loading clsql. Then try to figure out how > the other DB backends do that for CFFI on MacOS with CLISP. Well there are two ways to indicate to CFFI to load libraries. (cffi:define-foreign-library <libname> (:unix (:or "<libname>.so.<version>" "<libname>.so")) (t (:default "<libname>"))) (cffi:use-foreign-library <libname>) http://common-lisp.net/project/cffi/manual/html_node/Tutorial_002dLoading= .html#Tutorial_002dLoading and also directly cffi:load-foreigh-library (which is used by cffi:use-foreign-library): http://common-lisp.net/project/cffi/manual/html_node/_002aforeign_002dlib= rary_002ddirectories_002a.html#_002aforeign_002dlibrary_002ddirectories_0= 02a (With some variant on MacOSX for frameworks). The question is whether the libraries are re-open automatically when we reload a saved image? --=20 __Pascal Bourguignon__ http://www.informatimago.com/ This is a signature virus. Add me to your signature and help me to live.


text:
Following the path of Bhaag Milkha Bhaag, a film is being planned on a big scale to showcase the journey of 14 players who brought India's first cricket World Cup home, both on and off the field.
There were rumours that Vishnu Vardhan Induri, the founder and MD of Celebrity Cricket League was planning a movie on the league. However, he recently revealed the details of the movie to Mumbai Mirror.
"The film will be the official biography of India's first big win that was instrumental in putting the country on the international map," he said.
Vishnu is already in contact with the players to get all the information about their life and it is planned that A-list actors will play the role of Kapil Dev and Sunil Gavaskar.
Vishnu said, "We want them to officially be on board and give their inputs on the script. I also want them to take part in the promotions. We will sign A-list actors to play Dev and Sunil Gavaskar."
As per sources, few of the roles in the movie will be done by the players themselves but major part will be played by the actors.
"If the script, which should be locked in a couple of weeks, requires real cricketers, you will see them on screen. But, this is a big-budget commercial film along the lines of Bhaag Milkha Bhaag. So, it's mostly going to be actors playing the cricketers," said Vishnu.
The film will be directed by national award winner director Sanjay Puran Singh Chauhan.
- Arts & Entertainment
- Movies
- Kapil Dev
- Bhaag Milkha Bhaag
- Celebrity Cricket League


text:
No products
Prices are tax included
+41 22 552 26 26
Tue. - Sat. : 15h - 19h
Beautiful penty in transparent red lace from the lovica collection.
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The sexy thong of Melodia collection.
The string for a romantic evening for two naughty.
Made roar the tigress in you through this beautiful thong with leopard.


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text: Asia Kitchen may be outside of your area. Please enter your delivery address to check availability Do you need any... Napkins & Utensils Continue to Checkout Asia Kitchen Back to Menu Your order refresh Asia Kitchen Your Shopping Cart is empty edit Delivery Address Please enter delivery address edit Delivery Time N/A* Add someone to this order Mongolian Three Delights Chicken, beef, shrimp. x $16.99 Special Instructions: Label Item For: or Cancel


text:
Olga from Russia
Verified Purchase
Olga from Russia
soft skin, nice smell, so I use little effect so far
Posted by 5661478682627321834 on Feb 11, 2013
Scar is still there(now a bit smoother) but it is a good replacement for SPF creams/lotions/mists.
Olga from Russia
soft skin, nice smell, so I use little effect so far
I think it works!
Posted by Reviewer1155879 on Jul 16, 2011
I bought this for the scars on my leg and feet I've gotten from evil shoes. Seems to work! Effects should be quicker and more obvious if I've used it daily instead of always forgetting. Smells like cocoa butter, convenient stick form. Texture is kind of sticky and oily - doesn't bother me as I'm applying on my legs, but if you're going to use it on your face as I've seen in some reviews, not sure how that will feel.
Seems nice, a little sticky and greasy though
Posted by Reviewer2002330 on Aug 19, 2012
I haven't used it regularly, so I don't know how it affects scars, but using it on my thyroid operation scar feels a bit sticky and I have to be careful not to leave too much of it on when wearing somethign around my neck. Smell is good and ease of use too, convenient size to have in my purse.
1-4 of 4 total


text:
Season 2 of the Overtime podcast is here! Listen to the first episode with Meg Robichaud now.
As you may know, I've been working a lot to redesign and rebuild my website, portfolio and overall brand image. About a week ago, I became aware that someone had copied my website, both in layout and text (almost word for word in some instances). Might I also add this person was on my University course...
In response, I had only one thing to do, get my blog back online, design it to fit my new site design, and then write this blog post.
Her site is now offline, but my post – filled with screenshots – will remain online.
Named and shamed.
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Show and tell for designers
What are you working on? Dribbble is a community of designers sharing screenshots of their work, process, and projects.
Copyright © 2009–2017 Dribbble LLC. All screenshots © their respective owners. Shipped from Salem, Mass. USA.


text: Soundcheck Edit this Published by Soundcheck Of Montreal Listen 21 min Queue Want to embed this player? Use the code below. More info. <iframe frameborder="0" scrolling="no" height="130" width="100%" src="https://www.wnyc.org/widgets/ondemand_player/soundcheck/#file=/audio/json/41643/&share=1"></iframe> Share on Facebook Share on Twitter Email a Friend On the latest album from Of Montreal, "Skeletal Lamping," songwriter Kevin Barnes summons his inner Prince and coos, "We can do it soft-core if you want, but you should know I go both ways." Barnes joins us to talk about his latest psychedelic sonic romp, and we'll hear an acoustic performance live in our studio.Video: Of Montreal performs on Soundcheck -- and talks about their outrageous stage costumes


text:
Family Disney Villa to Rent, Kissimmee, Florida
Over 2900 sq. ft. of single storey
luxury vacation home and private pool. Only 9 miles from
Walt Disney World and 24 miles from Orlando Airport. Local Area Information.
Accommodation comprises 4 bedrooms and 3 bathrooms (2 bedrooms ensuite) and sleeps up to a maximum of 8 people.
The Villa has an extended pool area to ensure everyone can enjoy the Florida sunshine and excellent facilities.
Situated in the beautiful gated community of Cumbrian Lakes amongst 122 acres of conservation land.
Fully airconditioned and all rooms have additional ceiling fans. All bedrooms have televisions.
The garage has been transformed "New for 2014" into a games room which includes a pool table, table tennis, electronic darts and table football.
Perfect holiday location. Lots to do for everyone.
Available to let all year round, this beautiful executive family villa is situated on a substantial corner lot, which ensures extra privacy when enjoying the spacious 1080 sq. ft. pool and lanai area.
"It's south facing aspect ensures that you benefit from the wonderful Florida sunshine with shade provided by the lanai."
It also ensures that you can enjoy the beautiful picturesque sunset while sat in or beside the pool. The Villa also backs onto a children's play area which allows the kids to unwind while you remain by the pool in complete view of your children. Enjoy also the new games room as described above.
Whilst being able to enjoy the surroundings of this quiet development you are also conveniently placed with many varied shops and restaurants only a short drive away on US Highway 192. A large Wal-Mart store is within a 5-minute drive of your holiday home.
Other facilities of interest in the immediate area are: horse riding, golf, helicopter rides, Kissimmee "Old Town", Balloon Rides, Fishing, Water Sports plus much, much more.
Detailed information on any of the above can be provided if necessary before you travel.


