text:
About Your Space Coast Symphony Orchestra
Our Home – The River House
Space Coast Symphony Chorus
Space Coast Symphony Jazz Orchestra
SCSO Blog
How to Purchase Tickets
18 & Under Club
Symphony for Everyone
Ticket Outlets
Host a Musician
Advertise with the Symphony
Celebrating Ten Years
Maestro's on Air
Ask Aaron
Volunteers Corner
Inside the Music
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Quartet Movement
Our Youth Orchestra
Symphony FAQ
Bundle up for 'Sinfonia Antartica'
Space Coast Symphony Orchestra concert invokes frozen imagery
(Vero Beach, FL) November 6, 2016 – As Florida temperatures grow blissfully cooler, Maestro Aaron T. Collins will lead the Space Coast Symphony Orchestra in a concert inspired by visions of glaciers and giant ice mountains, when they present Sinfonia Antarctica at the Vero Beach High School Performing Arts Center, 1707 16th Street, 32960. The program on Saturday, November 19 at 7:00 PM will open with Malcolm Arnold's rollicking Tam o' Shanter, based on the Robert Burns poem about the exploits of a hard drinker, and guest conducted by Professor Gareth Pritchard of the Central Florida Brass Band. International soprano, Mary Anne Kruger will then perform Richard Strauss' powerful Four Last Songs. Composed in the final year of his life, Strauss' musical farewell is among history's most moving compositions, and the culmination of his lifelong love affair with the soprano voice. The concert continues with Vaughan Williams' magnificent Seventh Symphony, Sinfonia Antartica, a by-product of the composer's film score for Scott of the Antarctic. One of the most important composers of the 20th century, Williams' imagination was captured by the heroic failure of Captain Robert Scott to reach the South Pole; his 1952 five-movement symphony reflects Scott's awe and changing emotions at the frozen polar landscape. $20 advance tickets are available through the orchestra website atwww.SpaceCoastSymphony.org or at Marine Bank & Trust (beachside and mainland branches). Tickets at the door are $25. The concert is free for those aged 18 and under or with a student ID. For more information, call toll free to 855-252-7276 or visit www.SpaceCoastSymphony.org.
For Space Coast Symphony Orchestra Conductor and Artistic Director, Aaron T. Collins, Sinfonia Antarctica is an opportunity to work with two respected Central Florida music professionals, in addition to performing a revered masterwork.
"Soprano Mary Anne Kruger has an international reputation and a vast operatic repertoire ," said Collins. "We are delighted that she will be joining us to perform the Strauss. And I'll be handing my baton over to Professor Pritchard to bring the Robert Burns' poem to raucous life. Gareth Pritchard is an incredible conductor, composer, arranger and musician. Sinfonia Antarctica is going to be a musical feast for the ears."
For more information about the Space Coast Symphony 2016-17 season or the Symphony for Everyone discount ticket program, please visit www.SpaceCoastSymphony.org or call toll-free (855) 252-7276.
ABOUT MARY ANNE KRUGER — The American soprano, Mary Anne Kruger, studied voice with Margaret Harshaw and first went to Germany in 1992 when she completed a Master of Music degree at Indiana University. From 1994 to 2005 Mary Anne Kruger was a member of the opera ensemble at the Hessische Staatstheater Darmstadt where she built a repertoire of over 40 major roles and concerts. She has also guested in over 20 European theaters including the Bayerische Staatsoper- Nationaltheater in Munich, Staatstheater Stuttgart, and Theater Basel.
Mary Anne Kruger's prizes include a nomination in the category "Best Singer" in the 1997 Opernwelt Jahrbuch for her portrayal of Alcina. She was awarded the role of Donna Elvira in the 1996 Don Giovanniproduction at the Athens Concert Hall directed by Ruggiero Raimondi and conducted by Gustav Kühn. She won the "Sonderpreis der Wiener Staats-und Volksoper" at the 1993 Belvedere Competition, which secured for her a performance as Hanna Glawari (Die Lustige Witwe) at the the Wiener Volksoper. She was also awarded a DAAD scholarship (Deutscher Akademischer Austauschdient) to study at the Hochschule für Musik in Munich 1992-1993. In Mary Anne Kruger's vast operatic repertoire, the roles of Madama Butterfly, Suor Angelica and Alcina have won her particular acclaim. Her concert repertoire includes the Missa Solemnis and 9th Symphony of L.v. Beethoven, Verdi's Missa di Requiem, and Benjamin Britten's War Requiem.
ABOUT PROFESSOR GARETH PRITCHARD — Gareth Pritchard entered Trinity College of Music, London in 1974 to study Trumpet with Norman Burgess (Principal Trumpet of the BBC Concert Orchestra), Composition with Vera Ayton & Conducting with Bernard Keefe (Assistant Conductor to Sir Georg Solti at Covent Garden Opera) graduating with an LTCL (Teacher's Diploma) by the 3rd year & being admitted a Fellow of the College FTCL at the end of his 4th year (an unprecedented feat still not surpassed today). Gareth was the Cornet Solo Champion of Great Britain in both 1977 & 1979. He has performed regularly as a soloist all over the world & on many radio/tv broadcasts & recordings. Gareth played Principal Cornet for Parc & Dare, Fairey Engineering, Brodsworth Colliery & GUS/Rigid Containers Bands winning the Granada Band of the Year in 1979 & the British Open in 1979 & 1988. He played under the influence of many eminent Conductors at this time such as Dennis Carr, Walter Hargreaves, Harry Mortimer, Roy Newsome, Major Peter Parkes & Bram Tovey. In 1991 Gareth moved to Norway as a full time Freelance Musician and in only seven attempts, he conducted a band to 1st prize at the National Finals there on an unprecedented 4 occasions in 3 different divisions, including the Championship Section with Sandefjord Brass Symposium in 1998. He also won SIDDIS Brass (the Norwegian "Brass in Concert") with SBS using 3 of his pieces in their programme. He is also regularly invited to conduct other Norwegian Championship Section Brass Bands & Professional Military Bands. He was also a Conductor & Teacher at 10 different School Bands & was the Musical Advisor & Conductor of the Telemark Wind Orchestra & Telemark Summer School from 1996-2005.
