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Need Help? Call Our Specialists
Luxury Cruises
Travel Search Box:
Tell us as much as you can about the luxury cruise you are looking for in the Search Box and we'll give you all of your money-saving options. Our database contains early-bird specials, two-for-ones, last-minute markdowns and other cut-rate promotions. Additional, unpublished discounts are available.
Exclusive promotions for Key Traveler guests are available for Oceania Cruises, Regent Seven Seas Cruises, Silversea Cruises, Seabourn Cruises, Crystal Cruises, Windstar Cruises and Paul Gaugin Cruises.
Call 1-888-539-1881 for details. If you already have a cruise vacation in mind, please CLICK HERE to email us and one of our travel advisors will contact you shortly.
We will beat any published prices on luxury cruises. Guaranteed. If you know the ship and sail date, call Key Traveler for a better deal.
Climb aboard a luxury ship and you'll immediately understand what sets the experience apart from life on a mass-market or contemporary cruise ship. Lavish attention to every detail. Ultimate convenience. A level of personal attention and pampering that leave guests delighted and amazed.
When you make the leap to luxury cruising, you'll enjoy the best of everything – the finest available accommodations, cutting-edge amenities, gourmet dining from the world's most celebrated culinary artists, an elegant array of enrichment programs and activities, and service that prides itself on knowing exactly what you need (and when you'll need it). Today's luxury cruise ships offer passengers every posh perk imaginable.
Accommodations & Amenities
All-suite accommodations are available on some luxury cruises. Spacious marble baths and generously-sized walk-in closets are some of the amenities you can expect from many of the luxury cruise ships. Expansive views of the sea from the comfort of large and private teak verandahs. Frette linens and feather pillows on many luxury cruises. Cruise on Azamara Cruises, Oceania Cruises Regent Seven Seas Cruises, Crystal Cruises, Silversea Cruises, The Yachts of Seabourn, Sea Dream Yacht Club, Paul Gauguin Cruises and Windstar Cruises. You can expect staterooms to be spacious and well-appointed. And a sophisticated selection of room amenities is the standard. Watch first-run films on a flat-screen plasma television. Order from a pillow menu designed to cater to the wishes of a discerning clientele, and expect nightly turndown service (and delicious surprises) when retiring to your cabin each evening.
Dining on a luxury cruise line equals imaginative gourmet creations prepared under the watchful eyes of award-winning chefs, a selection of vintage wines that even the most discerning sommelier would admire, innovative venues, and so much more. Open-seating policies are the rule on some luxury cruise lines, allowing guests the flexibility to dine as they choose, and mealtime options abound. Think tuxedo-clad waitstaff on most luxury cruise lines, multiple courses of truly inspired cuisine, and elegantly appointed tables set with fine china and even European crystal on many luxury cruise lines. Casually elegant choices are also on the menu – sumptuous buffets served along teak-lined pool decks, as well as opportunities to enjoy course-by-course dining experiences in the comfort of your suite, where available with complimentary butler service to cater to your every whim in suite staterooms.
Entertainment / Enrichment Programs
The freedom to do anything. Or nothing at all. Aboard a luxury cruise, you'll encounter the same simple pleasures you'd expect to find aboard any traditional cruise – sunbathing poolside, for example, or the chance to enjoy an invigorating workout. But a luxury cruise augments these activities with exclusive programming and special offerings that are unique to your voyage. Choice is paramount. Succumb to the tranquility and the exotic treatments that can only be found in the most elegant spa and wellness facilities at sea. Or treat yourself to the ultimate souvenir – exclusive partnerships with premier brands afford passengers some very luxurious shopping opportunities.
A luxury cruise will take you places you've only dreamed about. And onboard enrichment programs, a signature feature of the luxury cruise experience, help bring those places to life. World-renowned experts and celebrities set sail alongside passengers on select voyages, sharing anecdotes, insights, and a wealth of behind-the-scenes information with guests. Stimulating lectures and hands-on demonstrations illuminate and enhance your journey.
Exceptional Personalized Service
Highly personalized service is the hallmark of the luxury cruise experience. Favorable staff-to-guest ratios mean that there's always someone at your disposal, and you'll reap the benefits instantly. Carefully selected professionals greet you warmly by name, take pride in anticipating your needs, and fulfill your every whim. Personal butlers are also available (their services, in some instances, are already included in your fare) to attend to every detail of your stay, from special shipboard requests to shoreside arrangements. The service on luxury cruise ships is simply unrivaled, and the examples are legendary – uniform clad stewards that wade into the surf to serve caviar and champagne while you're ashore, and much more.
Your Key Traveler luxury cruise specialists will guide you to the best choice for your luxury cruise experience. Our valued suppliers offer a variety of destinations, one to suit every traveler's wishes.


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EBSCO Discovery Service (EDS) - Branding and Customization Questionnaire
The attached questionnaire is used during implementation to customize your EBSCO Discovery Service (EDS) with your library's colors, logos, links, and branding.
Looking for this resource in another language? Check the International Resources pages to see if a translation is available.
Download: eds_branding_customization_questionnaire.doc

subset: commoncrawl


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What We Do
Sunday Worship
First Time Guest
Sermon Videos
Prayer Request
Mission and Service
Get Involved
Young Adults
Small Groups
CORE Groups
Core Signup
United Methodist Women
United Methodist Men
Reconciling Ministries Team
About Us
Map & Directions
Contact Us
Church Resources and Links
Safe Church / Safe Sanctuaries Policy
E-news Sign up
Reconciling Ministries Team
The Reconciling Ministry Team at Bothell United Methodist Church affirms that all are welcome in Christ's church. The Team recognizes, however, that churches have often spent more time and energy building boundaries and placing restrictions on God's reconciling love than on breaking down barriers of race, ethnicity, socio-economic class, and gender identity. The Bothell UMC Reconciling Ministry Team is thus committed to embodying a "ministry of reconciliation" by "breaking down the walls that divide" (Eph 2:14).
"All these new things are from God, who reconciled us to himself through Christ, and who gave us the ministry of reconciliation." II Corinthians 5:18 (CEB)
Recognizing that the United Methodist Book of Discipline expressly excludes "self-avowed practicing homosexuals" from being "certified as candidates, ordained as ministers, or appointed to serve in The United Methodist Church," a group of Bothell UMC members began in 2011 to explore affiliation with the
Reconciling Ministries Network
, an organization within the United Methodist Church that is committed to working toward full inclusion of all people, regardless of gender identity. By 2014, a recommendation was presented to the Church Council which authorized a task force to craft a public statement welcoming all. As a result, Bothell UMC's Welcome Statement was unanimously adopted by Church Council in May 2014.
In March, 2016, the Bothell UMC Reconciling Ministry Team voted to join the Reconciling Ministries Network as a "Community," and the Team was formally added to the Reconciling Ministries Network on May 3, 2016. The Reconciling Ministry Team "community" acknowledges the diversity of viewpoints within Bothell UMC while affirming a prophetic call to full inclusion.
Since its founding in 1885, Bothell UMC has ministered to the those on the edges of social norms. Whether veterans struggling with the trauma of the Civil War, refugees from war-torn Europe after World War II, or South East Asian refugees after the end of the Vietnam War, Bothell UMC has been there to accept and support people as they move toward what God has called them to be.
While Bothell UMC is a community that reflects a broad spectrum of theological, political, and social commitments, it is genuinely committed to expressing God's boundless love to all through word and deed. If you are interested in learning more about the Reconciling Ministry Team or getting involved with this Team, please contact Jeff Staley, co-chair, Reconciling Ministry Team at
© 2017 Bothell United Methodist Church |
Contact Us
18515 92nd Ave. NE, Bothell, WA 98011-2299
Phone: 425.486.7132 Fax: 425.486.0303


text: THINGS NEEDED: -2 people - 1 Rail (powered,rail) - Minecart with Chest*** - 2 blocks of any kind (dirt, sand, etc.) - 1 person with a pickaxe (any kind; preferably iron or diamond pickaxe) - 1 person with the desired item to be duplicated Steps: 1. put the rail...anywhere you want...on the floor though LOL 2. with the 2 blocks put each block at EACH end of the rail, so the minecart WON'T MOVE** 3. Put the "minecart with chest" on the rail 4. The person with the desired item to be duped put it in the minecart with chest and HOLD A*** 5. The other person with the pickaxe breaks the "minecart with chest" 6. There should be 4 things that drops in the ground: minecart, chest, the glitched item, and the original item. 7. Then just duplicate the glitched item just like what you do with the Furnace duplication glitch. 8. If you don't know how to duplicate the glitched item...KEEP READING! How to Duplicate the Glitched Item: ***make sure you don't have the original item/another stacked of the original item in your inventory because you won't be able to pick up the GLITCHED Item*** 1. pick up the glitched item and dont stand on top of it make sure you pick it up and go far away from the glitched item. 2. Open your inventory and you should see a stack of 64 of the desired item that you want to be duped. 3. Press A on the stack and make it hover over an empty slot in your inventory. 4. Press and HOLD X so that it places that item in that empty slot one by one. 5. Once that slot is full...a stack of the desired item another should be ready for you to hover another empty slot and repeat the process. 6. Once you finished duplicating have a regular chest ready for you so that you can put those stacks in. 7. When you're putting the stacks in the chest make sure you do it ONE BY ONE by pressing A in your inventory to pick it up and Pressing A again when you're putting it in the empty slot in the chest. 8. DO NOT BE LIKE " THIS IS TAKING SO LONG SO I JUST PRESS Y AND YOLO" you will regret it...you'll see why. LOL 9. After that, go celebrate by duplicating TNT and blowing up EVERYTHING and make SURE YOU SAVE!<3...im kidding...well you can do that if you want. 10. after putting it all in the chest make sure you save or BYE BYE Duplicated stacks of desired item. Important NOTES*** PLZ READ if you're having trouble: - This glitch works with the latest patch...How do i know? cause i just did it with a friend just now. so dont give me messages saying it doesnt work please. - This doesnt work 100% of the time just like the Furnace Glitch. - In the event that this glitch fails 2 things might happen: 1. The mine cart with the chest broke with no duplicated item. Solution: Make the minecart with chest again NOTE***You do not need to have a crafting bench with you to make the Minecraft with chest. if you have both of these items (minecart and chest) just make them by pressing x and go to the transportation tab or w.e and make it. pick up the original item and repeat the process. 2. The original item dissapears. Solution: Make sure you have at least a back up or 2 of the desired item sitting inside another chest before doing this glitch...or the furnace glitch too. - This works with ANY STACKABLE ITEM doesnt have to be a block. TROUBLESHOOTING: - if you guys are still having trouble even after you read and repeated the process a billion times then send me a message or post here and i'll try my best to reply back to you as fast as i can. - SEND ME A MESSAGE IN XBOX saying you're having trouble and i'll pick a right time to join your party and you can invite me to you're game and we'll go through the process together like a nice *** couple...no homo and im kidding LOL GT: Steph Ly iLy LASTLY IF THIS HELPED YOU GUYS PLEASE +REP AND THANK ME CAUSE I'M STILL A LEECHER AND I NEED THOSE THINGS SO I CAN PUT FOOD IN MY TABLE. PLEASE AND THANK YOU PS: IF you guys are wondering that i dont have a video proof...because i dont have a recording device...im a hobo what do you expect? but seriously i dont have a recording device. if someone who's kind enough to post a video proof that would be awesome well PEACE and have fun DOING THIS BORING DUPLICATION GLITCH!


