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After a long time, Shankar-Ehsaan-Loy look in their full color. No, I won't say ZNMD was bad, but somehow it was patterned, without much 'new' really. But they are back again with Aarakshan, with their old mate Prasoon Joshi, this time not just writing but also composing a number, which surprisingly, almost beats SEL's compositions.
The album starts with 'Mauka' which is, in a way, the theme song of the movie. The song, that basically talks about giving a chaanas (chance) is a song with enthusiasm and a wish to do something. Sung by five singers led by Raman Mahadevan and Mahalaxmi, the song has an item'ish, addictive touch to the music and zeal in the lyrics. Will get promotion, has to work.
Next comes Achcha lagta hai, sung by Mohit Chauhan and Shreya Ghoshal. The song that reminds me of rut aa gayi re in the first few moments, is a peppy number with some nice and fresh arrangements and while Mohit Chauhan is as great as ever with his lovely voice, Shreya is completely into the conversation happening in the song. Liked it. Very much.
Still, Kaun si dor/ Saans Albeli, the composition of Prasoon Joshi, is the surprise of the album. While I had no idea what to expect from him in his first ever composition (at least to my knowledge, in and out of movies), he completely took me by wonder with the song that, sung by Pt Channulal Mishra (with Shreya in case of the duet version Kaun si dor), goes almost completely classical, and the best part is that once I started listening to the song, I did not feel like stopping it for even once in the quarter to six minutes' run of the song. Completely loved Prasoon's debut as a composer.
The last song of the album is Roshanee, sung by Shankar Mahadevan, which starts in a slightly 'Uff teri adaa' style, but shifts totally to a passionate, 'joshila' number very soon. Again some new arrangements from Shankar-Ehsaan-Loy with some good words by Prasoon.
In short, the album is a small treat of four songs from Shankar-Ehsaan-Loy and Prasoon Joshi. Prakash Jha, who got four different composers to create four songs in Raajneeti has once again got it right, in fact better this time.


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BDSM, Bondage, Domination, Pornstar, Sex and Submission, Submissive | December 30th, 2011 |
Trained and Degraded Pornstar – Pornstar Mckenzie Lee gets her first taste of real BDSM with James Deen.
This hot busty English girl explores a sexual world where she must give up all control to please others. She is tied up, made to come, humiliated, punished and ass fucked!
Tags: BDSM, big tit, Bondage, busty, domination, Mckenzie Lee, rough sex, submissive, submissive female

subset: commoncrawl


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Loan Watch: Campbell's Debut Delight
Written by Newcastle Utd
Adam Campbell scored on his St. Mirren debut but was unable to prevent his new side slipping to 2-1 defeat
By Anthony Marshall - Newcastle United Managing Editor
Adam Campbell scored on his St. Mirren debut but was unable to prevent his new side slipping to 2-1 defeat away at Kilmarnock on Thursday afternoon.
The teenage striker, who turned 19 on New Year's Day, fired the Buddies ahead just 12 minutes into his bow, swivelling in the box before finding the bottom corner with an impressive finish.
Campbell continued to threaten and went close on a number of occasions but Kilmarnock levelled just before the half-hour mark when Rory McKenzie fired home.
St. Mirren looked to be heading for a share of the spoils and a decent away point but, with time almost up, experienced front man Kris Boyd won it for the hosts.
Campbell completed the full 90 minutes at Rugby Park alongside fellow Magpies' loanee Conor Newton, who has been with the Saints since the summer.
Elsewhere over the New Year period, Newcastle's English-based loanees all ended up on the losing side in their respective games.
In the Championship, Dan Gosling's Blackpool suffered a 2-0 reversal at Sheffield Wednesday. Midfielder Gosling played the full match at Hillsborough but the Tangerines have now gone seven games without a win.
League One saw James Tavernier and Haris Vuckic both handed starts for Rotherham United.
However, Coventry came from behind, scoring three times in the final 20 minutes to run out 3-1 winners at the New York Stadium.
Tavernier played the full match while forward Vuckic was replaced midway through the second half.
In League Two, Michael Richardson's Accrington Stanley suffered a first defeat in four as they went down 3-1 at high-flying Fleetwood Town.
Richardson was involved throughout for Stanley but was unable to inspire his side to victory.
Photograph courtesy of Allan Picken Photography


text: Sometimes other women intimidate me. My husband asked me if I was nervous. I knew I shouldn't be, but I was. I knew it was silly, but I was just being realistic. I was headed to a two-day retreat with a bunch of other youth pastor's wives that I didn't know. I was about to … Continue reading Confession #10


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JOB RESPONSIBILITIES: Physical testing of aggregates, concrete and asphaltic products. Assist RTS team in ongoing research into mix design and testing for concrete, asphalt and aggregates. Perform field QA/QC sampling. Participate in Statistical Process Control (SPC). Provide continuous improvements based on the Company's Corporate Objectives.
EDUCATION: Current student in Civil Engineering, Physical Science, or related major.
SKILLS: Ability to work independently and as a team member. Excellent math and analytical skills; MS office skills for testing and reporting purposes; excellent customer service skills. Some knowledge of aggregates and Asphaltic Concrete helpful.
Must have a valid CA. Driver's License and clean driving record.
Please visit our Careers webpage to learn more about Intern and New Graduate positions www.graniterock.com/careers and to apply online.
Check out our Intern video @ www.graniterock.com/careers


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WASHINGTON — It speaks to the paralysis in the Middle East peace process that the most noteworthy development of the past week came when a mild-mannered analyst at a pro-Israel think tank unfurled three color-coded maps.
The analyst, David Makovsky of the Washington Institute for Near East Policy, wanted to show, in concrete terms, how negotiators could create a new Palestinian state in the West Bank, using the pre-1967 boundaries of Israel as a baseline, while taking into account the roughly 300,000 Jewish settlers who now live there.
The goal, Mr. Makovsky said, is to "demystify" the territorial hurdles that divide Israelis and Palestinians, and to debunk the notion that there is no way to reconcile the Palestinian demand for sovereignty over the West Bank with the Israeli demand for control over a majority of the settlers.
"In my view, it is definitely possible to deal with each other's core demands," he said. "There are land swaps that would offset whatever settlements Israel would retain. The impossible is attainable."
To be sure, Mr. Makovsky's maps are an academic exercise. Direct negotiations between Prime Minister Benjamin Netanyahu and the Palestinian Authority president, Mahmoud Abbas, broke down in September, just three weeks after President Obama inaugurated them at the White House. By all accounts, the two men scarcely confronted borders, let alone other thorny issues, and there is little sign that the impasse will end anytime soon.Continue reading the main story
Still, the maps are getting attention. Mr. Makovsky has given briefings to senior officials in Mr. Netanyahu's government and the Palestinian Authority, as well as to the administration's special envoy, George J. Mitchell, and members of the National Security Council. Mr. Makovsky is also close to Dennis B. Ross, a key Middle East adviser to Mr. Obama. "He has put forward some interesting ideas that could make a valuable contribution to a future agreement," Tommy Vietor, a White House spokesman, said in a statement.
To some seasoned observers, the significance of the maps is less what they show than where they come from.
The Washington Institute was founded in 1985 by scholars affiliated with the American Israel Public Affairs Committee, the influential pro-Israel lobbying group. While the institute has earned a reputation for solid scholarship, and has wholeheartedly supported the peace process, it has remained a staunch supporter of Israel.
Still, this latest effort to prod Mr. Netanyahu and Mr. Abbas back into negotiations resonates with a broader frustration at the impasse among many American Jews.
"There is an increasing trend toward an 'oy vey' angst over how to save the two-state solution from the settlement juggernaut and by extension how to save Israeli democracy," said Daniel Levy, a senior research fellow and co-director of the Middle East Task Force at the New America Foundation.
"The specifics of his proposed one-to-one border swaps are interesting," Mr. Levy continued, "but the subtext reads: 'Yikes! We need a border and an end to the settlement phenomenon now.' "
Mr. Levy, an unabashed liberal, said he was particularly struck by Mr. Makovsky's so-called maximalist map, which shows that Israel could absorb 80 percent of its settler population by swapping land with the Palestinians equivalent to less than 5 percent of the West Bank.
That is less than a previous Israeli prime minister, Ehud Olmert, proposed to Mr. Abbas in 2008, when the two discussed land swaps. Mr. Olmert wanted a swap equivalent to 6.3 percent of the territory Israel seized in the 1967 Arab-Israeli war; Mr. Abbas wanted to transfer only 1.9 percent. Mr. Netanyahu has not taken a position on the swaps. To some right-wing Israelis, any such maps are taboo.
Mathematical tradeoffs are sprinkled throughout these maps, which were a year in the making and draw heavily on statistical data. Mr. Makovsky said he was able to give the Palestinians quality land in exchange for what they would give up, and to ensure that a Palestinian state would not be cut up like a jigsaw puzzle.
To those unfamiliar with the ethnic and religious landscape of the West Bank, the maps can be bewildering — showing Israeli "fingers" jutting into Palestinian territory to absorb major settlements, with offsetting chunks of new Palestinian land, some along the Gaza Strip and the Egyptian border.
Critics point out that Mr. Makovsky's exercise does not take into account other issues, like Israel's security or the fate of Palestinian refugees. Nor does he contend with the status of Jerusalem, which some analysts believe will be the most contentious of all the issues that divide the two sides.
But Mr. Makovsky says that is up to the parties; his job is merely to stimulate their thinking. A former journalist at The Jerusalem Post, he is reluctant to be drawn into partisan arguments.
Of course, nothing in the Middle East can be divorced from politics. On Mr. Makovsky's maps, for example, Israel would not annex Hebron, a mostly Palestinian city that lies deep in the West Bank but has religious significance to both Jews and Muslims. Nor does he have a solution for Kiryat Arba, a settlement next to Hebron that is one of the most militant in the West Bank.
"It's up to the parties to decide what to do with these settlers," he said, conceding that when only 9,000 settlers were uprooted from Gaza after Israel withdrew in 2005, it caused years of political upheaval.
"I'm in the think-tank world to solve problems, not be polemical," he said. "The idea here is to bring the two-state solution down to earth."Continue reading the main story