text:
Need inspiration! Making cake for 26 people for mum's 70th...(14 Posts)
Wigeon Fri 14-Feb-14 18:28:20
livelablove Fri 14-Feb-14 18:47:22
MsAspreyDiamonds Sat 15-Feb-14 06:51:00
MikeLitoris Sat 15-Feb-14 07:50:25
bakingaddict Sat 15-Feb-14 08:04:35
MikeLitoris Sat 15-Feb-14 08:44:49
bakingaddict Sat 15-Feb-14 09:40:25
Electryone Tue 18-Feb-14 22:41:33
Electryone Tue 18-Feb-14 22:41:46
Wigeon Wed 19-Feb-14 17:41:59
PurplePidjin Wed 19-Feb-14 17:49:01
Wigeon Wed 19-Feb-14 17:52:44
PotteringAlong Wed 19-Feb-14 17:57:26
PurplePidjin Wed 19-Feb-14 18:15:50
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Applying Henna During Menses in Urdu | Urdu Sex Education
March 3, 2011 | Islamic QA Adult | No Comments|
Applying Henna during Menstruation
Question: what is the Islamic point of view regarding applying henna during menstruation? Will it be considered as impurity unless the color of henna remains on hands?
Answer: it is permittedfor women to apply henna on hands during menses period because her body is clean. This is why; it is admissible to shaking hand with a woman who is having menstrual cycle. It is proved that Prophet Muhammad (SAW) has drunk water after Hazrat Ayesha (R.A.) when she was suffering from periods.Prophet Muhammad (SAW) kept His mouth on the place where Hazrat Ayesha (R.A.) has kept. When Prophet Muhammad (SAW) has demanded from Hazrat Ayesha (R.A.) to bring a cloth, She has told Him about Her menstrual cycle. He has narrated:
"Your menstruation is not in your control."
Henna is sacred. It is been applied on pious parts. There is nothing in it which expresses the disapproval of applying Henna in menstrual cycle.
……..Sheikh ibn e Jabreen………
VN:F [1.9.22_1171]Applying Henna During Menses in Urdu | Urdu Sex Education,


text:
The Promise and Perils of Writing Program Administration
AUTHOR: Enos, Theresa and Borrowman, Shane
PUBLISHER: Parlor Press
Also available at Amazon.com
AUTHOR: Enos, Theresa and Borrowman, Shane
PUBLISHER: Parlor Press
Also available at Amazon.com
Note: Not guaranteed to come with supplemental materials (access codes, CDs, DVDs)
Product Description: Combining formal quantitative research with narrative-based scholarship, THE PROMISE AND PERILS OF WRITING PROGRAM ADMINISTRATION represents multiple voices from faculty balancing between the demands of teaching, writing, and administering writing programs in professional, ethical ways-often under circumstances that can be defined, at best, as difficult. In these pages, junior faculty tell their stories of triumph and trauma, while more firmly established composition scholars reflect upon the changing and challenging profession we all share. CONTRIBUTORS Chris Anson, Shane Borrowman, Stuart C. Brown, Nita Danko, Suellynn Duffey, Ernest J. Enchelmayer, Theresa Enos, Megan Fulwiler, Ann E. Green, Jeanne Gunner, Douglas Hesse, Elizabeth Hodges, Lauren Sewell Ingraham, Emily Isaacs, Patti J. Kurtz, Claire C. Lamonica, Camille Langston, Andrea A. Lunsford, Randall McClure, Susan H. McLeod, Richard McNabb, Thomas P. Miller, Cynthia Nearman, Erin O'Neill, Melissa Nicolas, Christine Norris, Chere L. Peguesse, Louise Wetherbee Phelps, E. Shelley Reid, Stephanie Roach, Duane Roen, Shirley K Rose, David Schwalm, Jillian Skeffington, Matt Smith, Martha A. Townsend, John Trimbur, Victor Villanueva, Margaret E. Weaver, Edward M. White, Kathleen Blake Yancey, and Art Young. ABOUT THE EDITORS THERESA ENOS is Professor of English at the University of Arizona, the founder and editor of Rhetoric Review, and a past president of the Council of Writing Program Administrators. She has edited or coedited twelve books, including the Encyclopedia of Rhetoric and Composition and The Writing Program Administrator's Resource: A Guide to Reflective Institutional Practice. SHANE BORROWMAN is Assistant Professor of English at the University of Nevada, Reno, a former writing program administrator, and the author of articles appearing in Rhetoric Review, College English, and WPA: Writing Program Administration. JILLIAN SKEFFINGTON is a PhD candidate at the University of Arizona.
CATEGORY: Language Arts & Disciplines, Reference
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text:
- {{matchup.date.fullday}}{{matchup.date.month + ' ' + matchup.date.day}}
Packers surprise Falcons
Samkon Gado rushed for 103 yards and two touchdowns on 25 carries Sunday to give the Green Bay Packers a 33-25 win over the Atlanta Falcons.
The Packers were a 9-point underdog in the game. The combined score was OVER the posted over/under total (42.5).
Brett Favre completed 26-of-39 passes for 252 yards with a touchdown and an interception for Green Bay. Ryan Longwell booted four field goals.
Michael Vick completed 20-of-30 passes for 209 yards with two touchdowns, while also running in for a pair of scores for Atlanta. Warrick Dunn rushed for 76 yards on 17 carries.


text:
Current production time is 30 days.
Please read our policy section before placing an order.
Like us on Facebook: https://www.facebook.com/hookedonhudson
Current production time is 30 days.
Please read our policy section before placing an order.
Like us on Facebook: https://www.facebook.com/hookedonhudson
Tina Taylor on Oct 31, 2015
Gorgeous, superbly made hat. So cute, I adore it. Good seller communication, and it arrived well before I thought it would so it was a lovely surprise. Delighted.
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text:
Health and medicine , Mental health , Education , Volunteering , Human services , Economic development , Poverty and hunger , Youth , Community development , Philanthropy , , , , , , Autres
Community Concerns Uganda
À l'heure actuelle, nous n'avons pas d'ouvertures d'emploi, de possibilités de bénévolat, ou d'événements prochains.
S'identifier ou adhérer pour être parmi les premiers à savoir quand de nouvelles ouvertures ou des événements sont ajoutés!
À propos
Community Concerns Uganda (CCUg) is a Christian community based organization founded on the pillars of love, hope and a desire for change. CCUg works to address the needs and aspirations of the underprivileged using a holistic approach directed specifically towards poverty reduction. The organization was started in 2010, registered in 2011 and currently has projects in Nakalanga Village (Bukatube Sub county), Magamaga (along the Jinja-Iganga Highway) and in Wairaka Town (Jinja district) where our offices are located.
1. To increase the household income of the community members and improve their standard of living so that they can move towards a self sustainable future.
2. To provide assistance through education in reducing diseases caused by poor sanitation and personal hygiene.
3. To increase the literacy rate among the Orphans and Vulnerable Children (OVC) through the provision of scholarships for school education.
4. To increase on school retention among adolescent girls and boys by improving the general awareness about sexual/reproductive health and HIV/AIDS.
5. To provide psycho-social support to men and women affected by domestic violence
6. To promote government programs especially those related to health and enhancing community development.
7. To be the voice that helps persuade policy makers to be more responsible towards the needs of the community.
Mission: To improve the social, economic and psychological welfare of the community.
A self - sustaining community with healthy, educated members living in safe and clean households.
Core values:
Sharing, Serving, Transparency, Respect, Hope, Love and concern for others.
Date de fondation : April 29, 2011
Adresse email : Identifiez-vous pour voir ces adresses email
Site(s) web:
Numéro de téléphone : +256774936122
Adresse :
Jinja, Eastern Region, P.O. Box 196, Ouganda


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The pictures are accurate, and the size feels ok. However the construction is pretty cheap. I went to go push the pegs into the pre-drilled holes on the puck part of the hanger, and the wood is chipping all around the pegs because the holes are slightly too small for the pegs.
- Sizing:
- Feels true to size


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Travel from Partinico to Marsala and discover Sicily city of 77.9 Thousand inhabitants. The distance from Partinico to Marsala is 58 kilometers; with Wanderio you can compare the available treno so that you can organize your trip choosing the fastest route, the most comfortable or the cheapest one. For example, by choosing a treno from Partinico to Marsala, you'll get to your destination in about 2 hours on average; the average treno ticket prices are 8 €.
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In case you missed the Nightline segment.
In case you missed the Nightline segment.
Absolutely amazing! Made my hubby watch, too…he's encouraging me to start NOW. Thanks, Colleen!
Congratulations on the Nightline interview. You looked amazing and I am proud to have read your wonderful books. Enjoy your success and please keep writing!
Thank you for that segment! I love the New Adult genre and you are officially my new favorite author! Slammed and Point of Retreat were great but Hopeless took it to a whole new level and completely blew me away! I can't wait to read your upcoming books. Please don't EVER stop! Congratulations on all your success and for sharing with us a part of your personal life.
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Alpine FIS Ski World Championships - Opening Ceremony
SCHLADMING, AUSTRIA - FEBRUARY 04: General view as fireworks explode during the opening ceremony for the Alpine FIS Ski World Championships on February 4, 2013 in Schladming, Austria. (Photo by Clive Mason/Getty Images)
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We will promote constructive and witty comments to the top, so that everyone sees them!


