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Programme Updates. Programme Overview and - Video Symposium on Challenging Cases now available.
In the long-term, does an on-axis incision cause significantly lower surgically induced astigmatism than an off-axis incision?
(results will display both Free Papers & Poster)
Session Details
Session Title: Cataract Surgery Outcomes / Femto
Session Date/Time: Monday 15/09/2014 | 08:00-10:30
Paper Time: 09:39
Venue: Auditorium
First Author: : S.Perera UK
Co Author(s): : H. Ali
Abstract Details
Cataract incisions affect the corneal curvature and placing the incision on the steeper axis (on axis) has shown to reduce surgically induced astigmatism (SIA). Most of these studies have short term follow up. Corneal curvature has been shown to change with time. This study aims to ascertain if in the long term, an on axis incisions cause less SIA than an off axis incision.
Hospital Practice
30 consecutive patients who had cataract surgery by the same surgeon, and had automated corneal keratometry done by the IOL Master device both pre and post operatively 1-7 years (mean 4yr.) following the cataract procedure was included in the study. The same surgical technique was used for all eyes with a 2.75mm. primary clear corneal incision placed centred on the horizontal axis and enlarged to 3.00mm for lens insertion. Eyes with the steeper axis between 75-105 degrees were considered to be off axis while those between 0-15 and 165-180 degrees were considered as on axis. SIA was calculated using the vector analysis method from the pre operative and post-operative K readings.
The mean SIA in the on axis group was 0.567D (SD0.32) with a range of (0.18-1.03D) while that of the off axis group was 0.635D (SD0.34). range (0.11-0.95D) The difference of 0.068D in the SIA seen between the two groups was not statistically significant. (p>0.05) The range of SIA was also similar in the two groups.
This study suggest that in the long term no statistically significant difference is seen in SIA between on axis and off axis incisions.
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Earthquake swarm in Nevada
Belski's Blog
Earthquake swarm in Nevada
Belski's Blog
Updated Saturday afternoon.......
Over the last few months there have been a large number of small earthquakes in the northwest corner of Nevada. In the last 24 hours there have been 13 earthquakes of 2.5 magnitude or greater. The strongest today was 3.3
Since the beginning of August, there have been more than 550 quakes greater than a 2.0 magnitude with many more less than 2.0.
42 quakes have been stronger than 3.0 in the last 3 months.
The strongest one occurred yesterday with a magnitude of 4.7.
This is a fairly remote area with two towns about 40 miles away. Both have a population of about 2,500.
In the past, other earthquake swarms in Nevada have led up to the main quake of 5.0 which resulted in some minor damage.


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Horse racing is considered one of the most popular sports in Australia. It comes in third for the most attended sport in all of Australia. Close to two million people are registered to the 350 plus race courses all over the Australia area. It was first commenced after the European settlement and is now one of the most popular sports in the world. Well over 12 billion dollars has been wagered annually on gambling and horse racing in Australia.
There are two types of horse racing in Australia. There is flat racing and there is races over fences or hurdles. The racing over fences and hurdles tends to take place in Victoria and more of South Australia rather than anywhere else. Flat racing is typically the most popular style of racing in most of Australia. It is a form of horse racing in which horses run on an even level race course. The race course often consists of a certain amount of miles and features no obstacles. Racing over fences and hurdles is a form of horse racing in which horses follow a track and must jump over hurdles and other obstacles to complete the race. Both forms of racing are extremely popular in Australia. Australia does not offer harness racing which is typically popular in other parts of the world.
Horse racing first became popular in Australia way back in 1700 and 1800. Ever since then Australia has a deep rooted history in horse betting and racing. There are many different horse meetings and clubs all over Australia. One of the first organized racing events was held in Tasmania in 1814. There are also a large number of breeders and stud farms located throughout Australia. Horses are often bred and exported to America where they become very successful. Some of the most notable and popular horses in horse racing history have been bred, studded and raced in Australia. Australia is also home to some of the top jockeys in the world. Some of the most notable jockeys include Edgar Britt, Roy Higgins and Nash Rawiller just to name a few.
Most consider Victoria to be the home of racing in Australia. It hosts many international races like the Melbourne Cup and the Cox Plate. The Melbourne cup is a very popular annual horse race that takes place in Melbourne, Australia. It is considered one of the richest races and turf in the world. It is conducted by the Victoria Racing Club and draws in millions and millions of viewers and visitors. It first started back in 1861 and is considered one of the oldest and most respected races. Cox Plate is a thoroughbred horse race that is specifically for horses that are aged three years old and over. It is also for underweight horses as well as horses with other conditions. It also takes place in Melbourne, Australia. It was first introduced way back in 1922 and is still incredibly popular today.
Horse racing is also very popular in New South Wales and often viewed through online casino Australia apps. There are many different notable tracks and they host some of the largest carnival races in Australia. Racing in New South Wales is typically governed by racing NSW. Racing in South Australia is typically governed by thoroughbred racing SA Limited. South Australia typically hosts a very popular Easter racing Carnival during Easter weekend every year that is fun and exciting and offers large prize payouts.
Racing throughout Queensland is typically governed by the racing Queensland. They have one principle Club named Brisbane Racing Club. The Brisbane Racing Club is in Australian organization that is based in Brisbane. It was founded in 2009 and has conducted more than 95 race meetings just one year. Racing in Western Australia is actually governed by the racing and wagering Western Australia. It is a government-owned body that follows their own unique rules and guidelines. They hold races at Belmont Park as well as Ascot race course. One of the most popular Western Australia races is the Perth Cup. It is held each New Year's at Ascot. It draws in hundreds and thousands of attendees. Tasmania also offers horse racing and is governed by the Tasmanian thoroughbred racing Council. Some of the race meetings take place at the King Island Racing Club. Recently racing in the Northern Territory has been governed by the thoroughbred racing NT. Australian Capital territory racing is governed by the principal Club the Canberra Racing Club.
Betting in Australia is fun and unique. There are three or four main ways to bet on a horse race in Australia. There are licensed on track bookmakers who offer fixed odds for betting. They mostly pay on wins and places. There is off track betting which is usually controlled by the individual state government but must adhere to a general guideline. Many license on-course bookmakers now accept bets over the phone as well as over the internet. Betting may also be done online with person to person exchange betting. Through this members set their own prices and will pay a percentage of their winnings in commission after the race. Ever since 1913 Australia has used the automatic totalisator which can automatically calculate the race odds. As time goes on many horse racing facilities in Australia are beginning to operate through online and mobile apps. It makes it easier and much more convenient for betting.
The automatic totalisator was invented in Australia by a man named George Julius. George Julius was a creator who invented a way of automatically calculating the odds of a race by using a series of betting patterns. It is still used till this day.
One of the many reasons why horse racing is so popular in Australia is because of the big annual races that are held there. Aside from the Melbourne Cup and the Perth Cup other top races include the Victoria Derby, the VRC Oaks meet, the Golden Slipper Stakes and the Caulfield Cup. Each event attracts hundreds of thousands of attendees and millions and millions of viewers. Horse racing will continue to be one of the most popular sports in all of Australia. Not only do they host some of the top name horses and jockeys but Australia offers some of the most notable trainers in the business. Bart Cummings is one of the most notable Australian trainers and has been the trainer of 12 different Melbourne Cup winners over the years. Overall with all of the great trainers and jockeys who hail from Australia and all of the great deep rooted history it is easy to see why horse racing is one of the top watched and attended sports in all of Australia.


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Update to Burst Main – St. Joseph
Date(s) - 13/04/2017
All Day
Cleavers Hill
The repair of the burst main at Cleavers Hill, St. Joseph is complete.
The immediate surrounding areas may continue to be impacted for a short time while the system returns to normal.
The BWA regrets any inconvenience this disruption in service may have caused.
Original Report
The Authority has sustained a burst on a 3-inch main at Cleavers Hill, St. Joseph
work continues on a burst 3-inch main at Chimborazo in the same parish.
Only the immediate surrounding districts may be affected by low pressure and water outages.
Crews have been assigned to the affected areas.
The BWA regrets any inconvenience this disruption in service may cause.


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SANTA CLARITA (KABC) --A 46-year-old Santa Clarita woman has pleaded guilty to unleashing two dogs on an Internal Revenue Service employee.The IRS worker was trying to serve a summons to Lisa Blechman's husband. She taped the summons to their front door and then started walking back to her car when Blechman released the two dogs.
The employee was not hurt, but prosecutors say she was scared the larger dog would maul her.
Blechman is scheduled to be sentenced March 31.