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We heard plenty from Brett Favre during Sunday night's preseason game between the Vikings and the 49ers. Peter King of SI.com got a few words after the game from coach Brad Childress, and they appear in King's latest edition of Monday Morning Quarterback.
Criticized for the perception that he begged Brett to play again and the subject of a much-discussed report that Favre believes Childress "has no clue" about offense, Childress says he had one motivation: to win.
"There are no sacrosanct rules in this business," Childress told King. "You do what you have to
do to win, and I've got no problem with that. You can't get a hit if
you don't swing the bat."
As to that much-publicized Hattiesburg hunt, with Childress sending three players to Mississippi to track, tag, and bag, the elusive albino lion, King thinks that Favre was going to show up on his own in due course, but that Ryan Longwell, Steve Hutchinson, and Jared Allen were allowed to miss meetings and practice with the goal of accelerating the process.
"The hardest thing we had to do, the hardest thing by far, was getting
him down that long driveway in Hattiesburg," Childress said. "Once we
got him to go down that long driveway, we had him. He was in."
That's the most palatable explanation, far better than the notion that it was staged for dramatic effect or that Favre never would have mustered on his own the desire to return, and that he needed to be cajoled into playing by the simultaneous application of three sets of lips to his rear end.


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Veteran NASCAR driver Mike Harmon denied to reporters that he knew the whereabouts of Jennifer Jo Cobb's racing trailer. However, the story that the 55-year-old Harmon gave to the Rowan County Sheriff's Office was considerably different.
Cobb's hauler which transports the Camping World Truck Series team's racing equipment and vehicles, was stolen May 11. The theft is part of an ongoing dispute between Cobb and former business partner David Novak. Cobb and Novak were romantically involved before they stopped working together.
"I guess her hauler is gone but she does not have no video of me because I was on my way back from Darlington," Harmon said after the theft of Cobb's trailer was reported. "The truth has got to come out. It's not me."'
As Sporting News reports, when Harmon met with the Rowan County Sheriff's Office he indicated he knew where the trailer was and would return it if it was not returned to Cobb according to a probable cause affidavit.
Harmon never delivered the trailer.
The detective in the affidavit says "I called Mr. Harmon and he stated he had changed his mind and was prepared to be arrested if necessary."
According to the affidavit, Carl Long told the detective that he drove Harmon's semi back from Darlington arriving at Harmon's shop at 1:30 a.m. and that Harmon instructed him to leave the keys inside the truck.
Survalience video released from Cobb's shop shows a black semi which matches Harmon's according to the detective.
The hauler and its contents are worth $279,000.
Cobb did not compete in the most recent truck series race at Charlotte Motor Speedway after she noticed a problem with her engine in practice. Harmon finished 36th in the race.


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these are some of the things ive been doin
Thursday, September 18, 2008
It's football season and I've been helping my bud Tommy, work on the coach we use to tailgate. You would think we would do this before the season started,but NOOOO, we do the same thing each year. This time the water holding tank sprung a leak and fixing it snowballed into a 2 week project. The seat over the tank was broken , the electronics under there needed to be moved and rewired . We had to completely replum the water lines. The floor needed reworked. had to run a new electrical outlet to the side etc etc.
these are some of the things ive been doin
these are some of the things ive been doin
Posted by Greg "COOP" Cooper at 11:57 PM
2 comments:
- Ken Mitchroney said...
well, this explains why i have not heard from you since the neolithic period. Wow! I will have a list of repairs for you when you come back to Martinez.
It's okay, I pay out in " In and Out " burgers.
The fink can looks great by the way.
- September 25, 2008 at 1:18 PM
- Greg "COOP" Cooper said...
Woo hoo , I'll be right over NUK NUK
- October 1, 2008 at 8:29 PM
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Today's CVN Engle progeny trading card features one of the leading defense attorneys in Florida's landmark litigation.
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- Has won 9 of 12 verdicts.
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The Castle -
Alpine Luxury Suites
It is immediately apparent to any visitor that our Castle is a house of the highest quality, in which luxury is the highest law. Even on long stays, our guests are sure to feel at home here. Ideal for families or couples that prefer separate sleeping rooms.
An underground tunnel connects the castle with the Hotel Bismarck Main House, so that all of the hotel's facilities are easily accessible without having to go outdoors.
Lavish Apartments
In the Hotel Bismarck Castle, you'll stay in richly appointed apartments and lavish suites (64-66m²) decorated in the traditional Austrian cottage style. These rooms are also ideal for families with children. Additionally, our guests can find two large family apartments (80m²) with your choice of either one or two double rooms with two baths. All of our suites and apartments are situated so that the living rooms and balconies have access to direct sunlight.
For especially refined guests we suggest:
- Our brand-new Alpine Luxury Suite (rooms 101, 103, 201,
203) with two separate bedrooms, lavish
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- Our brand-new Panorama Luxury Suite (room 301, 302 303) on the third floor with luxurious bathrooms, walk-in closet and glorious view from the southwest-facing balcony.
This autumn we rebuilded our Schlössl for
There will be two more "Alpine Luxury Suites" each with 2 separate bedroom / 1 living room / 1 bathroom
as well as another "Panorama Luxury Suite" on the third floor.
Our middle apartments 102 and 202 will each turn into 2 double rooms with a connecting door. These are perfect for families with children or couples who want to have separated bedrooms.
Also our Penthouse Suite will be completely newly renovated and turned into a real Crown Jewel of the house. A lot of special extras will fascinate you: a shower with integrated steam bath, bath tub with beautiful view to the mountains, … and more, so treat yourself to a nice surprise next winter!