text: Ok, so as some of you know I posted a while back that my son was going to a WC in Hawaii for Severe Anxiety Social and General, Depression, Mood Disorder (Intermittent Explosive Disorder (IED)). Well we used a transport co. on the 11th. He wasn't happy but he went. I haven't gotten to talk to him yet, I have spoken to his counselor and they said he is settling in well and being open to treatment. They did send me some pictures so I can atleast "see" that he made it. A couple of questions..... 1. Since he is going to go on to a longer term Residential Treatment Center (RTC) (therapy school), is it necessary to attend the 2 day parent workshop. They said it is optional.........the Easy Child that we hired said that the experience is invaluable. She isn't the one paying for this though. 2. Is it normal to not be able to talk to your child after a week? Is it usually 2 or 3 weeks? 3.Back to the parent workshop it isn't until April 26/27th, at this point it will be over 6 weeks since he has been there. The Easy Child is hoping that he will only be at wilderness camp 8 weeks. I am sure there will be workshops that the longer treatment centers. What would you do? Money isn't falling off of trees, but I don't want my son to think I don't care if I don't attend a 2 day workshop. We will have airfare, hotel, car, etc. and this is money we can use to pay for his treatment. I guess I am feeling guilty, confused and just need to vent. On top of this I still have a 13yr old that needs to be treated fairly. UGH!!!!!!!


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Drury Lane's "West Side Story" is the most explosive production in years.The production is getting rave reviews (including 3.5 stars from the CHICAGO TRIBUNE and a "Highly Recommended" from the CHICAGO SUN-TIMES) and is the fastest selling production in the theater's 30 year history. I also gave this production a MUST SEE!
Tickets are scarce, BUT, here is an opportunity for you to win a pair so you can be an audience member.
Here are three questions about the show. The answers can be found in my review of course at www.aroundthetownchicago.com and all you need to do is look at Theater Reviews to find it.
Answer the three questions and send them by e-mail to firstname.lastname@example.com by February 28th at 5 p.m. or you can also mail them to the paper ATTN: Al Bresloff.
All correct entries will have the opportunity to be selected as the grand prize winner and awarded the tickets for this memorable experience.
1. Tony and Maria and their love is based on what Shakespeare play?
2. Who were the ladies that perform "America" the satirical look at what they have found?
3. What Latina actress sings the solo in the beautiful rendition of "Somewhere"?
answer these questions correctly and have the opportunity to win a pair of tickets to this experience.
e-mail address_____________________@_____________
address_____________________________city____________________ zip___________
Good Luck
You can also go to www.theatreinchicago.com, go to Review Round-up, click at "West Side Story" and find the review at www.aroundthetownchicago.com


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Campaign 2016
2016 is an exciting year for DMIA, because this year we turned five!! Please see the anniversary letter from our President describing DMIA's work in promoting Dhrupad to date (http://dhrupad.com/about-dmia/annual-letters/).
Your support made all of that possible. Help Dhrupad continue to bloom and flourish. Make a generous donation today!! (http://dhrupad.com/donate/)


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Diploma in Information ScienceHINDUSTAN AVIATION ACADEMY
Price on request
Important information
|3 Years
- Diploma
- Bangalore
- Duration:
3 Years
Course programme
Diploma in Information Science
Courses conducted by HEA polytechnic leads to Diploma in Engineering awarded by the Directorate of Technical Education, Govt. of Karnataka and All Example content imageIndia Council for Technical Education, Govt. of India. The semester examinations are conducted by the Board of Technical Examinations, Govt. of Karnataka. After successful completion of 3 year Diploma courses, students become eligible to get admission directly to 2nd year Bachelor of Engineering (B.E.) courses conducted by approved Universities.
Scope for Higher Studies
* A Student finishing his/her diploma can appear for selection for lateral entry in to B.Tech/B.E Second year directly. The greater benefit from this scheme is that a student would be studying almost all same subjects of diploma course with higher syllabus content. Hence he/she will be definitely in a better position to do very well as compared to a direct B.Tech/B.E entry after +2/PUC.
Jop Opportunities
* With globalization of the Indian economy, vast opportunities of growth have been thrown open for the computer software and hardware industry. Software being one of the largest growing sectors of the Indian economy, the Department of Information Science Technology &Engineering focuses in guiding the students to learn how information and computing system supports business, research and communication needs. Needless to say, this is the most sought after area in the rewarding career scenario.


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Centenary College linebackers
Think about what career is best suited for you, and where you want to study. Request more information now.
Think about what career is best suited for you, and where you want to study. Request more information now.
Think about what career is best suited for you, and where you want to study. Request more information now.
Information about Centenary College linebackers, and registering for elective seminars and discussion sections. Not all recruiting is done like NCAA Division I teams. College coaches are constantly looking for the best players to fill next year's open positions, and rely on many different methods to finding the right recruits. If you haven't been noticed, cold-calling coaching staff can get you an initial interview. Finally, check with your major department for assistance with prerequisites, tuition payment deadlines, and Centenary College Football information. Please use the form above to request admissions info for Centenary College.
You don't have to quarterback the university football team to a state championship to earn an athletic scholarship. Most major sports that field a team in intercollegiate competition have sports scholarships available. From women's gymnastics to men's track and field, coaches are seeking players to build a team around. Further, that leadership role often is accompanied by a full-scholarship.
The best time to begin searching for scholarships is before your senior year, in order to meet deadlines. Schools confirm that it is important to file financial aid forms such as the CSS Profile early in the school year. There are many sports scholarships that focus on less tangible qualities, such as the ability to lead a team, motivate others, and learn new skills rapidly. Take stock of your athletic abilities as you mature through high school. Bicol University roster may indicate that it's an option to delay college entrance by a year. An extra year in high school or prep school can allow players to mature mentally, bring grades and SAT test scores up, put on weight and gain strength. Beyond consideration for Centenary College linebackers, you may think about Western Piedmont Community College short courses for complementary electives to your primary course of study.
Athletic Training Fellow
University of Colorado
STUDENT ATHLETIC TRAINER PROGRAM. Years experience as a certified athletic trainer. Work in conjunction with staff athletic trainers in the prevention and care..
2 days ago Texas Athletic Coaches (2017-2018)
Great Hearts Academies
Basketball and Soccer. Basketball, Track and Field, Tennis. ATHLETIC COACHES - Great Hearts Academies in Texas (2017-2018).
1 day ago Assistant Coach - High School - Boys Lacrosse
Cherry Creek School District
Has previous successful teaching, coaching and/or sport participation experience. Possess or has the ability to acquire a valid Colorado Coaching Certificate.
2 days ago Athletic Trainer Fitness
Riverview Hospital
Athletic Trainer Certification. Athletic Trainer Job Functions:. The Athletic Trainer encompasses all duties relating to providing exemplary patient care at..
1 day ago Assistant Athletic Coach I-Football
Wayne State University
Three years previous football coaching experience required. Assist in all aspects of the football program, including coaching, sports camps, recruiting..
2 days ago


