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The Duke and Duchess have left the building.
Prince William and wife Catherine, Duchess of Cambridge, left the U.S. yesterday after a whirlwind tour of North America -- but they met one more huge star before their flight home.
The two hopped back to Santa Barbara on Sunday to attend a reception to mark the launch of Tusk Trusts's US Patron's Circle, a wildlife conservation effort, where they met actress Reese Witherspoon. The royal couple was spotted speaking to Reese and Catherine Keener at the event.
The Santa Barbara event wasn't their only stop of the day though, as William & Kate also stopped by a inner city arts event on Skid Row in Los Angeles and helped pack care packages for military children in Culver City.
The two left for the U.K. after. Check out their California tour photos above!
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Rapid City Student Arrested in Gun Incident
RAPID CITY, S.D. (AP) — A teenager is in custody for allegedly bringing a BB gun that resembled a handgun onto school property in Rapid City.
Mitchell Man Falls Asleep, Shoots Self by Accident
MITCHELL, S.D. (AP) — Authorities say a Mitchell man suffered minor injuries when he accidentally shot himself after falling asleep on his back porch while holding a loaded handgun.

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Letter: Voting number for integrity
D. Cobb, Powell
In response to a letter on Nov. 11, I find that the writer is completely misunderstanding the ballot system. Yes, we do check each voter for their photo ID and address against a list of eligible voters. That is to ensure that you have not previously voted in the early election. When you go to the machine to get your number to open the ballot on the machines, the four-digit number is randomly picked by a computer. There is no order in these numbers. As a precinct officer I could collect all the numbers processed during Election Day and put them into a supercomputer and it would not be able to find any pattern.
As far as being assigned to a specific voting machine, the only assignment is the next machine that becomes vacant. The number you are issued is only for the computer to open the ballot when entered. You cannot enter that number a second time as it cannot be used again during the voting period. At this time the system we currently are using is the best one to prevent fraud and ensure the privacy of the voter.
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The Skin Food Lettuce and Cucumber Essence Mask Sheet Review
Here's one you can actually avoid, The Skin Food Lettuce and Cucumber Essence Mask Sheet, DO NOT WANT!
I'm currently in lust with The Skin Food Lettuce and Cucumber Skin Care Collection. It's slowly becoming my HG (holy grail) skin care of choice for daily day care. Loves it!
Of course, when I seen they had a sheet mask available in the Lettuce and Cucumber range I was all about it. This had to be an awesome mask right? Hells the Lettuce and Cucumber Skincare line has been changing my skin life so the sheet mask must be just as incredible.
Not so much.
Cheapest sheet mask, not the good kind of cheap.
On occasion I've run into cheap sheet masks. These are the kind that fall off your face, don't properly fit, are made from rough paper (not as soft as most masks, and just aren't saturated enough with product. All of these failings apply to The Skin Food Lettuce and Cucumber Essence Mask Sheet.
One of my biggest pet peeves are face masks sold separately. You've no idea how irritating this is. I wanna buy a box of five, a box of ten, a box of twenty, but please don't sell me one. In this case I'm glad I got one because damn it's really a rotten mask.
I actually find it interesting that Skin Food excels at good skin care but fails at something as simple as a sheet mask. This particular mask I tried was really rough on my face. The material it's made from isn't soft and it's a bit harsh on, nothing crazy but it's def not in the realms of feeling lush on my face as most masks are. Secondly, it's BARELY wet. Essence? What essence? Did someone forget to add in the essence? The mask isn't moist at all and once it's on your face and left on for a while it truly feels like you don't even have a mask on at all. No incredible hydrating facial with this bad boy. And finally it slips, slides, and doesn't fit correctly at all. Grr! I have to keep patting it down because it peels up and off.
Overall, Skin Food Sheet Masks are so not making a guest starring appearance on my great list. I paid retail which is like a buck I think at most but don't go so far as to dish out a fiver on these as that's how much I've seen them for online.
No likey.
Needs a whole heck of alot of work. Hope the other masks I got are a bit better.
Anyone try The Skin Food Sheet Masks?
Good experience?
Do share!
Disclosure: The following item was purchased by me for review purposes (and because I like flinging my money at makeup products of any kind) on Musings of a Muse. For further information please see my FTC Guidelines or my Disclosure/Disclaimer posts.
2/18/10 18:45 Chung:
Haven't tried the mask but I have the emulsion. The lettuce & cucumber emulsion is great! I really love it. It seeps into your skin super fast. Like literally a min-2 min max later it's gone. It's non sticky and makes my skin feel pretty hydrated. It's really preps the skin well for applying moisturizer after. Love it, you should try it!
2/18/10 19:24 Melissa:
I have yet to make my skinfood order. I keep finding better prices on eBay but some items are yet to be seen in the flea world. I got samples of the Agave Cactus line and love it… has a very clean not floral smell. SO I just took the masks out of my cart, and i'm gonna go for the powder one you mix because it sounds so darn good. Have you tried Missha sheet masks? They have a smokin deal on their site right now 2 of each type so 20 masks for $19.99. I'm debating on doing that since I haven't had an allergy to any missha products but I have to a few skinfood.
2/19/10 0:20 Melissa:
Missha sheet mask review please
2/18/10 20:38 Agnes:
Hi, I am one of those who love using facial sheet masks and bought alot of Skin Food masks when I was in Korea last year.
I am surprised yours was dry because mine was pretty wet but I hate the scent and I also do not find it particularly moisturising hence this mask is not on my wanted list.
Thanks for all your honest reviews so far. Your concealer reviews are one of those which made me want to try those brands that you have mentioned, however I have difficulty in getting Kevin Beautymaker. Haha.
2/19/10 3:33 aquahartz:
hi there Muse,
its a shame reading that the Skin Food sheet mask didnt do any good to u.
i am a fan of some of some of their sheet masks especially the repairing mask, the black raspberry mask & the snow tea mask. havent tried others much, but definitely adore these variants. my skin is left soft supple & healthy after each use.
i suppose not all products in one brand are good, e.g. the cucumber essence mask. have u put in a feedback to Skin Food, so that they could improve it?


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Daily Announcements Feb. 8
The blood drive is TODAY from 9am to 3pm in the weight room. If you are interested please contact Maggie Schmaedick or Matt Binkerd or sign up in the front office. Save a life and enter to win a prize and get a free cookie!
Today is the last day to donate food to the Sheldon Food Drive. 72,962 emergency food boxes were distributed by Food for Lane County this past year. Without these food boxes, some people in our community, including families with children, would not be able to meet their nutritional needs. Please do what you can to help by donating nonperishable food items in one of the barrels located in the front office, scheduling office, and near the theater and gym. Every little bit helps!
It's time to pick up the state football apparel that you ordered! Your orders will be available from 4:30-6:30pm in the cafeteria on Monday the 11th. If you are unable to make it you can pick up your apparel in the office after the 11th.
Students interested in taking the LCC placement test… There will be an opportunity to do so February 12th at 12:30 in the Career Center. Please sign up with Kim in the Career Center before February 11th. Be sure to have your "L" number with you!
Drivers, the parking lot in the very back of the school, the KRVM lot, is a Staff ONLY parking lot! The KRVM lot is NOT open to students before school, during school, or after school. There is a sign stating this and it is also marked on the ground. Thank you for keeping this lot for staff only.
Spanish Club Members: It's time to make Buzones and get ready for el Dia de Amor y Amistad. Bring a friend and join us tomorrow during lunch, in room B15.
Are you sick of waiting for Prom? Well tonight is the Mardi Gras dance at the Boys and Girls Club at 8pm. Come buy your tickets at the coffee cart for $6 or tonight at the door for $7! See you there!