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Download Open: Mission Appeal 2014 [ENGLISH] in Scribd.com
ROME, October 15, 2015
To All the Members of the Congregation of the Mission
Dear Confreres,
May the grace of the Lord be with us always.
During this month dedicated to the missions, I extend to each and every one of you my heartfelt greetings.
Before continuing to read this letter, I invite you to reflect on the following words of Saint Vincent:
"Let's give ourselves to God, Messieurs, to go throughout the world to carry His holy Gospel and, wherever He may lead us, let's stand by our post and observe our practices until it's His good pleasure to withdraw us from it. We mustn't let difficulties shake us; the glory of the Eternal Father and the efficacy of the Word and Passion of his Son are at stake. The salvation of nations and our own is so great a good as to deserve to be won at any cost; it doesn't matter whether we die sooner or later, provided we die arms in hand; we'll be all the happier for it and the Company will be no poorer, "for sanguis martyrum semen est Christianorum". For one Missioner who has given his life for the love of God, the goodness of God will raise up many others to do the good he will have left undone (CCD:XI:366).
During the month of October the Church traditionally asks us to pray for missionaries and to reflect on the missionary dimension of the Church. During the same month the Congregation calls upon the confreres to reinforce our international missions and other mission territories that are in need of assistance. At this critical moment in the Church's history, Christians are being persecuted in many different parts of the world. At the same time many significant ecclesial events have occurred: the visit of Pope Francis to Cuba and the United States where he spoke words that filled people with hope, the Synod of Bishops where the delegates are reflecting upon the important theme of the family.
Significant and important events have also occurred in the Congregation: confreres have been involved in Provincial Assemblies that are preparing the way for the General Assembly of 2016 and the process of reconfiguration is moving forward, especially in some of the European Provinces. Many parts of the world are calling upon the Missionaries to meet urgent needs and these cries challenge our apostolic and Vincentian generosity.
In this letter I also want to share with you an important activity that was planned by the General Curia during the year 2015, namely, the First Encounter of Superiors of the International and Provincials Missions. The gathering took place from September 14-18 (Rome). Representatives from the international missions, as well as provincial were present (the only exception was the Solomon Islands). During that time the participants shared their experiences, reflected on the theology of the mission and studied and reviewed the documents, Ratio Missionis, the Statutes of the International Missions and the Practical Guide for the Local superior.
In light of Pope Francis' message for World Mission Day 2015 and in the context of the Encounter of the Superiors of the Missions, I want to share with you three thoughts. I share these thoughts in a special way with those Missionaries who might be motivated to respond to the missionary call this year.
1. In his message the Pope refers to a reality that is very much related to our charism: mission is part of the "grammar" of faith, something essential for those who listen to the voice of the Spirit who whispers "Come" and "Go forth". Those who follow Christ cannot fail to be missionaries, for they know that Jesus "walks with them, speaks to them, breathes with them. They sense Jesus alive with them in the midst of the missionary enterprise" (Evangelii Gaudium, #266). Our Constitutions state clearly that the purpose of the Congregation is to follow Jesus Christ evangelizing the poor. Therefore, a response to the missionary call is not simply the result of some sentimental desire but is part of our charism.
2. In another part of his message the Pope reminds us: there is an urgent need to reaffirm that the central ideal of mission is Jesus Christ, and that this ideal demands the total gift of oneself to the proclamation of the Gospel. On this point there can be no compromise: those who by God's grace accept the mission, are called to live the mission. For them, the proclamation of Christ in the many peripheries of the world becomes their way of following him, one which more than repays them for the many difficulties and sacrifices they make (Message of lope Pape Francis for World Day 2015). Those members of the Congregation who are called to live their missionary vocation in a radical manner must be willing to offer their lives to Jesus Christ and also willing to serve those who live in situations of poverty in the existential and geographical peripheries of our world.
3. Finally, during the gathering of the superiors of our missions I was touched by the fact that the Missionaries are able to adapt to the reality of the place where they have been sent and where they are ministering. They are able to do this despite the fact that many of those said realities have not previously been part of their own experience. At play here is not only the capacity to adapt to another situation but also an openness that enables one to respect and to value the culture of another (an openness that is revealed by a willingness to learn the local language and to assimilate the many other aspects of that new reality). I have seen this with my own eyes as I
have visited various missions. In this regard the Pope states in his message: Today, the Church's mission is faced by the challenge of meeting the needs of all people to return to their roots and to protect the values of their respective cultures (Message of lope Pape Francis for World Day 2015).
In the next part of my letter I want to present to you and remind you about some points that I have stated in previous years, matters that we must be very clear about: the reality and the needs of the missions, the criteria for the selection of the Missionaries, the steps that should be followed by the Missionaries wishing to volunteer for the missions and the manner in which you can contribute to the Vincentian Solidarity Fund.
This year it is the very Superiors of the International Mission who will speak and extend to you an invitation to engage in the missionary endeavor. Taking advantage of their presence in Rome during the gathering in September and with the assistance of the Communications Office, each one of these Missionaries has extended an invitation which you can view at (www.gospel- joy.org). There you will also find further information with regard to the theme, the various reports and documents relative to that meeting.
- Angola – (2012). Local language and Portuguese. Parish ministry, accompany local communities and the Vincentian Family. Missionaries presently ministering there: Marcos Gumieiro, José Ramirez Martinez, and Jason Christian Soto Herrera. They request the presence of more Missionaries in order to expand their missionary service.
- Benin – (2013). Local language and French. Pastoral ministry, assisting the Sisters of the Miraculous Medal and supporting the Vincentian Family. Missionaries presently ministering there: Stanislav Deszez and Rafael Brukarczyk. A Missionary is preparing himself to go there but there is need for more Missionaries. This mission is supported primarily by the Province of Poland.
- Chad – (2012). Supported by COVIAM (the Conference of Visitors of Africa and Madagascar). Local language and French. Parish ministry and teaching in schools. Missionaries presently ministering there: Roch Alexander Ramilijaona, Onyeachi Sunday Ugwu, and Esigbemi Ambrose Umetietie. Very soon one of the Missionaries will be leaving and another Missionary arriving in order to serve the diocese. More support is needed.
- Tunisia – (2011). Local languages, French and English. Parish ministry, charitable assistance through CARITAS and ministry with the Diocesan Office of Education. Missionaries presently ministering there: Firmin Mola Mbalo and Narcisse Djerambete Yotobumbeti. These Missionaries would like another confrere to join with them in their ministry.
New invitation to participate in this mission: in a recent letter the Bishop of Tunis told me: The members of the parish in Sousse are retired people, expatriates, many tourists, and mixed families (Christian and Muslim). It would be important to minister in this parish in order to continue to build bridges and dialogue with the Muslim community on behalf of serving the poor in this country. The Church in Tunis is positioned on the peripheries: it is not self-sufficient and there is no seminary to train future priests. The priests have to come from outside the country. The faithful in Tunis are composed of about thirty thousand foreigners, university students, and expatriates. The majority of the population is Muslim and as a result there is always an environment of suspicion and indifference and prejudice;workers, all of which we attempt to change through the witness of our life.
- Bolivia, El Alto – (1994). Local language and Spanish. Pastoral ministry with the indigenous peoples which implies rural pastoral ministry and the formation of the laity. Missionaries presently ministering there: Aidan Rooney, Cyrille de la Barre De Nanteuil and Diego José Pla Aranda. They request the assistance of one more Missionary.
- Bolivia, Cochabamba – (2009). Local language and Spanish. Pastoral and parish ministry. The Missionaries presently ministering there: David Paniagua, Jorge Manrique Castro, and Luis Miguel Montaño Flores.
New Invitations:
Missionaries are needed for a new mission in Beni, Bolivia. Sister Gerarda, DC, who has ministered in Bolivia for more than fifty years, wrote me a letter and stated: We are about to conclude three popular missions that were given by the Vincentian Missionaries and together with our bishop, we are extremely grateful and satisfied with their ministry. It is hoped that these missions will bring about even greater happenings. Two of the four Missionaries stated that these were the most difficult popular missions that they had engaged in during the two years that they have ministered together as an itinerant missionary team. The people were very happy to receive the Missionaries and are praying that they will return. The people bade farewell to the Missionaries with tears. The Daughters of Charity, who minister here as a team, are at a critical moment…
I responded to Sister's letter and told her that the confreres are promoting this mission in Beni and that I also support the idea of a shared mission, among all the branches of the Vincentian Family, especially during this Year of Collaboration in the Vincentian Family.
Territorial Prelature of Tefé Brazil: The Prelature of Tefé, located in the heart of the Brazilian Amazon, had been entrusted to the Spiritans. Bishop Fernando Barbosa dos Santos, CM, is the fourth bishop of Tefé and the first one who is not a member of the Congregation of the Holy Spirit. The Prelature extends for some 264,669 square kilometers and has a population of about 237,782 inhabitants. There are 10 cities, about 405 riverside communities and 40 urban communities. There are 88 Indigenous communities in 15 different villages with a total population of 8,500 persons. It is a poor missionary territory that needs more priests and evangelizers. The primary means of transportation is by boats (long hours of travel in precarious and difficult conditions). The language is Portuguese and the local language.
Chile, Punta Arenas – Tierra del Fuego (2013). The official language is Spanish. This mission would involve pastoral ministry in Punto Arenas. At the beginning of the present year we accepted the administration of a parish in Tierra del Fuego. The Missionaries presently ministering there are: Ángel Ignacio Garrido Santiago, Pablo Alexis Vargas Ruiz, and Margarito Martínez González. They request the support of one more Missionary.
Anchorage, Alaska – (2014). Languages: English and Spanish. The confreres ministering there are serving an increasing number of Latinos who live in the Diocese of Anchorage. The Missionaries presently ministering there are: Andrew Bellisario, Arnoldo Hernández Rodríguez, Pedro Nel Delgado and they are awaiting the arrival of Jesús Gabriel Medina Claros.
- Solomon Islands – (1993). Languages: English and Pidgin. The conferes are involved in formation ministry of candidates for the Congregation (major seminary) and rural parish ministry. The Missionaries presently ministering there are: José Manjaly, Raúl Eduardo Castro, Gregory Cooney, Varghese Ayyampilly, and Antony Punnoth. In 2016 Ricardo Padrhan and Paulus Suparmono will begin ministry there. There is always a need for Missionaries to teach philosophy and/or theology.
- Papua New Guinea – (2003). Language: English and Pidgin. The mission there includes ministry in various dioceses. In addition to formation ministry in the seminary at Port Moresby, the confreres are involved in rural parish ministry and also work at the center of missionary cultural formation. The Missionaries presently ministering there are: Homero Marín, Marceliano Oabel, Emmanuel Lapaz, Jacek Tendej, Neil Lams and Joel Bernardo Yco. Justin Eke continues his ongoing formation.
In addition to the needs of the International Missions, we also need Missionaries who are willing to support our missionary provinces:
- Province of China: The Province of China continues to provide for the pastoral needs of the people of Taiwan and mainland China. The language is Mandarin and the confreres are involved in a variety of ministries.
- Vice Province of Sts. Cyril & Methodius: This Vice-Province has evolved with the support of missionaries from outside the province. The number of native missionaries is increasing. Charitable works have been established there in order to provide for the material and pastoral needs of the poor. With ministry in Ukraine, Russia and Belarus the Vice-Province is willing to accept confreres who are willing to minister there (either with long-term or short-term commitments). At the present time there are missionaries from Africa, India and in a short time a Missionary from Vietnam will arrive there. The language is Russian.
- Province of Cuba: Even though this Province is searching for a way to reconfigure itself with another province, there is a lack of personnel in that province. The language is Spanish and confreres will be welcomed to minister there (either with long-term or shorn-term commitments).
- Vice Province of Mozambique: The members of the Vice-Province have been involved in ministry with men and women who are in great need, in formation ministry in seminaries and in a mission in South Africa. The Vice-Province is still dependent on Missionaries from other provinces in order to continue its ministry. They are most willing to accept new members. The language is Portuguese and the local language. There are 29 ministry positions but only 17 Missionaries.
Since I began this letter with a reflection on the words of Saint Vincent when he spoke about the missionary spirit, I would like to conclude this letter with some other words of our Founder referring to two Confreres, Nicolas Duperroy and Guillaume Desdames, missionaries in Poland:
What have they suffered in that country? Famine? It's there. The plague? Both of them have had it, and one of them has had it twice. War? They're in the midst of armies and have passed through the hands of enemy soldiers. In short, God has tried them by every form of scourge. And here we are, like homebodies without courage or zeal! We'll see others risking their lives in dangers for the service of God, and we'll behave like cowards! What a pity! What weakness! (CCD: XI: 365).
Your brother in St. Vincent,
G. Gregory Gay, C.M.
Superior General