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Type your paragraph here.
Type your paragraph here.
Dash and Daisy May
We know just how much you love your dog, and with us they are a member of our family. We love hearing from their new family on how they are doing.
Here at Country Setter Kennel we love all are dogs they are family to us. The Dame is Daisy May and is a great hunter. The Sire is Pat's Kentucy Dash, he is a 2 time UFTA Amateur Divison champion and 1 time Llewellin champion


text:
5 Reasons You Shouldn't Give Up on Church
May 13, 2015
Jayson is a God-botherer, writer, sinner, husband, determined dreamer, father, musician and audiophile. He blogs at jaysondbradley.com. Find him on Twitter and Facebook.
The early '90s were awash in books explaining why Generation X was abandoning church. In a similar vein, there's been no shortage of blog posts, books and conferences about how Millennials are leaving, too.
A portion of every generation has pushed their churches to grow in areas of sin and weakness. From monastics urging churches out of the distracting cities and into the deserts, to aggressive arguments over the sale of indulgences, fights for emancipation in Europe, women's suffrage, civil rights, Vietnam and so much more—there's been a prophetic portion of the Church seeking to realign churches with their larger purpose and roles in the world.
And, I'm sure there have also been those who walked away from churches out of frustration for their deficiencies.
I don't want to diminish this struggle. I know what it's like to wonder if it's all worth frequent headaches. For two years, I couldn't darken the doorway of a church; I was sure I was done. Many of the issues that continually come up in the "why millennials are leaving the Church" posts definitely played a part in my disenchantment.
But here are five reasons I am back and more committed to the local church than ever:
1. We're all part of the church's problems.
Despite Christ's prayer that the Church would model a trinitarian-like oneness (John 17:20–21), we can be frequently fractured and set against each other. This isn't just the Church on a macro-level—the local church models this behavior in similar ways.
I've gossiped, sown discord, manipulated events, fought for power, demanded my way, etc. And the truth is that if you're part of a church, you have, too.
There's nothing more Christlike than challenging the Church to be more sincere and full of grace and truth. If the Church is going to continue reforming, it will be because of the ones who stay— not the ones who leave.
It's been a problem since the Church's inception. This is why Paul had to warn the Galatians that, "If you bite and devour each other, watch out or you will be destroyed by each other" (Galatians 5:15).
These kinds of issues might seem small, but that's because we don't see how our behavior affects the entire organism that is the church. I'm sure that if we saw the full effect of our judgments, selfishness, backbiting and power-plays, we'd be surprised at how far and deep they reach.
2. Churches need reformers.
Despite glaring problems with the establishment's religious expressions and exclusive behavior at the time, Jesus started His reformation from within Judaism. These actions are what led to His crucifixion.
If I really want to identify with Jesus (and the prophets), I'll continue to love the Church from within while I push, cajole and shout her into Christlikeness. It would be much easier to leave.
Every voice that has called for reform (even the ones we celebrate now) experienced pushback, threats and misunderstanding. But there's nothing more Christlike than challenging the Church to be more sincere and full of grace and truth. If the Church is going to continue reforming, it will be because of the ones who stay— not the ones who leave.
3. There's still so much good in the church.
Jesus encourages us not to make a show of our goodness and promises us that the God who sees what is done in secret will reward us (Matt. 6:1–4). This means that many of the most faithful and hardworking people are doing good work that we know nothing about.
For every divisive news story about hot-button social issues involving Christians, there are many serving on the streets, in prisons, in soup kitchens and everywhere else there's need.
For every story of ministry corruption or a pastor's financial misdeeds, there are many sacrificing to keep people fed, clothed and cared for.
News websites and TV stations make their advertising money on outrage and fear. If you want to see the good that's being done, you're going to need to look a little deeper.
4. Church is a support system.
There are many areas I wish the Church at-large would grow in empathy and compassion. But when I stop to think about it, it's been people in church who've been there for me in my darkest hours.
When I look back on those dark times, I'm tempted to count the names of people who've betrayed me or hurt me in one way or another. But I often neglect to remember the people who have been there, cared, sacrificed and stood beside me.
Those people were there not only because they loved me, but because they loved Jesus. They were the Church to me, and it's disingenuous for me to ignore them to focus on the others (whose failures I am probably blowing out of proportion).
When I take a moment to think about it, I'm so thankful for the people who will meet me at a moment's notice and encourage me, cry with me, share Scripture with me, admonish me and remind me of what's important.
Sometimes they say stupid and hurtful stuff, but they've also loved me despite the stupid, sinful and hurtful stuff I've said and done.
5. Church is a spiritual discipline.
I have no doubt that I could abandon the local church and cherry pick some friends to meet with regularly who would make spirituality and theological discussions deep, challenging and fun.
But when I'm honest with myself, most of my growth has come from interacting with people I wouldn't choose. By handpicking my social circle instead of submitting to a local community of believers, I'll generally choose people who fall within my comfort zone.
I've grown in my ability to love by getting close to people with opinions I disagree with, different lifestyles, disabilities and all sorts of issues I had not been previously been exposed to.
I need people close to me who I can disagree with and challenge in a healthy way—while still loving and wanting what's best for them.
I need a multi-generational, ethnically and financially diverse community of people to mentor me and broaden my perspectives. I need people close to me who I can disagree with and challenge in a healthy way—while still loving and wanting what's best for them.
There's no question that every church has significant problems, and I've often daydreamed about quitting. But I truly believe we need each other.
One Caveat
I know that there are some reading this who've experienced abuse at the hands of a church. I don't intend for this post to gloss over, ignore or provide a glib answer to your legitimate pain. Sometimes there are people too traumatized by church to jump back into that relationship.
If that's you, I'm terribly sorry. I'm sincerely praying that you find healing and can come to a place where you are ready to give it another shot, but most of all, I don't want you to read this as condemnation for your experience.
I totally get why people would consider walking away from the Church, but I think we desperately need each other.
This post was originally published on jaysondbradley.com
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This article is one person's approach and I'm glad he has found he can work within the system for reform. Maybe he has a church that is open to change. I don't expect perfection from the church. I wish perfection wasn't expected of me either. The "drive" for excellence and being on mission to give God's best meant I had to be at a level I couldn't demonstrate. I haven't abandoned, but I need to find an expression of church that doesn't reject me or wear me out. I just want to love people and I want to have the freedom to do that imperfectly
What if you don't want to leave your church, but you want to change your church. Is that okay?
21 Comments
I think the biggest reason why Millenials are leaving the church is because they stopped believing traditional Christian theology. People increasingly do not believe that faith in Jesus is necessary for salvation, and they do not believe that good people can go to hell.
That's the biggest struggle that the church is up against.
Jesus prayed for unity between all believers but there seem to be so many new churches created every year, each a splinter from the church they came from with different theology and values. Does this mean Jesus' prayer wasn't answered?
That is a great question!
The Bible does tell us that in the end every knee will bow and every tongue will confess so I like to think there is hope. I also think that as the church becomes more persecuted we will become more united. There is data to confirm the correlation of the relationship between persecuted nations and church unity...
Maybe doesn't fully answer the question but I think the point I'm trying to make is it's not finished yet...
The "church" across the spectrum have planted the seeds of their demise on their own. Whether it's by the blanket, unloving rejection of entire groups of people or because churches have lost their center and made it hard to know what to believe in. The confusion and the lack of love make it easy to move on to a life minus the heartache of trying to make church work. There are exceptions, but they are few and far between.
Isn't it interesting that most of the articles like this (1) don't recognize the reality of the church as she lives and breathes beyond our institutions, and (2) are written by those who make their living from those institutions?
He is only stating the reasons why he's still in church... so im not sure what you're getting at...
Also (1) the "reality of the church" is up for interpretation and debate so the way you make it sound is that only people who have been "enlightened" to not join an "institution" can really have a legitimate opinion and (2) the worker is worth his wage and that wage does not disqualify his opinion
It's not that I'm leaving the church because I don't love God. I love my church to bits. The problem I have is when church picks and chooses what commandments to follow. Especially when the first is to 'love thy neighbor as you love yourself' yet they condemn gays (when I'm bisexual myself) and boycott them from church. Slightly unfair if you ask me.