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Cosmological history of stars and metals
Observatoire de Midi-Pyrénées, 14 avenue Édouard Belin, 31400 Toulouse, France
2 Institut d'Astrophysique de Paris, 98bis Boulevard Arago, 75014 Paris, France
3 Department of Physics, Astrophysics, 1 Keble Road, Oxford OX1 3NP, UK
Corresponding author: R. Sadat, email@example.com
Accepted: 15 January 2001
We study the evolution of stellar content and the chemical enrichment of the universe averaged over the whole population of galaxies by means of a series of chemo-spectro photometric models that take into account the metallicity and dust obscuration effects. We investigate various classes of cosmic star formation rates (CSFR) histories consistent with current estimations. We are able to reproduce a variety of observational constraints such as the emissivities at rest-frame 0.44, 25, 60 and 100 μm of the local universe and also the overall shape of the extragalactic background light from UV/NIR galaxy counts and the cosmic infrared background (CIB) from DIRBE/FIRAS measurements. We find that the CIB at 140 μm is crucial for discriminating between the CSFR histories. The best-fit model to this constraint seems to favour mid-infrared derived CSFR at low-z and a flat CSFR at higher z consistent with the most recent estimations, although the shape of the CSFR at high redshifts has little impact on the FIR/submm part of extragalactic background. We suggest that the bulk of the CIB energy is produced by a population of moderately obscured normal galaxies lying at . We then derive the global chemical enrichment and stellar content of the universe and find that this model predicts metallicities in good agreement with the metallicity in DLAs given that some outflow of metal-enriched gas from galaxies is assumed, but it overproduces the current present-day stellar mass density and NIR luminosity density.
Key words: cosmology: diffuse radiation / galaxies: evolution / galaxies / intergalactic medium


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Yes your decision is absolutely right. there are so many universities which offer distance learning courses and these courses are actually worth.
Yes your decision is absolutely right. there are so many universities which offer distance learning courses and these courses are actually worth.
Yes its worth doing an distance MBA, it gives you the same knowledge and also adds value, . But choosing a correct institute is important, its about how well-known & renowned the institute is, whether the institute is Govt recognized or not, whether they'd provide recruitment support for you to kick start you career after completion of your course. Distance MBA will give you flexibility of studying, continuing your present life as it is with an ease.
One such institute that I can suggest you is Welingkar Institute, Welingkar Institute of Management Development (WeSchool) was established in the year 1977 with a focus on giving a new dimension to Business Studies. It is part of the Shikshana Prasaraka Mandali, which has 43 prominent institutions across the country under its purview.
It has a strong educational legacy of over thirty years, WeSchool believes in the excellence of quality education catered in relevance to the current times and industry demands; thus giving students a complete educational experience.
WeSchool has gained the distinction of being one of the few institutes whose hybrid programs are recognised by UGC-DEB, Govt. of India.
If you wish to do a course you can visit Welingkar's Website @ Welingkaronline.org
Yes, If you are working then distance education is also the best option for MBA. You can continue with your job with the study.
Absolutely, with advanced means of communications like video calls etc.
Distance learning saves so much time.
Specially for working professionals it's a boon
and easy way to keep them upgraded.


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The health benefits of Dalia are known to all. That is why large numbers of people incorporate Dalia in their regular diet. It is because Dalia helps in keeping digestive system in good health and proper working order. Keeping in mind the same fact, Patanjali store of Baba Ramdev Ji has prepared and presented Patanjali Pushtahar Dalia.
Key features
a. Patanjali Pushtahar Dalia incorporates all the essential nutrients required by the human body. Hence it supports overall good health of the body naturally.
b. It is light and hence easily digestible by all.
c. It is very much delicious and hence liked by all.
d. It is rich in fibre content that is quite important for the digestive system.
e. It regulates bowel movements.


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July is coming to a close, and the Minor League season is hitting its homestretch, with Tim Locastro, L.B. Dantzler, Dalton Pompey, K.C. Hobson, Adam Loewen and Kevin Pillar turning in dazzling performances. Join the radio broadcasters of the Blue Jays' farm system as we take you inside the ballpark!
Starting Lineup
- Jesse Goldberg-Strassler, Lansing Lugnuts (A)
- Interview with Fred McGriff
- Trey Wilson, Bluefield Blue Jays (Rookie)
- Ben Wagner, Buffalo Bisons (AAA)
- Chris Shuker, New Hampshire Fisher Cats (AA)
- Tyler Murray, Dunedin Blue Jays (A-Adv.)
- Slavko Bekovic, Lansing Lugnuts (A)


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2512 NW Prairie View Rd, Platte City, MO 64079, United States
Service Is Our Passion
Enjoy our free airport shuttle and free WiFi
Our non-smoking hotel offers guests free shuttle service to KCI airport as well as local shuttle service, so that you and your companions can travel around town with ease. Pet-friendly guest rooms offer free high-speed WiFi, microwave, mini-refrigerator, iron, and ironing board. Onsite amenities include coin-operated laundry, business center, fitness center, indoor pool, and hot tub


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Within a shower, you will find that it would always be better when you have decided to make some considerations, which will mean that, you can always get to have the ability of making sure that everything which you might get to do can work to your advantage at all times, it will mean that you will be able to have something which can be appealing to you. Learn more about The Woodlands Frameless Shower Doors, go here.
When getting to consider on how you can make your shower to look amazing, you will have always to consider what it is that can work to your advantage, thus being able to know what it is that can be able to fit perfectly into your home or even with your shower thus being able to make sure that you can attain some value for your money. Find out for further details on The Woodlands Auto Glass right here.
Some of the doors which you can get to consider are the frameless doors, one of the things which you will note will be that they are beautiful, which means that they will get to add some beauty to your shower, this will, therefore, make sure that you can always have a shower which you can be contented at all times while in and also know of what it is that can be of some help to you at all times.
By conducting your assessment on the shower doors, there should always be a few things which you need to look for, meaning that, you have been able to know of something which can be of some help to you, this will make sure that everything which you might get to do will ensure that indeed you can be contented or appeased.
When making your consideration on the quality, you also should not forget about the budget, you have to make sure that you have been able to stick with your budget, when using the internet to look for the best shower doors, you will find that there can be a variety in which you can choose from, which will mean that, you can always get to make sure that you can find the door which you can afford and also the one which will be amongst the best quality.
On the other hand, you need to make sure that you can focus on the installation, this will mean that you have to look for a service provider who will be having the ability of making sure that you can attain the best services available, ones who can make sure that you can be appeased and also that you can have a good looking shower.


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The October. 71th Anniversary.. (1988)
Film-document №9619 1 part, Duration: 0:09:56 Price category A
Not Digitized
Availability of videosources
Needs clarification of possibility of digitizing in HD.
AnnotationThe film is about the parade and demonstration related to celebration of the 71th anniversary of the October Socialist Revolution.
Temporary descriptionCity of Moscow. The parade of troops of the Moscow garrison at Red Square on the 71-th anniversary of the October Revolution. Parade commander, Army General Kochetov gave a report to the Minister of Defense of the USSR D. T. Yazov. The passage of troops, military hardware on Red Square. Gymnastic parade, a workers' demonstration on Red Square. On the podium of the Mausoleum of Lenin: Mikhail Gorbachev, Ryzhkov, AP Biryukov, EM Primakov.
Reel №1
The film is dedicated to the parade and demonstration of workers on the occasion of the 71st anniversary of the Great October Socialist Revolution.