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Losing weight can be tough. Unfortunatley, there's no quick-fix. No magic pill. And no one can do it for us.
It all boils down to one simple equation: Calories in vs. Calories out.
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A new client told me today that she wants to only do aerobics because she wants to get toned and lose weight, FAST! I hear this SO often, and ladies, it's a MYTH!
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How should I spend training? What should my weekly routine look?
How much time you spend really comes down to how long you've been on a regular program and what your goals are. For example if it's been six months since you've worked out, you'll need to take it slowly. Start with a walking program of 3-5 days per week for 10-20 minutes, building up to 30 minutes, 5 times per week. After that, bump it up to 200 minutes per week.
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Does this sound familiar?
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'Tis The Season: Authors Talk Holidays with Jennifer Maschari
'Tis The Season: Authors Talk Holidays is a special seasonal feature on Pop! Goes The Reader in which some of my favourite authors help me to celebrate the spirit of the season and spread a little holiday cheer. So, pour yourself a cup of hot chocolate and snuggle in by the fireside as they answer the question: "What does the holiday season mean to you?" You can find a complete list of the participants and their scheduled guest post dates Here!
About Jennifer Maschari
Jennifer Maschari is a former classroom teacher who writes about kids who do brave things. Her novel for middle grade readers, The Remarkable Journey Of Charlie Price, is available in early 2016. Other things she finds remarkable include fried pickles, exclamation points, and Youtube videos of animals holding hands. She lives with her husband and two stinky bulldogs in Ohio.
Our Christmas is full of traditions, old and new.
The annual Christmas cheeseball with all of its olive-y, cream cheese-y goodness. Driving through the lights display at Sharon Woods. A white elephant gift exchange (from which I've received a fine fake rooster). Eating cinnamon rolls in our pajamas and slippers on Christmas morning.
But Christmas has never quite felt like Christmas without one thing:
Mechanical Santa!
Every year when we were younger, my sister, brother and I would rush into our grandma's house to her fireplace where this little Santa lived. We'd unzip his miniature jacket, hit the switch and suddenly, tinny sounds of Silent Night and Santa Claus is Coming to Town would burst forth out of the speakers on the top of Santa's head, hidden by his fuzzy hat. Santa would shuffle his feet forward and ring his bell. We'd open presents and sneak M&Ms from a little dish on Grandma's coffee table, all under the watchful eye of mechanical Santa.
As things go, my grandma eventually moved someplace smaller, and Santa was packed away in storage in his worn cardboard box.
Then, a year ago, I got the opportunity to bring Santa to my own home. I took him out of his box, unzipped his little jacket, and replaced his battery. He stared at me, ready to play his music. I hit the switch like so many years ago.
Santa was silent.
My face fell. I tried again. Nothing.
My husband took Santa out to his workbench, adjusting this and fixing that. He brought Santa back in and set him on the floor. He flipped the switch. Santa shuffled his feet. He rang his bell. The music played.
I was brought back to those Christmases so many years ago. So many things have changed since then. It's nice to have one thing that hasn't.
Mechanical Santa runs on batteries, sure. But maybe, also, a little bit of Christmas magic.
Title The Remarkable Journey Of Charlie Price
Author Jennifer Maschari
Pages 304 Pages
Target Audience & Genre Middle Grade, Contemporary
To Be Published February 23rd, 2016 by Balzer + Bray
Find It On Goodreads ● Amazon.com ● Chapters
Ever since twelve-year-old Charlie Price's mom died, he feels like his world has been split into two parts. Before included stargazing and Mathletes and Saturday scavenger hunts with his family. After means a dad who's completely checked out, comically bad dinners, and grief group that's anything but helpful. It seems like losing Mom meant losing everything else he loved, too.
Just when Charlie thinks things can't get any worse, his sister, Imogen, starts acting erratically — missing school and making up lies about their mother. But everything changes when one day he follows her down a secret passageway in the middle of her bedroom and sees for himself.
Imogen has found a parallel world where Mom is alive!
There's hot cocoa and Scrabble and scavenger hunts again and everything is perfect…at first. But something doesn't feel right. Whenever Charlie returns to the real world, things are different, and not in a good way. And Imogen wants to spend more and more time on the other side. It's almost as if she wants to leave the real world for good. If Charlie doesn't uncover the truth, he could lose himself, the true memory of their mother, and Imogen…forever.
2 responses to "'Tis The Season: Authors Talk Holidays with Jennifer Maschari"
Mechanical Santa sounds magical indeed! That's a tradition that could stick around for years 🙂
Alexa S. recently posted…Books in Real Life, Episode 10: Bunheads
I love old Mechanical Santa! It's amazing how something like this can carry so many memories. We had a Santa with a big red nose you pushed and it played music. It was pretty sad when it fell off the door and broke. But remembering it still puts me right back in my childhood home.
Thank you for sharing your story!
Janet Johnson recently posted…Top Five: Reasons for Lists