Since returning to the UK in 1998, Gareth has been Professional Conductor for Besses o' th' Barn, Bodmin Town, BTM, Tongwynlais & St Austell Bands, while also being Musical Director for EYMS, Lewis Merthyr, Woodfalls & YBS Bands. He has forged himself a reputation over the years of drastically improving & doing extremely well at contests with 'non-fancied' bands. This is especially true in Entertainment Contests where his innovative programmes have resulted in 1st Prizes at Yeovil (twice); 2nd Prizes at Butlins (three times) & Wychavon (once) plus 3rd Prizes at Wychavon (twice). This success has also transpired at Test Piece Contests with two 1st Prizes at the Senior Cup & numerous other Prize winning performances at The Areas, Grand Shield, Senior Trophy & many local contests from 2nd Section up to Championship Section. He has conducted many times at major concert halls all over Europe – Royal Albert Hall, London; Symphony Hall, Birmingham; Griehallen, Bergen, Norway; Stavanger Konserthus, Stavanger, Norway; Kulturzentrum Gasteig (Philharmonie), Munich, Germany; Jaarbeurs Congrescentrum, Utrecht, Holland. His arrangements & compositions are extensively played all over the world & they have also been recorded by Brass & Military Bands both in Norway & the UK. Gareth is in regular demand as an adjudicator worldwide having adjudicated at many competitions including the Lower Section National Finals of Great Britain, Belgian Nationals, SIDDIS Brass Entertainment Contest in Norway, the Welsh Area, Thameside Open, IBBC, SCABA, Isle of Man Festival & Pontins. He is a member of the exclusive Association of Brass Band Adjudicators.
ABOUT THE SPACE COAST SYMPHONY ORCHESTRA— The SCSO is a lively and passionate Central Florida orchestra comprised of professional musicians, music educators and mentored amateurs of exceptional ability. The Orchestra was formed by Conductor and Artistic Director Aaron T. Collins with a two-fold mission: to provide mentoring and performance opportunities for Florida's finest young musicians as well as to provide professional concerts at genuinely affordable prices. The ultimate goal of the organization is to help ensure the symphonic tradition for the enjoyment of future generations.
33-year old Aaron T. Collins is garnering recognition for his achievements. LEAD Brevard named the ambitious Collins one of their "4 under 40" Young Professionals, the youngest-ever recipient of the honor. Space Coast Business magazine listed him as one of Brevard's "100 Most Admired Businesspeople." In 2016, Collins won the Richard A. Stark Award for cultural leadership from the Cultural Council of Indian River County. Through his personal involvement with more than a dozen performing arts groups, pioneering reciprocal ad program and social media cross promotions, Collins has gained a reputation for generosity; championing other arts organizations throughout Central Florida for the cultural enrichment of the community.
The Space Coast Symphony Orchestra performs in several very comfortable venues in Brevard and Indian River counties, with advance general admission tickets priced at just $20. School-aged children 18 years and under are admitted free of charge, as are college students with a valid ID. In keeping with their mission, the symphony provides a unique Symphony for Everyone ticketing program, a "pay-what-you-can" option designed to provide substantially discounted admission to any Space Coast Symphony concert.
Relying on a non-traditional orchestra model and a slim budget, the Space Coast Symphony Orchestra is one of only 19 U.S. orchestras to perform year-round. In addition to maintaining the Space Coast Symphony JAZZ Orchestra and Space Coast Symphony Wind Orchestra, the non-profit organization provides numerous separate and free ensemble performances. More information about the Space Coast Symphony Orchestra's upcoming concerts, ticket sales, venues, directions and sponsorship opportunities is available at www.SpaceCoastSymphony.org by calling toll free (855) 252-7276.
By Space Coast Symphony Orchestra|2016-12-25T21:00:01-05:00November 11th, 2016|In The News, Press Releases|0 Comments
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Gulf Coast Region
Texas Point National Wildlife Refuge
Texas Point National Wildlife Refuge Keeping watch © Pat Butaud
Alligator sunning in April 2010.
Texas Point National Wildlife Refuge Partially submerged gator2 © Pat Butaud
Texas Point National Wildlife Refuge Alligator in April 2010 © Pat Butaud
Texas Point National Wildlife Refuge Texas Point © Pat Butaud
Sabine Pass - Texas Point
Texas Point National Wildlife Refuge Another gator © Pat Butaud
Texas Point National Wildlife Refuge Birds © Pat Butaud
These two birds kept up with me for about 400 yards...as though they were my tour guides
Texas Point National Wildlife Refuge Water Fowl © Pat Butaud
Texas Point National Wildlife Refuge Birds in Spring © Pat Butaud
Texas Point National Wildlife Refuge Water Fowl 2 © Pat Butaud
Texas Point National Wildlife Refuge Water Birds © Pat Butaud
Seagulls and ducks in the water.
Texas Point National Wildlife Refuge McFaddin Beach © Pat Butaud
McFaddin Beach Sabine Pass, Texas.
Texas Point National Wildlife Refuge Black Bird © Pat Butaud
Texas Point - Sabine Pass
Texas Point National Wildlife Refuge Birdwatching Boardwalk © Pat Butaud
Texas Point, Sabine Pass
Texas Point National Wildlife Refuge Pelican © Pat Butaud
Texas Point NWR Sabine Pass, Texas
Sabine Pass, Texas 77655
Bayous weave through a seemingly endless expanse of cordgrass, reptilian eyes at the waters surface witness the ever-changing variety of waterfowl, and the call of the clapper rail reverberates through the marsh. For hundreds of years, many of the sights and sounds within this dynamic eco-system have gone untouched. Under the protective umbrella of the U.S. Fish and Wildlife Service, the hope and expectation is that they will continue for hundreds more.
Welcome to the McFaddin and Texas Point NWRs, located on the upper Texas Coast. The two refuges supply important feeding and resting habitat for migrating and wintering populations of waterfowl. Established in 1980 and 1979, respectively, the 55,000 acre McFaddin NWR consists of the largest remaining freshwater marsh on the Texas Coast and thousands of acres of intermediate to brackish marsh, while neighboring Texas Point NWR encompasses 8,900 acres of fresh to salt marsh with some wooded uplands and prairie ridges.
McFaddin and Texas Point National Wildlife Refuges are two of the 540 refuges that comprise the National Wildlife Refuge System, a national network of lands and waters set aside for the benefit of wildlife, and you!