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Westminster Shorter Catechism
Q. 67. Which is the sixth commandment?
A. The sixth commandment is, "Thou shalt not kill [b]."
Westminster Shorter Catechism
Q. 69. What is forbidden in the sixth commandment?
A. The sixth commandment forbiddeth the taking away of our own life, or the life of our neighbour unjustly, or whatsoever tendeth thereunto [e].
- Westminster Larger Catechism
Q.136. What are the sins forbidden in the sixth commandment?
A. The sins forbidden in the sixth commandment are, all taking away the life of ourselves [c], or of others [d], except in case of
- • public justice [e],
- • lawful war [f], or
- • necessary defence [g];
- the neglecting or withdrawing the lawful and necessary means of preservation of life [h]; sinful anger [i], hatred [k], envy [l], desire of revenge [m]; all excessive passions [n], distracting cares [o]; immoderate use of meat, drink [p], labor [q], and recreations [r]; provoking words [s], oppression [t], quarreling [v], striking, wounding [w], and whatsoever else tends to the destruction of the life of any [x].
Be a "Berean" (Acts 17:11)
Why Important?10,000/yr vs. 10,000 per day. What is the tendency of legitimizing purposeful, orchestrated, intentional, organized, systematic, institutionalized killing?
Review: "Capital Punishment" - Atonement
- Genesis 9:4-6
- But flesh with the life thereof, which is the blood thereof, shall ye not eat. And surely your blood of your lives will I require. At the hand of every beast will I require it, and at the hand of man; at the hand of every man's brother will I require the life of man. Whoso sheddeth man's blood, by man shall his blood be shed; for in the image of God made He man.
- Numbers 35:33 - [In ContextRead ChapterOriginal Hebrew]
- So ye shall not pollute the land wherein ye are: for blood it defileth the land: and the land cannot be cleansed [kaphar] of the blood that is shed therein, but by the blood of him that shed it.
- Deuteronomy 21:8 - [In ContextRead ChapterOriginal Hebrew]
- Be merciful [kaphar], O LORD, unto thy people Israel, whom thou hast redeemed, and lay not innocent blood unto thy people of Israel's charge. And the blood shall be forgiven them.
lawful war [f], Deuteronomy 20 throughout. Ver.1 When thou goest out to battle against thine enemies, and seest horses, and chariots, and a people more than thou, be not afraid of them: for the LORD thy God is with thee, which brought thee up out of the land of Egypt, etc.
Ver.2 So it shall be, when you are on the verge of battle, that the priest shall approach and speak to the people.
Jeremiah 48:10 Cursed be he that doeth the work of the LORD deceitfully, and cursed be he that keepeth back his sword from blood.
- Micah 4:3
He shall judge between many peoples, And rebuke strong nations afar off; They shall beat their swords into plowshares, And their spears into pruning hooks; Nation shall not lift up sword against nation, Neither shall they learn war anymore.
Isaiah 2:4
He shall judge between the nations, And rebuke many people; They shall beat their swords into plowshares, And their spears into pruning hooks; Nation shall not lift up sword against nation, Neither shall they learn war anymore.
Swords Into Plowshares
- Joel 3:10
Beat your plowshares into swords And your pruning hooks into spears; Let the weak say, 'I am strong.'"
Jamieson-Fausset-Brown Bible Commentary
10. Beat your ploughshares into swords-As the foes are desired to "beat their ploughshares into swords, and their pruning hooks into spears," that so they may perish in their unhallowed attack on Judah and Jerusalem, so these latter, and the nations converted to God by them, after the overthrow of the antichristian confederacy, shall, on the contrary, "beat their swords into ploughshares, and their spears into pruning hooks," when under Messiah's coming reign there shall be war no more (Isa 2:4; Ho 2:18; Mic 4:3).
let the weak say, I am strong-So universal shall be the rage of Israel's foes for invading her, that even the weak among them will fancy themselves strong enough to join the invading forces. Age and infirmity were ordinarily made valid excuses for exemption from service, but so mad shall be the fury of the world against God's people, that even the feeble will not desire to be exempted (compare Ps 2:1-3).
War Cleanses the Nations
We see God's sovereignty in Isaiah 13 verses 3ff., where God judges Babylon using the Medes:
- The soldiers of the Meden army are His "sanctified ones." (v. 3)
- The Lord "commands" them.
- He calls His "mighty ones." (v. 3)
- He musters the armies together. (v. 4)
- The armies of the nations
- come from the Lord
- to destroy the earth. (v. 5)
- (Not the whole earth, just the land that pertains to Babylon.)
- The destroying armies are "the weapons of His indignation" (v. 5).
- This is the day of the Lord's "wrath and fierce anger" (vv. 6-9).
- The coming destruction is a terror for these evil doers (v. 11).
- God is sending a sword to cut them down (v. 15).
- Isaiah 10 is a prophecy against Israel. God is going to use Assyria in the same way He used the Medes against Babylon.
- Verse 5 says the Assyrian army is the rod of the Lord's anger. Their weapons are the Lord's indignation.
- Ultimately it is God Who is sending these unGodly barbarians. Their purpose is the judgment of a hypocritical nation, those who have incurred God's wrath. God gives the army "a command" (v. 6a) to steal, confiscate wealth and property, and in general to do the things the Assyrian army does best: to rape and pillage Israelites, treating them like sewage in the street (v. 6b).
- God says -- to Israel -- "I give him a command to spoil." A command to beat their plowshares into swords. It is truly evil and ugly, a violation of the Sixth Commandment. You would be sinning to voluntarily enlist in the Assyrian Army.
Somewhere between 50-90 million human beings were killed in World War II. The clear winner of that war was Marxism/Communism, which took Eastern Europe and Asia, over 1 billion slaves. Would Moses or Isaiah have considered the wars of the 20th century to be Godly, lawful wars? How about the wars we're already seeing in the 21st century? Will they make atonement and cleanse the land?
What is the tendency of the belief that we should be training for war?
End of Lesson, Day 124
For Further Study:
- War, Capital Punishment, and "The Sword"
- Blessed Are the Peacemakers
- Is War Ever "Just"?
- Would Jesus Celebrate Memorial Day?
- War is an Evil, Though Predestined by God


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Australian Cattle Council : Yearbook 2008
page 62 C aTTL e COUNCIL OF a USTR a LI a Y ea RBOOK 2008 The NT cattle herd grew to 2 million head in 2007 and cattle numbers shipped through the port of Darwin rose to 283,000 head which was considerably higher than last year. The pastoral industry maintained its status as the Territory's third largest GDP earner, raking in $400 million in direct benefits, and with a roll-on effect worth about $880 million for the local economy. Our members remain optimistic about the future, as they position themselves for predicted future expansion in both our domestic and export markets. NT Cattlemen's association remains strong, focused and vocal in lobbying for the rights of its members, and it also maintains its unique position of covering at least 98 per cent of the NT cattle herd on its membership books. Cattle export matters, Drought conditions, local government reform, unacceptable beef roads and environmental matters have been continuing key issues for Territory pastoralists during 2007. With 47% of the total NT production destined for the export trade in asia, the northern beef industry continues to focus on partnerships and programs to strengthen this trade. Once again the export industry program has delivered a successful market development and promotional program in key South east asian trading markets – congratulations to the MLA/Livecorp joint program. LOCaL gOveRNMeNT ReFORM In October 2006 the NT Minister for Local government announced a new local government reform package. This local government reform package in the Northern Territory will involve the establishment of an experimental framework of Municipal and Shire councils that will incorporate the whole of the NT by 1 July 2008. presently only 2% of the NT is incorporated in local government including the 4 major urban regions of the NT. Local government reform in the Northern Territory has far-reaching implications for the business community. Reforms of this magnitude will have long lasting effects on the Territory's economic and social sustainability. The pastoral Industry, as part of a large group of business and community groups, have a number of concerns about the reforms and the lack of consultation with the wider community. In particular, some of the reform proposals put forward by the NT government and other stakeholders would harm the pastoral Sector and business community, and as a consequence, the NT economy. The association has called on the NT government to slow down and get the process and reforms right, for the benefit all Territorians eNvIRONMeNTaL MaTTeRS The association called on the NT government to stop playing political games with the Douglas Daly Region and has requested that the NT government heed the hardship suffered by farmers of the Douglas Daly as a result of three successive moratoriums on land clearing. The association has asked that the pastoralists of the Douglas Daly be considered as a special case and exempted from the latest moratorium on land clearing on the basis of scientific evidence provided to government by its own departments, by two committees, and by an NTCa adaptive Management plan prepared in conjunction with the government that small scale farming and cattle grazing in the district can be carried out sustainably. Currently less than 5% of the Daly Region is cleared, which is hardly the 'destruction' portrayed by out of context pictures in environmentalists' advertising. What's more, cleared land is planted with improved pastures guided by significant scientific research from institutions such as the government's Douglas Daly Research Farm (DpIFM). The association will continue to call on government to allow land development for pasture production so that industry can meet its commitments to our Se asian customers. DROUgHT Central australian pastoralists are faced with the 4th and 5th year of severe drought conditions with little to no rain over the past 36 months. With only 12 properties declared in the 2nd year of drought under the NT Northern Territory Cattlemen's association as the president of the Northern Territory Cattlemen's association (NTCa), I am delighted to report that the northern industry is in good shape and getting stronger every year. Roy Chisholm, President, NTCA
Yearbook 2009
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In 1734, the General Assembly looked to the past when naming Orange County after William of Orange, who ruled initially with Queen Mary and then solo as King William III.
To encourage settlers to move to the western lands, those who arrived in Orange County by January 1 of 1734/5 (at that time, the "new year" started on March 1) were exempt from taxes for the next three years. The delay maid it difficult to start the new county's operations, limiting its ability to raise funds for three years.
The county boundaries extended "northerly by the grant of Lord Fairfax and westerly by the utmost limits of Virginia." The vague descriptions were common for counties on the frontier of settlement. The boundaries of the Fairfax grant had not been defined at this time.
Orange County originally was bounded by the Rappahannock River on the north, the Rapidan River on the south, and on the west by the "utmost limits of Virginia"
Source: Memoir of Col. Joshua Fry (p.26)
Gordonsville served as a hospital center for the Confederacy during the Civil War, because railroads could transport the wounded there from battlefields in northern and central Virginia
Source: Library of Congress, Atlas of the War of the Rebellion, Southeastern Virginia and Fort Monroe, Va. (1892)


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I picked up a book at the library a couple weeks ago that looked like something a bit different from the usual reference book on the writing craft. Writing With The Master by Tony Vanderwarker turned out to be amusing and both discouraging and encouraging, also to have a few gold nuggets in how one goes about writing a novel.
We follow Vanderwarker's journey as he writes a suspense novel with the help of author, John Grisham. They live near one another and had a friendship going before Mr. Grisham offered to be a mentor to help Tony write his book. Tony, a one-time ad writer, had already written several novels but none had been published. Self-publishing does not ever come into play with him, he wants to do it the way other famed authors have--through a publishing house via an agent.
Who wouldn't be excited if John Grisham offered to serve as a mentor? If it happened to be me, I'd give very serious thought to the offer before accepting. Acceptance means a hard road ahead, which is exactly what Tony found. Every time he thinks he's come up with a masterpiece of an outline that John will love, he's shot down by the myriad criticisms from his mentor. Again and again, Tony rewrites the outline. How can he give up and disappoint his mentor?
We watch as he follows one frustrating path after another, as he interacts with his wife about his writing, and also another important part of his life as a volunteer for the Piedmont Environmental Council, an environmental agency. Any writer will wince at the thought that Tony writes seven outlines for his book, receives critiques from author Grisham that send him back to the drawing board over and over. Despite the harsh critiques he receives, the two remain friends.
I'm not going to spoil the conclusion of the book for you, but any writer contemplating writing a novel would do well to read this book. It's entertaining, filled with humorous self-jabs, but honest in what it takes to become a published novelist. Like many other reader/reviewers, I didn't find enough about John Grisham in the book to satisfy my curiousity about him. But yes, I did get a picture of how hard the man works to turn out one bestseller after another. At this point, he doesn't need the money so he's got to do it for the love of birthing a book from conception to delivery.
There are little tips from Grisham throughout the text, things Tony derives from their lunchtime conversations and the notes scribbled by the author on Tony's outlines. I would have liked to see a compiled list of all of these tips somewhere in the final pages of the book but there wasn't one. If you read the book, I'd suggest keeping a pad and pencil handy so you can jot down a tip when you read it.
Is this book going to help you write a novel? I doubt it but it will help you decide if you want to give it a try or not. It will help you determine if you have the drive, the passion, the stamina it takes. I found Writing With The Master to be an entertaining read. Check out further info at Amazon.