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Active Member
Ikonija Connie Joy, MD FACS
Ikonija Connie Joy, MD FACS, is an ASPS Member plastic surgeon who is board certified by The American Board of Plastic Surgery® and trained specifically in plastic surgery. ASPS members operate only in accredited medical facilities, adhere to a strict code of ethics and fulfill continuing medical education requirements in plastic surgery, including training in patient safety techniques. As your medical partner, Dr. Joy is dedicated to working with you to achieve your goals.
Schedule A Consultation
Meet Dr. Ikonija Connie Joy
As director of Plasticare Surgery Center Dr. Joy is focused towards a trusting relationship with her patients. Dr. Joy's ultimate goal is to reconcile the patients aesthetic objectives and expectations with the reality of plastic surgical procedures.
Your Consultation
Learn all that you can to separate fact from fiction regarding cosmetic plastic surgery. Our experienced staff will ascertain your needs and send you a complete information package prior to your consultation. your consultation will be with Dr. Joy who will ascertain your needs and discuss your ultimate surgical results. The consultation will include review of patient photographs or slides. A digital computer image evaluation may be indicated. Educational videos are also available.
The first consultation with Dr. Joy will require approximately one hour. Information regarding surgical fees and financing will be available at this time.
Patient Testimonials
I have had multiple surgeries at Plasticare and I am absolutely pleased with the results and the TLC that I received from this office. rjc
Procedures Performed
Arm Lift
Body Contouring
Body Lift
Botulinum Toxin
Breast Augmentation
Breast Implant Removal
Breast Implant Removal
Breast Lift
Breast Reconstruction
Breast Reduction
Brow Lift
Buttock Lift with Augmentation
Chemical Peels, IPL, Fractional CO2 Laser Treatments
Chin Augmentation
Chin Surgery
Cosmetic Surgery
Cosmetic Surgery & Computer Imaging
Dermal Fillers
Ear Reconstruction Surgery
Ear Surgery
Endoscopic Technique
Eyelid Ptosis Repair
Eyelid Surgery
Facial Implants
General Reconstruction
Hair Replacement
Head and Neck Skin Cancer Reconstruction
Head-Neck Cancer Reconstruction
Injectable Fillers
Laser Hair Removal
Laser Skin Resurfacing
Lip Augmentation / Enhancement
Male Breast Reduction
Post Burn Reconstruction
Retin-A Treatments
Scar Revision
Skin Cancer
Spider Vein Treatment
Surgery for Genitourinary Diseases
Tattoo Removal
Thigh Lift
Tram Flap Breast Reconstruction
Transgender Surgery
Tummy Tuck
Vascular Malformations


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At the Lieber Institute for Brain Development, I am part of the Data Science team whose goals include better understanding and characterizing genomics signatures in the human brain, including DNA methylation and gene expression. I help mentor other team members, provide support for LIBD projects and I am advancing my academic career as part of Andrew Jaffe's lab.
As a quick background, I graduated from the Undergraduate Program on Genomic Sciences (the About page is in English) from the National Autonomous University of Mexico (UNAM) in 2009 and worked for two years at Winter Genomics analyzing high-throughput sequencing data. I then got a PhD in the Department of Biostatistics at Johns Hopkins Bloomberg School of Public Health thanks to a CONACyT scholarship awarded in 2011. I worked with Jeff Leek and Andrew Jaffe in developing derfinder, regionReport, recount as well as other applied projects.
I am a fan of using R and Bioconductor for data analysis, which I use with R Markdown for creating reproducible reports.
PhD in Biostatistics, 2016
Johns Hopkins Bloomberg School of Public Health
Bachelor in Genomic Sciences (LCG), 2009
National Autonomous University of Mexico (UNAM)
† indicates corresponding author, * indicates equal contribution
† indicates corresponding author, * indicates equal contribution
Tue, Apr 17, 2018, DennisDal2018
Thu, Aug 3, 2017, JSM 2017
Thu, Jul 27, 2017, BioC2017
Tue, Jun 27, 2017, ICSA 2017
Download my cv or view it at GitHub.
While working at Winter Genomics I taught two courses for students of the Biomedical Sciences PhD Program (PDCB) from the National Autonomous University of Mexico (UNAM).
While I was at the Institute of Biotechnology (UNAM) working with the Winter Genomics crew I organized two courses. One was a series of various bioinformatics and biology mini-courses and another one involved members of different institutions.
I taught three courses during my undergrad stage at the Undergraduate Program on Genomic Sciences (LCG). Each of these courses has its own website organizing the material. These are:
If you have questions about the Bioconductor packages I maintain, please read this post. If you send me an email, I'll simply refer you to the same post.


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Diane Davies of the University of Maryland talks about a global fire alert system that detects wildfires from space and then sends land managers urgent information.
This resource is audio only.
© 2018 PBS & WGBH Educational Foundation. All rights reserved.
You must be logged in to use this featureLog In Enter Code Need an account?


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Most Helpful Guy
I met my best friend in middle school, though we really became close in high school. I have known him for thirty three years and God willing will know him another 33 more. I officiated at his wedding (no, I'm not a minister, but you can be authorized by the state to perform wedding ceremonies). I was there when his children were born. They call me uncle. He was there for me during my wife's illness. He was there when I buried my wife and my father, as were his children. I love them. They love me.00|00|0


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The Melburnian
104/250 St Kilda Road
It's a very rare occurrence to find an apartment with two bedrooms on the ground floor of the prestigious Melburnian, so when a home like this enters the market, potential buyers are going to want to act decisively and promptly to make it theirs. A sanctuary of sophisticated style and space, the luxurious home showcases an elegant living room, an open yet delineated area for formal dinner parties and a casual meals area for more informal get togethers. The granite kitchen is superbly equipped with the full complement of high-end Miele appliances. Flowing from the pampering master ensuite bedroom, second bedroom and living room is an exceptional entertaining terrace stepping out to the gorgeous park-like gardens. Soul-soothing sounds of the surrounding water promote a peaceful calming atmosphere. The apartment also features a second bathroom, separate laundry, 2 car spaces ( Tandem ) storage cage, ducted heating and cooling and video-monitored security.2 2 2
Obligation-free expert advice.
If you would like any advice or information regarding any aspect of Melbourne's prestige apartment or unique property market.We'd love to hear from you »
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With our specialist knowledge of the prestige market we select only the very best apartments on offer.Tell us your dreams - we'll find it »
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Looking to invest ? Our expert local knowledge helps you make the most of your investment with great returns and friendly service.Discuss your options »
Getting you better results
Our success is attributed to the exceptional level of personalised service provided, a service specifically designed to meet your needsContinue »