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Gloucester County Sheriff's Office
In the 8100 block of Roaring Speings Road
On Aug. 28 a report was received of an unauthorized use of a vehicle, taken from a residence in the 8100 block of Roaring Springs Road on Aug. 27. A 2005 Toyota tundra, Virginia tag XFE1732 was taken without the owner's permission. Warrant pending service, vehicle has not been located.
In the 7500 block of Hospital Drive
On Aug. 28 a report was received of a domestic assault, which occurred in the 8000 block of Cook Drive between adult father and son. Male transported to Walter Reed Riverside Hospital where a Physician's assistant was also...


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The Writings of Bertell Ollman
to the Editor
|Class Struggle
Board Game
Dance of the Dialectic: Steps in Marx's Method
Reviews of Ollman's Books
Featured article -
America Beyond Capitalism: A Socialist Stew Prepared for Liberals and Conservatives
Featured speech -
McCoy Award Acceptance Speech
Video: Marxism and Progress
Marxism (the cartoon version)
From Theory to Practice
Radical Jokes
Recommended Web Sites
NYU Course Bibliographies
ETF Site
Not To Dare
Butcher Shop
Kiki & Bubu explain the neoliberal shift in labor relations
Of Marxism and Universities
By Bertell Ollman
The role of Marxism in universities is only slightly less obscure than the role of universities in Marxism, but perhaps some light can be shed on both subjects by examining Marx's little known response to an ancient Roman myth.
Cacus was a Roman mythological figure who stole oxen by dragging them backwards into his den so that the footprints made it appear they had gone out from there. After quoting Luther's account of the story, Marx exclaims, "an excellent picture, it fits the capitalist in general, who pretends that what he has taken from others and brought into his den emanates from him, and by causing it to go backwards, he gives it the semblance of having come from his den."
Capitalists present themselves as producers of wealth, providers of jobs, donors and public benefactors. The press (their press) usually refers to them as "industry." Is this an accurate description of who they are and what they do? What stands out clearly from the example of Cacus is that what Marx and Marxists call bourgeois ideology does not so much falsify the facts as misinterpret them so as to reverse what has taken place: The footprints are there for all to see, but if we limit ourselves to what is immediately apparent (the subject matter of "empirical" social science) we will arrive at a conclusion that is the exact opposite of the truth.
Only if we examine what led up to the event in question and its surrounding circumstancesthat is, its real history and the system of events in which it residescan we hope to understand what really happened and why.
In the case of the capitalists, only by examining how they got their wealth from the surplus labor of previous generations of workers (history) and how our laws, customs and culture are biased in their favor (structure) can we see it is not the capitalists who are serving society but the rest of society that is serving them.
Though many have criticized Marxism as one-sided because of its emphasis on economic processes, Marxism is really our only all-sided analysis of capitalism as a social system, including the perspective provided by this analysis, the different events disconnected and arbitrary, and often acquire a meaning that is the exact opposite of how these events function inside of capitalism.
Yet some people continue to ask: Should Marxism be taught in a university? If we let our eyes wander away from the footprints left by Cacus's oxen, then we can see the correct question: Does an educational institution that does not teach Marxism deserve to be called a university?
Serious non-Marxist scholars in every field appreciate the contribution Marxism makes in posing the bigger questions, at least. And enough have come to accept its holistic explanations to make Marxism the major alternative to orthodox approaches in history and economics (political science is soon to follow).
And the capitalists, and those Marx called their "ideological handmaidens," who are protesting that teaching Marxism constitutes "indoctrination"...? Well, Cacus, too, had an interest in keeping people from finding out what went on in his cave.
|Copyright © Bertell Ollman 2004-2019. All rights reserved.


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Well, here you are. You happened upon the website of Kevin McCreary. Congratulations! You really did it. I think, if you look down there, you're going to see a few pictures or whatever. Those are the projects that Kevin has going on these days! They're pretty great, I PROMISE! But don't take MY word for it, check out these amazing quotes:
"They're pretty great, I PROMISE"
-The Guy Who Made This Website
"Sure I can throw something together real quick"
-The Guy Who Made This Website
"What do you mean, you can't pay me?
Fine, I'm not going to bother getting
those amazing quotes you asked for."
-The Guy Who Made This Website
Who can argue with that? So check out those links and spend the afternoon binge watching all of those videos. Then, if you want, share them with your friends! Kevin will love you for it....but don't expect him to pay you, because he won't. He never does.
Join Kevin for a movie night, in his ongoing quest to hold Christian Films up to the same standard as other movies
Obviously, Kevin loves movies, especially the behind-the-scenes aspect. Join him as he shares what he's discovered along the way!
Watch everything else! From politics, social commentary, and thoughts on media. Say Goodnight Kevin is the show where Kevin shares it all. It's fun, funny, and will keep you entertained for hours....or at least a few minutes!


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Seminar on Algerian literature by Prof. AOYAGI April 24th 2015
The Embassy of Algeria to Tokyo, Japan | 24 April 2015
Prof. Aoyagi made, on April 24th 2015 at the embassy, a lecture to the members of Algeria-Japan association about the literature of the Algerian writer Mouloud Ferouan, in particular, his book le fils du pauvre which she is preparing its Japanese translation. The lecture was followed by discussion about Algerian literature.
HEMr. Mohamed El Amine BENCHERIF expressed thanks to prof. Aoyagi to make Algerian literature known to Japanese readers affirming that her works will certainly contribute to introduce an important part of Algerian culture to Japanese public. The lecture was followed by a reception hosted by the Ambassador.


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Christmas Program
Christmas Sponsorship 2018
Click here if you would like to sponsor a low-income family this Christmas
Since 1995, LIFE*SPIN's Christmas Sponsorship Program has provided additional support to poor families in our community. This program relieves poverty for low-income families with children over the Christmas season. LIFE*SPIN matches low-income families with those wishing to help a disadvantaged family celebrate Christmas.
This program is critical for the young children who are impacted by the loss of income that leaves their parents struggling to provide even the most basic needs of shelter and food.
Donations from our community help to alleviate the pain and desperation experienced by those continuing to live in poverty and also the newly poor resulting from the dramatic downturn in the economies of the world and an aging work-force.Your support is deeply appreciated.
Volunteers accept Christmas "wish lists" from the recipient family and coordinate a match with a suitable sponsor.Sponsors provides a gift for each child in the family. We ask that gifts requested be limited to roughly $35-$40, so we can assist more children. Some sponsors provide a grocery store gift card for the family to shop together for the Christmas meal or a small gift for the parents, although these are at your discretion.
If you would like to receive a Charitable Tax receipt for the value of your purchases, please retain all your receipts and mail them to us by December 30th to P.O. Box 2801, London, ON N6A 4H4. This will ensure they don't get misplaced when you drop off your gifts. Please note that we cannot issue the receipt for the HST portion of your donations in-kind.
While this is a little dated... you can watch a video clip from 2011 by clicking here.
Christmas Lights & Sirens ~ Toy Drive
London Emergency Services welcomes donations of toys on Saturday, December 8 from Noon to 3:00 at Fire Station #1 on Horton Street. Help us fill the EMS vehicles and visit "Fire Santa"
All toys collected will support LIFE*SPIN's Christmas Family Sponsorship Program that ensures that local children, living in low-income families, each receive a new gift this holiday season. Last year, LIFE*SPIN assisted more than 1,100 families at Christmas and it is expected that this number will increase this year.
Help us make this a happy holiday season for those less fortunate while supporting local farmers and businesses. If you are unable to attend, please consider sponsoring a family of your own. You can do so by completing this form.