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The Horse Forum (http://www.horseforum.com/forumindex.php)
- Horse Colors and Genetics (/horse-colors-genetics/)
- - What color could their foal be? (http://www.horseforum.com/horse-colors-genetics/what-color-could-their-foal-224866/)
What color could their foal be?
I'm really curious to know what color could result in a foal between my mare and the horse.
The last pic is of the horse.
He looks buckskin. Depending on whether they are homo or heterozygous for black and agouti you would most likely see bay or buckskin but could see palomino, chestnut, smokey black or black.
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Fifty percent of the teams and the top 25 individuals in each section move on to State. If there are an odd number of teams in the race then the 50% is rounded up to...
Keep checking back to see who makes the Arizona Milesplit 'Watch List" in Track and Field.
It was the last big weekend to qualify for state and many athletes were taking a shot at it but the wind was just not cooperating. Some were even competing in two...
Desert Vista Juniors Shaylah Simpson and Matt Yalung on the podium after winning their events. Simpson place first in the pole vault with a vault of 12-3. Yalung won...
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Ephrata to celebrate girls section title
By Ephrata Review on February 27, 2013
There will be a Championship Celebration Night for the Ephrata girls basketball team, which recently captured the Section Two title. The celebration will be held Thursday, Feb. 28, at 6:30 p.m. in the Middle School Gym.
Coaches and players will be on hand for autographs and pictures.