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Early Decrease of XRCC1, a DNA Base Excision Repair Protein, May Contribute to DNA Fragmentation After Transient Focal Cerebral Ischemia in Mice
Background and Purpose—DNA damage and the DNA repair mechanism are known to be involved in ischemia/reperfusion injury in the brain. The x-ray repair cross-complementing group 1 (XRCC1) protein plays a central role in the DNA base excision repair pathway by interacting with DNA ligase III and DNA polymerase β. The present study examined the protein expression of XRCC1 and DNA fragmentation before and after transient focal cerebral ischemia (FCI).
Methods—Adult male CD-1 mice were subjected to 60 minutes of FCI by intraluminal blockade of the middle cerebral artery. XRCC1 protein expression was analyzed by immunohistochemistry and Western blot analysis. DNA damage was evaluated by gel electrophoresis and terminal deoxynucleotidyl transferase–mediated uridine 5′-triphosphate-biotin nick end-labeling (TUNEL). The spatial relationship between XRCC1 expression and DNA damage was examined by double staining with XRCC1 and TUNEL after FCI.
Results—Immunohistochemistry showed the nuclear expression of XRCC1 in all regions of the control brains and that it was predominant in the hippocampus. The XRCC1 level was markedly reduced in the caudate putamen at 10 minutes, further decreased in the entire middle cerebral artery territory at 1 hour, and remained reduced until 4 and 24 hours after FCI. Western blot analysis of the normal control brain showed a characteristic band of 70 kDa, which decreased after FCI. A significant amount of DNA fragmentation was detected by DNA gel electrophoresis 24 hours but not 4 hours after FCI. Double staining showed that the neurons that lost XRCC1 immunoreactivity became TUNEL positive.
Conclusions—These results suggest that the early decrease of XRCC1 and the failure of the DNA repair mechanism may contribute, at least in part, to DNA fragmentation after FCI.
The DNA protein x-ray repair cross-complementing group 1 (XRCC1) plays a central role in the DNA base excision repair (BER) pathway, which is responsible for repairing damage caused by various kinds of insults, including oxidative stress, by interacting with DNA ligase III and with DNA polymerase β.1 2 3 4 5 6 The critical role of XRCC1 in the cellular response to DNA damage was found when the repair defects of the mutant Chinese hamster ovary cell lines were corrected by transfection with the human XRCC1 gene.1 Such a role was confirmed in 4 other mutant cell lines lacking the XRCC1 protein.6 Although no catalytic domain was found in XRCC1 itself, it was reported to interact with DNA ligase III by its N-terminal domain containing a BRCA1 C-terminus (BRCT) module, which is a widespread motif in DNA repair and DNA damage–responsive cell cycle checkpoint proteins.3 5 7 XRCC1 also interacts with DNA polymerase β by its N-terminal half of the XRCC1 protein.3 In response to DNA-damaging agents, a preferential decrease in XRCC1 mRNA was found in vitro, suggesting its link to DNA damage.8 The predominant expression of XRCC1 mRNA and/or protein was found in rat brains8 and in baboon brains.9 However, detailed cellular/subcellular distribution of XRCC1 in the central nervous system and its role in pathophysiological conditions in vivo are totally unknown.
DNA damage and repair are drawing more attention in the field of acute central nervous system injury, including cerebral ischemia and brain trauma.10 Liu and colleagues11 have suggested that free radicals could attack the nuclear genes and cause genetic instability after mouse forebrain ischemia. As for the relationship of the DNA BER proteins to DNA damage after ischemia, we have shown that a decrease in apurinic/apyrimidinic endonuclease (APE/Ref-1) DNA BER protein preceded the occurrence of DNA fragmentation in the entire ischemic lesion after transient focal cerebral ischemia (FCI)12 and that loss of APE/Ref-1 was closely associated with the occurrence of DNA fragmentation in hippocampal CA1 neurons after transient global ischemia.13 These facts suggest that the reduction of DNA BER protein and a failure of the DNA repair mechanism may contribute to DNA damage after ischemia/reperfusion.
To further investigate the relationship between DNA BER protein expression and DNA damage after ischemia, we analyzed the expression of the XRCC1 protein, which is known to play a central role in DNA BER, before and after transient FCI. We further sought to clarify both temporal and anatomic relationships between XRCC1 alteration and the occurrence of DNA fragmentation.
Materials and Methods
Focal Cerebral Ischemia
Adult male CD-1 mice (weight, 35 to 40 g) were subjected to transient focal ischemia by intraluminal middle cerebral artery (MCA) blockade with a nylon suture, as described.14 The mice were anesthetized with 2.0% isoflurane in 30% oxygen and 70% nitrous oxide with the use of a face mask. The rectal temperature was controlled at 37°C with a homeothermic blanket. Cannulation of a femoral artery allowed the monitoring of blood pressure and arterial blood gases, with samples for analysis taken immediately after cannulation, 10 minutes after occlusion, and 10 minutes after reperfusion. After the midline skin incision, the left external carotid artery was exposed, and its branches were electrocoagulated. An 11.0-mm 5-0 surgical monofilament nylon suture, blunted at the end, was introduced into the left internal carotid artery through the external carotid artery stump. After 60 minutes of MCA occlusion, blood flow was restored by the withdrawal of the nylon suture.
Immunohistochemistry of XRCC1
Anesthetized animals were perfused with 10 U/mL heparin and subsequently with 4% formaldehyde in 0.1 mol/L PBS (pH 7.4) after 10 minutes, 1 hour, 4 hours, and 24 hours of reperfusion after ischemia. Brains were removed, postfixed for 12 hours in 4% formaldehyde, sectioned at 50 μm on a vibratome, and processed for immunohistochemistry. As a negative control, sections were incubated without primary antibodies. For histological assessment, alternate slices from each brain section were stained with cresyl violet. The sections were first reacted with mouse monoclonal antibody against XRCC1 (Lab Vision) at a dilution of 1:100. To avoid cross-reaction between the secondary antibody and mouse immunoglobulin in the tissue, immunohistochemistry was performed with the use of the DAKO ARK peroxidase kit. After development with diaminobenzidine (DAB), the sections were mounted on slides and counterstained with methyl green solution.
Western Blot Analysis
Whole cell protein extraction was performed. To confirm the early reduction of XRCC1 expression in the ischemic brain, we used the samples obtained from the ischemic core on the ipsilateral side and homologous tissue from the contralateral side. Approximately 30 mg of both ipsilateral striatum and homologous tissue from the contralateral side was cut into pieces after 4 hours of reperfusion and put into 10× volume of Tris-glycine SDS sample buffer (Novex). Samples were then gently homogenized 20× in a Teflon Dounce homogenizer (Wheaton). Equal amounts of the samples (10 μL) were loaded per lane. The primary antibodies were either 1:1000 dilution of monoclonal antibody against XRCC1 (Lab Vision) or 1:10 000 dilution of anti–β-actin monoclonal antibody (Sigma). For XRCC1 detection, Western blots were performed with horseradish peroxidase–conjugated anti-mouse immunoglobulin G with the Boehringer Mannheim chemiluminescent system. As the internal control, Western blot analysis of β-actin was performed with horseradish peroxidase–conjugated anti-mouse immunoglobulin G reagents (Amersham International).
Gel Electrophoresis
Animals were killed 4 and 24 hours after 60 minutes of MCA occlusion. Thirty to 50 mg wet weight of ischemic tissue was taken from the third 2-mm section along with homologous tissue from the contralateral side after the brain was cut coronally. Samples were incubated overnight in 0.6 mL lysis buffer (0.5% SDS, 10 mmol/L Tris-HCl, and 0.1 mol/L EDTA) with 0.6 mg proteinase K (Boehringer Mannheim) at 55°C. The DNA was extracted with equal volumes of phenol and phenol-chloroform-isoamyl alcohol (25:24:1) and precipitated overnight in 0.2 mol/L sodium chloride in 100% ethanol at −80°C. The DNA was washed with 75% ethanol 2 times, air dried, and resuspended in DNase-free water (Sigma). The DNA concentration was measured with To-Pro-1 dye (Molecular Probes). Gel electrophoresis for detecting DNA laddering was performed according to the manufacturer's instructions (Trevigen). Before electrophoresis, 1 μg of DNA was incubated with 50 μg/mL of DNase-free RNase (Boehringer Mannheim) for 30 minutes at 37°C. Then the samples were reacted with Klenow enzyme (Trevigen) and dNTP (Trevigen) in 1× Klenow buffer (Trevigen) for 10 minutes at room temperature. Samples were mixed with loading buffer and subjected to electrophoresis on 1.5% agarose gel. Then the gel was washed with 0.25 mol/L HCl, 0.4 mol/L NaOH/0.8 mol/L NaCl, and 0.5 mol/L Tris buffer (pH 7.5). DNA was transferred to a nylon membrane overnight in 10× SSC. The membrane was first blocked with 5% powdered milk (BioRad) in PBS for 30 minutes and incubated with streptavidin–horseradish peroxidase conjugate (Trevigen) for 30 minutes. Finally, the bands were visualized by the chemiluminescence method with the use of PeroxyGlow (Trevigen), and the films were exposed to x-ray film.
In Situ Labeling of DNA Fragmentation
Frozen brain sections at the level of the caudate putamen that showed typical infarction were stained by an in situ technique (terminal deoxynucleotidyl transferase–mediated uridine 5′-triphosphate-biotin nick end-labeling [TUNEL] reaction) to detect the DNA free 3′-OH ends, as described.15 Briefly, frozen brain sections were fixed for 30 minutes in 3.7% formaldehyde in 0.1 mol/L PBS, pH 7.4. The slides were placed in 1× terminal deoxynucleotidyl transferase (TdT) buffer (Life Technologies) for 15 minutes, followed by reaction with TdT enzyme (Life Technologies) and biotinylated 16-dUTP (Boehringer Mannheim) at 37°C for 60 minutes. The slides were then washed 2× in SSC (150 mmol/L sodium chloride, 15 mmol/L sodium citrate, pH 7.4) for 15 minutes, followed by a washing in PBS 2× for 15 minutes. Avidin-biotin horseradish peroxidase solution (ABC kit, Vector Laboratories) was applied to the sections for 30 minutes, then the slides were washed for 15 minutes with 0.175 mol/L sodium acetate. Staining was visualized with the use of 0.025% DAB and 0.075% H2O2 in PBS with 0.4 mg/mL nickel sulfate. The slides were rinsed with water, stained with methyl green for 10 minutes, dehydrated, and mounted.
Double Labeling With XRCC1 and DNA Fragmentation
To clarify the spatial relationship between XRCC1 expression and DNA damage, we performed double staining of XRCC1 antibody and TUNEL, as previously described.13 After transcardiac perfusion, fixed sections were immunostained with XRCC1 antibody as described above, the sections were mounted on glass slides, passed through ethanol (70%, 95%, and 100%), and then immersed in chloroform for 5 minutes. The sections were rehydrated by passage through a decreasing ethanol series, rinsed with water, and processed to TUNEL staining as described above. Staining was visualized with the use of 0.025% DAB and 0.075% H2O2 with nickel sulfate. The slides were rinsed with water, stained with methyl green for 10 minutes, dehydrated, and mounted.
Quantification and Statistical Analysis
The number of XRCC1-immunoreactive cells and methyl green–positive cells was counted in a high-power field (×400) and expressed by the percentage of XRCC1-positive cells. The number of TUNEL-positive cells was also counted, as previously described.15 The quantitative analysis of these cells was evaluated by factorial ANOVA between each group. Significance between groups was assigned at P<0.05.
Physiological Data and Cerebral Infarction
Physiological parameters showed no significant differences in mean arterial blood pressure and arterial blood gas analysis between each time point (Table⇓). The time-dependent increase of infarction in the mouse brains with the use of intraluminal suture blockade is consistent with previous reports that employed the same focal stroke model in mice.14 15
Immunohistochemistry Showed the Constitutive Expression of XRCC1 in Normal Adult Mouse Brain, Which Decreases After Ischemia
The XRCC1 protein was constitutively expressed in the entire region of the normal mouse brain (Figure 1A⇓ through 1D). It was mainly expressed in the nucleus and had a regional predominance in the hippocampus, including the CA1 (Figure 1C⇓) and CA3 regions and the dentate gyrus (Figure 1D⇓), compared with the MCA territory cortex (Figure 1A⇓) and caudate putamen (Figure 1B⇓). After 10 minutes of reperfusion that followed 1 hour of FCI, reduction of XRCC1 was observed in the lateral caudate putamen (Figure 1E⇓) but not in the cortex (data not shown). After 1 hour of reperfusion, XRCC1 expression was reduced in the entire MCA territory, including the caudate putamen (Figure 1F⇓) and cortex (data not shown). This reduction was sustained at 4 hours (Figure 1G⇓) and 24 hours (Figure 1H⇓) in the caudate putamen, as well as in the MCA territory cortex. In contrast, the contralateral brain including the caudate putamen (Figure 1I⇓) showed no significant alteration of XRCC1 expression until 24 hours after FCI. There was no immunoreactivity in the control specimens, which were treated without a primary antibody (data not shown).
Western Blot Analysis of XRCC1 Protein Expression After Transient MCA Occlusion
XRCC1 immunoreactivity was evident as a band of molecular mass 70 kDa of the whole cell fraction from the nonischemic hemisphere (Figure 2⇓, lane 1, top panel) and was significantly decreased 4 hours after ischemia (Figure 2⇓, lane 2). A consistent amount of β-actin immunoreactivity between each lane is shown in the bottom panel, suggesting that the amount of the loaded protein was consistent. These data not only confirm the specificity of the antibody for XRCC1 used in this study but also suggest that XRCC1 decreased after transient focal ischemia.
DNA Laddering Was Detected by Genomic DNA Gel Electrophoresis
To detect the occurrence of apoptosis as characterized by intranucleosomal DNA fragmentation, we analyzed DNA from both the ischemic brain and the homologous sample on the contralateral side. DNA laddering was absent in both the control tissue and ischemic tissue 4 hours after ischemia (Figure 3⇓, lanes C and 4). A significant amount of DNA laddering was detected 24 hours after ischemia (Figure 3⇓, lane 24).
Early Decrease of XRCC1 Preceded the Occurrence of DNA Fragmentation Detected by TUNEL Staining After FCI
To elucidate the spatial and temporal profile of DNA fragmentation, we examined in situ labeling of DNA breaks in the infarcted brain sections (Figure 4⇓), as described.15 TUNEL staining did not label normal neuronal cells in the noninfarcted area (Figure 4A⇓) or the cells in the infarcted area 4 hours after ischemia (Figure 4B⇓). In contrast, 2 different patterns of staining were observed in the neuronal cells in the infarcted area 24 hours after ischemia (Figure 4C⇓). Some neuronal cells in the infarcted area were densely labeled in their nuclei, accompanied by small particles around the nuclei that resembled apoptotic bodies (Figure 4C⇓, arrows). These cells are compatible with those in the apoptotic cell death process, as previously described.15 16 17 Besides these typical apoptotic neuronal cells, slightly TUNEL-stained cells were also observed (Figure 4C⇓, arrowheads). These cells showed diffuse nuclear and cytoplasmic TUNEL staining, which is consistent with necrotic cells.15 16 17 To quantify the apoptotic neurons, the number of TUNEL-positive cells was counted 4 and 24 hours after FCI. As illustrated in Figure 5⇓, only a small number of TUNEL-positive cells were detected 4 hours after ischemia. They were significantly increased 24 hours after ischemia both in the caudate putamen (Figure 5A⇓; P<0.0001) and the cortex (Figure 5B⇓; P<0.01). As shown in Figure 5⇓, the reduction of XRCC1 expression preceded the occurrence of DNA fragmentation, which was detected by gel electrophoresis and TUNEL at 24 hours but not at 4 hours after FCI. In addition, there was no significant difference in the number of XRCC1-positive cells between nonischemic controls and the contralateral side after FCI (data not shown).