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Saving Has Never Been This Easy For Car Insurance Customers
- link: http://cheapcarinsurance.uk.com/
- Listed: January 23, 2015 4:47 am
- Expires: This ad has expired
Auto insurance, in its simplest forms, seeks to protect the buyer from liability and loss during an auto accident. Coverage can be expanded to provide a replacement vehicle, cover medical costs, provide roadside car insurance service and protect against uninsured motorists. There are other coverages available as well. This article seeks to help you understand the nature of insurance and help you decipher which coverages are best for you.
The year and options on your vehicle will play a large roll in how much your insurance premium is. Having a newer car with lots of safety features can save you some, but if you have it financed you will also have to pay more for full car insurance coverage.
Asking for an age discount can help save a bundle on car insurance for older drivers. If you have a clean driving record, insurance companies are likely to offer you better rates as you age. Drivers between the ages of 55 and 70 are most likely to be eligible for such discounts.
To get the most for your money when pricing automobile insurance, be sure to consider the extras that are included with some policies. You can save on towing costs with emergency roadside assistance that is included by some insurers. Others may offer discounts for good drivers or including more than one car on your policy.
If you are considering signing up with another insurance provider, you should allow your policies to overlap rather than going a single day without any coverage. There's never a time when you want to have no car insurance, even if it's for a very short period of time.
When dealing with car insurance, you need to know what type of coverage you think you need. You can just get the bare minimum, and pay a lower out of pocket cost, though beware, if you are in a bad accident, you will end up paying more for it. You can go onto any car insurance website and see what they offer, and pick and choose what you think you will be needing.
If you are serious about saving money on automobile insurance, think about downsizing the number of vehicles you use. If your family can get by on only one car, you will save a substantial amount of money. Not only will you pay less for your insurance policy, but you'll also see savings in your monthly car maintenance bills.
Take a driving course to get a discount on auto insurance. Young drivers who have taken a driver's education course are usually eligible for discounted auto insurance rates. Similarly, older drivers who take a defensive driving course are also eligible for discounts on auto insurance. Check with your insurance company to see if they offer these discounts.
One thing that many people overlook when reviewinging car insurance is that they may not be covered for other "uninsured" drivers. Consider the fact that it does increase your premium. This is also the insurance that reimburses you should a driver without insurance be at fault for an accident you're involved in.
So, after reading and applying the helpful tips listed above, you should feel a bit more at ease in the world of auto insurance plans. You have the tools, and you need to use them. You should feel confident and ready to begin looking for auto insurance plans to better protect your vehicle.
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Knoxville,Tennessee (1983 – )
Floating down a lazy river of beautiful women and acid encrusted dreams, Olga Noes was hatched as the godchild of filth in a sweeping east Tennessee town located on the outskirts of mischief. Her most prolific hobbies include routine beer drinking, solo dance parties and the ritual of dousing the divine with the atrocious, using pencils and paintbrushes as the viscosity. Currently, she resides on the sullied streets of southern Germany.


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Superfeet aim to create affordable orthotics for the general public.
They deliver a premium insole with a shape and design based on the proven biomechanical principles of podiatric medicine.
Superfeet insoles are made of the highest quality materials, providing customers with superior comfort and stability at an affordable price.
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text: Industrial facilities rely on a clean and sturdy infrastructure to conduct business. Has your industrial property recently fallen victim to flood, fire, or mold damage? Then it's essential to seek restoration services as soon as possible. For an industrial restoration team with top-notch industry knowledge, hands-on experience, and proven tactics, look no further than Restore Force Inc. Call us to schedule an appointment today.


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About Cambia
Cambia by NFP® is a venture formed by Northland Forest Products to manufacture and market thermally modified (TM) lumber as an environmentally responsible choice over tropical hardwoods or petrochemical-based wood alternatives.
The wood that Cambia thermally modifies is manufactured in the United States by local sawmills with logs harvested from sustainably-managed forests and is available with FSC certification.
NFP's lumber concentration yards are located in Kingston, New Hampshire and Troy, Virginia.


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For the Pesdjentiu (/pes-jen-ti-oo) yesterday (Saturday 11/18/2017), I started putting together an observance of the New Moon Feast and a self-initiation as a shemsu-Anpu and a hem-Netjer. A new moon marked the beginning of a new month (LaBorde, Sharon, Circle of the Sun: Rites and Celebrations For Egyptian pagans and Kemetics. 2012). Now according to the Kemetic Orthodox Festival Calendar, yesterday was IV Akhet 16, Regnal Year 25. In The Ancient Egyptian Prayerbook, by Tamara L. Siuda, IV Akhet 16 is the first of twelve-day festival of the Mysteries of Sokar-Wesir at Abdju.
The following is a combination of the Pesdjentiu (New Moon Feast), the Self-Initiation Rite, and Long Iru found in Circle of the Sun: Rites and Celebrations For Egyptian Pagans and Kemetics by Sharon LaBorde.
Lighting your shrine Candle, say:
Come in peace, bright Eye of Horus, come in peace. Receive the light. The Eye of Horus shines, like Ra in the twin Horizons, and evil hides before it. Receive the light. The Eye of Horus comes, and I am purified with it. Receive the light.
Invocation to the Gods, say:
Hail to You, Great God! I have come to You, my Lord, that I may see Your beauty, for I know You and I know Your name. Behold, I have co e before You, oh Anubis, I bring you ma'at and I repel isfet for You.
Pesdjentiu Invocation, say:
Open, Clouds! The dimmed Eye of Ra is covered, and Horus proceeds daily in joy, Great of Shape and Weighty of Power, to dispel dimness of the eye with His fiery breath. Behold, Oh Ra, I have come sailing, for I am one of those four gods at the corners of the sky, and I show you He who is present by day. You rigging fast, for there is no opposition to You.
Anointment of Head and Hands with Water for Purification, say:
I have cast out all evil that is within me. I am pure, I am pure, I am pure, I am pure.
Offering rite and Eating Bread, say:
(Water, pour water into water cup, say:)
Take these, Your cool waters that are the Inundation.
(Milk, pour milk into the milk cup, say:)
Milk – may you taste it in Your shrine.
(Food, place your food offering on its dish; hold up the dish before the icon statue, say:)
Take this, Your bread/food offering on which gods live.
(Eating Bread/the offering, say:)
Of what shall I eat? I shall eat the bread which Renenutet provides for me in the shrine of the gods. I shall eat of the bread and partake of truth [ma'at]. (Eat the bread and drink the milk, but save the water for the final step.)
Consecration, (anoint your forehead with oil; if you will be using a piece of jewelry or vestment to ritually signify your initiation, place it on at this time), say:
Hail to You, oh Anubis, in all Your names and in all Your places! Grant this pure office of Hem-Netjer upon me. May You make me it beneficial to me, and may You protect me from evil. Receive me and place Your arms over me, for I am Your shemsu (if using a Kemetic name, use it here) Anpuhotep upon this earth. May I be beneficial to You and to humankind, as You are beneficial to me.
Closing the observation of the New Moon and Self-initiation, drink the water from the offerings to signal the end of the rite, say:
In-un-ma'a [Truly it is].