Texas Point National Wildlife Refuge is located near Beaumont, Port Arthur
Sabine National Wildlife Refuge
Share Your Texas Point National Wildlife Refuge Photos (click here)
The best way to get to the refuge is through Sabine Pass. Highway 87, washed out by coastal erosion, no longer connects High Island with the refuge. The refuge is located west of Sabine Pass on Highway 87.

redpajama_set_name: RedPajamaCommonCrawl


Free Search Filters - Filtering programs by H1B/J1 visa, Specialty, Location, Prematches, University/Community, Non-ERAS, Non-NRMP, Application deadline, State licensing eligibility, Unfilled positions, Percentage of IMGs. No subscription required.
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Requirements or preferences matching. Example: If you do not have USCE, you will match 'US clinical Experience preferred' but won't match 'US clinical Experience required'.This is helpful in determining what programs to apply first.
We provide a list of IMG friendly residency programs individually matched with your profile. We have qualification residency requirements for more then 2000 programs. We individually called and e-mailed each program to verify its requirements. The residency lists are compiled from answers provided by Program Coordinators, Program Directors and faculty members. Each residency program card contains formalized requirements, preferences along with the 'raw information' provided by the program. We are constantly updating our lists. Our last verification was finished in the beginning of February 2018. You can rely on getting the most up to date information.
In spite of shortage of physicians in the U.S. only 30% of IMGs match successfully. This leaves about 6,500 foreign trained doctors without a position every year. Even though IMGs are submitting on average 63 ERAS® applications, the average number of interviews is just 2.5. A "rule of thumb" is to have 7-10 interviews to have a successful match. Following these statistics an average IMG should increase number of applications 3 times – 189. ERAS® pricing schedule makes this impossible for most applicants. The key is to apply wisely and rule out the programs that will not consider your application anyway. We called all the programs in most IMG-oriented specialties and will tell you where to apply. We honestly believe that the list provided by the Residency Place will help you focus on the programs that you qualify for, thus significantly reducing the cost associated with submitting ERAS® applications.