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Job Overview
Civilian employees serve a vital role in supporting the Army mission. They provide the skills that are not readily available in the military, but crucial to support military operations. The Army integrates the talents and skills of its military and civilian members to form a Total Army.
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This position is located in the General Pediatrics Service, Department of Pediatrics at the Tripler Army Medical Center, Hawaii.
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Who May Apply: U.S. Citizens
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Travel Required
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We chose Rimini to realize these big dreams. We chose the city of elegance and warm atmosphere in order to live big emotions.
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On-Airport Parking
Call 512-530-3300 for the parking status
Ground Transportation
Earn Points and Park Free
Earn 20 points for each dollar you spend with our ParkPerk Program™.
Family Friendly Valet: $18/day (tax included) New Location
24 hour minimum stay required
Drive to the upper level of the garage and leave your car with the attendant. Directions to the garage upper level.
Premium Valet Parking: $27/day (tax included) New Location
24 hour minimum stay required
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Drive to the upper level of the garage and leave your car with the attendant. Directions to the garage upper level.
Let us know if you'd like your car cleaned. Services range from exterior wash to high end detail. We'll take it from there.
Parking Garages (Garage 1 and 2): $23/day (tax included)
The On-Airport Parking Garage 1 is located directly across from the terminal and now has parking on the upper level. Garage 2 is a single level parking area adjacent to Garage 1 and within walking distance of the terminal. What could be easier? Zip in, zoom out. Hourly rate is $3/hour (or any part of), up to the daily maximum.
Lot A: $11/day (tax included)
Next to the garage and a short walk to the terminal. $3/hour (or any part of), up to the daily maximum.
Economy (Lots B - G) $7/day (tax included)
Free shuttle service between On-Airport economy lots and the terminal, 24 hours/day, in clean and comfortable shuttles that run on natural gas.
Credit Card Express Lane
Swipe the same credit card on the way in and on the way out. No parking stub to keep track of. Details here.
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To access the new Cell Phone Lot, from SH 71, take the Spirit of Texas Drive "Airport Cargo/Service Entrance" exit. Turn right onto Spirit of Texas Drive. Go straight at the four-way stop and at the second four-way stop go straight. The next right is Rental Car Lane. Turn right onto Rental Car Lane and the cell lot will be on the left. One may also access the cell phone lot from the main airport entrance road, Presidential Blvd. via Spirit of Austin Lane. From SH 71, take Presidential Blvd. to Spirit of Austin Lane and take a right. Spirit of Austin Lane comes to a four-way stop at Spirit of Texas Drive. Take a left onto Spirit of Texas Drive. The next right is Rental Car Lane. Turn right onto Rental Car Lane and the cell lot will be on the left.
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Take a picture of the aisle with your cell phone. Our friendly drivers will also give you a reminder card upon request.
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Complimentary Parking
Vehicles displaying the following plates with disabled veteran (DV) or without disabled veteran (DV) designation may receive up to five (5) days complimentary parking in the garage. Any day(s) beyond the five (5) will be charged at regular pricing.
- Medal of Honor
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Please note: Complimentary Parking for disabled veterans is not applicable to Valet Parking Services, Hourly Parking Services on the garage or surface lots where fee collection is automated. The eligible license plate must be on the vehicle exiting the parking operation to receive complimentary parking. Vehicles occupying more than one space will be charged the daily maximum for each space used.
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The spore-forming bacterium Bacillus thuringiensis produces intracellular inclusions comprised of protoxins active on several orders of insects. These highly effective and specific toxins have great potential in agriculture and for the control of disease-related insect vectors. Inclusions ingested by larvae are solubilized and converted to active toxins in the midgut. There are two major classes, the cytolytic toxins and the δ-endotoxins. The former are produced by B. thuringiensis subspecies active on Diptera. The latter, which will be the focus of this review, are more prevalent and active on at least three orders of insects. They have a three-domain structure with extensive functional interactions among the domains. The initial reversible binding to receptors on larval midgut cells is largely dependent upon domains II and III. Subsequent steps involve toxin insertion into the membrane and aggregation, leading to the formation of gated, cation-selective channels. The channels are comprised of certain amphipathic helices in domain I, but the three processes of insertion, aggregation and the formation of functional channels are probably dependent upon all three domains. Lethality is believed to be due to destruction of the transmembrane potential, with the subsequent osmotic lysis of cells lining the midgut. In this review, the mode of action of these δ-endotoxins will be discussed with emphasis on unique features.
The spore-forming bacterium, Bacillus thuringiensis (Bt) is closely related to two other important spore-forming Bacilli, Bacillus cereus and Bacillus anthracis, and is differentiated largely on the basis of containing several plasmid-encoded protoxin genes. There are hundreds of Bt subspecies and most produce, primarily during sporulation, one or more parasporal inclusions, each comprised of either one or several related insecticidal protoxins, the so-called δ-endotoxins. Each of these toxins is active on a subset of insect larvae from at least three orders of insects [1]. Because of unique but overlapping specificity profiles, Bt subspecies are generally effective against a broad range of insects, usually within a particular order of insects. Many also produce, during growth, less well characterized insecticidal proteins, the so-called vegetative insecticidal proteins [2] as well as secreting other pathogenic factors [3].
Since a substantial fraction of the post-exponential metabolism of these bacteria is devoted to the synthesis of these highly specific insecticidal proteins, there must be some survival benefit to their production. While Bt strains have been isolated from many environments, some or all of the natural habitats must involve interactions with insect larvae. Epizootic infections are rare, however, so it is not likely that the primary reason for toxin production is to allow Bt to exploit nutrient-rich insect larvae. The bacterium is more likely to have a more subtle symbiotic interaction, perhaps with plants, to account for the extensive production of these highly specific and efficacious proteins. In fact, the commercial use of suspensions of spores and inclusions has been limited in part due to the need to spray at rather frequent intervals in order to sustain an effective level of the biopesticide. Not only are the δ-endotoxins readily inactivated, but the number of spores and/or vegetative cells decreases rather rapidly in the sprayed area. This problem has been circumvented by engineering plants (cotton, corn, potato, rice, etc.) to produce the toxin [1].
When insect larvae ingest inclusions (and the spores, which can be synergistic), the protoxins are solubilized and processed to toxins by midgut proteases. On the basis of in vitro studies, it is most likely that the toxin binds to receptors on cells lining the larval midgut, inserts into the cell membrane and forms ion channels (which are somewhat cation-selective) resulting in loss of the transmembrane potential which leads to osmotic cell lysis [4]. While this is the generally accepted mode of action, it should be noted that at sublethal concentrations of toxin, there are larval behavioral changes such as avoidance of the toxin during feeding and paralysis of feeding. These effects may be due to the formation of ion channels but a different or additional mechanism of toxin activity has not been excluded.
The mode of action of these toxins has been well reviewed [1,4–7], including a recent thorough presentation of the specificity of toxin-binding to receptors [1]. In this review, we shall discuss recent studies on the steps involved in toxin-binding, insertion and organization within the membrane which are unique features of these very effective biological insecticides.
An overview of toxin structure
Most of the research has been done with toxins active on Lepidoptera, designated Cry1 (see [8] for the terminology). The structures of a Cry1 toxin, Cry1Aa, and of a toxin active on Coleoptera, Cry3A, have been determined and are very similar despite only about 25–30% amino acid sequence identity [9,10]. There is another major group of toxins, designated Cry4 and Cry11B, which are active on Diptera. There may be additional toxins which are selective for other orders of insects as well as binary toxins including the dipteran-active toxin produced by Bacillus sphaericus [11]. The Cry3A and Cry1Aa toxins have a three-domain structure (Fig. 1) consisting of a seven α-helix bundle for domain I with six of the helices surrounding a central helix α5. Domains II and III are comprised of β-sheets but in different conformations, with domain III containing two antiparallel β-sheet sandwich structures. The helices in domain I are largely amphipathic, with the α4–loop–α5 region being the most hydrophobic [10]. There are numerous studies, primarily mutagenesis, implicating domain I, especially helices α4 and α5, in the formation and function of the ion channel. Domains II and III have functions in specific binding to receptors and for domain III, in modulating ion channel activity [1].
The structure of domain I is very similar to pore-forming regions of the E1 group of colicins and the B subunit of diphtheria toxin, despite little sequence similarity among these toxins [12]. The monomeric form of colicin E1 may be sufficient to form ion channels in bacterial membranes. The state of oligomerization of the diphtheria B subunit is not known and ion channels formed by δ endotoxins appear to be oligomers (see below). In contrast, toxins comprised of β-sheets, such as the pore-forming aerolysin and the α hemolysin of Staphyloccus aureus, form heptamers at the membrane surface prior to insertion [12].
Toxins initially bind reversibly to larval membrane receptors
Inclusions are solubilized by the high pH and reducing conditions in the midgut of lepidopteran and dipteran larvae. The protoxins of ca. 130 kDa are then proteolytically converted to active toxins of 55–65 kDa, primarily by removal of the carboxyl halves. Active Cry1 toxins bind reversibly and with high affinity to receptors on the surface of larval midgut cells [1]. In several insects, the receptors are aminopeptidase Ns. There appears to be more than one toxin-binding site on these molecules on the basis of kinetic studies [13–15] and peptidase digestion of the receptor [16]. A larger cadherin-like molecule has also been implicated as having other membrane proteins.
Binding has been measured by adding labeled toxins in the presence or absence of unlabelled toxin to membrane vesicles prepared from the midguts of insect larvae, so-called brush border membrane vesicles (BBMVs). In this system, there is reversible binding to a receptor(s) as well as insertion of the toxin into the membrane (non-reversible binding), complicating the calculations of the rate constants. More accurate determinations of the binding constants have been obtained by anchoring purified toxin-binding proteins in a surface plasmon resonance system for the determination of the on and off rates [13–15]. The former would reflect the initial reversible binding and the latter provides a measure of the irreversible insertion into the membrane. More qualitative assessments of the initial binding, especially of the variety of proteins involved, have been obtained by immunoblotting.
In many cases, there is a good correlation between a so-called binding constant measured as described above with BBMV and toxicity. This value, which has been designated Kcom, really reflects the ratio of the initial reversible binding and a second irreversible binding [1]. Even then, there are several exceptions to the correlation. The reason for high-affinity binding with low or no toxicity is not known. It has been suggested that the initial 'on rate' may not be indicative of the function of the receptor(s) in toxicity and that the membrane insertion step may be better correlated with toxicity [1]. There are also cases of several binding proteins in certain insects, some or all of which may function in toxin insertion into the membrane. In addition, there is evidence that the Cry1Ac toxin binds to more than one site on the aminopeptidase N receptor [13,15]. Site I is located within domain III and displays fast association and dissociation kinetics. Site II within Domain II possesses slower kinetics yet tighter affinity [15]. This dual binding may reflect a two-step process with an initial relatively non-specific reversible association followed, in cases of toxins active on the particular insect, by binding to a second site on the same receptor or on a receptor in close proximity. Binding at the second site may lead to a conformational change of the toxin necessary for membrane insertion. Alternatively, binding may be important for mobilizing toxin at the membrane surface, either for oligomerization or for a concentration-dependent insertion step. This possibility would probably depend upon the localization or mobilization of the toxin receptor(s) within the membrane which has yet to be established.
Despite the uncertainty of the function of the initial toxin–receptor interaction, there is evidence for its importance. In some cases, there was a correlation between resistance of an insect to a particular toxin and reduced binding (Kcom value) of that toxin to vesicles prepared from BBMVs of the resistant insect [1]. The decreased binding may be due to an alteration of the receptor itself or of a membrane component other than the toxin receptor, which results in a change in the accessibility of the receptor for toxin-binding. There is evidence for a decreased fitness of toxin-resistant Plutella xylostella [1] which may reflect such pleiotropic membrane changes. In several cases, a correlation between resistance and reduced toxin-binding was not found. Many of these insects were resistant to a number of toxins, including some to which they had never been exposed. The basis for resistance in these cases is not known but could involve such pleiotropic changes.
An important role for the receptor was also demonstrated in a variety of in vitro experiments. The concentration of toxin required to obtain ion channel activity in synthetic phospholipid vesicles was reduced at least 100-fold by the inclusion of toxin receptor [7]. Such experiments included the fusion of BBMVs with artificial phospholipid membranes and measurements of conductance by patch-clamping. The release from vesicles of voltage-dependent fluorescent dyes such as calcein [7] as well as the release of 86Rb+–K+ from vesicles reconstituted with purified aminopeptidase N [17] have also been used to assay ion channel activity. The latter results imply that the only membrane protein required for ion channel formation by toxins is the aminopeptidase N receptor. However, the relation of the 86Rb+ release activity to ion channel function in vivo or in BBMVs has not been established. As a result, interaction of the toxin with other membrane components is not excluded. In fact, there were significant differences between the conductance of the ion channels formed in synthetic phospholipid vesicles and in fusions to BBMV implying a role for some undefined membrane components in ion channel function.
Clearly, more studies of toxin–receptor interactions are needed. The genes encoding several receptors have been cloned so in vitro mutagenesis can be done in order to study receptor function in reconstituted vesicles. It should also be possible to compare the deduced sequences of receptors from resistant and sensitive insects to determine if such a change were the basis for resistance. This is obviously an important issue to resolve, especially for dealing with insect resistance.
It should be noted that, in general, there is surprisingly little known about the interaction of bacterial toxins with receptors. In contrast, the binding of δ-endotoxins to specific membrane receptors, especially aminopeptidase N, has been extensively analyzed. There is considerable potential, therefore, for developing relatively simple in vitro systems comprised of phospholipid vesicles containing one or a few membrane proteins (including the receptor) for exploring toxin-binding and the subsequent formation of ion channels. These systems could serve as a paradigm for understanding such interactions for other toxins.
δ-Endotoxins insert irreversibly into membranes
The so-called 'irreversible binding step', i.e. insertion of the toxin into the larval membrane, is not yet understood. Other bacterial pore-forming toxins utilize a variety of mechanisms for toxin-binding to and insertion into the membrane. Pore-forming colicins and perhaps the B subunit of diphtheria toxin (both comprised of α-helices in the pore-forming region as are the δ-endotoxins) depend upon an acidic environment for a conformational change of the membrane insertion domain of the toxin resulting in a charge interaction of the toxin with membrane phospholipids [12]. Aerolysin and the S. aureus α-hemolysin are comprised of β-sheets in the pore-forming region. For these toxins, aggregation to heptamers at the membrane surface results in a conformational change exposing hydrophobic regions for membrane insertion [18,19].