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- Research article
- Open Access
Conservation of intron and intein insertion sites: implications for life histories of parasitic genetic elements
© Swithers et al; licensee BioMed Central Ltd. 2009
- Received: 19 December 2008
- Accepted: 31 December 2009
- Published: 31 December 2009
Inteins and introns are genetic elements that are removed from proteins and RNA after translation or transcription, respectively. Previous studies have suggested that these genetic elements are found in conserved parts of the host protein. To our knowledge this type of analysis has not been done for group II introns residing within a gene. Here we provide quantitative statistical support from an analyses of proteins that host inteins, group I introns, group II introns and spliceosomal introns across all three domains of life.
To determine whether or not inteins, group I, group II, and spliceosomal introns are found preferentially in conserved regions of their respective host protein, conservation profiles were generated and intein and intron positions were mapped to the profiles. Fisher's combined probability test was used to determine the significance of the distribution of insertion sites across the conservation profile for each protein. For a subset of studied proteins, the conservation profile and insertion positions were mapped to protein structures to determine if the insertion sites correlate to regions of functional activity. All inteins and most group I introns were found to be preferentially located within conserved regions; in contrast, a bacterial intein-like protein, group II and spliceosomal introns did not show a preference for conserved sites.
These findings demonstrate that inteins and group I introns are found preferentially in conserved regions of their respective host proteins. Homing endonucleases are often located within inteins and group I introns and these may facilitate mobility to conserved regions. Insertion at these conserved positions decreases the chance of elimination, and slows deletion of the elements, since removal of the elements has to be precise as not to disrupt the function of the protein. Furthermore, functional constrains on the targeted site make it more difficult for hosts to evolve immunity to the homing endonuclease. Therefore, these elements will better survive and propagate as molecular parasites in conserved sites. In contrast, spliceosomal introns and group II introns do not show significant preference for conserved sites and appear to have adopted a different strategy to evade loss.
- Insertion Site
- Host Protein
- Conservation Score
- NADH Dehydrogenase Subunit
- Homing Endonuclease
Inteins are intervening polypeptide sequences that are translated as part of a protein [1–4], and are removed in the maturation of the final protein product. Some inteins contain a homing endonuclease [5] that has a large specific recognition site (12-40 base pairs). The intein-encoding DNA is inserted in frame within a host gene; after translation the intein catalyzes its own excision resulting in removal of the intein (internal protein) and splicing of the extein (external protein, the mature active host protein) (see [6] for detailed review) [2, 7, 8]. Comparative analyses have shown that all inteins are homologs; however, their sequences are so divergent that phylogenetic analyses of inteins inserted into different host proteins remains largely unresolved [4, 9]. Inteins that are found in different insertion sites of the same host protein are not necessarily closely related to each other, and often highly divergent. However, inteins inserted into the same site in orthologous proteins are closely related to each other and share a common ancestor, but their molecular phylogeny does not always reflect the history of the host protein or of the host organism [4, 10, 11], indicating transfer of the intein between divergent hosts.
Introns are defined as non-coding regions of a gene that are excised during post-transcriptional processing. Since their discovery in 1977 [12] three major groups of introns have been identified: group I, group II, and spliceosomal introns. Group I and group II introns have distinct structures that facilitate their self-splicing activity (see [13, 14] for detailed review), and they often encode an open reading frame (ORF) or contain an internal ORF [15–17]. The internal ORF of the group I introns encodes a homing endonuclease and the ORF of the group II introns encodes proteins with one to four of the following functionally defined domains: reverse transcriptase (RT), maturase, DNA-binding protein, and endonuclease [17, 18]. These proteins serve two functions for the intron: assisting in splicing and folding, and allowing the intron to act as a mobile element and invade intron-free alleles via retrohoming or retrotransposition [13, 17–19].
Endonucleases [4] provide mobility to some inteins and introns, through a process called "homing" [15, 16, 20]. These endonucleases are known as homing endonucleases (HE). The HEs initiate homing by cleaving the HE free allele. Similar to traditional restriction endonucleases, the HE makes a double strand break, and the HE containing element is copied during repair into the intein/intron free allele [15]. The HE recognition site is accessible in the host gene when flanking regions of the intron or intein integration sites are joined. Presence of the HE containing element makes the allele resistant to HE digestion. Free-standing HE genes also function as molecular parasites/symbionts [21–24], and can provide mobility for neighbouring HE-less group I introns through a collaborative homing mechanism [23].
Previous work found inteins and group I introns in conserved parts of their host genes [4, 25–27], while group II introns were shown not to target conserved genes [28, 29]. Here we provide quantitative support and extend previous analyses to other proteins that host inteins, group I, group II and spliceosomal introns across all three domains of life.
Where are inteins and spliceosomal introns located in their host sequence and structure?
Our earlier analyses of three host proteins, ATPase catalytic subunit, replication factor C (RFC), and cell division control protein 21 (CDC21), confirmed the notion that inteins appear at highly conserved sites within their host proteins [4]. Since publication of [4] the number of inteins discovered in these three proteins has increased substantially, including some found in new insertion sites.
In addition, we analyzed all proteins that were reported in InBase [30] as containing an intein for conservation of the insertion site. With one exception all of these additional 30 proteins harbor their inteins in conserved regions of the respective host protein. The fillamentous hemmagglutinin protein was annotated by InBase as being an allele for an intein; however, this element has been shown to be a bacterial intein-like protein domain (BIL) [32]. BILs are found in non-conserved regions of hypervariable proteins and our analysis supports this notion as the BIL was not found to be in a conserved site. (See additional file 1 for profiles and p-values.) Using Fisher's combined probability method to calculate the overall significance level for inteins (omitting the BIL) inserting into conserved sites is p < 0.0001 (in calculating the combined probability, p-values for individual proteins p < 0.01 were considered as equal to 0.01).
Where are group I and group II introns located in their host sequence and structure?
The cytochrome C oxidase subunit I (cox1) gene found in the mitochondria of eukaryotes accommodates self-splicing group I and group II introns in more than forty different sites (See additional file 2 for intron insertion sites.). The number of introns found in different species varies. The cox1 gene of Podospora anserine, an ascomycete fungus, is more than 24 kb long and harbours 14 group I and two group II introns [34]. In Saccharomyces cerevisiae different strains host a total of six group I and three group II introns [18, 35, 36]. Similar to inteins, group I introns appear to target more conserved sites (p = 0.0003), and similar to spliceosomal introns, group II introns were found not to have a significant preference for conserved sites (p = 0.4176).
A linear regression analysis was performed to determine if there is a correlation between the number of fungal species that harbour the introns in a particular site and the conservation of the intron insertion site. A weak negative correlation between site conservation and number of species with group I introns is found at each site (p = 0.0609), while there is no correlation between site conservation and number of species with group II introns at each site (p = 0.2907). These findings suggest that for fungi, either group I introns tend to target conserved DNA sites more frequently, or group I introns survive in conserved sites for longer periods of time, while group II intron targeting is not strongly influenced by site conservation.
An additional eight proteins which host group I and group II introns were analyzed. These represent all protein families reported in the comparative RNA web site database [37] as containing group I and group II introns. These proteins followed a similar pattern where the group I introns showed a strong preference for conserved sites and the group II introns did not. Two exceptions were Chlorophyll alpha apoprotein A2 and NADH dehydrogenase subunit 3 where neither intron type showed a preference for conserved sites (See additional file 1 for p-values and profiles.).
The combined significance levels for group I and group II introns targeting conserved sites is p < 0.0001 and p = 0.49, respectively.
Introns have played a role in gene and genome evolution [40]; most or all of them may be later invaders of the genes in which they are currently located [41]. Interesting questions remain regarding the origin and evolution of introns, including: (1) How often did they arise? (2) How are they transmitted between divergent species? (3) How has their activity been maintained by natural selection? [16, 17, 20, 42–44].
Regardless of their origin, many of the extant introns and inteins, especially the ones utilizing homing for survival, can be considered molecular parasites [20, 45, 46]. Some of these molecular parasites have acquired additional roles that are adaptive to the host [20, 45, 46]. While we discuss HE genes and self-splicing elements as molecular parasites that have their own life-cycles, this does not negate the fact that many of these elements adapted to play a beneficial or at least a necessary role for their host [47], and these roles may play a role in the long term survival of these elements [20].