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NBCU
Top 10 Vacation Moments
Check out the top ten vacation moments from Todd, Julie, Chase, Savannah, Nanny Faye and the rest of the Chrisley family.
Five Hong Kong pro-democracy protesters seek asylum in United States
Five Hong Kong protesters who reportedly fled to Taiwan by boat six months ago have arrived in the United States and intend to seek asylum, according to an activist group. The activists, all under the age of 30, faced protest-related arrests or charges in Hong Kong, and escaped the city by boat in July last year, the Washington, DC-based Hong Kong Democracy Council (HKDC) said. They are the latest protesters to seek sanctuary in the West after Beijing imposed a national security law on Hong Kong in June that makes it easier to crack down on dissent in the Chinese city. In recent weeks, police have arrested dozens of pro-democracy activists and former lawmakers in Hong Kong. The HKDC statement didn't confirm that the five that are now in the US had been in Taiwan - a popular destination for protesters, but which closed its borders last year because of the pandemic. Simon Chu, managing director of the nonprofit HKDC, said he had welcomed the five to the US last week after a "perilous journey to freedom". "The dangerous trip made by these activists illustrates how it remains difficult for Hong Kongers to seek safe harbour in the US and that options for safe passage are limited," he said. "Their desperate effort exemplifies the rapidly deteriorating human rights condition and growing humanitarian crisis Hong Kong is in right now," he said. Media in Taiwan had reported last year that the five had been intercepted by the Taiwanese coast guard as they tried to flee to the self-ruled island in late July. Officials in Taiwan have repeatedly declined to comment on the case. Activists say the five were held incommunicado in Taiwan as negotiations were carried out to help them leave for the US. In August, another group of Hongkongers attempted to flee to Taiwan in a speedboat, but were caught by the Chinese coastguard. Ten of the 12 Hongkongers were sentenced to up to three years in prison for organising and participating in an illegal border crossing last month after a "secret" trial that was condemned by Britain and others.
GitHub apologizes for firing Jewish employee who warned of Nazis in U.S. Capitol: TechCrunch
After a probe found "significant errors of judgment and procedure" in the termination of the employee, GitHub's head of human resources resigned, GitHub Chief Operating Officer Erica Brescia said on Sunday. "In light of these findings, we immediately reversed the decision to separate with the employee and are in communication with his representative," Brescia said in a blog https://bit.ly/2KnkVhI, adding that the company apologized to that employee.
In taking charge of a Pentagon battered by leadership churn, the Biden administration will look to one holdover as a source of military continuity: Gen. Mark Milley, chairman of the Joint Chiefs of Staff. President-elect Joe Biden will inherit Milley as his senior military adviser, and although Biden could replace Milley, he likely won't. A Princeton-educated history buff with the gift of gab, Milley has been a staunch defender of the military's apolitical tradition even as President Donald Trump packed the Pentagon with political loyalists.
Iran asks watchdog not to publish 'unnecessary' nuke details
Iran urged the United Nations' nuclear watchdog to avoid publishing "unnecessary" details on Tehran's nuclear program, state TV reported Sunday, a day after Germany, France and Britain said Tehran has "no credible civilian use" for its development of uranium metal. The report quoted a statement from Iran's nuclear department that asked the International Atomic Energy Agency to avoid publishing details on Iran's nuclear program that may cause confusion.
Brazil's health regulator is seeking further data on Russia's Sputnik V coronavirus vaccine before considering its approval for emergency use. Documents supporting drugmaker Uniao Quimica's application for emergency use of the vaccine have been returned to the company because they did not meet its minimum criteria, the watchdog said on Saturday. In a statement on the Health Ministry's website, regulator Anvisa said the request failed to provide adequate assurances on Phase III clinical trials and issues related to the manufacture of the vaccine.
How 'bewildered' Trump campaign aides would reportedly discreetly escape election challenge meetings
The pandemic devastated airlines in 2020 and forced many out of business - here's the most notable that didn't make it to 2021
How Princess Eugenie's royal baby will shift the line of succession
Biden plans dozens of exec orders in first 10 days of presidency
In final Trump administration days, Israel approves new settler homes