The Cry1 toxins function in the alkaline environment of the larval midgut (at least for Lepidoptera and Diptera) which appears to be important for inclusion solubilization and conversion of the protoxin to toxin. There are indications, however, that a charge interaction with the membrane may be important at some stage for the insertion of the Cry toxins. A positive charge in the region of the loop connecting helices α2 and α3 in domain I is essential for activity [1]. A number of mutations of R93 in this region to non-charged or negatively charged residues or an A92D mutation resulted in toxins which still bound reversibly to BBMV but did not insert into the membrane. An alternative explanation for these results is that an intramolecular salt bridge, known to involve R93 in the Cry1Aa toxin [10], may be critical for toxin conformation and thus for insertion.
It is likely that prior to such a charge interaction, toxin insertion involves a conformational change, perhaps analogous to the conversion of toxins to a 'molten globule state'[12]. The initial reversible binding involves certain loops within domain II of the toxin which are in close proximity to helix α7 of domain I [1](see Fig. 1). A swinging out of domain I with exposure of helix α7 to the membrane surface has been postulated on the basis of disulfide cross-linking of various domain I helices with each other and with loops within domain II [7,20]. Cross-linking resulted in the loss of activity which could be restored by reduction of the disulfide bonds. A reasonable explanation for these results is the requirement for a conformational change involving these helices.
Experiments with synthetic peptides of the domain I helices also support a role for helix α7 in enhancing the insertion of the critical pore-forming region, helix α4–loop–α5, into the membrane. Among the seven synthetic peptides of the α-helices in domain I, peptide α7 bound most rapidly to phospholipid vesicles [21], consistent with its primary role in membrane insertion. This peptide also enhanced the penetration of peptides α4 and α5 into such vesicles. For example, synthetic peptides corresponding to helices α5 and α7 from two different α-endotoxins were incubated with phospholipid vesicles and only the former was found to permeate the membrane and oligomerize to form ion channels [21], Furthermore, the α5 peptide specifically associated with the partially inserted α7 peptide as well as with itself in a parallel manner within the membrane as expected if the pore were formed from several such helices.
In a more detailed study, peptides of the seven helices of the pore-forming domain of the Cry3A toxin were synthesized and compared as to their membrane interactions, structure within the membrane, orientation relative to the membrane plane and the network of peptide–peptide interactions in the membrane between all pairs of peptides [21]. Attenuated total reflectance-Fourier transform infrared spectroscopy was used to study the structure and orientation of the helices when bound or inserted into the membrane. All of the helices, except α1, interacted with the lipid membrane and only α4 and α5 were in a transmembrane orientation. Remarkably, a network of recognition of all the possible combinatorial pairs of membrane-bound helices showed that only α4 and α5 self-assembled within the membrane. Moreover, α4 and α5 coassembled in an antiparallel manner, an orientation expected if they had inserted as a hairpin.
Peptides of helices α4, α5 and α6 recognized α7 in the membrane-bound state. Since helix α7 is located at the interface between the pore-forming domain and the receptor-binding domains (Fig. 1), its ability to coassemble with α5 and α6 may assist the insertion of the α4–loop–α5 hairpin into the membrane. Following the binding of the toxin to the receptor, the affinity of α7 for the membrane surface and for the other helices could lead to the unpacking of the pore-forming domain and facilitation of the insertion of helices α4 and α5 into the membrane. Overall, these results are consistent with a model in which only helices α4 and α5 insert into the membrane as a helical hairpin in an antiparallel manner, while the other helices lie on the membrane surface like ribs of an umbrella (the 'umbrella model') as proposed earlier by Li et al. [9] (Fig. 2). Alternatively, toxin monomers may aggregate at the membrane surface prior to insertion as do the pore-forming aerolysin and the S. aureus α-hemolysin. The resulting changes in toxin properties apparently enhance exposure of hydrophobic regions and thus toxin insertion.
In the case of the membrane-bound Cry3A toxin, there may be a proteolytic cleavage between helices α3 and α4 which could trigger a conformational change leading to the formation of functional channels [1]. The Cry4A toxin can also be cleaved between helices α5 and α6 to fragments of 20 and 45 kDa apparently without a substantial loss of activity for mosquito larvae [22]. This cleavage is not essential, however, since mutant toxins lacking the cleavage site were active. Deletion of the region encoding helix α5 from the smaller fragment did not alter toxicity when it was mixed with the 45-kDa proteolytic fragment whereas a further deletion into the region encoding helix α4 resulted in loss of toxicity of the reconstituted fragments. It appears that helix α5 but not α4 is dispensable for toxicity in reconstituted Cry4A toxin. It is likely that the structure of this toxin is very similar to that of the Cry1Aa and Cry3A toxins given the presence of conserved sequence blocks in all cases [1] and the extensive structural similarity of these two toxins despite limited sequence identity. If so, the Cry4A toxin probably has the same mode of action implying that helix α5 is not essential for ion channel formation, at least in such reconstitution experiments. Mutagenic studies of cry1 toxin genes, however, clearly implicate helix α5 in toxicity [1] and as discussed below, probably in the aggregation of toxins within the membrane. The interaction of two proteolytic fragments of the Cry4A toxin may have resulted in the exposure of other regions of the toxin which compensated for the absence of helix α5.
To date, there have been no definitive reports of a conformational change of the toxin in part because of the difficulty of detecting a signal from a fluorescently labeled toxin (or from tryptophan residues) in the presence of BBMV. While toxins can form ion channels in synthetic phospholipid vesicles, the efficiency is very low and a large excess of toxin is required [7] so experiments to look for such changes have not been feasible. Definitive studies on toxin insertion may be possible with vesicles constructed from phospholipids (perhaps selected ones) containing the purified protein(s) (such as aminopeptidase N) required to mimic in vivo conditions. As previously mentioned, there is evidence that aminopeptidase N reconstituted into such vesicles may be sufficient [17] but further experiments are needed to compare the functional properties of ion channels formed in such vesicles to those due to toxin interaction with BBMV. Correlations of the behavior of various toxins in vivo and in such systems would be helpful as would the study of various mutant toxins. There is a large collection of Cry1 mutant toxins which could be exploited for this purpose [1].
Most of the toxin molecule is intimately associated with the membrane and forms aggregates therein
Whatever the mechanism for inserting δ-endotoxins into the membrane, most of the molecule is protected from digestion with protease K [23]. The only susceptible part is about 30 amino acids from the amino end which comprise helix α1. As mentioned above, a synthetic peptide of this helix is the only one of the seven fluorescently labeled peptides from domain I which did not bind to phospholipid vesicles [21]. In general, there is a good correlation between the membrane-binding and insertion properties of domain I synthetic peptides and those of the intact toxin. Synthetic peptides of helices α4 and α5 with specific amino acid changes known to inactivate the Cry1Ac1 toxin have altered membrane-binding and insertion properties.
In a random mutagenesis study of the regions encoding the domain I helices in the Cry1Ac toxin, the most significant loss of toxicity was due to mutations in helices α4 and α5 but not in the loop connecting these helices [1,24]. Among the synthetic peptides of the domain I helices only these two helices penetrated into the plane of the membrane [21,25]. Both helices are long enough to project through the membrane into the cytosol [9,10] and could thus potentially functionally interact with a cytoplasmic component. In order to examine whether any region of the toxin projected into the cytoplasm, BBMV were preloaded with protease K. Neither the Cry1Ab nor the Cry1Ac toxins were altered in size or function when inserted into these preloaded BBMV [26]. Most of the toxin molecule must remain within the plane of the membrane and function in such an environment. The helices within the membrane may be tilted or perhaps altered in conformation in order to enhance the interactions necessary for pore formation.
The retention of toxicity for Manduca sexta larvae by Cry1Ac toxins with mutations within the region encoding the loop connecting helices α4 and α5 [24] also indicates that a specific interaction between this region of the toxin and a cytoplasmic component is not essential for toxin activity. While this sort of functional interaction is unlikely, the overall orientation of the toxin within the membrane must be critical to its function. Fluorescent studies with domain I peptides indicate that specific domain I helices exist in different environments within or on the membrane. In particular, the helix α4–loop–α5 region is buried within the membrane with a more superficial location for the other helices (Fig. 2) [21,25]. Domains II and III are intimately bound to the membrane (based on resistance to protease K) but their exact orientations are not known.
Aggregated forms of the Cry1A toxins could be extracted from BBMVs by employing relatively mild conditions for membrane solubilization [23]. Active Cry1Ab or Cry1Ac toxins formed aggregates, about the size expected for trimers, in M. sexta and Heliothis virescens BBMVs. No such aggregates were formed by toxins which were inactive due to mutations either within helix α5 or of the R93 residue implicated in an ionic interaction required for toxin insertion into the membrane. The aggregates did not contain the aminopeptidase N receptor although interactions with other membrane components have not been excluded. While possible trimers were detected with the extraction conditions employed, larger functional aggregates, perhaps even heptamers as found for the β-sheets in aerolysin [18] and the S. aureus α-hemolysin [19] may be present.
In contrast to the results with the helix α5 mutants, inactive toxins with mutations encoding residues within helix α4 did aggregate in the membrane, but did not form functional ion pores as determined by light scattering [24]. In comparable experiments, an inactive Cry1Ac toxin with a mutation in helix α4 (N135Q) was examined by surface plasmon resonance for binding to purified aminopeptidase N from M. sexta anchored in a lipid environment [14]. The initial binding rate was the same as for the wild-type toxin but the off rate was more rapid suggesting a lack of interaction with the lipids by the mutant toxin. These results suggest that this particular α4 mutant toxin may not be able to insert into the membrane and aggregate there. Differences in lipid environments and the presence or absence of membrane proteins in the two protocols may account for the apparent discrepancy. Alternatively, only a subset of inactive α4 mutant toxins may be able to insert into the membrane and aggregate.
As discussed above, synthetic peptides of helices α4 and α5 were the only ones among the seven comprising domain I which inserted and oligomerized within the phospholipid bilayers [21]. A synthetic peptide of the α4–loop–α5 region was much more effective than either of these helices alone or as a mixture of the two helices in promoting the release of calcein from phospholipid vesicles [25]. While the α5 but not the α4 peptide aggregated in the membrane and formed a channel, it did so much less efficiently than the α4–loop–α5 peptide. These results and the mutagenic studies of the cry1Ac toxin gene form the basis for the models presented in Figs. 2 and 3 in which most of the toxin molecule is at or close to the surface and only helices α4 and α5 project into the plane of the membrane but not onto the cytoplasmic face. The exact relative locations of the helices and of domains II and III are not known. Since helix α5 is involved in aggregation, it must be available to interact with this helix from other toxin molecules in order to initiate ion channel formation.
This difference in aggregation properties of the mutants altered in α4 or α5 as well as the peptide results imply separate functions in ion channel formation for these two helices. Perhaps in analogy with the formation of the heptameric ion channel by the S. aureus α-hemolysin, there is a two-step process [19]. An initial aggregation due to interactions among the hydrophobic α5 helices would be followed by the formation of functional ion channels lined by the α4 helices. Separate functions for these two helices were also postulated on the basis of a different set of mutations of critical anionic residues within helix α4 [27]. In contrast, a synthetic peptide resembling helix α5 aggregated and formed calcein-releasing channels in phospholipid vesicles [21,25]. Caution must be exercised, however, in relating measurements of channel activity to the true function of a protein. In the case of the α5 peptide, channel activity was used as an additional indicator of oligomerization in the membrane rather than mimicking the function of this helix in the intact toxin. There are examples of proteins forming ion channels, such as the B subunit of the diphtheria toxin, which do not have a direct bearing on their primary function.
Aggregation may occur at the surface of the membrane or after integration of toxin monomers within the membrane. The results with fluorescently labeled synthetic peptides favor the latter possibility. The kinetics of fluorescent emission by peptide α5 were consistent with self-aggregation in the membrane in contrast to the binding kinetics of the other peptides of the domain I helices [21]. Since virtually the entire toxin molecule is intimately bound to the membrane, it is likely that the other helices of domain I, as well as domains II and III, are important for toxin function. For example, a highly conserved arginine cluster within domain III appears to be involved in the voltage-dependent regulation of the ion channels [1]. Mutation of the two outer R residues to K in this region resulted in lower insecticidal activity and reduced ion channel function as measured by light scattering or voltage clamping. This part of domain III is likely to be in close proximity to the ion channel within the membrane.
Conclusions and outlook
Some of the unique features of these δ-endotoxins as related to function have been discussed above but are worth emphasizing for future experiments. Despite a distinct three-domain structure of these toxins, there is considerable evidence for extensive functional interactions. (a) Specific toxin-binding to membrane receptors seems to involve residues in both domains II and III, perhaps in sequential steps leading to toxin insertion [13–15] (Fig. 3). The latter could require an induced conformational change for the insertion of toxin monomers or an enhancement of the concentration of the toxin at the membrane with the conformational change occurring during oligomerization. This step and the formation of ion channels could involve interactive functions among the three domains. (b) Regulation of the conductance properties of the ion channels almost certainly involves an interaction between domains I and III [1]. (c) A number of domain swapping experiments among cry1 genes support such functional interactions for receptor-binding specificity and for the properties of the ion channels [28]. Some of the hybrid toxins had enhanced activity for certain insects which could be due to improved binding or perhaps more efficient insertion, aggregation or ion channel-forming processes.
Another consideration is that these toxins may have function(s) in addition to ion channel formation. Behavioral changes of larvae such as avoidance of the toxin and paralysis of feeding, may be more rapid than the formation of ion channels. If so, these behavioral changes could be due to effects of the toxin prior to ion channel formation, perhaps within the membrane but before there is extensive aggregation. Possibilities are alteration of the properties of pre-existing ion channels or of a membrane ATPase. These speculative modes of action may involve parts of the toxin not yet implicated in its function(s).
There are still uncertainties as to the role of receptor-binding, the process of insertion of the toxin into the membrane and the extent and mechanism of oligomerization to form ion channels. The progress in these areas has been considerable, however. There are now a number of experimental techniques and tools available to resolve many of the basic issues regarding the mode of action of these toxins. There is an extensive number of well characterized mutant toxins, especially the Cry1 class, purified and cloned toxin-binding proteins and the potential to develop relatively simple lipoprotein vesicles or preparations for investigating toxin insertion, ion channel formation and perhaps other functions of these toxins. There are indications that the mechanism of action of these δ-endotoxins, especially functional interactions among the domains, may be unique. Given the promising experimental systems and the wealth of information already available, novel insights into the mechanism of action of a multidomain toxin should be forthcoming. With the refinement of the experimental systems for investigating their function(s), it should be possible to develop a complete picture of δ-endotoxin action. Such studies have very broad implications for dealing with resistance and for designing even more effective and specific biological control agents.
We thank our numerous colleagues who provided reprints and preprints and to Dr. Sarjeet Gill for his insightful comments. Dr. Jeff Bolin graciously provided Fig. 1 using MolScript and Raster3D. We apologize to all of the researchers in this field for not being able to cite more extensive references because of editorial limitations.