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Krallice kicked things off with an older song ("Dimensional Bleedthrough" I think) which was surprisingly good. I've seen the band 3 (or 4? I can't recall) times now and they've done an excellent job in sounding tighter and more energetic since I caught them two years ago with A Storm of Light and Wolves in the Throne Room. Unfortunately, the rest of the set suffered as their newer songs showcase more attention to vocals (deeper, less interesting ones), heavier and more technical riffs and structures that really detract from the hypnotic melodies that were so absorbing from their s/t. They got a good response from the crowd and I heard mixed things while waiting at the bar.
Thou, as I expected, dominated the show, putting Krallice in their place as openers, and distracting me from the headliner as they couldn't be lived up to. Beyond the cohesion between the band, their spot on execution and excellent sound, their set-list was worth it alone. Starting with my favorite song "They Stretch out Their hands," and eventually "Burning Black Coals And Dark Memories," "Here I Stand Head In Hand" (possibly my third favorite of theirs) and ending with a very honest and energetic cover of Black Sabbath's "Into the Void." While the stage was too big for them (they're the kind of band that thrives in small settings) they lived up to my expectations and easily stole the show.
Thou - Burning Black Coals And Dark Memories from (((unartig))) on Vimeo.
Wolves in the Throne Room are a far cry from the band I obsessed over with their first release. Whether it's the relative stardom they've accomplished, or maybe my own shifts in taste, the band bored me this time around. I fondly recall them playing Brooklyn nearly 4 years ago (right around the release of Two Hunters) and thought they were one of the best acts I had ever seen. That show was such a clash compared to this one as they practically played in the crowd and let their music speak for itself that time. However, in recent years, the band's employed incense, smoke, banners, candles, and other gimmicks that feed into this "natural aesthetic" that just seems a bit silly in such a big venue like The Bell House. Rumored to be a possible fairwell tour, I'm happy I was able to catch them, but I'm happier to think of a time when they were a small outfit with only two releases and no gimmicks.
Wolves in the Throne Room | NYC @ The Bell House | 12 Sep 2011 from (((unartig))) on Vimeo.
Thou and WITTR continued on two days later to Death By Audio (this time with support by the excellent Mutilation Rites). Unfortunately I missed the show, but I assume Thou really shined in the small death trap that is Death By Audio.


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We are proud to be providing the music for the Peoria Ballet's Nutcracker presented on December 11th and 12th at the Peoria Civic Center.
Get tickets for the 11th at 7:30pm – Get tickets for the 12th at 2:00pm
A recent article in the Peoria Journal Star
Interview of Academy Director Rebekah von Rathonyi on WCBU 89.9's "Out and About"
A message from our Artistic Director David Commanday:
"The Heartland Festival Orchestra and I are thrilled to collaborate with the Peoria Ballet as we make LIVE music part of the Nutcracker this holiday season. We will bring Tchaikovsky's masterpiece to life in this all-new production. I have been the conductor for countless Nutcrackers in my career – none will be more special or more meaningful to me than this one"