Sattvik food has it's own health benefits. It detoxifies the body and improves one's stamina. At times, it is often referred to as the yogic diet as it is the preferred diet of ascetics who can endure long periods of meditation without food and water. To enable the body to adjust to such extreme conditions, it needs to be brought to a state of balance by following pure eating habits. Because the emphasis is more on the seasonal produce, ghee, nuts, whole grains and dairy products, consuming a Sattvik diet ensures that less amount of preservatives make their way into the body.
While it is tough to make the switch for people who are used to non-vegetarian food on a regular basis, one must try and adopt a Sattvik diet on certain days of the month. It is best when done on a periodic basis and also in moderation. The 'Kartika' and 'Margasira' months in the Odia calendar offers have earmarked days when one has to stick to a Sattvik diet. On these days, the usual dishes prepared in most Odia homes are Dalma (dal with vegetables), sagaw bhaja (stir fried green leafy vegetable), khatta ( sweet tangy chutney) and a few stir fried vegetables like raw banana, yam, radish or drumsticks . No onion or garlic is used in any of these preparations.
However, it sometimes happen that guests do come over on such days. And it becomes necessary to cook some universally accepted dishes for them. Especially for those folks who prefer North Indian food. This 'Makhana Malai Matar' is one such light gravy based dish that goes well with puris or rotis.
Preparation - Heat 1 tsp of ghee in a pan. Add the Makhana and roast to a golden. Remove and keep aside.
Add 1 tsp oil to a wok. Add the whole spices and chilis. Wait for 20 seconds. Once fragrant, throw in the ginger and chopped coriander stem. Fry for 2-3 mins or till the raw smell of ginger goes off.
Finally add the tomato and sprinkle the salt over it. Cover for 2 mins to soften a bit. Once the tomatoes are mushy, add the cashews and fry for 1 minute. Remove from flame and keep aside.
Transfer to a blender jar once it has cooled down. Add 1/5 cup of warm water and blend into a smooth paste.
Cooking - Heat the remaining ghee in the wok. Throw in the green peas. Fry then for 2-3 mins.
Then add the masala paste and cook till the oil separates.
Add 1 cup of warm water, turmeric, garam masala and Kasuri methi. Adjust the salt. Bring to a slow boil.
Let it simmer for a while before adding the malai or cream.
Add the roasted Makhana just before switching off the flame.
Serve hot with rotis or puris. Goes well even with jeera rice.


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Q: (android, JAVA) How to implement 3 or 4 depth Json with GSON? I have a question about GSON lib.
My JSON data is HERE :
 {
    "UserID":"",
    "Data1":{
        "Data1_1":[{
            "name":"name1",
            "test":"test2"
        },
        {
            "name":"name2",
            "test":"test2"
        }],
        "Data1_2":{
            "Data1_2_1":[{
                "addr":"addr1",
                "phone":"phone1"
            },
            {
                "addr":"addr2",
                "phone":"phone2"
            }],
            "Data1_2_2":[{
                "country":"country1",
                "sex":"M"
            },
            {
                "country":"country1",
                "sex":"F"
            }]
        }
    }
}

My GSON Object Class is HERE :
    public class GsonClass {
    public String UserID;
    public ArrayList<Data1> Data1;

    public class Data1 {
        ArrayList<Data1_1> Data1_1;
        ArrayList<Data1_2> Data1_2;

        public class Data1_1 {
            public String name;
            public String test;
        }

        public class Data1_2 {
            ArrayList<Data1_2_1> Data1_2_1;
            ArrayList<Data1_2_2> Data1_2_2;

            public class Data1_2_1 {
                public String addr;
                public String phone;
            }

            public class Data1_2_2 {
                public String country;
                public String sex;
            }
        }
    }
}

But,
GSON library returned JSONParserException..
When I test without Data1_2, GSON Parser was OK.
But, with Data1_2. GSON library return exceptions.
How set the GSON class in 3 or 4 depth JSON?

A: Try this .
Firstly , we must know about how to deal with [] and {} in the Gson .

*

*If we find {} in the JSON , we must use DataBean in the Gson .