In accordance with other studies [4, 25, 26], we find that inteins and group I introns are found in conserved regions of their respective host proteins. This same trend was also seen for 37 other protein families that host inteins and group I introns (see additional file 1). We also find that group I introns within highly conserved positions are more likely to be found in a broader range of species of fungi. Group II introns, BILs and spliceosomal introns do not reveal a significant bias toward conserved sites.
The splicing elements utilize a small portion of the flanking extein/exon region for splicing. Inteins are inserted before one of the following amino acids in the host protein: C, S, or T [6]. The amino acid is required to complete the self-splicing reaction. For the group I intron an internal guide sequence binds to 6 - 12 nucleotides of the exon. This internal guide sequence is not conserved between different group I introns and exact base pairing between the guide sequence and exon is not required [48]. Similarly, in the case of the group II introns two exon binding sites of the intron interact with six or more nucleotides each at the 5' and 3' flanking exon [13, 49]. For both group I and group II introns this base pairing between intron internal sequence and the flanking exon restricts the possible sites in which an individual intron can retain its splicing activity. However, this does not restrict the locations of the introns as a group, because most mutations in the exon binding sites change the site specificity without impacting catalytic activity. Therefore, the splicing mechanism can be ruled out as a reason group I introns are found in conserved sites at the amino acid level. Exon splicing enhancer and silencer nucleotide motifs have been characterized surrounding individual spliceosomal introns [50, 51]. In the case of spliceosomal introns these requirements for exon sequence motifs surrounding the intron have not led to a detectable preference of insertion sites that are conserved at the amino acid level.
Parasitic elements have likely evolved two different strategies to propagate and survive. Inteins and group I introns utilize homing endonucleases to target conserved sites. Conserved sites tend to be in functionally important; consequently, precise excision of the intein or group I intron is required to maintain functionality of the protein. Furthermore, the functional importance of the residues limits the range of substitutions that can modify the target sequence so that it no longer is recognized by the homing endonuclease. Although group I intron and intein insertions are targeted to a DNA sequence, this is merely a proxy for sequence conservation at the protein level, upon which purifying selection can act. Targeting conserved sites will also facilitate transfer of the intein or group I intron to new intron-less alleles, as they will likely contain near identical amino acid sequences in these regions. In contrast, group II introns use a more random and less specific retrohoming or retrotransposition mechanism (see [52] for review). This suggests an alternative strategy evolved by these elements, relying on frequent propagation to outpace more rapid loss. If group II introns are the ancestor of spliceosomal introns [28, 53], it would not be unexpected to find a similar site preference for group II and spliceosomal introns.
Both of these strategies have successfully ensured the survival and propagation of inteins, and group I and group II introns [15, 18, 20]. These genetic elements can be considered molecular parasites that have their own life cycle, only occasionally evolving functions that contribute to the fitness of the host organism, or that increase the complexity of the host in an irreversible manner, without necessarily increasing the host's fitness [47, 54].
In most eukaryotes the coxI gene (encoding cytochrome C oxidase subunit I) is found in the mitochondrion, and hosts many introns representing both group I and group II introns. Several studies provide evidence for horizontal gene transfer of this element between distantly related groups of eukaryotes, confirming an intron homing model of evolution [38, 55, 56].
As shown in figure 5, the cox1 gene introns were found in both conserved and variable regions of the host; however, introns that are found in numerous species are more often found in highly conserved sites. The only group I intron found in the cox1 gene of several vascular plants [39] is inserted in the most conserved site of the host protein. The intron present in plants was reportedly acquired from a fungal donor [38]. As the conservation of the insertion site is the key for homing process, the observed preference for conserved sites may reflect the mode of propagation of the group I introns.
It is possible that inteins and group I introns target all sites but only the ones that end up in a conserved site are retained. The two exceptional proteins in this study, NADH dehydrogenase subunit 3 and Chlorophyll alpha apoprotein A2, may be an example of this. This mechanism would result in a preference for conserved sites even in the absence of a site specific homing mechanism. Such independent preference for conserved sites may have caused splicing elements (introns and inteins) and homing endonucleases to target the same sites, resulting in the fusion of these elements [57]. Free-standing HE genes are found in intergenic regions, their survival is dependent on the homing cycle [23, 24] and they might be more easily eliminated as compared to HE associated with self-splicing elements since their removal does not need to be precise. Upon fixation of the HE in the population, homing is no longer possible, because the HE is already present in all target sites. If the HE has not acquired another function that can create a selection pressure to maintain the HE gene, it is likely to decay and be lost [20].
The fact that BILs do not show a conserved site preference [58] suggests that the conserved site preference is associated with the life-cycle of the homing endonuclease containing molecular parasite. This life cycle was first formulated as a homing cycle involving movement of the molecular parasite across population or species boundaries [4, 59], but it has been suggested that this life cycle, with its succession of empty target sites, sites invaded by a molecular parasite with functioning homing endonuclease, sites containing a dysfunctional homing endonuclease, can operate within in spatially distributed population [20], and for some values of fitness reduction of individuals carrying molecular parasites may also operate continuously in homogenous well mixed populations [60]. As discussed above, both the invasion phase and the deletion phase of life cycle may cause a conserved site preference. The targeting of a conserved site will make it more difficult for the host protein to evolve immunity towards the homing endonuclease, and it will make deletion of the molecular parasite more difficult.
The rate and tendencies to gain and lose introns vary considerably between lineages of eukaryotes [42, 43], with the number of spliceosomal introns per gene and the degree of sequence conservation at spliceosomal intron boundaries also varying greatly [61]. Spliceosomal introns can be gained via homologous recombination with intron-containing genes unrelated to the conservation of the site, and through retrohoming [42, 62, 63]. The intron removal processes also appears to be unrelated to the sequence conservation at the insertion sites. The best characterized model for intron loss in multicellular eukaryotes is via homologous recombination between intron containing genes and spliced cDNAs produced by reverse transcription [42, 62, 63].
We have provided statistical support for the notion that inteins and group I introns target conserved protein sites for survival. This may also provide evidence for the homing cycle that describes the life cycle of these two molecular parasites. Furthermore, our findings suggest that group II and spliceosomal introns persist in their host genes using a different evolutionary strategy.
Construction of the Conservation Profiles
To calculate conservation profiles along a protein sequence, BLASTP [64] was used to detect homologous protein sequences from the NCBI protein database. Each dataset was aligned using the CLUSTALW program version 1.83 [65], and inspected for alignment accuracy.
To construct archaeal/vacuolar-type ATPase catalytic subunit A protein sequence alignments sequences from 29 Eukaryotic species, 13 Archaeal species, and four Bacterial species were aligned. Ten Archaeal species and fifteen Eukaryotic species were aligned to construct the Replication factor C protein sequence alignments. Nine Archaeal species and 21 Eukaryotic species were aligned to construct the cell division control protein 21 protein sequence alignment. Sequences from two Eukaryotic species, 18 Bacterial species and four phage sequences were used to construct the DNA polymerase I protein sequence alignments. 21 Eukaryotic species and nine Archaeal species were use for the Cytochrome C Oxidase Subunit I protein alignment. All other proteins found in the InBase database [30] (data retrieved November 2009) that host inteins were also analyzed. And all other proteins found in the comparative RNA web site database [37] (data retrieved November 2009) were analyzed for group I introns and group II introns positions. (See additional file 3 for accession numbers.)
The conservation profiles were calculated from the protein alignments using an in house PERL script (Olga Zhaxybayeva, Dalhousie University, see additional file 4). This program calculates the number of substitutions over a sliding window of 11 aligned positions and the window is moved through the alignment one position at a time. Inteins, introns, and sites where more than 50% of the sequences had a gap inserted into the alignment are omitted in the calculations. The lower the conservation score the more conserved the position is and the higher the conservation score the less conserved the positions is. These conservation scores for each protein were mapped on the protein structures using MacPyMOL [66].
Statistical Analysis
Statistical Support for Inteins and Introns Targeting Conserved Sequences.
Group I Intron
Group II Intron
Spliceosomal Intron
Simple linear regression was used to correlate site conservation with intron/intein penetrance (the number of species infected with a specific element). Significance was then calculated using standard methods based upon the resulting correlation coefficient (r), and the degrees of freedom in the sample (n-1).
The authors thank Olga Zhaxybayeva for providing perl scripts. This work was supported in part by grants from the UConn Research Foundation and NASA Exobiology Program (NNX08AQ10G, NNX07AK15G, NNG05GN41G).
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