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Female germ cells are essential for organogenesis of the ovary; without them, ovarian follicles do not form and functional and structural characteristics of the ovary are lost. We and others showed previously that when either Wnt4 or β-catenin was inactivated in the fetal ovary, female germ cells underwent degeneration. In this study, we set out to understand whether these two factors belong to the same pathway and how they maintain female germ cell survival. We found that activation of β-catenin in somatic cells in the Wnt4 knockout ovary restored germ cell numbers, placing β-catenin downstream of WNT4. In the absence of Wnt4 or β-catenin, female germ cells entered meiosis properly; however, they underwent apoptosis afterwards. Activin βB (Inhbb), a subunit of activins, was upregulated in the Wnt4 and β-catenin knockout ovaries, suggesting that Inhbb could be the cause for the loss of female germ cells, which are positive for activin receptors. Indeed, removal of Inhbb in the Wnt4 knockout ovaries prevented female germ cells from undergoing degeneration. We conclude that WNT4 maintains female germ cell survival by inhibiting Inhbb expression via β-catenin in the somatic cells. Maintenance of female germ cells hinge upon a delicate balance between positive (WNT4 and β-catenin) and negative (activin βB) regulators derived from the somatic cells in the fetal ovary.
Citation: Liu C-F, Parker K, Yao HH-C (2010) WNT4/β-Catenin Pathway Maintains Female Germ Cell Survival by Inhibiting Activin βB in the Mouse Fetal Ovary. PLoS ONE 5(4): e10382. https://doi.org/10.1371/journal.pone.0010382
Editor: Laszlo Orban, Temasek Life Sciences Laboratory, Singapore
Received: November 5, 2009; Accepted: April 7, 2010; Published: April 29, 2010
Copyright: © 2010 Liu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This research is supported by NIH HD046861 and Basal O'Connor Starter Award to H. H. Yao. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Female germ cells not only are essential for the propagation of species but also play an important role in ovarian organogenesis. Presence of germ cells is required for the formation of follicles, the functional unit of the ovary. Disruption of germline-specific genes, such as Dazla and Factor in the germline α (Figla) led to degeneration of oocytes and failure in folliculogenesis [1], [2], [3]. In addition, if germ cells are lost after follicle formation, characteristics of the ovary vanish [4], [5], [6]. Therefore, defects in female germ cell survival have detrimental impacts on fertility and reproductive health of the affected individuals.
Once they have migrated to the gonad, germ cells start to differentiate by following their intrinsic programs as well as responding to instructions from the somatic environment [7], [8]. In mouse fetal ovary, female germ cells enter meiosis around 14.5 dpc (day post coitum) as a result of the action of mesonephros-derived retinoic acids [9], [10] and then immediately arrest at the prophase of meiosis I [8], [11]. Ideas of the influences of somatic cells on female germ cell development have long been proposed. Gene screening schemes have yielded putative candidate genes that may play roles in this process. Among these candidates, the Wingless-type MMTV integration site (Wnt) family of genes, including Wnt4, Wnt5a, Wnt6, and Wnt9a, are found expressed in the somatic cells of the fetal ovary [12], [13]. WNT proteins are known to be involved in cell fate decision and cell cycle regulation [14]. These ovarian WNTs may work synergistically or redundantly in regulating female germ cell development. Previous studies by our lab and others have shown that inactivation of Wnt4 or β-catenin resulted in degeneration of female germ cells starting at 16.5 dpc [15], [16], [17], [18]. In this study, we provide evidence that β-catenin lies downstream of WNT4 to suppress expression of activin βB. When the Wnt4/β-catenin pathway is inactivated, upregulation of activin βB leads to loss of female germ cells.
Effects of somatic cell-specific inactivation of β-catenin on female germ cell apoptosis and meiosis
In our previous study, we generated a somatic cell-specific β-catenin conditional knockout (cKO) mouse by introducing the Steroidogenic factor 1-cre (SF1/cre) transgene into an embryo carrying floxed and null β-catenin alleles (Ctnnb1f/−) [18]. The SF1/cre mouse line starts to show Cre recombinase activity in the somatic cells of fetal gonads around 10.5–11.5 dpc [19]. The Cre recombinase removes the DNA sequence between the two loxP sties that flank the β-catenin gene, therefore producing a null allele of β-catenin. Inactivation of β-catenin in the SF1-positive somatic cells of fetal ovaries resulted in a progressive loss of female germ cells starting at 16.5 dpc [18]. Double staining of the germ cell marker TRA98 and the apoptotic marker cleaved caspase 3 revealed an increase in germ cell apoptosis in the β-catenin cKO ovaries compared to the control (SF1/cre; Ctnnb1f/+ or Ctnnb1f/−) starting at 17.5 dpc (Fig. 1A–B). On average, only one or two germ cells underwent apoptosis per section in control ovaries (Fig. 1A) but more than 5 apoptotic germ cells per section were observed in the β-catenin cKO ovaries (Fig. 1B), indicating that β-catenin in the SF1-positive somatic cells is involved in regulation of germ cell apoptosis.
(A–B) Immunohistochemistry for germ cell marker TRA98 (red) and apoptotic marker cleaved caspase 3 (green) in control (SF1/cre;Ctnnb1f/+) and β-catenin conditional KO (cKO; SF1/cre;Ctnnb1f/−) ovaries at 17.5 dpc. The arrows indicate cells that are double-positive (yellow) for TRA98 and cleaved caspase 3. The inset in B is an image of a higher magnification of cells double-positive (yellow) for TRA98 and cleaved caspase 3. (C–F) Analysis of the meiotic status of germ cells via immunohistochemistry for SCP3 on germ cell smear obtained from the control and β-catenin conditional KO ovaries at 15.5 dpc. The samples were counterstained with nuclear DAPI (blue). Scale bar represent 100 µm in A–B and 10 µm in C–F.
Defects in the meiotic machinery are a major cause for loss of female germ cells [20]. To evaluate whether progression of meiosis was compromised in the absence of β-catenin, we performed immunostaining for synaptonemal complex protein 3 (SCP3) on chromosome smear obtained from female germ cells at 15.5 dpc and 16.5 dpc. Staining of SCP3, a scaffold protein formed on prophase I of meiosis, allowed us to monitor the progression of meiosis in female germ cells. In the absence of β-catenin, most female germ cells entered and progressed through prophase I of meiosis (zygotene and pachytene stages according to [21]) indistinguishable from the control (Fig. 1C–F). We previously analyzed the presence of another meiotic marker, phosphorylated-Histone2AX to detect double strand breaks in the DNA recombination events and found no differences between control and β-catenin cKO ovaries [18]. These findings together indicate that β-catenin in the somatic cells of fetal ovaries is not required for meiosis entry and progression of female germ cells; however, β-catenin is indispensable in the SF1-positive somatic cells for meiotic germ cell survival.
Establishment of the connection between β-catenin and Wnt4 in female germ cell survival
The germ cell loss phenotype in the β-catenin cKO ovary shares striking similarities with that in the Wnt4 KO ovary [16], [18], [22], suggesting these two factors belong to a common pathway. To test whether β-catenin is a downstream mediator of WNT4, we introduced a constitutively active form of β-catenin (Ctnnb1fl.(ex3)) specifically in the SF1-positive somatic cells in the Wnt4 KO ovary. Ctnnb1fl.(ex3) mice contain a genetically engineered β-catenin gene that loxP sequences are inserted in either side of the exon 3. The peptide encoded by the exon 3 is responsible for degradation of β-catenin. Once the exon 3 is removed by the Cre recombinase, the mutant β-catenin becomes resistant to degradation and therefore constitutively active in the SF1-positive somatic cells [23].
We hypothesized that if β-catenin is a downstream effecter of WNT4, introducing active β-catenin to the Wnt4 knockout ovaries should restore normal germ cell development. We examined the total germ cell numbers in the newborn ovaries from controls (Wnt4+/−;SF1/cre and Wnt4+/−;SF1/cre;Ctnnb1fl.(ex3)), Wnt4 KO (Wnt4−/−;SF1/cre), and Wnt4 KO plus active β-catenin (Wnt4−/−;SF1/cre;Ctnnb1fl.(ex3); Fig. 2A–D). To obtain the total germ cell number per ovary, we sectioned the entire ovary, stained the sections with germ cell marker TRA98, and counted TRA98-positive germ cells in sections that were 30 µm apart. The total germ cell number in the Wnt4 KO ovary was significantly lower than the controls (Fig. 2E), consistent with previous findings [22], [24]. However, presence of active β-catenin in the Wnt4 KO ovaries increased the total germ cell numbers to the level similar to the controls (Fig. 2E). Although the size of ovaries in the female with active β-catenin (Fig. 2C & D) was larger than that in the female without the active β-catenin (Fig. 2A & B), the difference in ovary size did not contribute to the difference in total germ cell numbers.
(A–D) Whole mount light field images of Wnt4+/−;SF1/cre, Wnt4−/−;SF1/cre, Wnt4+/−;SF1/cre; Ctnnb1fl.(ex3), and Wnt4−/−;SF1/cre; Ctnnb1fl.(ex3) ovaries at birth. (E) Average total germ cell number was obtained from Wnt4+/−;SF1/cre, Wnt4−/−;SF1/cre, Wnt4+/−;SF1/cre; Ctnnb1fl.(ex3), and Wnt4−/−;SF1/cre; Ctnnb1fl.(ex3) ovaries at birth (n = 3 embryos for each genotype). Tukey tests revealed that the average germ cell number in Wnt4−/−;SF1/cre was significantly different from that in Wnt4+/−;SF1/cre (P = 0.017), Wnt4+/−;SF1/cre; Ctnnb1fl.(ex3) (P = 0.002), and Wnt4−/−;SF1/cre; Ctnnb1fl.(ex3) (P = 0.023). The asterisk represents statistical significance. (F–I) Immunohistochemical staining for androgen-producing enzyme CYP17 (green) was performed in Wnt4+/−;SF1/cre, Wnt4−/−;SF1/cre, Wnt4+/−;SF1/cre; Ctnnb1fl.(ex3), and Wnt4−/−;SF1/cre; Ctnnb1fl.(ex3) ovaries at birth. The inset in G represents a higher magnification of the CYP17-positive cells. Scale bar = 100 µm.
In addition to the restoration of female germ cells, the ectopic CYP17-positive cells in the Wnt4 KO ovary (Fig. 2G) were no longer present in the Wnt4−/−; SF1/cre; Ctnnb1fl.(ex3) ovary (Fig. 2I), indicating activation of β-catenin in SF1-positive Wnt4 KO somatic cells were able to prevent the ectopic appearance of CYP17-postve cells. This genetic evidence places β-catenin downstream of WNT4 in a somatic cell-specific pathway responsible for female germ cell survival and preventing ectopic appearance of CYP17-positive cells in the fetal ovary.
Exclusion of the involvement of androgens in germ cell loss phenotype in the β-catenin cKO ovary
In addition to germ cell loss, inactivation of Wnt4 or β-catenin resulted in ectopic appearance of androgen-producing CYP17-positive cells in the ovary (Fig. 2F & G) [18], [25]. These ectopic CYP17-positive cells produce sufficient androgen that maintains androgen-dependent male reproductive organs such as epididymis and vas deferens in the Wnt4 and β-catenin cKO female embryos [18], [22]. To examine whether ectopic androgen production is responsible for the loss of germ cells, we injected the anti-androgen flutamide daily from 12.5 dpc to birth into pregnant female mice carrying β-catenin cKO embryos. Flutamide is a potent androgen antagonist that has been wildly used to block androgenic effects for clinical treatment of prostate cancer and for basic research on androgen action during embryogenesis [26]. Flutamide injection efficiently blocked the masculinizing effects of androgens in the β-catenin cKO female embryos based on the fact that male reproductive characteristics such as the epididymis were inhibited (Fig. 3D & H) compared to the cKO female without flutamide treatment (Fig. 3B & F). To further confirm that androgen functions were properly inhibited, we examined control male embryos exposed to flutamide in utero. We observed underdeveloped testis and other male reproductive organs compared to the vehicle-treated control (data not shown). These results were similar to what was reported in the literature [27], indicating that the flutamide treatment was sufficient to block androgen action in our system. However, regardless the presence or absence of flutamide treatment, loss of female germ cells was still observed in the β-catenin cKO ovaries at birth (Fig. 3J & L). Flutamide treatment had no effects on development of the female reproductive systems and female germ cells in the control females (SF1/cre; Ctnnb1f/+; Fig. 3A, C, E, G, I, & K). These results demonstrate that loss of germ cells in the β-catenin cKO ovary does not result from ectopic androgen production.
(A–H) Whole mount images of reproductive tracts and ovaries and (I–L) immunohistochemistry for TRA98 were performed on the ovary of the control (SF1/cre;Ctnnb1f/+) and β-catenin cKO (SF1/cre;Ctnnb1f/−) female with or without flutamide treatment. o = ovary; arrow = epididymis, arrowhead = oviduct. Scale bar represents 500 µm in A–D and 100 µm in E–L.
Genetic identification of activin βB as the factor downstream of WNT4/β-catenin that is responsible for inducing female germ cell loss
The TGFβ superfamily has been shown to play a role in inducing apoptosis [28], [29]. In the case of freemartins, where female embryos were exposed to anti-Müllerian hormone, a member of the TGFβ superfamily, germ cell loss was observed [30], [31]. Therefore, we screened various TGFβ family members that showed an increased expression in the fetal ovary lacking either Wnt4 or β-catenin. We found that mRNA expression of activin βB (Inhbb) was significantly elevated in the Wnt4 KO ovaries [32] and β-catenin cKO ovary compared to the control (Fig. 4A–B). In addition, introduction of the active β-catenin to the Wnt4 KO ovary decreased Inhbb mRNA expression to the level similar to that in the control (Fig. 4C). We therefore hypothesized that if elevated Inhbb is indeed responsible for female germ cell loss in the absence of Wnt4, removal of Inhbb in the Wnt4 KO background should reverse this phenotype. Indeed, in the Wnt4−/−; Inhbb−/− double KO ovary, female germ cell number was significantly increased compared to the Wnt4 single KO at birth (Fig. 4D–F, n = 3). To monitor the status of germ cell meiosis, we examined the expression of SCP3, a meiosis marker, in control, Wnt4 single knockout, and Wnt4−/−; Inhbb−/− double KO ovaries at 15.5 dpc the time before the germ cell demise occurred in the Wnt4 KO ovary. Female germ cells in the Wnt4−/−; Inhbb−/− double KO ovary entered meiosis properly as evident by double immunostaining for SCP3 and TRA98 (Fig. 4J–L). Similar to the Wnt4 single knockout ovary, ectopic CYP17-positive cells and epididymal structure were found in the Wnt4−/−; Inhbb−/− double KO ovary but were absent in the control female at birth (Fig. 4G–I), further supporting that ectopic production of androgen is not responsible for the loss of female germ cells.
(A–C) Whole mount in situ hybridization for Inhbb was performed on control ovary (SF1/cre;Ctnnbf/+ or Ctnnb1f/−) (A), β-catenin cKO ovary (SF1/cre;Ctnnbf/−) (B), and Wnt4−/−;SF1/cre; Ctnnb1fl.(ex3) ovary (C) at 13.5 dpc. (n = 2–3 for each genotypes). o = ovary, m = mesonephros. (D–F) Immunohistochemistry for TRA98 (red) and CYP17 (green) was performed on ovary sections from control (Wnt4+/−; Inhbb+/−), Wnt4 single KO and Wnt4; Inhbb double KO ovary at birth. (G) Light field microscopic images of the reproductive tract were taken from control female, Wnt4 single KO, and Wnt4; Inhbb double KO females at birth. Arrowheads indicate oviduct and arrows indicate epididymal structure. o = ovary. (J–L) Immunohistochemistry for TRA98 (red) and SCP3 (green) were performed on ovary sections from control, Wnt4 single KO and Wnt4; Inhbb double KO ovary at 15.5 dpc. At this stage, the female germ cells have not lost yet in the Wnt4 KO ovaries, allowing us to monitor the status of meiosis. The insets are the images of a higher magnification of cells double-positive (yellow) for TRA98 and SCP3. Scale bar represent 250 µm in A–C, G–I and 100 µm in D–F, J–L. (M) A proposed model for the somatic cell-derived pathway on female germ cell survival: In the mouse fetal ovary, WNT4 signals via β-catenin to decrease the expression of activin βB or Inhbb, which causes loss of meiotic germ cell. WNT4 also stimulates the production of follistatin (Fst), which acts to antagonize the activity of Inhbb. The WNT4/β-catenin pathway also prevents the ectopic production of androgens in the fetal ovary, which is not responsible for the germ cell loss. WNT4 could possibly regulate its own expression via β-catenin.
The WNT4/β-catenin pathway, operating in the SF1-positive somatic cells in fetal ovaries, is essential for maintaining the survival of meiotic germ cells. Although the initiation and progression of meiosis are not affected by the absence of WNT4/β-catenin in the fetal ovary, the meiotic germ cells undergo apoptosis and are lost at birth. WNT4 maintains female germ cell survival by activating β-catenin in the SF1-positive cells, which in turn suppresses expression of activin βB. Our results provide the genetic proof of somatic cell contribution to female germ cell survival via a delicate balance between positive (Wnt4 and β-catenin) and negative regulators (activin βB or Inhbb, Fig. 4M).
Somatic cells of the fetal ovary are the supporting cells that nurture the germ cells and provide them proper environment to grow. In vitro experiments using human and mouse ovarian tissues demonstrated that factors such as Kit ligand, leukemia inhibiting factor, bone morphogenetic factor 4, basic fibroblast growth factor, and activin A stimulate folliculogenesis and survival of germ cells in culture [33], [34]. However, in vivo evidence is lacking to support a functional role of these factors in female germ cell development in the fetal ovary. In the Wnt4 knockout and β–catenin cKO ovary, female germ cells undergo apoptosis and are progressively lost. Despite entering meiosis properly, most germ cells disappear around the time of birth. The ability of constitutively active β-catenin to restore germ cell numbers in the Wnt4 KO ovary indicates that β-catenin is involved directly or indirectly in the downstream pathway of WNT4. Evidence of WNT4 signaling via β-catenin is also found in nephron induction, kidney epithelial cells, and renal fibrosis [35], [36], [37], [38]. These observations collectively support the model that β-catenin operates downstream of WNT4 in the fetal ovary. In addition to serving as an intracellular signaling molecule of WNT4, β-catenin also has a possible role in regulating the expression of Wnt4. We found that expression of Wnt4 was lost in the absence of β-catenin in the SF-1 positive-somatic cells of the fetal ovary; however, R-spondin 1 (Rspo1) expression was not altered [18]. These results suggest that RSPO1 or other WNT proteins including WNT4 itself could stimulate Wnt4 expression via β-catenin.
Ovaries with active β-catenin in the somatic cells are larger than the controls, suggesting that the activation of β-catenin promotes proliferation of somatic cells. WNT/β-catenin signaling pathway is known to regulate genes that are involved in cell proliferation and cell fate decision during embryogenesis [14]. Mis-regulation of WNT/β-catenin signaling pathway results in various types of cancers [39], [40], [41], [42]. Introduction of activated β-catenin to fetal testes also increases the size of affected testis [43]. Further experiments are needed to investigate the impact of active β-catenin on somatic cell proliferation.
Germ cell loss still occurred in Wnt4 KO and β-catenin cKO fetal ovaries after anti-flutamide treatment, therefore excluding the involvement of androgens. Furthermore, androgen receptors are not present in germ cells at fetal stages [44], supporting our conclusion that ectopic androgen is not responsible for the death of germ cell in Wnt4 KO and β-catenin cKO fetal ovaries. Rescue of the germ cell loss phenotype in the Wnt4−/−; Inhbb−/− double KO ovary provides a genetic link implicating Inhbb as the gene responsible for germ cell demise. Inhbb encodes the subunit for inhibin B and activin B. Germ cells are known to express receptors (Acrr-IB and ActR-IIB) for activins [45]. The ectopic production of activin B from somatic cells of the fetal ovary in the absence of Wnt4 and β–catenin could therefore act directly on female germ cells and cause their death. Using Transcription Element Search System (TESS) program, we found several putative β-catenin LEF/TCF response elements in the promoter region of Inhbb. β-catenin could bind to these response elements and suppress the expression of Inhbb in the fetal ovary.
The germ cell loss phenotype is also observed in the follistatin (Fst) knockout fetal ovaries [24]. Based on a genetic epistasis experiment, Fst acts downstream of WNT4 [24]. Furthermore, expression of mouse Fst is dependent upon a consensus β-catenin LEF/TCF binding site in the promoter region [46], [47], and expression of Fst was lost in the β-catenin cKO ovary [18], placing Fst downstream of β-catenin. FST is known to bind activins with high affinity, therefore preventing activins from activating their receptors [48]. Expression of Inhbb mRNA is present in mouse gonads of both sexes at 11.5 dpc. Its expression is down-regulated but remains detectable in the ovary at 12.5 dpc [32]. Interestingly, in contrast to the Wnt4 and β-catenin cKO ovary where Inhbb expression is upregulated, Inhbb mRNA expression levels are not altered in the absence of Fst [32]. We speculate that the function of FST is to antagonize and inhibit the action of the residual activin B to prevent it from affecting female germ cell survival. WNT4/β-catenin acts at two levels to block the effects of activin B on germ cells, by downregulating transcription of Inhbb and by activating FST to antagonize activin B protein (Fig. 4M).
It is known that oocytes start entering apoptosis prenatally, therefore leaving a finite number of oocytes for the rest of the reproductive life of female individuals. At least two hypotheses have been proposed for the cause of female germ cell demise during embryogenesis [49], [50]. The first possible mechanism is that abnormal oocytes with defects on their chromosomes or mitochondrial genomes are eliminated from the oocyte pool via intrinsic check-point mechanism [51]. Another possibility is that the somatic cell environment controls the numbers of female germ cells. In this study, we found that the upregulated of Inhbb resulted in the death of female germ cell in the absence of Wnt4/β-catenin signaling from somatic cells. We propose that the balance between somatic cell signaling (WNT4/β-catenin) and Activin B (the protein product of Inhbb) is critical for the maintenance of female germ cells during embryonic stage. It is possible that increasing germ cell apoptosis close to birth is the result of a shifted balance toward action of activin B. If our hypothesis is correct, one would predict that loss of Inhbb should lead to decrease in germ cell apoptosis and presumably more oocytes in the ovary. Inhbb knockout females are fertile despite an increase in length of gestation and a decrease in ability of nursing [52]. It remains to be determined whether more oocytes are present in the Inhbb knockout ovary.
When either Wnt4 or β-catenin is inactivated, female germ cells enter meiosis and progress to meiosis prophase I normally, based on the time course analysis of chromosome smears of germ cells and examination of expression of SCP3 and γH2AX [16], [17], [18], [24]. These findings suggest that the retinoic acid (RA) pathway that regulates meiosis entry is probably not affected by the absence of Wnt4/β-catenin. Studies on the R-spondin1 (Rspo1) KO mice show that Rspo1 is the upstream regulator of WNT4 and β-catenin in ovarian development [15], [17]. In the absence of Rspo1, components of the RA pathway are not significantly affected [15]. Although a decrease in germ cell numbers is reported in the Rspo1 KO ovary at 14.5 and 16.5 dpc, germ cell entry into meiosis appeared to be normal based on the SCP3 staining [17]. The possibility of a direct action of RSPO1 on female germ cells remains to be determined.
Involvement of WNT/β-catenin pathway in regulating proliferation of primordial germ cell (PGCs) is evident in Drosophila and mouse. Activation of β-catenin in PGCs promotes proliferation in Drosophila whereas it delays cell cycle progression in mouse [53], [54]. In this study, we report an essential while indirect role of β-catenin in somatic cells in controlling female germ cell numbers. β-catenin, operating downstream of WNT4 in the somatic cells, acts as a suppressor of activin βB, which is a negative regulator of female germ cell survival. In summary, our data provide genetic identification of a molecular hierarchy in ovarian somatic cells that is essential for maintenance of female germ cell survival.
Materials and Methods
Ethics Statement
All procedures described were reviewed and approved by the Institutional Animal Care and Use Committee at University of Illinois, and were performed in accordance with the Guiding principles for the Care and Use of Laboratory Animals.
SF1/cre;Ctnnb1floxed/− embryos were derived from breeding of the Ctnnb1+/−; SF1/cre, which carrying Cre recombinase under the control of the Sf1 promoter and its regulatory elements [19], and the Ctnnb1floxed/floxed parental strains. Wnt4+/− mice were obtained from Jackson Laboratory (strain 129-Wnt4tm1Amc/J). Ctnnb1fl(ex3) mice were obtained from Harada et al. [23]. To generate mice expressing the stabilized form of β-catenin specifically in the somatic cells of Wnt4 knockout ovary, Wnt4+/−; SF-1/cre were mated with Wnt4+/−; Ctnnb1fl(ex3) mice to obtain Wnt4−/−;Sf-1cre; Ctnnb1fl(ex3) embryos. To obtain the Wnt4 and Activin β B double knockout mice (Wnt4−/−; Inhbb−/−), Wnt4+/− mice were mated to Inhbb+/− mice to generate Wnt4+/−; Inhbb+/− double heterozygotes. Wnt4+/−; Inhbb+/− double heterozygotes were mated to generate Wnt4−/−; Inhbb−/− double knockout mice. The day when the vaginal plug was detected in the mated female was considered as 0.5 dpc. Genotypes were determined by PCR using gene specific primers. The primers were: SF-1/Cre genotyping: 5′-GTGTGAACGAACCTGGTCGA AATCAGTGCG-3′ and 5′-GCATTACCGGTCGATGCAACGAGTGATGAG-3′; Ctnnb null allele: 5′-AATCACAGGGACTTCCATACCAG-3′ and 5′-GCCCAGCCTTAGCCCAACT-3′; Ctnnb1 wild type and floxed alleles: 5′-AAGGTAGAGTG ATGAAAGTTGTT-3′ and 5′-CACCATTGTCCTCTGTCTATTC -3′; Wnt4 null and wild type alleles: 5′- CTG AGGAAGAGCAGGGTCAC -3′, 5-ATGGTCACC CCCATTTTACA -3′ and 5-TGGATGTGGAATGTGTGCGAG-3′; Ctnnb1fl(ex3) alleles 5′-GGTAGTGGTCCCTGCCCTTGA CAC-3′ and 5′-CTAAGCTTGGCT GGACGTAAACTC-3′; Inhbb null allele: 5′-CTTGGGTGGAGAGGC TATTC-3′ and 5′ - AAAGCTGATGATCTCGGAGACG - 3′; Inhbb wild type allele: 5′-ATGGTC ACGGCC CTG CGC AA-3′ and 5′-GCGGAT CCC TCT GCA AAG CTG ATG ATT TC-3′; Sry genotyping: 5′-TGAAGCTTTTGGCTTTGAG-3′ and 5′-CCGCTGCCAAATTCTTTGG-3′. All experiments were performed on at least two to five animals for each genotype.
Samples were fixed in 4% paraformaldehyde overnight at 4°C and then washed in PBS for 5 minutes (3 times) Samples were put through a sucrose gradient (10%, 15% and 20%) and incubated in 1∶1 20% sucrose and OCT freezing media (Tissue-Tek) overnight at 4°C. Samples were embedded in 1∶3 20% sucrose and OCT mix and cut to 10 µm thick frozen sections. Sections were washed with PBS and then blocked in the blocking solution (5% heat-inactivated donkey serum and 0.1% Triton X-100 in PBS) for 1 h at room temperature. Primary antibodies were added to the blocking solution and incubated with sections at 4°C overnight. Sections were then washed with the washing solution (1% heat inactivated donkey serum and 0.1% Triton X-100 in PBS) followed by incubation in the blocking solution with the corresponding secondary anybodies. Sections were then washed with the washing solution and mounted with DAPI antifade reagent. The sources and dilution of primary antibodies were the rat monoclonal antibody against germ cell nuclear fraction (TRA98, 1∶000, a gift from H. Tanaka), the rabbit polyclonal antibody against cleaved caspases-3 (1∶200; Cell Signaling), and the rabbit polyclonal antibody against CYP17 (1∶100, a gift from B. Hales). All the secondary antibodies were purchased from Jackson Immunochemical and a 1∶200 dilution was used.
Chromosome smear and immunostaining
Fetal germ cell chromosome smear and immunostaining were performed according to the protocol described in [55]. Briefly, ovaries from 15.5 dpc embryos were incubated in a 24-well dish with the hypoextraction buffer (15 mM Tris,pH 8.2, 50 mM sucrose, 20 mM citrate, 5 mM EDTA, pH 8.2, 0.5 mM DTT, 0.09 mg/ml PMSF, collagenase, 0.5 mg/ml) for at least 30 min. Then each ovary was placed in a 10 µl drop of 0.1 M sucrose on the slide and another 10 µl drop of sucrose was added. The ovaries were dispersed by repetitive pipetting. Cell suspension was then placed onto the slide coated with the fixative (0.1% paraformaldehyde, pH 9.2, 0.1% Triton-X 100). The slides were placed in a humid chamber for 4 h and then were gently washed three times (5 min each) in 1∶250 photo-flo (Kodak) in water. Slides were air-dried and stored in −20°C.
For immunostaining of spread chromatin, slides were washed three times (10 min each) in 10% antibody dilution buffer (10% donkey serum, 3% BSA, and 0.05% Triton-X in phosphate-buffered saline or PBS). Then slides were incubated with anti-SCP3 antibody (1∶500, Abcam) in a humid chamber overnight at 4°C. Samples were washed three times (10 min each) in 10% antibody dilution buffer. Slides were then incubated with secondary antibody for 2 h at room temperature in the dark followed by three washes (5 min each) in PBS. Slides were air-dried and mounted with DAPI antifade reagent.
Germ cell counting
Newborn ovaries were obtained from 3 animals for each genotype. Samples were fixed in 4% paraformaldehyde in PBS at 4°C and processed according to the immunohistochemistry procedure described above. Germ cell count was obtained by counting TRA98-positive germ cells in sections (30 um apart) from the entire ovary. Data were analyzed using one-way ANOVA followed by Tukey test for pair wise comparisons.
Flutamide treatments
Flutamide (F9397, Sigma-Aldrich) was dissolved in 1∶1 (vol/vol) mixture of absolute ethanol and sesame oil. Ten pregnant Ctnnb1floxed/floxed female mice that were plugged by SF-1/cre; β-Ctnnb1+/− male were injected daily with flutamide subcutaneously (100 mg/kg/daily) from 12.5 dpc until birth [27]. Five pregnant mice from the same breeding scheme were treated with the vehicle (sesame oil) from 12.5 dpc until birth (control group).
Whole mount in situ hybridization
Tissues were fixed overnight in 4% paraformaldehyde in PBS at 4°C and dehydrated through a methanol gradient (25%, 50%, 70%, and 100%) in PTW (0.1% Tween20 in DEPC-PBS). Samples were stored in 100% methanol at −20°C up to 6 months. In situ hybridization was processed according to the standard non-radioisotopic procedure using digoxigenin-labeled RNA probes for Inhbb. Gonads with mesonephroi attached were rehydrated through a methanol gradient then wash with PTW. The rehydrated gonads were treated with proteinase K (10 mg/ml) at 37°C for 12 minutes and then post-fixed in 4% paraformaldehyde/0.1% glutaraldehyde at room temperature for 20 minutes. Samples were pre-hybridized in the hybridization buffer (5x SSC pH 5.0, 50% formamide, 0.1% CHAPS, 0.1% Tween20, 1 mg/ml Yeast tRNA, 50 µg/ml Heparin, and 5 mM EDTA pH 8.0) at 65°C for 1 hour. Then digoxigenin-labeled Inhbb RNA probe was added into the solution and samples were rotated in an oven at 65°C overnight (12–16 hours). On the Next day, samples were washing with pre-warmed hybridization buffer followed by washing with MABTL (5% MAB, 0.1% Tween20 and 0.05% Levamisole). Samples were incubated in 20% sheep serum in MABTL blocking solution at room temperature for 2 hours followed by incubating in alkaline phosphatase-conjugated anti-digoxigenin at 4°C overnight on a shaker. On the third day, after washed in MABTL three times for 1 hour each, samples were incubated in alkaline phosphates substrate in NTMTL (0.1 M NaCl, 0.01 M Tris-HCl pH 9.5, 0.05 M MgCl2, 1% Tween 20, 0.05% Levamisole) for color development.
We would like to thank Dr. Buck Hales for the CYP17 antibody, Dr. H. Tanaka for the TRA98 antibody, Dr. Martin Matzuk for the Inhbb plasmid for generating the RNA probe, Dr. Hsin-Yi Weng for statistics analysis and all the Yao Lab members for their assistance and support. We also appreciate the critical comments from Dr. Blanche Capel.
Author Contributions
Conceived and designed the experiments: CFL HY. Performed the experiments: CFL. Analyzed the data: CFL HY. Contributed reagents/materials/analysis tools: KP HY. Wrote the paper: CFL HY.
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I recently purchased a HP Pavilion 15-ak099nr Signature Edition Gaming Laptop from the Microsoft store. It comes with the Windows 10 Home OS. I have Windows 10 Pro running on another system that I want to retire. How can I migrate the Windows 10 Pro to the New Laptop.
I thought about doing a clean install of Windows 8 Pro then doing the free Windows 10 Pro upgrade. Can anyone think of why this shouldn't work?