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What's the most calorically dense and well rounded meal, barring trail mix and bars, that you can think of?
Edited by firstname.lastname@example.com on 04/20/2009 15:05:22 MDT.
that would be pretty rich ...pesto sauce: olive oil, parmesan cheese, pine nuts, basil
Food energy comes from fat, carbohydrates, and protein. Fat provides 9 calories per gram, and carbohydrates and protein provide 4 calories per gram. Pure fat gives you the "densest" calories. So basically, just fatten up your carbohydrates and protein with whatever form of fat you like.
Lard > butter.
Matt is right. A quick google search showed 1 Cup of lard has 1,849 calories, and 1 Cup of butter has 1,628 calories, probably because of the added water. The concept holds, however, that the more fat in your food per weight, the more calories. Popular fat additives for backpacking are cheese, nuts, and packets of olive oil. About the pesto sauce above - check Sarah's freezerbagcooking website to see one of her latest recipe posts for backpacking pesto sauce.
As others have said you want a good mix of protein, carbs, and fats. Get the highest fat content and the lowest water content for the most calorie dense foods. I shoot for a 40/40/20 ration of fats, carbs and protein respectively. My foods are Carbs----dried fruitnutsinstant ricecouscouspancakes/biscuits made on the trailProtein----Nuts/peanut buttercheesesalamijerkyFats----olive oilghee or clarified buttercheesesalaminutsAny of these foods can be eaten on their own and taste pretty good (even the oil and ghee). For meals I combine a grain or pasta with some nuts, dried fruit, and salami with a spice mix (I like garam masala or italian seasoning + garlic and onion powder) and maybe some sun dried tomatoes or dried mushrooms. Add in some extra fat and I have a pretty tasty well balanced, calorie dense meal.
Thanks for the replies. I'm looking for the most calorically dense, well rounded meal so I'm guessing eating just lard is out.I've always added olive oil to my meals when hiking to boost the fat calories and flavor of the dish. One of my favorite backcountry meals is mac n cheese, chorizo and olive oil, though the chorizo is heavier than I would like. I'm trying to find good meals that can beat Big Sur Bars for taste, calories and weight. On short hikes, food is more function over form, but longer hikes are a balance of the two--especially when the hyper sensitive gag reflex has kicked in.
I'll go out on a limb here - the best for calories/fat are not always food you can eat long term in large amounts. Carbs have their place even if they are not as big in calories. They often take up more space as well.Still, I'd rather cut weight in my gear than in the food I take. Food will get lighter over time, the gear won't ;-)
I don't know the exact weight / calorie ratio, but Mike's Super Spackel recipie in the Bio-Groovy article has got to be up there. Tastey too, you can actually eat a decent amount of daily calories that way. I can't stomach the concept but I've heard dog racers up north use sticks of butter rolled in brown sugar.....
One of the Packit Gourmet set menus focuses on calories:http://www.packitgourmet.com/Calorie-Packed-2-People-p344.htmlCheck out the dessert (molten chocolate lava cakes). 1160 calories per two muffin cup sized servings!
I have the same problems! But it looks to me that if you try to mix in various nuts this helps. Also some people like taling bagels along for the first few days. But here is a link to way to much info for me but maybe you will find it helpful!http://thru-hiker.com/articles/trail_foods.phphttp://thru-hiker.com/articles/pack_light_eat_right.php
"I can't stomach the concept but I've heard dog racers up north use sticks of butter rolled in brown sugar....."Also do not forget traditional pemmican -- the real thing is very high in fat (as opposed to some stuff that is sold as pemmican, but is mainly carbs).As to butter up North -- in my experience, I can tolerate a *lot* more fat in really cold weather than I can in the summer. When cooking supper for four at -10* - -20*F it is no big deal to drop a stick of margarine into the dinner pot -- something I would not seriously consider doing in the summer.--MV
Edited by blean on 05/07/2009 21:19:40 MDT.
they sell it in singles. 3oz for 250 calories, 200 calories from fat. it's already cooked so can be eaten a variety of ways. mmmm
could be fun, but isn't hard to store? i can already imagine a disgusting greasy spam mess. also that doesnt seem very efficient, due to the high water content of spam. i like to bring parmesan cheese and summer sausage for fat. i sometimes bring sesame oil, can make an otherwise boring meal very tasty. a nice all purpose food to bring along is pea soup mix. its light, high in protein decent amount of carbs and fat. pea soup, dry refried beans and quinoa seems to me to be the most well rounded and light staples to bring.
There are many trail foods that are over 100 calories/ounce versus the 84 calories/ounce for SPAM.
The Spam Singles are actually good if used right. Is Spam gourmet? No, but it is awesome in lo mein, fried rice, etc. It is very fatty and high in protein. For meat cravers it work well and the Singles pack well.Sure, legumes and grains are better, but lets face it....not everyone is into them. For every person who swears by say lentils there will 10 guys who want 'man food'.
"There are many trail foods that are over 100 calories/ounce versus the 84 calories/ounce for SPAM" Such as?
ramen: 3oz, 300 calories.wont clog arteries.
Pffftttt....you just need to combine the ramen and the Spam for Spam-tastic Lo Mein. Just saying......
Canned meat. Disgusting.Crushed potato chips are pretty good calories/weight.
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|10 Litres
|10 Litres
|Dishwasher Safe
|Model Number
|Product Type
|Mop Buckets
|0.7 kg
Delivered from £2.95
by Monday 12 October
by Monday 12 October
|Standard: 3-5 days
|Next day
|Named day
|Named day and time
|More delivery info
Returns for UKIf for any reason you are not satisfied with your George Clothing or George Home product, simply return it within 100 days with proof of purchase for an exchange or refund. George branded products only. Excludes watches, jewellery, hair accessories, part assembled flat pack furniture and mattresses. The 100 Day Guarantee only applies to George branded products. Please refer to our terms and conditions for details of our returns and refund process for non-George branded products. Your statutory rights are not affected. UK Mainland stores only. Excludes franchises.
Rated 4.9 out of 5 by 21 reviewers.
Rated 5 out of 5 by soubriquet innovative design Before buying this mop bucket we had never seen a design quite like it, although admittedly we bought the last one about 40 years ago! As you push the mop into it for wringing out,the mop head is squeezed by a....er....gripper. Wringing out is, therefore, a doddle. Very reasonably priced and very speedy delivery, too. 07 February 2012
Rated 5 out of 5 by maggieminx value for money Vileda Supermocio Bucket and Wringer is manufactured from recycled material to help protect the environment and contribute towards long-term sustainability. With power press wringer for improved cleaning performance. Plastic manufactured from recycled materials. A nice looking bucket, bright color and easy to use. The website is also easy to use and access, they have a good range of products that are all reasonably priced. The bucket and mop are of a good size, I was also able to find somewhere to store it without too much difficulty. The mop head is good for getting in-between my tiles to clean and it doesn't soak my floor too much either. Very easy to use and effective at cleaning. I have wooden and tile floors that I mop and found it excellent on both. Great quality, this bucket feels like it would last long without breaking, the mop is high quality too and feels very sturdy. I found this mop and bucket a high quality, affordable product. I would buy it myself and will continue to use it for years to come 04 February 2013
Rated 5 out of 5 by NEWDOGOWNER best thing i've ever bought This bucket is great for wooden floors and for cleaning up after new puppy. Wish i bought one ages ago and stopped strugging with my old one for so long. Best price on the market, excellent product, raving to my friends about it.Floors near dry after using it 19 August 2011
Rated 5 out of 5 by pussin great purchase this product is sturdy a top grade product lives up to is name, i would recommend this product for any one hows husband is going to use it as my husband has broke ever other ringer in my mop bucket so its definitely a must in my house. 24 March 2014
Rated 5 out of 5 by ricecake vileda super mocio bucket and wringer I think this bucket and mop is great It is great value and looks great to .My last bucket was a lot more expensive but was not as strong or looked as good as this . I would recommend this product to everyone . mrs ross 07 August 2012
Rated 5 out of 5 by jontyrhodes Great Buy This is a really good quality product that goes perfectly with the Vileda super mop. Would have cost £9.55 if I bought it from Amazon - so really happy that I got it for just £4 from ASDA! 24 April 2012
Rated 5 out of 5 by MissTrunchball Best mop and bucket you can buy This bucket with the Vileda Supermoccio mop is the best thing I've bought in a long time. It is so well made and takes the hard work out of mopping. Fantastic! 14 March 2014
Rated 5 out of 5 by Lutia Excellent Product, Even better Price Best mop buckets you can buy. I own a cleaning company and wouldn't use any other mop or bucket :) Also fantastic service and even better price. 26 March 2012


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Liverpool have reportedly overtaken Manchester United in the race to sign Southampton's Adam Lallana.
United have identified Lallana and his full-back team-mate, Luke Shaw, as key components in their planned £200million rebuilding project this summer.
However, the Sunday People claim the Anfield club have offered the £25million-rated Lallana £50,000 per week, with the Reds confident the added lure of Champions League football next term can trump anything United can offer.
Tottenham and Arsenal have also been linked with the 25-year-old, who is expected to be named in Roy Hodgson's World Cup squad.
Lallana put in another eye-catching display as Saints beat Everton 2-0 at St Mary's yesterday.


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About Us
Unicod4 is one of the leading software houses in Karachi, playing an important role in breakthrough of e-commerce industry of Pakistan. Unicod4 was a result of a passionate idea of its CEO, with eagerness to accomplish an aim and run a successful business. With many known local and international clients, Unicod4 has been contributing to IT industry for the past few years with its professionals having a combined experience of over 15 years.