Double Staining With XRCC1 and TUNEL Showed the Spatial Relationship Between XRCC1 Loss and DNA Damage
Double staining with XRCC1 and TUNEL 24 hours after ischemia showed a significant amount of TUNEL-positive cells with the characteristic features of apoptosis (densely labeled in their nuclei, accompanied by apoptotic bodies) in the entire MCA territory on the ipsilateral side, including the cortex (Figure 6A⇓, arrowheads). None of these cells showed XRCC1 immunoreactivity. On the other hand, some cells had a faint expression of XRCC1 (Figure 6A⇓, asterisks), but none were TUNEL positive. No TUNEL-positive cells were seen in the contralateral side at this time point (Figure 6B⇓). There was no immunoreactivity in the control specimens, which were treated without a primary antibody (data not shown).
The present study provides the first evidence that the early decrease of the DNA BER protein XRCC1, which is constitutively expressed in normal mouse brain, may play a role, at least in part, in the DNA-damaged cell death pathway in transient FCI. The immunohistochemistry of the normal mouse brain showed for the first time that the XRCC1 protein was constitutively expressed in the entire mouse brain and has a nuclear distribution and a regional predominance in the hippocampus. As early as 10 minutes after reperfusion, a significant reduction of the XRCC1 expression was seen in the lateral caudate putamen (Figure 1E⇑), extended to the entire MCA territory after 1 hour of reperfusion (Figure 1F⇑), and remained reduced until 4 and 24 hours after transient FCI. This reduction was confirmed by Western blot analysis of whole cell extract (Figure 2⇑). DNA fragmentation was not detected 4 hours after FCI, while it markedly appeared 24 hours after FCI, as shown by DNA gel electrophoresis (Figure 3⇑) and TUNEL staining (Figure 4⇑). Finally, the spatial relationship between the loss of XRCC1 expression and DNA fragmentation was shown by the double staining with XRCC1 and TUNEL (Figure 6A⇑). These results indicate that early reduction of XRCC1 precedes the occurrence of DNA fragmentation in the ischemic MCA territory that is destined to show necrosis and/or apoptosis and that the reduction of XRCC1 may contribute to DNA-damaged cell death after FCI. Although XRCC1 has no known catalytic activity for undergoing DNA repair, it is believed to play a central role in the DNA BER pathway by interacting with DNA polymerase β and DNA ligase III.1 2 3 4 5 6 Furthermore, using the same FCI model, we previously reported that the DNA BER enzyme APE/Ref-1 was reduced in the entire MCA territory on the ischemic side before the peak of DNA fragmentation.12 Thus, it is suggested that the tissue in the ischemic MCA is deficient in at least 3 major BER enzymes, such as APE/Ref-1, DNA polymerase β, and DNA ligase III, which can be extremely detrimental for the cells in overcoming DNA damage caused by ischemia/reperfusion injury.
Programmed cell death and the DNA-repairing mechanism are both assumed to play important roles in cerebral ischemia.10 13 15 16 18 19 However, little is known about the interaction of these processes, except the data suggesting a link between apoptosis and the cleavage of the DNA repair enzyme poly(ADP-ribose) polymerase (PARP).20 The interleukin-1β–converting enzyme family caspases are the human homologues of the nematode Caenorhabditis elegans, Ced-3,21 and are considered to play a critical role in programmed cell death.20 One substrate for caspase is PARP, whose proteolytic cleavage results in a dysfunctional PARP that is unable to contribute to repair or genomic maintenance.20 Furthermore, the Ca2+/Mg2+-dependent endonuclease that generates internucleosomal DNA cleavage characteristic of apoptosis is negatively regulated by poly(ADP)-ribosylation. Therefore, inactivation of PARP could increase DNA cleavage and contribute to programmed cell death. On the other hand, excessive activation of PARP is believed to also be deleterious because it might cause energy depletion and ultimate cell death.22 In fact, most recent studies with PARP knockout mice implicate the deleterious role of PARP in FCI.23 24 Although these studies did not provide the data that suggested its role in DNA damage, they clearly show the marked reduction of the infarct volume in PARP knockout mice after transient cerebral ischemia,23 24 suggesting that PARP activation plays a detrimental role in neuronal damage after ischemia. Interestingly, a recent study showed that XRCC1 interacts with PARP by its central region containing a BRCT module.7 In this study, overexpression of XRCC1 in Cos-7 and HeLa cells resulted in the marked decrease of PARP activity in vitro, which suggests that XRCC1 may negatively regulate the activity of PARP. On the basis of these findings, the early reduction of XRCC1 during reperfusion in the present study could contribute to the early activation of PARP and subsequent energy depletion and then could be extremely detrimental for the cells in overcoming ischemia/reperfusion injury.
The selective reduction of XRCC1 after FCI was implied by our following findings. Western blot analysis showed the significant loss of XRCC1 4 hours after FCI (Figure 2⇑, top panel), while there was no alteration of β-actin after FCI (Figure 2⇑, bottom panel). Furthermore, immunohistochemistry showed the marked reduction of XRCC1 in the entire ischemic area 1 hour after FCI (Figure 1⇑), when the immunoreactivity of the nuclear neuronal marker NeuN showed much less alteration in the ischemic area (data not shown). The exact mechanism by which the selective reduction of XRCC1 occurs after FCI is unclear. It is conceivable that XRCC1 reduction is caused by a selective decrease of XRCC1 protein synthesis, an increase of protein degradation, a difference in the half-life of the protein, or a difference in posttranslational regulation. Further examination is warranted to clarify this issue. Nevertheless, we have previously reported that APE/Ref-1, another DNA BER protein, decreased rapidly after focal cerebral ischemia/reperfusion in mice,12 and our preliminary study using the same stroke model demonstrates that mice overexpressing the endogenous antioxidant enzyme superoxide dismutase-1 (SOD1) show less reduction of APE/Ref-1 after ischemia/reperfusion compared with wild-type mice.25 Therefore, it is conceivable that the mechanism of the selective loss of XRCC1 could also be linked to oxidative stress. In fact, our transient FCI model is known to be associated with reactive oxygen species, as shown by our previous study using SOD1 transgenic mice and knockout mice deficient in SOD1.14 15 26 27 28
With regard to programmed cell death after ischemia, we have reported that mitochondrial cytochrome c was released to the cytosol after transient FCI in rats17 and in mice.29 Since translocated cytochrome c is known to be apoptogenic because of its role in caspase activation and subsequent DNA fragmentation,30 these results suggest that cytosolic release of cytochrome c may cause downstream caspase activation and subsequent DNA fragmentation after FCI. Consistent with this finding, the activation and cleavage of caspase 3 has been shown after FCI in mice,31 and inhibition of caspase family proteases resulted in significant protection against focal ischemia/reperfusion injury.32 From the point of oxidative stress and programmed cell death, our recent study showed that a 50% reduction of manganese SOD in knockout mice resulted in a marked increase of cytochrome c release and subsequent DNA fragmentation after FCI,29 suggesting that mitochondrial production of reactive oxygen species may contribute to the release of mitochondrial cytochrome c, thereby exacerbating DNA fragmentation after FCI. The correlation between the mitochondrial cascade and the postischemic modification of DNA repair proteins such as XRCC1 and APE/Ref-1 is totally unclear. Since it is well known that reperfusion increases mitochondrial production of reactive oxygen species,28 mitochondrial oxidative stress could contribute indirectly to the early decrease of DNA BER proteins. The future examination of DNA BER using mutant mice deficient in and/or overexpressing mitochondrial manganese SOD is warranted to address this important issue.
In conclusion, we have shown that XRCC1 rapidly decreased after transient FCI in mice and that this reduction preceded the occurrence of ischemic apoptosis and infarction. Furthermore, a spatial relationship between XRCC1 loss and DNA damage was shown by double staining with XRCC1 immunohistochemistry and TUNEL. These results indicate that the early decrease of XRCC1 and the failure of the DNA repair mechanism may contribute, at least in part, to DNA-damaged cell death after transient FCI.
This study was supported by National Institutes of Health grants NS14543, NS25372, NS36147, NS38653, and N01NS82386. Dr Chan is a recipient of the Jacob Javits Neuroscience Investigator Award. We thank Cheryl Christensen for editorial assistance and Liza Reola, Bernard Calagui, and Jane O. Kim for technical assistance.
- Received May 25, 1999.
- Revision received August 2, 1999.
- Accepted August 5, 1999.
- Copyright © 1999 by American Heart Association
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Thompson LH, Brookman KW, Jones NJ, Allen SA, Carrano AV. Molecular cloning of the human XRCC1 gene, which corrects defective DNA strand break repair and sister chromatin exchange. Mol Cell Biol. 1990;10:6160–6171.
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Caldecott KW, Aoufouchi S, Johnson P, Shall S. XRCC1 polypeptide interacts with DNA polymerase β and possibly poly(ADP-ribose) polymerase, and DNA ligase III is a novel molecular "nick-sensor" in vitro. Nucleic Acids Res. 1996;24:4387–4394.
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Chopp M, Chan PH, Hsu CY, Cheung ME, Jacobs TP. DNA damage and repair in central nervous system injury: National Institute of Neurological Disorders and Stroke Workshop summary. Stroke. 1996;27:363–369.
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Liu PK, Hsu CY, Dizdaroglu M, Floyd RA, Kow YW, Karakaya A, Rabow LE, Cui JK. Damage, repair, and mutagenesis in nuclear genes after mouse forebrain ischemia-reperfusion. J Neurosci. 1996;16:6795–6806.
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Kawase M, Fujimura M, Morita-Fujimura Y, Chan PH. Reduction of apurinic/apyrimidinic endonuclease expression after transient global cerebral ischemia in rats: implication of the failure of DNA repair in neuronal apoptosis. Stroke. 1999;30:441–449.
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Yang G, Chan PH, Chen J, Carlson E, Chen SF, Weinstein P, Epstein CJ, Kamii H. Human copper-zinc superoxide dismutase transgenic mice are highly resistant to reperfusion injury after focal cerebral ischemia. Stroke. 1994;25:165–170.
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Kondo T, Reaume AG, Huang T-T, Carlson E, Murakami K, Chen SF, Hoffman EK, Scott RW, Epstein CJ, Chan PH. Reduction of CuZn-superoxide dismutase activity exacerbates neuronal cell injury and edema formation after transient focal cerebral ischemia. J Neurosci. 1997;17:4180–4189.
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Fujimura M, Morita-Fujimura Y, Narasimhan P, Copin J-C, Kawase M, Chan PH. Copper-zinc superoxide dismutase prevents the early decrease of apurinic/apyrimidinic endonuclease and subsequent DNA fragmentation after transient focal cerebral ischemia in mice. Stroke.. 1999;30:2408–2415.
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Editorial Comment
Normal cellular metabolism, particularly mitochondrial respiration, produces reactive oxygen species (ROS) (eg, hydrogen peroxide and superoxide anion and hydroxyl radicals) as end products.1 Under physiological conditions, excessive ROS are neutralized by endogenous antioxidants (eg, ascorbate, α-tocopherol, β-carotene, and glutathione) and antioxidant enzymes (eg, superoxide dismutase, catalase, and glutathione peroxidase). Severe oxidative stress may overwhelm the antioxidant mechanisms and lead to oxidative DNA damage.2 Among the DNA lesions caused by ROS are base modifications, single-strand breaks, double-strand breaks, strand scissions, and the crosslinking of bases.3 The second line of defense against oxidative DNA damage is the DNA repair machinery. Oxidative DNA damage can be rapidly and efficiently repaired.4 However, this second line of defense against oxidative damage of DNA may be compromised in the setting of ischemia-reperfusion.5
Individual bases in DNA may be oxidized to form adducts such as 8-hydroxyguanine (8-OH-Gua), 8-hydroxyadenine (8-OH-Ade), 4,6-diamino-5-formamidopyrimidine (FapyAde), 2-hydroxyadenine (2-OH-Ade), 5-hydroxycytosine (5-OH-Cyt), 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua), 5-hydroxy-5-methylhydantoin (5-OH-5-MeHyd), and 5-hydroxyhydantoin (5-OH-Hyd).6 These oxidative DNA adducts are potentially mutagenic or lethal. The XRCC1 protein is involved in contributes to base excision repair and therefore, may contribute to enhance genome integrity and stability after ischemic insult. In the preceding article by Fujimura et al, the expression of the XRCC1 protein was reduced in the ischemic region in a mouse MCAO model. Suppressed XRCC1 expression preceded DNA fragmentation and cell death. This finding raises the possibility that a defective DNA repair machinery caused by reduced expression of DNA repair proteins such as XRCC1 may contribute to irreversible DNA damage and ultimate cell demise after focal cerebral ischemia-reperfusion.
- Received May 25, 1999.
- Revision received August 2, 1999.
- Accepted August 5, 1999.
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Chan PH. Role of oxidants in ischemic brain damage. Stroke.. 1996;27:1124–1129.
Halliwell B, Gutteridge JMC. Free Radicals in Biology and Medicine. Oxford, UK: Clarendon Press; 1989.
Chopp M, Chan PH, Hsu CY, Cheung ME, Jacobs TP. DNA damage and repair in central nervous system injury: National Institute of Neurological Disorders and Stroke Workshop Summary. Stroke.. 1996;27:363–369.
Liu PK, Hsu CY, Dizdaroglu M, Floyd RA, Kow YW, Karakaya A, Rabow LE, Cui JK. Damage, repair, and mutagenesis in nuclear genes after mouse forebrain ischemia-reperfusion. J Neurosci.. 1996;16:6795–6806.
Jaruga P, Dizdaroglu M. Repair of products of oxidative DNA base damage in human cells. Nucleic Acids Res.. 1996;24:1386–1394.
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- Early Decrease of XRCC1, a DNA Base Excision Repair Protein, May Contribute to DNA Fragmentation After Transient Focal Cerebral Ischemia in MiceMiki Fujimura, Yuiko Morita-Fujimura, Taku Sugawara and Pak H. ChanStroke. 1999;30:2456-2463, originally published November 1, 1999https://doi.org/10.1161/01.STR.30.11.2456
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- Early Decrease of XRCC1, a DNA Base Excision Repair Protein, May Contribute to DNA Fragmentation After Transient Focal Cerebral Ischemia in MiceMiki Fujimura, Yuiko Morita-Fujimura, Taku Sugawara and Pak H. ChanStroke. 1999;30:2456-2463, originally published November 1, 1999https://doi.org/10.1161/01.STR.30.11.2456Permalink:


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It certainly is the last leg of a tour you'll never see again, and more importantly it may be the farewell tour of Angus Young. The make up dates for the Axl/DC (AC/DC plus Axl Rose) tour begins tomorrow in Greensboro N.C., and makes a stop here on September 6th.
Here's a bit of Shoot To Thrill
It's possible the Brian Johnson may recover enough to do a final show or two for a DVD, but the chances of it becoming a world tour seem remote at best. With so much changeover in the band in the last 6 months it may be time for Angus to consider hanging up the schoolboy outfit for good. But that's what people were saying in 1980 after the death of Bon Scott.


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The worship ministry at The Ark serves to host the presence of God and desires to continually be a dwelling place for Him. They serve in our weekly services as well as many of our additional meetings. This ministry is dedicated to raising up sons and daughters who know who they are in God and who know the power of personal and corporate worship, as well as being excellent in music and performance. The team rotates singers and musicians weekly for the purpose of training and equipping.
For more information contact Jake Clemons
Men's Group
Our Men's Group brings both the young and older men together for a time of hanging out and doing what men like to do. Keep an eye on the Events page for the latest info.
Women's Group
We'd love for all the ladies to come out and join together for times of celebrating, growing, and loving each other right in the midst of every season. Beautiful Pieces is designed to be a safe place for women to be unapologetic about being real, honest, and vulnerable. All women of all ages are encouraged to come! This will take place every third Thursday of every month at 6:30 pm. Childcare is provided. Contact Marsha Clemons with further questions.
Little Praisers
AGES: 0-3yrs
TIMES: 10:15a – 12:45p on Sundays & 6:15p – 8:15p on Wednesday nights.
LOCATION: Directly to the right when entering the Main front Entrance.
Little Praisers provides childcare for infants through pre-school (0 months – 3 years) during all church services. While your child is in our care, dedicated workers share the love of God by singing songs and teaching simple lessons from God's Word. We are glad to provide friendly, loving workers to care for your baby.
For more information contact Michelle Wilkinson
226 Kids
AGES: 4-6 and 7-12 years
TIMES: They will stay in worship and go to class at the conclusion of worship or at 11:30 and will be assisted by a team leader!
LOCATION: 4-6 classroom is downstairs directly to the right if you're coming from the main entrance. 7-12 classroom is upstairs. *Stairs located to the right of main entrance.
"Where we are reaching for the stars!"
226 Kids (Psalm 22:6) is a place where children from kindergarten to 6th grade can grow in the Lord by creative teaching, Godly lessons, and solid leadership. Our prayer is to lead this generation into a passionate love for Christ and to empower them with the knowledge of God.
In "226 Kids", children are taught their importance in the Kingdom of God! We are implementing Shawn Bolz curriculum "Growing up with God" and this series is teaching all of them who God is to them on a personal level, how to connect with God more, and how to help other connect to Him, too! It's fun and exciting! This generation is destined to see all of the promises of God fulfilled in their lifetime and we are passionate about equipping them to do so.
"226 Kids" meets Sunday mornings after worship or 11:30 am (whatever comes first) and Wednesday nights after worship. **Check-in for children of all ages begins at 10:15. Parents, please see the leader at the check-in desk prior to service to ensure we know your child will be going to class! Thank you! We greatly appreciate you and your family!
Contact Michelle Wilkinson with any questions
Converge Youth Ministry
AGES: 6th-8th Grade
TIME: Wednesday nights @ 6:30p
LOCATION: Upstairs
AGES: 9th-12th Grade
TIME: Sunday nights @ 6:30p
LOCATION: Upstairs
Converge Student Ministries is an exciting, relevant ministry that meets students right where they are and brings them together, no matter their differences. Converge will also be meeting upstairs on Wednesdays at 6:30pm.
For more information contact Darien Harris
Our simple goal is to create an environment where students are taught how to live a lifestyle of Revival, Prayer, Purity and Love. Our desire is to help students discover just-how-great God has made them; connecting them to their true identity as beloved sons and daughters of God. We believe this is a generation of revivalists who can and will make a difference in the world.
In addition to our weekly gatherings, Converge also offers special events, student leadership programs, and community service projects. If you're a student grades 6th-12th, we'd love to have you join us!
Life Groups
Sunday and Wednesday| 6 pm | Located off-site
Life Groups are simply a great way for everyone to connect outside of a church service while engaging in a book study that is anchored in the revelation that GOD IS GOOD! Sunday and Wednesday nights, groups will meet in the comfort of someone's home (locations listed below) and together dive into Bill Johnson's book God is Good. If being connected with others who love you as you are and finding your place of belonging is what you're after, joining one of these groups is the right thing for you!