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Picking up from right where the first film ends, The Raid 2 follows Rama (Iko Uwais) as he goes undercover and infiltrates the ranks of a ruthless Jakarta crime syndicate in order to protect his family and uncover the corruption in his own police force.
Cant wait for The Raid 2 ! Posted this on all my accounts, Twitter, Facebook, LinkedIn, Google, Pinterest !
The images look slightly blury, but only slightly..Love the Hammer Girl still..
Enough with the news when do we get the freaking reviews ⊙_ʘ
Curious to know if Gareth Evans seen The Act Of Killing yet. Living in Indonesia I hope he's okay.
I love Hammer Girl, but she HAS to be a reference to Oldboy, I mean, that pioneered the 'beating-the-shit-out-of-someone-with-a-hammer' fighting style. God, I can't wait until this movie comes out though, it looks so awesome.
We are all live in peace. The Act Of Killing is just about the story of 1965
I'm glad to hear that my brother. When I watch that movie, I was sadden and very emotional. Why is the people not yet held responsible for killing?
Because the operations was sanctioned by Presiden Soeharto and they were acting under orders. No, that doesn't absolve them from the crimes but that kind of dirty secret is usual in Indonesia. To explain everything would be too long, but as a full blooded Indonesian whose family include a former member of DPR, close friends of the husband of a former president and a white collar mafia, I'm not really surprised by what was portrayed in The Killing.
cos after 1965, Gen. Suharto became the President of Indonesia and has rewritten the history according to his version, and it has been inserted into national education system for more than 30 years.
Me, among many millions, are the product of this system.
Only in recent years (since Suharto downfall in 1998) that people started to talk openly about what really happened during 1965 era.
edit: plus the fact that the whole Act Of Killing was sanctioned by Gen. Suharto to usurp the power from President Soekarno.
It kinda reminds me of my country Cambodia where Pol Pot and his stupid ass Khmer Rouge regime overthrew the gov't, then creating a puppet gov't and massacring over 2 million citizens mostly well educated and movie stars. What's ironic is that if it wasn't for the Vietnam War, Pol Pot stupid ass would have murdered even more citizens in that country, very sad stuff. So I'm very blessed and thankful that I'm living in America today. I'm sure it's different lifestyle now in Cambodia but still, what a sad and heartbreaking situation that stuff like this happened just 40 yrs ago.