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Review Details
ULTRA 6" tapering to 4" Hot Tub Cover 1.0# (R24-R34)
- Product Review (submitted on October 10, 2012):
- What a great hot tub cover, I love how its tapered for the water to run off. My factory cover collapsed in the winter due to heavy snow. this cover is everything that was advertised. It also looks great. I am looking forward to a reduction in my energy bi


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The Disability Income Plan was established January 1, 1988 in order to provide equitable replacement income for eligible state employees who become temporarily or permanently disabled for the performance of their duty prior to retirement. The determination of disability and eligibility for short-term benefits is made by the DOC Review Board and/or the Retirement System Medical Board. There are three types of disability benefits available to employees:
Short Term Disability
Short-Term Disability allows an employee to receive replacement income while temporarily or permanently disabled for the further performance of their duty prior to retirement. Short-Term disability also encourages disabled employees who are able to work to seek gainful employment after a reasonable period of rehabilitation. Eligible employees will receive 50% of their annual salary (plus 50% of longevity, if applicable), accrual of retirement service credits, and other benefits. The duration of the Short-Term benefits are from the 61st day of disability continuing for a period of up to one year.
The state will pay for the employee's health insurance coverage (the employee will have to pay for health insurance coverage of dependents) while the employee is on Short-Term Disability IF the employee has five (5) years of state service as of the date of the disability. Employees with less than five (5) years of service will have to pay for their health insurance coverage (the amount the state normally pays each month) as well as pay for coverage of any dependents while out on Short-Term.
Extended Short Term Disability
Extended Short Term Disability benefits are payable to eligible employees after the conclusion of the Short Term benefit and may be approved by the Plan's Medical Board for up to a maximum period of one (1) year for as long as the employee remains temporarily disabled. The Disability Income Plan's Medical Board makes the determination of eligibility for these benefits. Extended Short Term Disability allows an eligible employee to continue to receive 50% of their annual salary (plus 50% of longevity, if applicable).
Long Term Disability
Long-Term Disability benefits are payable to eligible employees after the conclusion of the Short-Term benefit period for as long as an employee is permanently disabled but not after he/she becomes eligible for an unreduced service retirement. The Disability Income Plan's Medical Board makes the determination of eligibility for these benefits. Long-Term Disability allows an eligible employee to receive 65% of their annual salary (plus 65% of longevity, if applicable), until such time as they are eligible for full service retirement. There are additional requirements for eligibility for long-term disability that are detailed in the booklet Your Retirement Benefits published by the Department of State Treasurer.
Click Here if you want to know How to Apply for Disability
|FAQ's - Short -Term Disability
A. If you and your doctor determine that your disability will keep you out of work for more than 60 continuous days, you should begin the application process for Short -Term Disability.
Q. How long will it be after submitting a complete application will I hear of a decision?
A. You will receive written notice of the Medical Board's decision. This process can take anywhere from six to twelve weeks.
Q. What if I am denied?
A. If the Medical Board denies an application an employee has ninety days to provide additional medical documentation for further review.
Q. Must I exhaust all my leave prior to receiving Short-Term Disability?
A. You must exhaust any available sick leave during the sixty day waiting period. After that, you may choose to continue to exhaust sick, vacation, or bonus leave.
Q. My Short-Term Disability is ending and I don't know whether to apply for Extended Short-Term or for Long-Term Disability. What should I do?A. You and your doctor should make this decision. Reminder: Extended Short-Term is for disabilities that are temporary and likely to end. Long-Term Disability is for disabilities that are permanent. You must have five years of contributing membership in the Retirement System to be eligible for Long-Term Disability.
|FAQ's - Long-Term Disability
Q. How soon can I make application for Long-Term Disability – my doctor says I'm permanently disabled?
A. You must exhaust a Short-Term Disability benefit period prior to making application for Long-Term Disability.
Q. I did not have five (5) years of service as of my date of disability but exhausted leave during the Short-Term benefit period giving me five (5) years of service at the end of the benefit period. Am I eligible to apply for Long-Term Disability?
A. Yes, if the exhaustion of leave during the Short-Term benefit period gives you five (5) years of creditable service in the Retirement System at the end of Short-Term, you are eligible to apply for Long-Term Disability.
Q. What if I am denied?
A. You will have ninety (90) days to provide additional medical documentation for further review.
Q. If approved, do I have to resign my position?
A. If approved for Long-Term Disability, you must terminate your employment (resignation due to approval of Long-Term Disability).
DOC Personnel Home |
DOC Home | NC General Assembly | State Personnel | State Personnel Manual
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about Todd FlowerdayA Roman Catholic lay person, married (since 1996), with one adopted child (since 2001). I serve a parish in music ministry.
about John Donaghy
John is a lay missionary since 2007 with a parish in western Honduras. Before that he served in campus ministry and social justice ministry in Iowa. His ministry blog is http://hermanojuancito.blogspot.com
He also blogs reflections on the lectionary and saints/heroes/events of the date at http://walktheway.wordpress.com
He'll be a long-term contributor here analyzing the Latin American bishops' document from their 2007 Aparecida Conference.
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Daily Archives: 12 May 2012
Ave Maria, Schubert
Last year, various people involved in our parish music ministry prepared a cd for our previous pastor who was just diagnosed with pancreatic cancer. It was a scramble to get various songs recorded before the semester's end, so we took … Continue reading
Posted in Music Leave a comment
More Trouble for the Legion
The AP released the news, hopped on by just about every Catholic outlet in the world over the past twenty-four hours, that the curia is pondering a small stack of dossiers in its possession: (T)he Legion (of Christ) confirmed it … Continue reading
Posted in Church News, Commentary, Politics, sex abuse 1 Comment
At the Core of the Paschal Mystery
This should be an intriguing match-up. Most of the NPM top-25 songs have been going down in polling in either the first round or in the second. Today we have the penultimate Theological Thirty-Two pairing, in which one will survive … Continue reading
Posted in 2012 Dance, Liturgical Music Leave a comment
GDC 263-264: Basic ecclesial communities
And lastly in this chapter covering "Loci and means of catechesis," let's look at base communities. Perhaps not so big in the US, but a significant part of the ecclesial experience in other countries, especially in mission lands: 263. Basic … Continue reading
RDCA II, 7: Choosing a Dedication Day
The name is important. I've outlined the text here to bullet points: 7. A day should be chosen for the dedication of the new church when the people can be present in large numbers, especially a Sunday. Since the theme … Continue reading