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A leading retailer is banning imported white veal and calves' liver from its stores, it announced today.
Animal rights campaigners complain that animals bred for this type of meat are reared in small, cramped crates and fed a poor diet.
Marks & Spencer today pledged to sell only rose veal bred in the UK to decent welfare standards.
It will remove all imported white veal from its stores from Wednesday and stock the UK-reared alternative from January.
Calf liver
Calves liver will also be banned from M&S because it comes from animals bred for white veal.
Animal rights campaign group People for the Ethical Treatment of Animals (Peta) says calves bred for white veal are forced to live in cramped crates which restrict their movement and muscle growth.
They are fed a milk diet for their entire lives, which gives their meat its white colour, and are slaughtered at around eight months old.
M&S said the UK-produced rose veal comes from animals reared in well-lit barns and fed a natural diet which meets the welfare standards of both Compassion in World Farming and the RSPCA.
Welfare standards
The chain's director of food Steven Esom said: 'We have banned white veal as we were increasingly unhappy with the animal welfare standards associated with the rearing of calves.'
M&S will source its rose veal from a family-run farm based in Lothian.
The Press Association, All Rights Reserved.


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The Book
YOU ARE MADE IN THE IMAGE OF GOD Take a journey through the systems of the body-explore how the body performs and how the Body of Christ should perform, as explained by a Christian physician. This insightful and beautifully written book draws parallels between our physical and spiritual needs from the unique perspective of a medical and spiritual professional. Just as all of the systems of the body work together for the function of the physical body, members of the global church should work together to carry out the function of the Kingdom of God here on earth. Learn how we are fearfully and wonderfully made…in the Body of Christ. "Without controversy, great insights are being released by the Holy Spirit today concerning the mysteries of the Kingdom set apart and reserved for the Last Days. With great insight and intellectual understanding,
"Dr. Melita Williams provides valuable teaching concerning our physical bodies and the parallel with the Body of Christ in her book The Body of Christ: The Spiritual Revealed Through the Natural. The apostle Paul spoke of this as one of the mysteries involving the Lord Jesus as our Head and His people joined to him as His body. These insights will clearly aid us in fulfilling 1 Thessalonians 5:23 and our admonition to be fully sanctified spirit, soul and body by the workings of the Holy Spirit and the revelation of the Scriptures."
-Paul Keith Davis White Dove Ministries


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Room for Love and Faith Paperback
The Shunammite's Blessing
by Betty H Smith;
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Room for Love and Faith
Full Product Description
Room for Love and Faith: The Shunammite's Blessing explores the unusual generosity of the great woman of Shunem.
This anonymous, biblical character was penned "the Shunammite" by the Prophet Elisha, who prophesied an extraordinary blessing for her.
Smith brings the Shunammite to life with serving hands, a lively hope, and a determination bigger than life. Discover the Shunammite's blessing-give and be blessed, hear God's voice and believe, walk with faith and love to receive God's favor.
Is there a Shunammite's blessing waiting for you? Is there room for love and faith in your life?n
Room for Love and Faith by Betty H Smith was published by WestBow Press in August 2015. The ISBN for Room for Love and Faith is 9781512704839.
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Details for Room for Love and Faith
|Author / Artist
|Betty H Smith
|Book Format
|Paperback / softback
|WestBow Press (August 2015)
|Number of Pages
|9 mm
|216 mm
|140 mm
|Product Code
|Page last updated
|25th October 2017


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Plastikos and Micro Mold (Erie, PA) are investing $1 million in capital expenditures this year to boost cleanroom molding and precision mold making capacity.
The company is adding an 88-ton Arburg in its clean room, effectively increasing molding capacity in that area 50%. Capacity additions are also being made in its whiteroom medical and electronics. Additional inspection equipment is also being purchased.
"All these investments come back to better quality, better efficiency, increased capacity and increased capability," says Philip Katen, Plastikos president. "The skill and experience of our team are immensely important, but as time goes on, strategic investments in the latest technology and equipment are equally important to maximize the performance of our Team."
Plastikos' sister company, Micro Mold, plans to increase the efficiency and quality of tooling production through an investment in automation and robotics. This will enable the company to provide more 3D EDM work and lights-out operations.
"At Micro Mold, we already have a very large investment in technology. We consider the current infusion of capital as 'doubling down' on our mold shop," says Ryan Katen, general manager of Micro Mold. "At a time when many others are exiting the domestic tool making business because of its low margins and ever growing requirements for capital investments, we see it as a critical component to the ultimate success of our customers' end product in the market. These investments in automation allow us to provide high quality, while remaining price competitive so that an overseas option is less and less attractive to our customers."


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Crossing lines : season one / Tandem Films.
Available copies
- 2 of 2 copies available at Evergreen Indiana.
Current holds
0 current holds with 2 total copies.
|Call Number / Copy Notes
|Shelving Location
|Due Date
|Culver-Union Twp PL - Culver
|DVD TV CRO 1 (Text)
|Adult - DVD
|Jefferson Co PL - Hanover Branch
|TV DVD CROS (Text)
Record details
- Physical Description: 3 videodiscs (425 min.) : sound, color ; 4 3/4 in.
- Publisher: [Canada] : Entertainment One, [2014]
- Copyright: ©2014
Content descriptions
|General Note:
| Title from container.
Originally broadcast on television in 2013.
Special feature: "Behind the lines" featurette.
|Participant or Performer Note:
| Donald Sutherland, William Fichtner, Marc Lavoine, Moon Dailly.
|Summary, etc.:
| Tapping into the charter of the International Criminal Court (ICC) to mandate a special crime unit to investigate serialized crimes that cross over borders, hunt down criminals and bring them to justice, a global FBI is born. Carl Hickman is a wounded New York cop pulled from the edge by a group of unlikely saviors, Michael Dorn, an inspector in the ICC, and Louis Daniel, the head of an elite cross-border police unit charged with hunting down the world's most brutal criminals.
|System Details Note:
| DVD, region 1, widescreen (1.78:1) presenation ; Dolby Digital.
|Language Note:
| English dialogue; English or Spanish subtitles ; subtitled for the deaf and hard of hearing.
Search for related items by subject
|International Criminal Court > Drama.
Criminals > Drama.
|Thrillers (Television programs)


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Another Leap Forward for Hybrane
Polymer Factory is pleased to announce another successful sale of a large quantity of Hybrane® polyesteramide products to a large German company for the development of novel surface coatings. Hybrane® is a hyperbranched polymer that has long been popular among customers due to its ease of post-modification and affordability which allows large scale use in a variety of fields. In particular, it has already shown great potential in coating applications as well as water purification. With our in-house expertise, it is no surprise that companies and research institutions look to Polymer Factory for quality solutions and products.
To learn more about our Hybrane® products, click here .
In addition to catalog products, Polymer Factory also offers customized solutions and partnerships to produce products for specific fields. We do love challenges!
If you wish to work on a customised project with us, contact us here .
- Posted by polymerfactory
- On February 20, 2017
- 0 Comments


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Does Shakespeare present love as a positive or a negative force in A Midsummer Night's Dream?
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Shakespeare does not really present love itself as either a positive or negative force. However, he does present the fact that love is guided by the irrational mind in a negative light. We know that he does not put a negative slant on love itself because all of the couples are happily united at the end, even Oberon and Titanius make up in the end. Also, Shakespeare gives us Theseus and Hippolyta as the one loving couple that remains strong in their relationship all throughout, showing us that love itself is not at fault, but the irrational mind.
We first see Shakespeare present the theme of the irrationality of the mind concerning love in the first scene with Helena's soliloquy. In this speech, Helena points out that all over Athens she is recognized as being as fair as Hermia, indicating that Demetrius's choice to pursue Hermia instead of Helena is irrational (I.i.232). To emphasize her point, she speaks her most important lines, "Love looks not with the eyes, but with the mind; / And therefore is wing'd Cupid painted blind" (239-240). In these lines Helena is arguing that love is an illusion; love is all in the head, rather than having anything to do with objective reality, such as beauty. These lines very clearly portray Shakespeare's theme that love is guided by irrational thought. Helena also points out that Demetrius made proclamations and vows of love to her before he saw Hermia, which is also central to the unfolding of the play as well as to the themes, as we see in her lines, "For ere Demetrius look'd on Hermia's eyne, / He hail'd down oaths that he was only mine" (247-248). By having Helena point out Demetrius's unfaithfulness to Helena, Shakespeare is pointing out how fickle love can be. Shakespeare is also telling us that had Demetrius made the more rational decision of remaining faithful to Helena, the characters would have been spared from a great deal of grief and hardship. Hence, Shakespeare created the moral ending by having Demetrius decide to remain with Helena at the end of the play, explaining that he was engaged to her before he saw Hermia and is now ready to be faithful (IV.i.173-177). Having Demetrius make the moral decision by the end of the play shows us Shakespeare's message that love can indeed be guided by rational thought, showing us that love should not be viewed negatively, but rather the irrational human mind.