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Several local South Florida junior tennis players fared well in the recent United States Tennis Association Florida State Doubles Championships last month at the Florida Tennis Center in Daytona Beach.
The 59th annual doubles tournament featured 368 of the top junior tennis players.
The tournament was held Sept. 15-16.
The finals of the Mixed 12s, 14s, 16s and 18s were not determined due to time constraints because of the large number of entries (252 matches on Saturday and 236 on Sunday).
In matches involving Broward and Palm Beach county players, the top-seeded team of Zach McCourt, of Sunrise, and Kosuke Hasegawa, of Wesley Chapel, won the Boys' 14 Doubles championship over the third-seeded Michael Rinaldi, of Palm City, and Gordon Watson, of Naples, 9-8.
Second-seeded Lindsay Graff, of Fort Lauderdale and Nicole Robinson, of West Palm Beach, defeated top-seeded Courtney Colton, of Davie, and Laura Wiley, of Fort Lauderdale, 8-5.
M'balia Bangoura, of Bradenton, and Kahalauamalunonala Companion, of Bradenton, defeated two other teams, including Sarah Dreyfuss, of North Miami Beach, and Katherine Wiley, of Fort Lauderdale, in the Girls' 10 division to claim the title.
In the Mixed-10 Doubles championship, top-seeded M'balia Bangoura and Zachary Bessette, both of Bradenton, defeated second-seeded Andrew Heller, of Weston, and Katherine Wiley, of Fort Lauderdale 8-1.
The first-year Broward Lady Canes recently placed second in the eighth-grade (14-and-under) Elite Division I at United States Specialty Sports Association Nationals in Tampa.
The Broward Lady Canes, an eighth-grade girls' travel basketball team, finished with a 24-10 record. In addition to the second place in Tampa, the Lady Canes were fourth at the USSSA State Tournament, first at the USSSA Auburndale Hoopfest and first at the USSSA Treasure Coast State Qualifier.
Gary Curreri can be reached via e-mail at email@example.com.