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Diversity among human leukocyte antigen (HLA) molecules has been maintained by host-pathogen coevolution over a long period of time. Reflecting this diversity, the HLA loci are the most polymorphic in the human genome. One characteristic of HLA diversity is long-term persistence of allelic lineages, which causes trans-species polymorphisms to be shared among closely related species. Modern humans have disseminated across the world after their exodus from Africa, while chimpanzees have remained in Africa since the speciation event between humans and chimpanzees. It is thought that modern humans have recently acquired resistance to novel pathogens outside Africa. In the present study, we investigated HLA alleles that could contribute to this local adaptation in humans and also studied the contribution of natural selection to human evolution by using molecular data.
Phylogenetic analysis of HLA-DRB1 genes identified two major groups, HLA Groups A and B. Group A formed a monophyletic clade distinct from DRB1 alleles in other Catarrhini, suggesting that Group A is a human-specific allelic group. Our estimates of divergence time suggested that seven HLA-DRB1 Group A allelic lineages in humans have been maintained since before the speciation event between humans and chimpanzees, while chimpanzees possess only one DRB1 allelic lineage (Patr-DRB1*03), which is a sister group to Group A. Experimental data showed that some Group A alleles bound to peptides derived from human-specific pathogens. Of the Group A alleles, three exist at high frequencies in several local populations outside Africa.
HLA Group A alleles are likely to have been retained in human lineages for a long period of time and have not expanded since the divergence of humans and chimpanzees. On the other hand, most orthologs of HLA Group A alleles may have been lost in the chimpanzee due to differences in selective pressures. The presence of alleles with high frequency outside of Africa suggests these HLA molecules result from the local adaptations of humans. Our study helps elucidate the mechanism by which the human adaptive immune system has coevolved with pathogens over a long period of time.
Keywords:Allelic lineage; Balancing selection; HLA; Out-of-Africa; Pathogen; Trans-species polymorphism
Modern humans (Homo sapiens) live in a wide variety of environments, ranging from polar to tropical regions. Physiological anthropologists have long addressed the issue of 'human adaptation' to a variety of environments (that is the ability of humans to survive in a changing environment). Molecular evolution and population genetics also focus on the adaptation of humans to environmental changes. The approach of physiological anthropology is mainly to investigate differences in physiological modifications among individuals or ethnic groups in various environments ('physiological polymorphism') in order to understand human adaptation. On the other hand, molecular evolution or population genetics seek indications of natural selection by comparing nucleotide sequences of a target gene. If a new mutation at a target locus confers advantage for fitness in a certain environment, such a mutation is expected to rapidly spread throughout a population because of positive natural selection. Methods to detect such a signal of natural selection have been developed. For instance, in a protein coding gene, an excess in the number of non-synonymous substitutions (that change the amino acid sequence) over synonymous substitutions (neutral mutation) suggests that positive selection or balancing selection has occurred during the evolution of the target gene. In addition, the relationship between an allelic frequency and the extent of linkage disequilibrium (LD) around the selected mutation help us to find an allele that has rapidly spread in a population [1]. The advantageous allele is expected to dramatically increase its frequency in a short time so that recombination does not substantially break down the LD around the selected site.
Humans live in various environments around the world. The endemic pathogens that humans are infected by in these areas differ and humans have evolved to deal with these pathogens. In the present study, we focus on polymorphisms in the major histocompatibility complex (MHC), which plays an important role in triggering immune reactions in response to pathogens, and we discuss the possibility that a human-specific MHC allele is involved in the immunological adaptation to a human-specific pathogen.
The MHC is a set of cell-surface molecules that are responsible for presenting antigens from pathogens to lymphocytes in jawed vertebrates. As such, it is an important genetic system for protection against infectious disease [2]. In humans, the MHC is termed human leukocyte antigen (HLA). The HLA genomic region is located on the short arm of chromosome 6 at 6p21.3, spanning approximately 4 Mbp and comprising 224 genes [3]. The region is classified into three subregions: class I, class II, and class III regions. Among HLA molecules, six class I and II molecules (HLA-A, B, and C of class I and HLA-DR, DQ, and DP of class II) are important for antigen presentation to T lymphocytes. Class I molecules mainly bind to peptides from cytosolic proteins and the HLA-peptide complex is recognized by CD8+ T cells. Class II molecules present extracellular antigens to CD4+ T cells. Class I molecules consist of two polypeptide chains, an α heavy chain encoded in the class I region, and a β2-microglobulin light chain encoded on chromosome 15. Class II molecules are composed of two polypeptide chains, α and β chains, encoded in the class II region. For instance, the DRA and DRB1 genes in the class II region encode the α and β chains, respectively, of the DR molecule. A peptide-binding region (PBR) was characterized with crystallography by Bjorkman et al. [4] for class I HLA-A and by Brown et al. [5] for class II HLA-DR. Molecular evolutionary studies of this region have revealed an enhancement of non-synonymous substitutions in the PBR, suggesting that the PBR is a target for balancing selection, which is responsible for the maintenance of HLA polymorphisms [6-10].
Polymorphisms in HLA genes have three unique features: (1) a large number of alleles, (2) a high degree of heterozygosity, and (3) remarkably long persistence time of the allelic lineage. These features are maintained by balancing selection but not by an increased mutation rate [11,12].
The chimpanzee (Pan troglodytes) is the closest extant relative of humans. Interestingly, chimpanzees appear to have resistance to several pathogens to which humans are susceptible, including HIV type 1 and human hepatitis B virus [13]. This indicates that the two species differ in their immune responses to these pathogens, and that possibly the pathogen recognition repertoire for MHC is different between the two species. Chimpanzees share some class II DRB1 allelic lineages with humans [14-16]. In humans, genetic variation and selective intensity on DRB1 are the greatest in the class II genes [17]. In humans, there are 13 DRB1 allelic lineages (HLA-DRB1*01, *03, *04, *07, *08, *09, *10, *11, *12, *13, *14, *15 and *16), while there are only four allelic lineages (Patr-DRB1*02, *03, *07 and *10) in chimpanzees [14-16].
Chimpanzees have stayed in Africa since their divergence from humans approximately six million years ago (MYA). On the other hand, modern humans have dispersed across the world from Africa from 100,000 to 50,000 years ago and have adapted to regions with various exogenous pathogens. This begs the question of how modern humans have acquired resistance to a variety of pathogens in different environments. Therefore, the present study investigated the evolution of HLA-DRB1 alleles that confer resistance to novel pathogens in humans. For this purpose, we studied nucleotide sequences of HLA genes using the IMGT/HLA database (http://www.ebi.ac.uk/imgt/hla/ webcite, [18]).
Materials and methods
Nucleotide sequences of humans, chimpanzees, rhesus monkeys (Macaca mulatta), and crab-eating macaques (Macaca fascicularis) were used for phylogenetic analyses. A dataset of human DRB allele sequences, including DRB1 and other functional DRB (DRB3, DRB4, and DRB5), was obtained from the IMGT/HLA database. The dataset of non-human primate DRB1 alleles was obtained from the IPD MHC NHP database (http://www.ebi.ac.uk/ipd/mhc/nhp/ webcite, [19]). In the database, there were many partial coding sequences (CDS) (mainly exon 2 sequences). Using incomplete sequences is likely to be misleading in analysis of the phylogenetic relationships among sequences; therefore, we performed phylogenetic analyses only for full-length DRB1 CDS. Because only partial sequences were available, we also excluded sequence data of the gorilla (Gorilla gorilla) and orangutan (Pongo pygmaeus) from the present analysis. We used two HLA-DQB1 alleles as outgroup sequences. Next, we removed sequences of potential recombinant alleles according to a method that assumes a binomial distribution of the ratio of substitutions in a particular region to that in the entire region [17,20-22]. For phylogenetic analyses, we used 104 complete CDS: 56 HLA-DRB1, 6 HLA-DRB3, 4 HLA-DRB4, 2 HLA-DRB5, 11 chimpanzee Patr-DRB1, 22 rhesus monkey Mamu-DRB1, and 3 crab-eating macaque Mafa-DRB1 alleles.
Brown et al. [5] identified 24 amino acids in the PBR of HLA-DRB1 genes. In addition to the defined PBR, we included three amino acid sites (positions of 57, 67, and 90; for a total of 27 amino acids), because Brown and collaborators have subsequently shown that the three sites are involved in the formation of peptide-binding grooves and peptide binding [23].
Multiple sequence alignment of nucleotide sequences and phylogenetic-tree construction were performed using the MEGA v5.10 software [24]. A maximum likelihood (ML) tree for the non-PBR region was constructed based on the Hasegawa-Kishino-Yano (HKY) substitution model [25] with the nearest-neighbor-interchange (NNI) ML heuristic search. The best-fit substitution model was estimated by MEGA. Bootstrap analysis was performed using 1,000 replications. The numbers of non-synonymous substitutions per non-synonymous site (dN) and synonymous substitutions per synonymous site (dS) were calculated using the modified Nei-Gojobori method [26] with a Jukes-Cantor correction [27]. The transition/transversion bias used in this calculation was estimated with the ML method in MEGA. The average divergence time of DRB1 alleles was estimated by the average of all pairwise dS values, and the time to the most recent common ancestor (TMRCA) of alleles was estimated from the maximum number of synonymous substitutions per site (dSmax). The divergence time was estimated by the following formula:
where μ is the neutral substitution rate of 10−9 per site per year at the MHC loci [9]. Pathogens recognized by HLA-DRB1 molecules were examined using the Immune Epitope Database (IEDB) (http://www.immuneepitope.org webcite, [28]). Information about HLA-DRB1 allele frequency among different human populations was collected from the NCBI dbMHC database (http://www.ncbi.nlm.nih.gov/gv/mhc webcite, [29]).
Results and discussion
Two phylogenetic groups of HLA-DRB1 alleles and human-specific HLA Group A
To examine phylogenetic relationships among DRB alleles in four primate species (HLA-DRB1/3/4/5, Patr-DRB1, Mamu-DRB1, and Mafa-DRB1), an ML tree was constructed from nucleotide sequences of the non-PBR region (Figure 1). Nucleotide sequences in the PBR were excluded for construction of the tree because they had an approximately ten-fold higher amino acid-altering (non-synonymous) substitution rate than synonymous substitutions due to balancing selection (Hughes and Nei [6,7]; Takahata and Nei [11]). When we focused on HLA-DRB1 alleles, we identified two distinct clades in the ML tree. We refer to these two groups as HLA Group A and HLA Group B. Of the 13 known HLA allelic lineages, seven lineages, including DRB1*03, *08, *10, *11, *12, *13, and *14, were assigned to Group A, while the remaining six lineages, DRB1*01, *04, *07, *09, *15, and *16, were assigned to Group B.
Figure 1. Maximum likelihood tree for nucleotide sequences (690 bp) in the non-peptide-binding region (PBR) of MHC DRB alleles. The sequence data of MHC DRB alleles, including those of humans, chimpanzees, and macaques, were obtained from IMGT/HLA and IPD databases. HLA-DRB1 alleles are indicated in bold. Arrow indicates the Patr-DRB1*03 lineage, which is a sister group of HLA Group A alleles. Only bootstrap values > 80% are shown. Two HLA-DQB1 sequences were used as an outgroup. The evolutionary distances were computed using the Hasegawa-Kishino-Yano (HKY) model. HLA Group A and HLA Group B indicate two major phylogenetic groups of HLA-DRB1 alleles. HLA, humans; Patr, chimpanzees; Mamu, rhesus monkeys; Mafa, crab-eating macaques.
In the ML tree, the Group B alleles showed trans-species evolution of polymorphisms with those in the chimpanzee (Patr-DRB1*02 and *07). Interestingly, 31 Group A alleles formed a monophyletic clade distinct from other primate DRB1 alleles, although the bootstrap value for supporting this cluster was not particularly high, suggesting that the Group A alleles are human-specific. Previous studies [14-16] have not identified this DRB1 monophyletic group in humans, because the nucleotide sequences used in those studies were limited to exon 2.
Both the mean and maximal dS values were larger in Group B (mean dS, 0.041; dSmax, 0.082) than in Group A (mean dS, 0.018; dSmax, 0.057) (Table 1). This indicates that most allelic lineages in Group B have been maintained for a longer time than those in Group A. Additionally, Group A alleles may have diverged more recently than Group B alleles. Based on these results, we propose two hypotheses for the monophyly of Group A: (1) Group A alleles specifically expanded in the human lineage or (2) the orthologs to Group A alleles were lost in chimpanzees. We estimated the divergence time for alleles in each group in order to test these hypotheses.
Table 1. The divergence time of the two HLA groups, HLA-Group A and HLA-Group B
Divergence time of alleles in HLA Groups A and B
The phylogeny showed a difference in divergence time between Groups A and B. The mean divergence times for Groups A and B were approximately 9 and 21 MYA, respectively, and TMRCAs were approximately 29 and 41 MYA, respectively (Table 1). These values suggest the presence of specific trans-species polymorphisms [10,30,31] in both groups, because the mean divergence time exceeded the speciation time of humans and chimpanzees [32-34]. Based on this result, we rejected the hypothesis that the HLA Group A allelic lineages specifically expanded in humans. However, the tree revealed that alleles in Group A did not intermingle with other non-human primate DRB1 alleles (Figure 1). The closest was the Patr-DRB1*03 lineage cluster (indicated by an arrow in Figure 1).
Furthermore, we estimated the TMRCA of Patr-DRB1*03 cluster to be 4.6 MYA (Figure 2), suggesting that the alleles in this cluster diverged in chimpanzees after their divergence from humans. Accordingly, only one allelic lineage leading to the cluster in extant chimpanzees existed in the common ancestral population of humans and chimpanzees. On the other hand, in humans, pairwise dS distances between HLA-DRB1 alleles suggested that seven allelic lineages existed in the ancestral population (Figure 2). Therefore, the common ancestral population likely possessed at least eight allelic lineages.
Figure 2. Divergence times of HLA Group A and Patr-DRB1*03 alleles. The dashed line represents the speciation event of humans and chimpanzees. Times to most recent common ancestor (TMRCAs) were estimated based on the maximum genetic distance at synonymous sites (dSmax).
Although alleles in Group A formed a single clade in the ML tree of primate DRB alleles, the TMRCA was 29 MYA, which is significantly older than six MYA (that is the speciation time of humans and chimpanzees). Thus, the molecular clock for DRB1 alleles may have been skewed by various factors, such as back or parallel mutations (multiple mutations) or recombination/gene conversion. Indeed, in the Group A allele sequences, there was segregation of 21 synonymous sites. Among them, ten were singletons with a unique nucleotide observed only once in the sampled alleles, and 11 were phylogenetically informative sites. Among 55 pairs of 11 informative sites, 13 pairs were phylogenetically incompatible with each other. This incompatibility was likely the result of either recombination/gene conversion or multiple mutations at a single site. In the event of recombination/gene conversion, however, double recombination in a relatively small region or a conversion tract with a small size should be considered. Multiple mutations are a more likely cause of this incompatibility. To examine whether the presence of multiple substitutions masked an accurate estimate of the TMRCA, we tested the accuracy of the correction for multiple substitutions in the calculation of dSmax.
For this purpose, we estimated the maximum number of synonymous substitutions in a different way. First, we placed synonymous substitutions observed in the Group A alleles on each branch of the ML tree parsimoniously (Figure 1 and Additional file 1: Figure S1) and re-counted the number of synonymous substitutions (KS) in each pair of Group A alleles. The maximum KS was thirteen (KSmax = 13). TMRCA was calculated from this KSmax divided by the mean number of synonymous sites (LS = 223). As a result, the TMRCA of the Group A alleles was estimated to be 29 MYA. This showed good agreement with the TMRCA estimated by the Jukes-Cantor correction (29 MYA). Because there was no bias in our method of estimating TMRCA, we considered it to be reliable.
Additional file 1: Figure S1. Synonymous substitutions occurring on each branch of the ML tree for HLA Group A alleles in Figure 1. HLA-DRB1*09:01:02 was used as an outgroup sequence. The asterisk represents a synonymous substitution at a singleton.
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Probability of maintaining seven human-specific HLA Group A allelic lineages over six million years
A method for calculating the probability, gnk(t) [35], that there were k allelic lineages among n extant lineages for t in N generations under balancing selection is available. In the present study, we tried to calculate the probability gnk(t) for seven ancestral allelic lineages being maintained since approximately six MYA among a sample of 31 Group A alleles (n = 31). However, because HLA-DRB1 also contains the 25 Group B alleles, the 31 Group A sequences are only a part of the samples in the entire HLA-DRB1. There were no means to determine the effective population size (Ne) of these subpopulations, which was required for the calculation of gnk(t); therefore, we could not calculate the probability of maintaining the current Group A alleles for six million years.
The effective population size Ne of modern humans is smaller than that of chimpanzees [36-38], and the eight allelic lineages in the ancestral population have likely been lost more frequently from the human lineage than the chimpanzee lineage. Nevertheless, the number of allelic lineages in humans is seven times larger than that in chimpanzees. This supports the hypothesis that natural selection selectively maintained Group A alleles in humans. It is important to understand the biological reasons why these seven lineages have been maintained only in humans.
Specific peptides bound to HLA Group A alleles
It is possible that the HLA Group A allelic lineages have been because they bind to peptides derived from human-specific pathogens. Thus, we examined pathogens and their specific peptides recognized by each of the Group A and B allelic lineages based on information of experimental data from the IEDB database (Table 2). There were ten pathogens that produced peptides bound only by Group A alleles (for example, human papillomavirus type 11 (HPV-11) and influenza B virus (IBV)), and some of them were candidates for human-specific pathogens. In fact, in addition to HPV-11, Bordetella pertussis and measles viruses have been reported to be human-specific pathogens [39,40] (Table 2). Moreover, the IBV is restricted to humans with the exception of an infection identified in seals stranded on the Dutch coast [41]. At present, however, the repertoire of peptides bound by each allele is limited in the experimental data. Future studies will establish whether chimpanzees and macaques MHC are able to bind HLA Group A-specific peptides.
Table 2. The comparison of specific pathogen bound by HLA-DRB1 molecules between Group A and Group B
In HLA Group B, although some pathogens infect not only humans but also other animals (for example, Brucella ovis and Burkholderia mallei), candidates for human-specific pathogens (for example, Helicobacter pylori) were included. This suggests that some Group B alleles might be also involved in local adaptation in humans.
The frequency distributions of eight HLA-DRB1 alleles (HLA-DRB1*0301, *08:02, *11:01, *11:02, *11:03, *11:04, *12:01, and *14:01) that recognize Group A-specific pathogens were investigated using information in the NCBI dbMHC database (Additional file 2: Figure S2). The frequency distributions of HLA-DRB1*08:02, *12:01, and *14:01 were high outside Africa, suggesting that the frequency of the DRB1 molecules might have increased since the human species disseminated outside Africa.
Additional file 2: Figure S2. Allelic frequency of HLA-DRB1 molecules that bind HLA Group A-specific pathogens. The allelic frequency data was obtained from the NCBI dbMHC database.
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Chimpanzees appear to have lost a relatively large number of alleles from the Group A allelic lineage while humans have maintained several allelic lineages since their speciation. The examination of genetic variation in MHC class I Patr-A, Patr-B, and Patr-C loci suggested that the genetic variations in chimpanzees have been severely reduced [42]. In this previous study, it was hypothesized that a selective sweep caused the loss of genetic diversity at MHC loci in chimpanzees in order to avoid widespread viral infection, such as that with chimpanzee-derived simian immunodeficiency virus, prior to a subspeciation of the common chimpanzee and bonobo (Pan paniscus) approximately two MYA. Although it is not known whether such selective sweep resulted in the loss of some DRB1 allelic lineages in chimpanzees, reduced genetic variation at the three class I loci in chimpanzees may have been linked to the relatively small number of Patr-DRB1 allelic lineages.
A phylogenetic analysis of the HLA-DRB1 gene identified two major groups of alleles, Groups A and B. Our findings suggest that Group A is human-specific and has been maintained by balancing selection in humans, while chimpanzees may have lost their counterparts to these allelic lineages due to different selective pressure. Some Group A alleles can bind to peptides derived from human specific pathogens and these showed a high frequency in populations outside Africa. Therefore, these alleles may have increased in frequency after the Out-of-Africa event. Our results imply that some of HLA Group A alleles may have contributed to local adaptation of humans.
In the present study, we identified a candidate human-specific HLA-DRB1 allelic group. However, the sample size of chimpanzees was smaller than that of humans. Specifically, there were at least 88 chimpanzees used in published studies [14,15,43-45], while the HLA-DRB1 alleles were detected in thousands of human individuals. Therefore, there is possible sampling bias among chimpanzees. The common chimpanzees are classified into at least four subspecies, which are, Pan troglodytes troglodytes, P. t. verus, P. t. ellioti, and P. t. schweinfurthii, in Mammal Species of the World[46]. In addition to the common chimpanzees, bonobo samples should also be included in the phylogenetic analyses of DRB1 alleles. To exclude the possibility that our finding is an artifact of sampling bias, we plan to increase the sample size of chimpanzees in future studies, which will help validate the present estimates.
In the present study, DRB1 alleles of rhesus monkeys and crab-eating macaques formed a taxon-specific clade with the exception of HLA-DRB4*01 sequences. All sampled alleles in the two macaques formed a sister clade with HLA Group A alleles in the ML tree but not with HLA Group B alleles (Figure 1). In the future, the reason why the DRB1 alleles of macaques formed a large monophyletic group should be investigated.
It is difficult to verify that a molecule in HLA Group A can recognize human-specific pathogens. In recent years, there has been increasing information on peptide-HLA binding. Future studies must examine the relationships among HLA alleles, binding peptides, and pathogens in order to elucidate the mechanisms by which modern humans have adapted to a variety of environments around the world.
The contribution of natural selection to local adaptation in humans was evaluated from genomic data. The genomic data provide a universal framework for understanding human evolution and enable quantitative analysis of the operation of natural selection. We believe that molecular genetics techniques can shed light on some important issues in physiological anthropology.
bp: base pair; CDS: coding sequence; dN: number of non-synonymous substitutions per non-synonymous site; dS: number of synonymous substitutions per synonymous site; dSmax: maximum genetic distance at synonymous sites; HKY: Hasegawa-Kishino-Yano; HLA: human leukocyte antigen; HPV-11: human papillomavirus type 11; IBV: influenza B virus; IEDB: Immune Epitope Database; KS: number of synonymous substitutions; KSmax: maximum number of synonymous substitutions; LD: linkage disequilibrium; LS: mean number of synonymous site; MHC: major histocompatibility complex; ML: maximum likelihood; MYA: million years ago; Ne: effective population size; NNI: nearest-neighbor-interchange; PBR: peptide-binding region; TMRCA: time to most recent common ancestor.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
YY carried out data analysis, and wrote the draft of manuscript. YS was the primary supervisor. Both authors have read and approved the final manuscript.
This work was supported by Grant-in-Aid for Scientific Research on Innovative Areas from the Ministry of Education, Culture, Sports, Science and Technology of Japan (22133007). We owe special thanks to Drs. Naoyuki Takahata and Jun Gojobori for providing valuable comments.
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Science 2000, 288:1051-1053. PubMed Abstract | Publisher Full Text
De Groot NG, Otting N, Doxiadis GGM, Balla-Jhagjhoorsingh SS, Heeney JL, van Rood JJ, Gagneux P, Bontrop RE: Evidence for an ancient selective sweep in the MHC class I gene repertoire of chimpanzees.
Proc Natl Acad Sci U S A 2002, 99:11748-11753. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Gyllensten UB, Sundvall M, Erlich HA: Allelic diversity is generated by intraexon sequence exchange at the DRB1 locus of primates.
Proc Natl Acad Sci U S A 1991, 88:3686-3690. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Bak EJ, Ishii Y, Omatsu T, Kyuwa S, Tetsuya T, Hayasaka I, Yoshikawa Y: Identification and analysis of MHC class II DRB1 (Patr-DRB1) alleles in chimpanzees.
Tissue Antigens 2006, 67:134-142. PubMed Abstract | Publisher Full Text
De Zorzi M, Caggiari L, Ahlenstiel G, Rehermann B, De Re V: Description of two new major histocompatibility complex (MHC) class II DRB1 [Pan troglodytes (Patr)-DRB1] alleles.
Tissue Antigens 2008, 71:490-492. PubMed Abstract | Publisher Full Text
Wilson D, Reeder D: Mammal Species of the World: A Taxonomic and Geographic Reference. 3rd edition. Baltimore: Johns Hopkins University Press; 2005.