*If we find [] in the JSON , we must use List<DataBean> in the Gson .
And this is the class .
public class GsonClass {

/**
 * UserID :
 * Data1 : {"Data1_1":[{"name":"name1","test":"test2"},{"name":"name2","test":"test2"}],"Data1_2":{"Data1_2_1":[{"addr":"addr1","phone":"phone1"},{"addr":"addr2","phone":"phone2"}],"Data1_2_2":[{"country":"country1","sex":"M"},{"country":"country1","sex":"F"}]}}
 */

private String UserID;
private Data1Bean Data1;

public String getUserID() {
    return UserID;
}

public void setUserID(String UserID) {
    this.UserID = UserID;
}

public Data1Bean getData1() {
    return Data1;
}

public void setData1(Data1Bean Data1) {
    this.Data1 = Data1;
}

public static class Data1Bean {
    /**
     * Data1_1 : [{"name":"name1","test":"test2"},{"name":"name2","test":"test2"}]
     * Data1_2 : {"Data1_2_1":[{"addr":"addr1","phone":"phone1"},{"addr":"addr2","phone":"phone2"}],"Data1_2_2":[{"country":"country1","sex":"M"},{"country":"country1","sex":"F"}]}
     */

    private Data12Bean Data1_2;
    private List<Data11Bean> Data1_1;

    public Data12Bean getData1_2() {
        return Data1_2;
    }

    public void setData1_2(Data12Bean Data1_2) {
        this.Data1_2 = Data1_2;
    }

    public List<Data11Bean> getData1_1() {
        return Data1_1;
    }

    public void setData1_1(List<Data11Bean> Data1_1) {
        this.Data1_1 = Data1_1;
    }

    public static class Data12Bean {
        private List<Data121Bean> Data1_2_1;
        private List<Data122Bean> Data1_2_2;

        public List<Data121Bean> getData1_2_1() {
            return Data1_2_1;
        }

        public void setData1_2_1(List<Data121Bean> Data1_2_1) {
            this.Data1_2_1 = Data1_2_1;
        }

        public List<Data122Bean> getData1_2_2() {
            return Data1_2_2;
        }

        public void setData1_2_2(List<Data122Bean> Data1_2_2) {
            this.Data1_2_2 = Data1_2_2;
        }

        public static class Data121Bean {
            /**
             * addr : addr1
             * phone : phone1
             */

            private String addr;
            private String phone;

            public String getAddr() {
                return addr;
            }

            public void setAddr(String addr) {
                this.addr = addr;
            }

            public String getPhone() {
                return phone;
            }

            public void setPhone(String phone) {
                this.phone = phone;
            }
        }

        public static class Data122Bean {
            /**
             * country : country1
             * sex : M
             */

            private String country;
            private String sex;

            public String getCountry() {
                return country;
            }

            public void setCountry(String country) {
                this.country = country;
            }

            public String getSex() {
                return sex;
            }

            public void setSex(String sex) {
                this.sex = sex;
            }
        }
    }

    public static class Data11Bean {
        /**
         * name : name1
         * test : test2
         */

        private String name;
        private String test;

        public String getName() {
            return name;
        }

        public void setName(String name) {
            this.name = name;
        }

        public String getTest() {
            return test;
        }

        public void setTest(String test) {
            this.test = test;
        }
    }
}
}

And use in your code like this .
And how to parse JSON in the code .
1.Use gson.fromJson(response,GsonClass.class); to convert JSON TO DataBean class .
2.Use get and set method .Like this gsonClass.getUserID() .
3.When we find List in it , we can Use the for loop .And it the for loop , we can use like this gsonClass.getData1().getData1_1().get(i).getName(); .
Gson gson = new Gson();
GsonClass gsonClass = gson.fromJson(response,GsonClass.class);

String UserID = gsonClass.getUserID();

for (int i = 0; i < gsonClass.getData1().getData1_1().size(); i++) {
    String name = gsonClass.getData1().getData1_1().get(i).getName();
    String test = gsonClass.getData1().getData1_1().get(i).getTest();
}

for (int i = 0; i < gsonClass.getData1().getData1_2().getData1_2_1().size(); i++) {
    String addr = gsonClass.getData1().getData1_2().getData1_2_1().get(i).getAddr();
    String phone = gsonClass.getData1().getData1_2().getData1_2_1().get(i).getPhone();
}

for (int i = 0; i < gsonClass.getData1().getData1_2().getData1_2_2().size(); i++) {
    String country = gsonClass.getData1().getData1_2().getData1_2_2().get(i).getCountry();
    String sex = gsonClass.getData1().getData1_2().getData1_2_2().get(i).getSex();
}

And you can use GsonFormat in your Android Studio to convert JSON TO class .
It will be quick for you .
1.Add GsonFormat