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Reports: Eric Bailly set to return from injury in December
Manchester United are set to be boosted by the return of Ivorian international Eric Bailly after reports confirmed that the defender is back in training. The news comes as a huge relief to Jose Mourinho and the Red Devils who were expecting Bailly to miss out on a majority of the remainder of the season. The Ivory Coast centre-back was one of Mourinho's first signings at United and cost them around £30m.
Bailly has quickly become a fan favourite after his arrival at Old Trafford. The defender started off the season with some brilliant performances to back his big money move from Villarreal. However, he suffered an injury in United's 4-0 loss against Antonio Conte's Chelsea last month and was expected to be side-lined at least until January. The 22-year-old will not be available for selection during January due to the African Cup of Nations but could feature in United's league game against Tottenham Hotspur on December 11th.
Also read: Jose Mourinho lining up Danny Drinkwater's return to Old Trafford
United have conceded five goals ever since Bailly's absence and have not looked convincing enough at the back. Mourinho was quick to point out that the Ivorian is well on track for recovery and added, "He is working hard and will be back by the end of the year, for sure." Another source close to United added, "The medical staff were impressed with his progress over the recent international break. They are refusing to set a comeback date, but Bailly hinted the game on December 8th (against Zorya Luhansk) will be his return."
Troubled times for Jose Mourinho and Manchester United
United have won just one out of their last seven league games and are currently 6th in the table, 11 points away from league leaders Chelsea. Mourinho is expected to look for reinforcements come January after it was clearly evident that he does not fancy Chris Smalling or Marcos Rojo to be a part of the United back four.
United face West Ham in the EFL Cup tomorrow and face a tough challenge in the three games against Tottenham, Crystal Palace and West Brom leading up to the festive season.
Mourinho's recent sending off against West Ham could also mean that he might miss out on two league games and things can go from bad to worse if United keep dropping points leading up to the second half of the season. The Red Devils have been linked to Virgil van Djik as well as Victor Lindelof of Benfica.
More Football news, transfer rumours, analysis and features at Sportskeeda.com. Also if you are attending a football game anywhere in the world and/or have a news tip for us drop us an email at football (at)sportskeeda (dot) com


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Here's some frustrating news for lots of parents (including me): A recent study found that children of "math-anxious" parents are more likely to develop "math anxiety" themselves. However, there is something you can do to prevent this from happening -- or not do, as the case may be.
According to the study, this trans-generational transmission of math anxiety only happens when parents help their kids with their math homework. In a sort of twisted snowball effect nightmare, parents who stress out about math make their kids stressed out about math, which makes them get bad grades in math, which makes everybody even more stressed out about math. And the more parents try to "help," the worse everything gets. Other exacerbating factors include the constant introduction of "new" methods of learning math (Common Core, anyone??), which makes already math-anxious parents downright math-phobic.
More from The Stir: Frustrated Dad Writes Epic Response to Kid's Common Core Math (PHOTO)
Hey, I totally get it. Just looking at my kids' math textbooks gives me hives. I haaaaaaated math growing up, even though I managed to get decent grades, and retained next to no memory of anything having to do with algebra or geometry (or even fractions or decimals, if I'm honest). But I'm relieved to hear that I'm at least doing one thing right: I never help my kids with their math homework. They figured out pretty early on that I didn't know what I was doing ("Mom, that's not how you do long division anymore!!"). Instead, either my much less math-anxious husband helps, or we turn to the Internet for answers. Neither of which method has turned my kids into math lovers, but at least they get good grades and don't lose any sleep over the subject.
Oh yeah, and neither do I!
Image via Madhavi Kuram/Flickr


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Processed Foods and Depression
Okay, we all fall from time to time be tempted to eat any processed food. Cookies, ice cream, and even junk food are all tripping over our respective diets, and the truth is not always regrets.
But sometimes, we forget that these foods not only provide us with too many calories, but can also cause a disorder in our blood sugar levels. Processed foods are high in refined sugar, and that leads to a slight feeling of joy once consumed. But, when the sugar rushes down to our body, it implies the opposite.
This is demonstrated by a team of scientists who published their study in The British Journal of Psychiatry, who have observed a group of volunteers who eat a lot of processed foods. The volunteers were shown to have an increase risk of depression.
Processed foods like ice cream, cookies, chocolates, burgers and just about everything at first glance obviously escapes the guidelines of our diet.
If you like to consume sugar, it is best to have a cereal bar.If you're hungry, go for a fruit or simply wait for an hour until the lunch or dinner. Keep the order, and avoid foods that not only hurt your diet, but also your mental health.


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On the first installment of his nightly video blog promoting his upcoming series, Jimmy Fallon confirmed the rumors that critically acclaimed Grammy winning hip-hop band The Roots will become Fallon's house band when Late Night with Jimmy Fallon makes its broadcast debut.
"We are really excited to be working with Jimmy on the new 'Late Night,' said Roots founder Ahmir "?uestlove" Thompson, "It's also an honor to be working with an icon like Lorne Michaels. We are looking forward to becoming part of the history of 30 Rock".
Executive produced by Lorne Michaels, Fallon's Late Night will depart from David Letterman and Conan O'Brien's Studio 6A and cross the hall to Studio 6B in NBC's 30 Rockefeller Plaza headquarters in New York.
Late Night with Jimmy Fallon will debut in 2009.