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We Take All Trades (Boats, Cars, Etc?)
These came out of freshwater Outerlimits.
2 – Merlin II BBC Blocks
Current 4.40" Bore
*Blocks Need Machining
4 – Dart Iron Eagle Cast Heads
Intake Valves – 2.25"
Exhaust Valves – 1.88"
Valves / Springs / Retainers
*Heads should be inspected before use
2 – Eagle 4340 Crankshafts
Factory Balanced
Chamfered Oil Holes – Rods + Mains
*Should be inspected prior to use
2 – Crane Camshafts
Lifters / Push Rods
15 – JE 757F Pistons / Eagle I Beam Rods 6.535"
*Should be inspected prior to use. We cleaned up about 1/2 of them and they look good.
2 – Holley EFI Systems
2 Cast Fuel Injected Manifolds
16 Injectors
2 Throttle Bodies
16 Ignition Coils
2 PCM's
2 Wiring Harness
2 Timing Wheels
2 Stainless Flame Arrestors
4 – Polished Aluminum Valve Covers
2 – Deep Sump Oil Pans
2 – Sea Water Pump Assy / With Brackets / Polished Pulleys
2 – Alternators / With Brackets / Polished Pulleys
2 – 'Dummy' Distributor Plugs
2 – Timing Covers / Timing Chains
2 – Crank Pulleys
All marine vessels, motors, drives or related parts and or equipment, vehicles, cars, trucks, motorcycles, trailers or any other goods or services depicted on this website (goods) for sale are being sold as is. Pier 57 offers no warranties, expressed or implied as to the quality or condition of any goods sold. Pier 57 makes no representation as to the quality or condition of the goods. Though some goods described on this website may be described as new or used, some components of the goods may be new and others used. Pier 57 encourages all buyers to obtain a Marine Survey prior to purchase, of both new or used goods (structurally & mechanically) because, Pier 57 does not represent and/or warrant the quality or condition of the goods listed for sale on this website, new or used. Pier 57 does not warrant accuracy of the description of the goods listed for sale on this website.