2.Use in the code

redpajama_set_name: RedPajamaStackExchange


text:
Classic Postmortem: People Can Fly's Bulletstorm
February 22, 2016 | By Adrian Chmielarz
More: Console/PC, Audio, Design, Production, Business/Marketing, History, GD Mag Exclusive
On the fifth anniversary of the release of this unique and underappreciated shooter, Here's an in-depth postmortem that first ran in Game Developer magazine in 2011. It was written by Adrian Chmielarz, who was creative director of People Can Fly.
What could go wrong if your first project is an unassuming old-school PC shooter, and your second project is a big, multiplatform AAA title? Why would things be different if you grew from 15 to 70 employees in a couple of years? How does it feel to go from e-mail interviews to standing in front of the entire world as it watches your live E3 presentation?
Yeah, this could be a book.
Ten thousand things went right and ten thousand things went wrong during the production of Bulletstorm. Obviously there's not enough space to talk about them all, so this article is a very subjective selection, mostly filtered through the design side. It took three and a half years to make Bulletstorm. We've learned, we've grown, and we hope you'll find something for yourself in this story as well.
DEVELOPER: People Can Fly/Epic Games
NUMBER OF DEVELOPERS: Around 80 at PCF
LENGTH OF DEVELOPMENT: 3.5 years
SOFTWARE: Unreal Engine 3, 3ds Max, Maya, Photoshop, Modo, ZBrush, Motion Builder, MS Office
PLATFORM: Xbox 360, PlayStation 3, PC
1) Focus on core combat loop
///Our philosophy is that it's better to have a great game that's just about spitting than a mediocre game about spitting, screaming, and playing the banjo.
If you have ever sighed at a game's attempt to offer variety (e.g., not every game needs a driving section), you already know what I mean.
Also, stop thinking about that banjo game. It's not going to happen.
We spent months polishing our core combat loop: shooting, kicking, sliding, leashing. A millisecond here, a 1% more transparent particle there. Improve, playtest to death, rinse, repeat. We knew it all worked together well when we started missing elements of our CCL while playing other games. I cannot count the amount of times I wanted to kick or slide into an enemy when enjoying a competitor's FPS.
Of course, modern games cannot rely on the CCL alone, but we also don't think that having myriad core features is the right solution. You will never have the time to polish them all properly.
We decided to focus only on the crucial ingredients, get them to 100%, and have the gameplay variety come from a wide palette of contexts to the CCL. For example, the kick is always the same thing—the same animation, the same sound, the same button—but its purpose can change depending on what the player needs to do. Kick to push an enemy away. Kick to destroy an enemy's armor. Kick to open a door.
2) Pacing and balancing pass
/// As with most games, Bulletstorm was built by multiple level designers, each working on their own fragment of the game. At a certain point we felt that every level had really great pacing: a good warm-up, varied encounters, and an interesting cliffhanger.
Most of the levels were done with "big picture" pacing in mind. For example, the Cave level started slow, because the ending of the preceding level was a six-minute boss fight, and we wanted to let players relax a little.
But as all game designers know, theory is one thing, and the actual implementation is another. When we put the game together, some pacing problems immediately crawled out of the woodwork. The Cave's relaxed opening was a good idea, but lasted far too long. The Underground and Ulysses sections were great on their own, but both exclusively featured one type of enemy (Burnouts), and that was tiring in the long run. The Dam level felt too long, even though in isolation it was one of the best levels in the game.
We made pacing and balancing our priority. We removed and added battles. We shortened cinematics. We fine-tuned all the values for ...everything.
In the end, we got a really well-paced game. Unfortunately, it was a little too well-paced for its own good. Wait, what?!
Imagine a shooter or a horror game that takes eight hours to finish, but where the experience is so intense that you only play one to three hours daily. It means it will take you three to five days to finish the game, which is how we imagined our game would be.
However, with Bulletstorm, we have seen time and time again that people got so engaged in the experience that they finished the game in one day. It was still eight hours, but played in only one or two sessions. That allows us to enjoy the fact that over 50% of players finished Bulletstorm, which is higher than the industry's standard (this is easily trackable by checking "Finished the game" achievements in services like Raptr—see References for more). But at the same time, opinions started floating around that the game was short.
It was not short by current standards. But to gamers, two four-hour sessions are not the same as five hour-and-a-half sessions.
We are still proud of the pacing and balancing work, but intensity versus time is something we'll have to think about in future projects.
3) Finding the fun through early focus tests
/// We never planned to offer a unique gameplay hook. We just wanted to evolve the genre a tiny bit. Maybe we'd do it by unmuting the main hero, even though it's a first-person game. Maybe we'd differentiate by making sure that the sidekicks are not automatons spewing context-sensitive comments, but actual people we can have strong feelings about. Our intentions were nothing more than that. We just wanted to offer a fun high adventure—that's it.
However, the aforementioned focus on the core combat loop, constant iterations, and—most of all—internal focus tests led to the invention of Skillshots.
I am a big fan of games that offer multiple pseudo-independent systems. That is the core of any emergent gameplay. Bulletstorm has a few systems like that: multifunctional weapons (e.g., the flail chain can wrap around enemies or objects), an interactive environment (e.g., the explosive trash cans of the future can be moved around) and the tools of war (kicking, sliding, leashing).