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How does Odysseus change from excessive pride to humility?
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This is a good question. The key moments for this change, I would say, appear in Book 9, Books 5 and 6, and Book 13.
In Book 9, Odysseus makes a crucial mistake that causes him to incur the wrath of the god Poseidon. That mistake is revealing his real identity to the Cyclops Polyphemus, who is the son of Poseidon. The key quotation appears near the end of Book 9:
So they argued, but could not daunt my ardent spirit, and I shouted to him again in anger: "Cyclops, if any man asks how you came by your blindness, say that Odysseus, sacker of cities, Laertes' son, a native of Ithaca, maimed you." (A.S. Kline translation)
Odysseus' angry boast here leads to Polyphemus' curse against him, which eventually leads to Poseidon's stormy attack on him when he sails away from Calypso's island (see Book 5).
By the time Poseidon is done with Odysseus in Book 5, Odysseus is naked and sleeping in a bed of leaves. This is literal humility as Odysseus is sleeping on the ground in the dirt (our word 'humility' comes from the Latin word for dirt, humus).
When Odysseus awakens in the land of the Phaecians, he is naked. The hospitable Phaeacians clothe him and return him to his native land, but upon arriving in Ithaca goddess Athena disguises him as a beggar:
She wrinkled the smooth skin on his supple limbs, and thinned the fine hair on his scalp, and gave him the body of an old man. She dimmed the beauty of his eyes, and dressed him differently, in a wretched cloak and ragged tunic, of tattered filthy smoke-grimed cloth. Then she flung a large deerskin, devoid of hair, over his shoulders, and handed him a staff, and a sorry-looking leather pouch, punctured here and there, hanging from a piece of braided cord.
Thus, by the end of Odyssey 13, Odysseus has become further humbled, dressed as a beggar, and this is the clothing he will wear until he strings his bow at the end of Odyssey 21. Accomplishing this feat, results in his transformation back into his true hero self.
We might also add that between Books 13 and 21, Odysseus experiences further humiliation as he begs for food in his own house and is even abused by another beggar, Irus, in Odyssey 18.


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Rev. Nancy Brink
More about Rev. Nancy Brink to come
Rev. Susan Chamberlain
More about Rev. Susan Chamberlain to come
Rev. Barbara Grear
The Rev. Dr. Barbara Grear is a member of our congregation. She recently celebrated her 40th ordination to the Christian Ministry through the Christian Church (Disciples of Christ).
- Bachelor of Arts in Religious Studies, Chapman College 1972
- Doctor of Ministry, School of Theology at Claremont, CA 1976
Board Certification as Chaplain through the Association of Professional Chaplains
During the past 40 years, Barbara has served congregations as the senior pastor, and has served specialized ministry on a university campus as well as in the hospital setting.
During the year 2015-2016, she is serving as co-chair of the Spiritual Life Ministry.
Her ministry in retirement includes study of California Natural Habitat and environmental concerns. Specific actions for these concerns are in process.
She writes, please meditate on these words written to the Church at Philippi: "Whatever is true, whatever is noble, whatever is right, whatever is pure, whatever is lovely, whatever is admirable–if anything is excellent or praiseworthy–think about such things."
Rev. Kel Henderson
Five quotes that inspire me
"You are the light of the world…..let your light shine." – Matthew 5
"We need to be the change we wish to see in the world." – Gandhi
"What we think ….We become." – Buddha
"Never place a period where God has placed a comma." – Gracie Allen
If you are not living on the edge, you are taking up too much room……. and if you end your journey in the same place you begin you have not traveled very far.


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Green Frill Hem Bralet
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llona wears a UK size 8 / EU size 36 / US size 4 and her height is 5'9"
Product number: TJ411268
Hand Wash
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text:
(Isaiah 58, 1-9a; Matthew 5, 13-20)
"What are we here for?"
Over the last couple of weeks the Gospel readings have been telling us about the beginning of Jesus's ministry, and the calling of his first disciples. And now, as we approach Lent, we turn to readings which set out the programme of action for his disciples, passages which begin to answer that question for them – and for us – "What are we here for?"
The programme of action is based on the covenant with Israel's God set out in the Old Testament. Jesus says he has not come to do away with the Law of Moses, but to fulfil it, that is to go beyond legalism to a way of expressing the underlying intention of the Covenant Law. His programme, he says, asks his followers to go beyond what was previously demanded, to meet a higher standard of righteousness (which, as he has already taught them, encompasses not just acting in accordance with the law, but also justice, integrity, charity and a particular concern for the weak and vulnerable members of society). Unless they do even more than the teachers of the Law and the ultra-religious Pharisees, he tells them, they won't really be under the rule of God (which is what entering the Kingdom of Heaven means).
The reading from Isaiah shows that the sort of life Jesus is asking his followers to live was already being spoken of as God's will by the prophets of the Old Testament. It is from the prophecy of Third Isaiah, writing to the Jews who have returned to Judea after the exile in Babylon. Isaiah observes that they are trying to earn God's approval by being ultra scrupulous about ritual and fasting – making a great fuss about being seen to follow all the rules. But, Isaiah tells them, this is not what God actually wants. The sort of 'discipline' God actually wants them to follow is not just about worship, but also about their everyday lives. It's about what we now call 'social justice' – feeding the hungry and poor, providing shelter for the homeless, and freeing those who are oppressed by the structures of society, ensuring justice for all.
When we hear this passage, we inevitably hear echoes of the agenda for his ministry which Jesus proclaimed in the synagogue at Nazareth, recounted in the Gospel of Luke; and with the story of the Last Judgement, the separating of the Sheep and Goats, in Matthew 25. This again confirms that the gospel of social justice is not a trendy modern invention. It is what Third Isaiah said discipleship was all about in the 8th century BC; it is what Jesus said discipleship was all about in the 1st century AD. It is still what Christian discipleship is all about.
It is the answer to the question "What are we here for?"
We are often told there is a hunger for spirituality in today's world. These passages tell us Christian spirituality is not to be discovered in withdrawal from the world, but in daily engagement with its realities. Christian discipleship is about politics and economics, about healing and housing human bodies, and removing the 'yoke' from the shoulders of the oppressed, whether that yoke be poverty, sickness or prejudice. It means working to oppose anything which prevents human flourishing.
So, our discipleship is to be lived out in service to the community. But how are we to carry out that service?
Jesus answers using two metaphors – light and salt. Both are ordinary common substances, both are God-given, not of human manufacture, both can be used to transform what surrounds them, to bring out the colour or flavour that is already there.
We have heard a lot about light in the readings over the last couple of months. It is a major theme of Advent, Christmas, and Epiphany. Light stands for vision, showing the right way, for promoting growth and health, even when it is not obvious how that can be done.
Our attitude towards light hasn't changed much in the years since these words were spoken by Jesus, but our attitude to salt may have done. In a world before refrigerators and antibiotics, salt was important for flavouring food, but also for preserving it, and as an agent of purification and healing. Salt was a common substance then, but at the same time so valuable that Roman soldiers were paid with it. Nowadays it is still a common substance, but with little value, and we are very much aware that too much of it can not only spoil the taste of food, but also damage our health.
Nevertheless, the metaphor of salt warns Christian disciples to keep their faith sharp and alive. It also indicates the values of their faith may have a different taste, which contrasts with those of the world around. For us today, that might mean that Christians value the small rather than the big, the spiritual as well as the material, but also value community cohesion above individual satisfaction, and watch out for the vulnerable rather than having increasing personal wealth as their highest ambition. A comment made on the last day of the football transfer window highlights this: "I live in a world where £35 million is paid for a footballer, yet 60 disabled people lose their day centre because it costs £200K to run". That is not a world where Christian salt is being effective.
But the metaphors of salt and light also warn Christian disciples that their values should permeate society, rather than dominating it. Too much salt makes food inedible. Too much light means that plants die, rather than flourish, and human beings cannot sleep. Christian dominance of society has not always proved beneficial either for society or for Christian discipleship. Particularly in today's world, to insist that so called Christian values should be the only guide to law making and enforcement, to the exclusion of other values which people think are important, actually damages the Kingdom of Heaven rather than helping it to grow. Just as in cooking, the balance between adding to little salt and adding too much is a difficult judgement, so in community life, we need much prayer and wisdom to decide how far our Christian beliefs and practices need to be defended or even worse, imposed by law, or whether their best defence is the difference they make to society.
Christ summarised the Covenant Law in two simple phrases – love God and love your neighbour as yourself. Keeping this summary in mind should keep all of us from turning the Sermon on the Mount, or our commitment to social justice, or our devotion to certain paths of Christian discipleship into just another tick list of standards by which we judge people to be inside or outside the Kingdom or justify separating ourselves from those who make different judgements. Christ's living out of the values of the Sermon on the Mount involved offering love and forgiveness to everyone, even to those who thought differently from him, and even to those who opposed and persecuted him. No matter what, the light of God shone through him, and showed up the limitations of legalism and separation. It is that different quality of life this passage urges us to seek.
Ultimately, it is up to each of us, both as individuals and in community, to decide how we can best be disciples of Jesus who bring the salt and light of God's rule to whatever situations we find ourselves in.
Because that, today's readings tell us, is what we are here for.


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