text: Our parts are of the same quality as our products - superior. Toro manufactures and stocks only the best parts – blades, belts, bedknives, reels, filters, hoses and more. We make sure to supply the highest-quality part for your equipment and each part meets specific standards - standards that are higher than industry norms. Toro uses the same parts for service that we use in each machine that comes off the manufacturing line. The reason for this is clear. Our customers expect Toro machines to perform every day. Replacement parts must work for the first time, every time. That isn't the exception - it's the main part of the complete Toro commitment to quality.


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Ms. Colleen Madalene Donovan and Mr. Leif Erik Munkel, both of Hattiesburg, MS, would like to announce that they are engaged!
Ms. Donovan is the daughter of Mr. Daniel Donovan and the late Mrs. Carol Donovan of Sterling, MA. Colleen Donovan is a 1999 graduate of Boston College in Chestnut Hill, MA, and she received her Master's degree in 2001 from Syracuse University in Syracuse, NY. She currently works for WDAM television as the station's weekday co-anchor of their Live at Five Broadcast.
Mr. Munkel is the son of Mr. Robert and Mrs. Mary Munkel of Branson, MO. Leif Munkel is a 2001 graduate of Concordia College in Moorhead, MN. He currently works as the communications coordinator for the Hattiesburg Convention Commission.
The couple is planning an October 2008 wedding in Hattiesburg.
Originally published September 14, 2008