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Mental Health Care, China Style
- Categories: International, News
- Date: Aug 26, 2011 16:14 JST
- Tags: BDSM, Bizarre, China, Chongqing, Crime, Medical, Mental Illness
The plight of a mentally ill man kept chained in a barn for 23 years by his elderly mother has lately been upsetting Chinese and non-Chinese alike.
- Anonymous commented on May 11, 2012 04:28
Those sort of actions, and his apparent normalcy before ~15 seem to suggest schizophrenia, which usually develops in teenage years. The crime is that with treatment one can live a normal, productive life. TBH that... More
- Anonymous commented on May 11, 2012 01:25
Prisons these days are actually very nice compared to the past.... More


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|Property type
|Minimum Stay
|3 - 7 nights
Our property is a classic 1925 Folly Beach oceanfront cottage. Harks back to simpler times with wood floors, ceilings, and walls. Screen porch along the length of the beach side provides outdoor living space with a hammock, rocking chairs, and wicker overlooking the beautiful Atlantic Ocean. Open and spacious floor plan has ocean views from every bedroom affording ocean breezes and gentle wave sounds to lull you to sleep. When there is no breeze, the house has central heat and air conditioning with ceiling fans in every room. The one bathroom has a shower but no bathtub.
Two bedrooms have queen beds and two have twin beds. TV in Living Room and Master Bedroom. Short walk to beach directly in front of the house. Outdoor shower with hot water. The house is completely furnished with bath towels, bed linen, and pillows. Underneath the house is the laundry room with washer and dryer or clothes line to hang up your swim suits.
Bring your beach towels, bikes, surf boards, kayaks, and beach toys. The Edge of America awaits you. No smoking. No parties. Will entertain pets. Please inquire.
Historic Charleston is just 20 minutes away with museums, site seeing, fine dining, and shopping. The business district on Folly, however, has good restaurants, bars with music, and shopping. Owner is a native Charlestonian and can help you plan your visit. Let me know if I can help you in any way.
Keywords: beach cottage, beachfront, oceanfront, ocean front, beach front, Folly Beach, 4 BR
Just returned from a week-long stay at the "Chicken House" and have nothing but great things to say. The house is charming and comfortable, especially if you like a vintage, retro feel -- it's all of that in the best way. Yes, only one bathroom, but our party of 6 really didn't have any problems rotating through. Our party was two couples and two individuals and the bedroom setup was perfect for everyone to have their own space, with a comfortable common room in between and a big kitchen table. The screened porch was the star of the show -- we almost had to arm-wrestle for hammock privileges. Not isolated at all, but far enough off the main drag of Center St. to be quieter in the evenings. The house really did have everything we needed and the views and beach access were stunning. Amanda was a thoroughly gracious host, and we were able to work out a last-minute addition of a couple extra nights to our stay because we weren't ready to leave yet! We've vacationed on Folly for years but have never stayed in the same place twice - but in this case we truly fell in love and can't wait to repeat!
We booked with Amanda Griffith after phone and email exchanges with her and were looking forward to our trip. After we already provided full payment and 4 days prior to our arrival, Amanda cancels on us without any explanation of any kind. She did return our full payment but refused to answer our calls or emails. We had already made all the arrangement to take the trip and received no details as to what prompted the cancellation.
We stayed at the Chicken Lady's house in early June and had a great time. The beach is easily accessible and the screened porch was fabulous. There were seven of us and we didn't have any problem with the one bathroom. The outdoor shower has hot water. Several of us took turns sleeping in the hammock. Our host even shared her fresh cherry tomatoes from the garden with us!
The kitchen renovation is great. We especially liked the retro refrigerator and gas stove. The dining room seats 8 easily and we used the outdoor grill most of the time to cook our meals. The house decoration is very distinctive just like a beachfront house should be. Everything we needed was there. I would bring your own beach chairs but there is a variety of beach toys under the house.
Charleston is only a half hour drive from the house. The restaurants were wonderful. So many, so little time.
Parking was convenient and the block is a little less crowded as there isn't any public parking in the block. We will be back again. Thanks, Folly Beach.
This is the second year that we have rented this house right on the beach. We fell in love with it last year and it did not disappoint this year. The cottage is a nice size - we are a family of 5 and everyone has their own space with room to spare without feeling lost. All of the necessary ammenities were available.
For those who are not familiar with the area, it is a lengthy walk to get to the center where the stores/restaurants are (so bike or drive a cart instead). The benefit of this is that you really do feel like you have your own semi-private beach!!!!!
The owner and the property manager are very friendly and make every efforts to ensure your stay is pleasant.
Can't say it enough - love this place!
This little house is in the perfect location and is really comfortable. It's only 20 minutes to downtown Charleston, provides easy access to the main streets of Folly Beach, and the ocean is literally just steps away. A little bit rustic and a little bit charm, the cabin is well maintained. We had a group of seven people and the single bathroom was a non-issue. It is also located towards the back of the house, so there is no traffic near that area constantly during the morning or evening hours - and we never ran out of hot water! We also thought we were going to have some refrigerator problems, but the property manager was quick to call a repairman - and even offered to purchase us a new fridge so we would enjoy our stay. Now that's service! We would love to come back to this charming little cabin for a future stay on Folly Beach!
Love the relaxed atmosphere of this beach cottage! Loved the rocking chairs and rope hammock out on the porch where I could relax and meditate on the ocean waves. Very comfortably furnished and perfect for a relaxing get away or family vacation with young children. Close enough to the end of the island that you feel you have your own private stretch of beach. Plenty of nice places to eat just a bike ride away. Can't wait to be able to go back!
|Rate Period
|Weekend Night
Early Spring 2017
Mar 1 2017 - Mar 25 2017
3 night minimum stay
Spring 2017
Mar 26 2017 - Apr 7 2017
3 night minimum stay
Spring Break
Apr 8 2017 - Apr 15 2017
7 night minimum stay
Apr 16 2017 - Jun 2 2017
3 night minimum stay
Summmer 2017
Jun 3 2017 - Aug 26 2017
7 night minimum stay
Late Summer
Aug 27 2017 - Sep 2 2017
7 night minimum stay
My Standard Rate
3 night minimum stay
|Refundable damage deposit
|$160 + Tax
|Pet Fee
|Tax Rate
Please ask about dogs.
50% of total rental to reserve your dates and balance 60 days before arrival.