text: DSpace Collection: http://hdl.handle.net/10204/890 2015-10-13T17:02:45Z Comparison of two detection algorithms for spot tracking in fluorescence microscopy images http://hdl.handle.net/10204/7858 Title: Comparison of two detection algorithms for spot tracking in fluorescence microscopy images Authors: Mabaso, M; Withey, D; Twala, B Abstract: Research in biological image analysis plays an important role in understanding the underlying mechanisms of cellular processes, which may lead to better knowledge of certain aspects of the cell function. The primary analysis of biological images requires the detection and tracking of hundreds of spots. In this paper we presents an approach for the tracking of spots in microscopy images based on the modification of the algorithm presented by Feng et al. The improved algorithm consists of replacing the original detection algorithm, Feature Point Detection(FPD) with Isotropic Undecimated Wavelet Transform (IUWT). The tracking algorithm based on Interacting Multiple Model (IMM) remains fixed. The performance of the presented method, IMM-IUWT along with two others, MHT, based on Multiple Hypothesis Tracking (MHT) and IMM-FPD, were validated on numerous challenging realistic synthetic image sequences, and their performance was evaluated using root mean square error (RMSE) metrics. The results indicate that the presented method outperforms the original method. At high level of signal to noise ratio (SNR), it is noted that the performance of the modified method, IMM-IUWT, is similar to that of MHT method. The quantitative comparative results demonstrated the importance of spot detection in tracking contexts. Description: Proceedings of the 2014 PRASA, RobMech and AfLaT International Joint Symposium, Lagoon beach, Cape Town, 27-28 November 2014 2014-11-01T00:00:00Z Graphene for improved femtosecond laser based pluripotent stem cell transfection http://hdl.handle.net/10204/7612 Title: Graphene for improved femtosecond laser based pluripotent stem cell transfection Authors: Mthunzi, P; He, K; Ngcobo, S; Khanyile, T; Warner, JH Abstract: Pluripotent stem cells are hugely attractive in the tissue engineering research field as they can self-renew and be selectively differentiated into various cell types. For stem cell and tissue engineering research it is important to develop new, biocompatible scaffold materials and graphene has emerged as a promising material in this area as it does not compromise cell proliferation and accelerates specific cell differentiation. Previous studies have shown a non-invasive optical technique for mouse embryonic stem (mES) cell differentiation and transfection using femtosecond (fs) laser pulses. To investigate cellular responses to the influence of graphene and laser irradiation, here we present for the first time a study of mES cell fs laser transfection on graphene coated substrates. First we studied the impact of graphene on Chinese Hamster Ovary (CHO-K1) cell viability and cell cytotoxicity in the absence of laser exposure. These were tested via evaluating the mitochondrial activity through adenosine triphosphates (ATP) luminescence and breakages on the cell plasma membrane assessed using cytosolic lactate dehydrogenase (LDH) screening. Secondly, the effects of fs laser irradiation on cell viability and cytotoxicity at 1064 and 532 nm for cells plated and grown on graphene and pure glass were assessed. Finally, optical transfection of CHO-K1 and mES cells was performed on graphene coated versus plain glass substrates. Our results show graphene stimulated cell viability whilst triggering a mild release of intracellular LDH. We also observed that compared to pure glass substrates; laser irradiation at 1064 nm on graphene plates was less cytotoxic. Finally, in mES cells efficient optical transfection at 1064 (82%) and 532 (25%) nm was obtained due to the presence of a graphene support as compared to pristine glass. Here we hypothesize an up-regulation of cell adhesion promoting peptides or laminin-related receptors of the extracellular matrix (ECM) in cell samples grown and irradiated on graphene substrates. By bringing together advances in optics and nanomaterial sciences we demonstrate pathways for enhancement of pluripotent stem cell biology. Description: Copyright: 2014 Wiley. This is an ABSTRACT ONLY. The definitive version is published in Journal of Biophotonics, vol. 7(5), pp 351-352 2014-05-01T00:00:00Z The potential for a fish ladder to mitigate against the loss of marine-estuarine-freshwater connectivity in a subtropical coastal lake http://hdl.handle.net/10204/7502 Title: The potential for a fish ladder to mitigate against the loss of marine-estuarine-freshwater connectivity in a subtropical coastal lake Authors: Weerts, SP; MacKay, CF; Cyrus, DP Abstract: Increasing water demand in coastal regions has resulted in the construction of weirs and barrages in coastal freshwaters. These form barriers to migrations of estuarine and euryhaline marine fishes and crustaceans. This study assessed the impact of loss of marine–estuarine–freshwater connectivity caused by a weir at the outlet of Lake Mzingazi, a coastal lake on the subtropical east coast of South Africa. Facultative catadromous estuarine and euryhaline marine fishes were most affected by the weir, which blocked their upstream migration. These fishes were absent in the lake and were prevented from accessing potential nursery habitat. Crustaceans were less affected by the outlet weir. Salinity tolerances of most penaeid prawn species occurring in southern African estuaries prevent penetration into freshwater reaches of connected systems. Movements of palaemonid prawns which either spawn in or have larvae dependant on estuarine water, and the marine spawning brachyuran Varuna litterata, were not completely impeded by the weir. These species have larvae and juveniles that can climb over the weir in its present form. Historically, the lake served as an estuarine nursery and its physico-chemistry and habitats still offer viable and valuable habitat to estuarine species. There is therefore great potential value in installing a fish ladder at the lake outlet that can be used by juvenile fishes and crustaceans. Re-establishing marine–estuarine–freshwater connectivity here will reinstate a natural migration route that has been impeded for over 70 years and partially restore the ecosystem to its original state. Description: Copyright: 2013 Water Research Commission. Thus is an abstract only. The definitive version is published in Water SA, vol. 40(1), pp 27-38 2014-01-01T00:00:00Z Defining pyromes and global syndromes of fire regimes http://hdl.handle.net/10204/6896 Title: Defining pyromes and global syndromes of fire regimes Authors: Archibald, S; Lehmann, CER; Gómez-Dans, JL; Bradstock, RA Abstract: Fire is a ubiquitous component of the Earth system that is poorly understood. To date, a global-scale understanding of fire is largely limited to the annual extent of burning as detected by satellites. This is problematic because fire is multidimensional, and focus on a single metric belies its complexity and importance within the Earth system. To address this, we identified five key characteristics of fire regimes—size, frequency, intensity, season, and extent—and combined new and existing global datasets to represent each. We assessed how these global fire regime characteristics are related to patterns of climate, vegetation (biomes), and human activity. Cross-correlations demonstrate that only certain combinations of fire characteristics are possible, reflecting fundamental constraints in the types of fire regimes that can exist. A Bayesian clustering algorithm identified five global syndromes of fire regimes, or pyromes. Four pyromes represent distinctions between crown, litter, and grassfueled fires, and the relationship of these to biomes and climate are not deterministic. Pyromes were partially discriminated on the basis of available moisture and rainfall seasonality. Human impacts also affected pyromes and are globally apparent as the driver of a fifth and unique pyrome that represents human-engineered modifications to fire characteristics. Differing biomes and climates may be represented within the same pyrome, implying that pathways of change in future fire regimes in response to changes in climate and human activity may be difficult to predict. Description: Copyright: 2013 National Academy of Sciences. This is an ABSTRACT ONLY. The definitive version is published in Proceedings of the National Academy of Science of the United States of America, pp 1-6 2013-04-01T00:00:00Z The fire-vegetation-climate system: how ecology can contribute to earth system science http://hdl.handle.net/10204/6895 Title: The fire-vegetation-climate system: how ecology can contribute to earth system science Authors: Archibald, S Abstract: Since the time of Darwin, Wallace and Schimper, botanists and ecologists have been interested in understanding the global distribution of vegetation. Such questions are becoming more pressing as Earth System modellers attempt to predict the current and future state of global vegetation. A key complexity that is currently not well captured by Earth System models is that vegetation is not always deterministically responsive to climate and soils. Feedbacks within the Earth System, top-down controls such as fire and herbivory, and historical contingencies related to the phylogenetic origins of the biota all contribute towards explaining the current distribution of vegetation. Here I discuss some of these aspects in more detail and, in particular, how this might shape the character and distribution of the global savanna biome. Specifically, I highlight the role that fire plays as both a driver and a response in the fire-vegetation-climate system. Description: Copyright: 2013 Elsevier. This is an ABSTRACT ONLY. The definitive version is published in South African Journal of Botany, vol. 86, pp 139 2013-05-01T00:00:00Z The soil and plant determinants of community structures of the dominant actinobacteria in Marion Island terrestrial habitats, Sub-Antarctica http://hdl.handle.net/10204/6697 Title: The soil and plant determinants of community structures of the dominant actinobacteria in Marion Island terrestrial habitats, Sub-Antarctica Authors: Sanyika, TW; Stafford, W; Cowan, DA Abstract: Marion Island is a Sub-Antarctic island made up of distinct ecological habitats based on soil physiochemical, plant cover and physical characteristics. The microbial diversity and ecological determinants in this harsh Sub-Antarctic environment are largely uncharacterized. Actinobacteria have diverse ecological functions related to soil and plant functioning. This study was aimed at characterizing the diversity and community structures of the dominant actinobacteria in the distinct habitats and to identify their determinant soil and plant characteristics. Using the 16S rRNA gene, the denaturing gradient gel electrophoresis patterns and clone library diversity were correlated with the soil and plant characteristics. Multivariate statistical methods were also used to identify determinant soil and plant characteristics. Salinity and pH were the most important soil determinants, and a number of important site-specific plant species may have been important. The Coastal Fellfield Habitat was dominated by sequences of the suborders Micrococcineae (44%) and Propionibacterineae (18%), with salinity identified as the principal determinant. The Cotula Herbfield Habitat was dominated by Frankineae (37%) and Streptosporangineae (38%), which were correlated with organic nutrient concentrations. The Wet Mire Habitat was dominated by Acidimicrobineae (61%), with moisture and organic carbon content as principal components. Culture-dependent studies were complementary to culture-independent studies with the majority of actinobacteria isolated not identified in 16S rRNA gene clone libraries. This study demonstrates how the soil physiochemical characteristics and plant species independently determine the community structures of the dominant actinobacteria in distinct ecological habitats. These factors subsequently influence their ecological adaptation, roles and functions. Description: Copyright: 2012 Springer Verlag. This is an ABSTRACT ONLY. The definitive version is published in Polar Biology, vol. 35(8), pp 1129-1141 2012-08-01T00:00:00Z


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Board Election Declarations Due
LACBA members seeking nomination by the Nominating Committee for LACBA officer and at-large trustee positions to be voted on in June are required to submit a declaration of intent to seek such nomination no later than March 17, 5:00 p.m. Applications not timely received will not be considered. Declarations of intent should be sent to Sally Suchil, Executive Director, Los Angeles County Bar Association, PO Box 55020, Los Angeles, CA 90055.
Members of the Nominating Committee may submit additional names of members for nomination to at-large trustee positions prior to the second meeting of the Nominating Committee. The Nominating Committee nominates one or more LACBA member(s) for each position of at-large trustee to be filled at the June election. (At-large trustees serve two-year terms.) It also nominates one or more current or former trustees for each of the offices of president-elect, senior vice president, and vice president. (Officers serve one-year terms. President-Elect Eric Webber assumes the presidency on July 1, 2011.)
The committee consists of LACBA's immediate past president, current president, president-elect, three members selected by lot from those who have represented LACBA sections on the Board of Trustees during the previous four years, three members designated by the Barristers president, three members selected by lot from those who have represented the affiliated associations on the board during the previous four years, and seven trustees-at-large selected by lot from those who have served on the board during the previous four years.
No member of the committee may be nominated, either by the committee or by a written nomination, for an elective office, including the position of trustee, during the year of service on the committee. Nominees for president-elect, senior vice president, and vice president must have served as a LACBA trustee for at least two years prior to July 1, 2011. Nominees for the six at-large trustee positions (term of service 2011-13) must 1) be a LACBA member for at least three years prior to July 1, 2011, and 2) have demonstrated service for LACBA through active membership on one or more LACBA or section committees or in LACBA public service projects for a total of at least two years.
The committee must complete its work by April 15. Nomination by the committee is not the only way for a member's name to be placed on the annual ballot for a trustee or officer position. A member meeting the eligibility criteria stated above may run for a trustee or officer position upon filing with the executive director a written nomination signed by at least 25 members. Nomination forms are available at the LACBA offices.
Election Deadlines
• March 15, 2011: Selection by lot of seven at-large, three affiliate, and three section trustee members of Nominating Committee
• March 17, 2011: Prospective nominees' submission of declaration of intent by 5:00 p.m.
• April 15, 2011: Selection of nominees
• May 2, 2011: Members' submission of written nominations to run for trustee and officer positions by 5:00 p.m.
• May 5, 2011: Candidates' statements submitted to the Executive Director by 5:00 p.m.
• May 31, 2011: Ballots distributed
• June 21, 2011: Election


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- Musician's Friend Best Selection, Price & Service. Guaranteed.
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Call our Gear Heads
at: 800-449-9128
12 timeless classics from the late Jim Croce: "Bad, Bad Leroy Brown," "I Got a Name," "I'll Have to Say I Love You in a Song," "Operator (That's Not the Way It Feels)," "Photographs and Memories," "Time in a Bottle," "Workin' at the Car Wash Blues," and more.
Our product catalog varies by country due to manufacturer restrictions. If you change the Ship-To country, some or all of the items in your cart may not ship to the new destination.


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I love this bag in every way.
Well made in Taiwan not China. Has quality parts. Filled easily. Pipes are good, especially the douche nozzle gave a great enema for my boyfriend.
Ladies, this is the best syringe made. I've searched long and hard and I believe this is the perfect size for the job. The nozzle fits just right and I love the clean feeling I have when used with Zonite douche concentrate. The best part is that my husband also loves this syringe as it does such a thorough job.
The nozzle is long, curved and thick.
Femine Compact Douche Enema Syringe Bulb By Cara, Luxury Size - 1Ea
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So April is all about lightening up. Enjoying everyday pleasures and recognising the small things that make your world a brighter place.
When Gretchen talks about this she talks about thinking beyond the obvious, not ignoring them but recognising some out of the ordinary things too, things that at the time might feel stressful and difficult – like throwing a party at home or being a parent or partner – things that may bring immediate stress but actually end up making you happier. She describes the happiness "fog" – that transparent thing that is there but not always that noticeable making you happy.
So I'm trying really hard to think of things because this is something I'm actually pretty good at particularly when I'm in a crisis. I count my blessings and recognise that things are pretty good in the grand scheme of things, it completely affects my happiness in a great way.
Well I'll start at the beginning…
Take Photographs
Taking photographs is my go to thing, from iphone to DSLR it soothes me, the process of snapping something I find interesting or beautiful, making it in to something cohesive or artistic later in editing, gives me such a buzz and I love it so much. I want to try and take a photograph every day in April, it will keep me focussed on something artistic.
Remember Today is Only 24 Hours.
No matter how great the day is, or how awful, then it will only last 24 hrs. I sometimes think too long term and forget that when things are awful that they won't last for ever. Particularly when it comes to food, I have to remember that one bad day is not a bad life.
Enjoy Planning
I see a lot of exciting things in my future, things that I need to have a plan to achieve fully and enjoy fully. I know this seems to contradict my previous resolution but stay with me. I like to plan, it's something that makes me happy even if the likely hood is I may never achieve those goals, they may be dreams but they are my dreams. In order to make the more achievable dreams come true, we as a family are going to have to have a plan of action, a savings plan and a commitment. Man and I also have a lot of stress in our lives right now and so a plan of what we want in the future will help keep us calm and focussed on the end prize.
Enjoy Who You Are
Blog When I Want To…And other things…
This is about doing more things that I enjoy because I enjoy them not because it's expected of me or because I feel obligated. If I don't want to cook I won't, if I want to cook a three course meal then I will! If I want to blog then I will, if I don't – guess what? I won't.
As part of my Enjoy who you are resolution I need to take time to be me, being me is pretty good if I'm honest so I'm going to do what I want.


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Anti Essays offers free essay examples to help students with their essay writing.
Below is a free essay on "Personally Identifiably Information" from Anti Essays, your source for free research papers, essays, and term paper examples.
"Personally Identifiably Information". Anti Essays. 10 Oct. 2015
Personally Identifiably Information. Anti Essays. Retrieved October 10, 2015, from the World Wide Web: http://www.antiessays.com/free-essays/Personally-Identifiably-Information-95973.html


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Interesting. Have you considered adding flocking behaviour to the fire imps so that a group which are friends will tend to flee in a similar direction? Or would this make catching them too hard if they are all breathing fire?
TinyKeep is an upcoming action roguelike game with procedurally generated dungeons, emergent monsters, deadly traps and mysterious secrets.
Learn how TinyKeep's AI handles monster retreat and defensive behaviours! Also a brief mention on the minor improvements over Part 1 - thanks to community feedback.
Posted by phi6 on May 19th, 2013
Before I crack on to Part 2 (Retreat and defense) I would like to quickly cover the minor improvements we made since Part 1 of this AI series. Thanks to community feedback we've added a few more features, removed others and hopefully will have created more realistic behaviour.
In short, the changes are:
1. If the Skeleton loses sight (raycasting towards hero) and smell (raycasting towards dropped breadcrumbs), he will still continue to head towards the location where the hero was last detected and investigate further.
2. If the Skeleton still cannot find the hero, it will make a best guess where it thinks he has gone and will set a patrol route to the nearest waypoint. Given these changes, we've removed the alert level as we've found this feature has become redundant and offers no extra intelligence. The "smell trails" have been reduced to balance the extra added behaviour - this also seems to help limit cheating in the AI.
Ok, let's bring on the fun stuff!
Fire Imps (and other monsters marked with the "Shy" personality flag) will run away from the player if he gets too close. Put him in this level of distress for too long, and he'll breathe fire on you.
The interactive demo is here on the TinyKeep website along with all of the previous demos:
As with all of these AI behaviours, what you see on the video and demo are the default settings for each of the personality flags (Aggressive, Cautious, Shy, Lumbering etc...). However we've designed our AI editor to be a lot more flexible than that - so our game designer Ben can choose from a multitude of variables to customize his monsters:
We know that this will give us the ability to create really unique and varied monsters, and part of the fun of the game is figuring out the behaviours of each one.
Hope you enjoyed the post, and let us know if you have any questions about the AI!
Interesting. Have you considered adding flocking behaviour to the fire imps so that a group which are friends will tend to flee in a similar direction? Or would this make catching them too hard if they are all breathing fire?


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No, no, no, certainly not. The following comes from academician Robert Kuttner writing about carbon (or so he thinks), markets (or so he thinks) and too much freedom (or so he thinks). BTW, ivory is 30% collagen, which contains the "C" word. Not sustainable. Unfortunately, gross ignorance of real science seems to be flourishing.
'But in the world where we actually live, markets do not produce the "right" price. There are many small examples of this failure, but also three immense ones that should have discredited the libertarian premise by now. Global climate change is the most momentous. The price of carbon-based energy is "correct"—it reflects what consumers will pay and what producers can supply—if you leave out the fact that carbon is destroying a livable planet. Markets are not competent to price this problem. Only governments can do that.'


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Houses - Then and Now
I love the old homes and buildings in Menard, though several have seen better days. This section is to provide information on where people lived in the past......and if the home still exists today.
In Pierce's book, "The Free State of Menard", the following drawing is said to have been the "first house" in Menard.
For sources on the "FIRST" home or store built in Menard....go here.
NOTE: While I strive for accuracy in all transcriptions, please be advised that typing errors may be present. I would suggest you always verify my online information with a copy of the actual record.
All contents of this web site are the property of Alicia Brown unless otherwise noted. You are free to copy or print out pieces of information for your personal genealogical research, but nothing on this site may be used in other ways without my permission.
Have a Question or Comment About My Web Site?
Due to the increasing amount of "spam" I'm receiving from email "harvesters", I only publish my email link in jpg form. I am sorry for any inconvenience this might cause you...but I'm really tired of the "advertising trash" that is filling up my email box..
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