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Thiago set for surgery
Bayern Munich will be without new signing Thiago Alcantara for almost two months after tore ligaments in his right leg on his starting debut.
- Related Content
The summer signing from Barcelona was included in the Bayern team for the 2-0 win over Nurnberg on Saturday, but was withdrawn after taking a knock.
He will now require surgery and has been ruled out for seven weeks, according to a statement on the Bayern website.
"Thiago picked up the injury during the second half of the game against Nurnberg," read the statement. "He will undergo surgery in Munich on Monday and is expected to be out for seven weeks.
Thiago therefore misses Bayern's games against Freiburg, Hannover, Schalke and Wolfsburg while he will also be absent from the Champions League holders' first European games of the season.
The Spain under-21 international has also been ruled out of their DFB-Pokal second-round tie with Hannover next month.
Sky Bet Free Bet Club: £10 free bet - £5 free every week


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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 instructable is a cheap way to incorporate abstract art into your home or office. This may also get rid of unwanted junk, in this case it's an ...
For this you will need as followed: An old picture frame, different sized washers (in this case seven sets of threes), and a hot glue gun.
First take apart you picture frame and lay out all the washers in the configuration that is wanted.
Glue all the washers in place and put the picture frame back together.
Inexpensive Flower Wall Art in Minutes .by SonikaJAnand
DIY Christmas decor! Easy&affordable ideas!by DIYlover89
Broken Mirror Art!by Meglymoo87
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1.Put the beans in the pressure cooker along with water and other spices like salt, cumin powder, onion powder, Mexico chili powder, ground pepper and liquid smoke.
2.Set the stove on medium high and cook till the pressure regulator starts rocking and pops.
3.Remove the pressure cooker from the stove and put it under running cold water. Once the air vent drops down, the beans are ready to be taken out of the pressure cooker.
4.Serve with rice with lemon juice sprinkled on top.
Serving size Per servingComplete recipe
Calories 115 Calories from Fat 12
% Daily Value*
Total Fat 1 g2.1%
Saturated Fat 0.2 g1%
Trans Fat 0 g
Cholesterol 0 mg
Sodium 117.7 mg4.9%
Total Carbohydrates 20 g6.5%
Dietary Fiber 4.9 g19.5%
Sugars 0.9 g
Protein 7 g13.1%
Vitamin A 3.4%
Vitamin C 3.9%
Calcium 4%
Iron 10.1%
*Based on a 2000 Calorie diet


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La Tamborrada: a guide to the drum festival of San Sebastian
This is an excerpt from Lonely Planet's A Year of Festivals.
Location: Begins in Plaza de la Constitucíon, San Sebastian, Spain, and spreads through the old city.
Date: 20 January
Level of Participation: 1 – it's about hearing not doing
Description: Can you hear the drums, Fernando? You'll be hard pressed to hear anything else for 24 hours in the Basque city of San Sebastian as it stages the unusual – and unusually loud – La Tamborrada. At the stroke of midnight a flag is raised in Plaza de la Constitucíon and regiments of soldierly drummers begin parading through the resort town, banging and thumping at drums and barrels to celebrate (or wake from the dead) the city's eponymous patron saint.
Throughout the day and into the next evening there's no pause in the drumming as up to 100 regiments of drummers take to the streets of the ritzy resort city. Even San Sebastian's children get a turn, with all of the city's schools marching and drumming in uniform during the day. The origins of La Tamborrada are unclear, with some stories crediting it to maids who banged their water buckets at one of San Sebastian's wells.
Essentials: Consider bringing earplugs if you want any sleep this day.
Local Attractions: After two long and loud nights, you'll probably appreciate some quiet time on the city beaches of La Concha and La Zurriola, which are among the best strands in Europe.
More Info: San Sebastian tourist office (www.sansebastianturismo.com)
See a list of other festivals in January here.