Players can manipulate and use these systems any way they want. That leads to a lot of emergent gameplay moments. You can kick an enemy into a trash can, which then explodes, which causes another enemy to go airborne and get pierced by a rocket fired by another enemy positioned on the rooftop.
But I would never have thought of naming these crazy actions—and indeed, I would never have thought of Skillshots at all—if not for internal focus tests.
To some developers, the focus test sounds like something to dread. They think: "There's a reason why I'm the creator, and they're the consumers." That's fair enough, and there is some truth to it; but on the other hand, there's also a reason why Blizzard wins so big after spending insane amounts of time playtesting and fine-tuning.
Our early focus tests were nothing official. We were just watching each other play. In casual conversations we debated what was fun and what sucked.
But it was during these monitored play sessions that I noticed people were using Bulletstorm's emergent combat not to be efficient, but to have fun. A headshot is a much faster kill than leashing an enemy toward you, then kicking him into a cactus. And yet still people kept on doing that, and even much more elaborate things. They were experimenting. Testing theories. Having fun.
"Hey, why don't we actually reward people for being creative?" I thought. "How about we call it a Skillshot system?" And one of our core selling points was born.
4) Many companies, one game
/// You can imagine the fragmentation challenges we faced with PeopleCanFly as the main developer and creative owner, Epic as co-developer and quality enforcer, and EA as the publisher. And there were a number of additional companies from all around the world helping us as well (in Germany, USA, China, Sweden, and Poland).
This stuff is not for the faint of heart.
Oddly enough, coordinating the outsourcing was not the hardest part. With good, dedicated producers in place, it's actually something I am sure to repeat in the future.
It was the PCF/Epic/EA cooperation that caused the most problems. Everybody was equally important, and everybody could influence any part of the development process: design, production, and so forth. For example, both EA and Epic were giving us independent feedback on the milestones and playable builds. It was a mess.
Fortunately, we all noticed that very quickly. We understood that the key to fluid cooperation is a proper distribution of roles and responsibilities. In other words, sometimes you have to let go.
We immediately streamlined all these processes. Using the feedback example, EA was no longer sending feedback to PCF but to Epic. Epic then merged EA's feedback with its own, empowered to remove whatever they disagreed with, and then sent one unified chunk of feedback to PCF. This way we only had to deal with one source of feedback, were no longer prey to multiple masters, and stopped worrying about the priorities of the feedback items.
If you are working with several partners, letting go is the key. One of you knows more about the marketing than the other, another is a better judge of quality. Clearly define who is responsible for what, and make life easier for everyone.
5) Unreal Engine 3
/// Ha! What a surprise: a guy from a company owned by Epic, creator of Unreal Engine, praising the Unreal Engine. Good one!
In 2004, we released Painkiller. The game took two years to make, with a 15-person team (on the average). We had a long single-player campaign, and a fun multiplayer mode that was a CPL (Cyberathlete Professional League) game of the year. And we did all that using our own engine, which we created from scratch.
Painkiller was really an extremely simple game at its core, though, and yet still we struggled. We had to outsource cinematics in order to finish the game on time, for example. If it had been any more complicated—and I am not talking about big stuff like going multiplatform, I merely mean things like featuring a sidekick—the game would never have happened.
Instead of merely listing the benefits of Unreal Engine—something you either already know or can easily google—let me just tell you three facts from PCF's past.
First, we decided to switch from our own engine to Unreal well before we had any idea that one day we might be a part of Epic. It was worth it to hear Mark Rein scream, "Finally!" into the phone. Second, from the moment we got the engine, it took us only a month to prepare a demo for publishers—a demo that Epic saw and said "Hold on for a second there, let's talk." Third, it took us two months to make two levels of Bulletstorm for a pitch demo that we showed in Leipzig to various publishers, and which got us a deal with EA. I have absolutely no idea how we could have achieved that without UE3.
1) Creative f-bombs
/// Bulletstorm was often critiqued for its seemingly endless stream of four letter words.
Do you know any swear word in a foreign language? German, French, Polish? When you say it out loud, no biggie, right? Not a problem to use it during a family dinner, I assume?
That is how all the f-bombs sounded to us. Being Polish, all the strong language in Bulletstorm was just exotic and fun to us. We did not feel its power. In other words, Epic thought this is what we wanted and respected our creative vision, while we had no idea this vision was a bit more than we really wanted.
It was only at the end of the development, when I read the Polish translation of the game, that I realized how dirty we were. I swear a lot. A LOT. Yet still I ... kind of blushed.
But, to be honest, the language would not have been a problem if not for its creative usage. There are games that use way more f-bombs than Bulletstorm, and yet somehow it's less of a problem for them. Why?
That's because most of the time the language is forgettable. If you hear a character say "You f***** scared me, you a******!" you forget about it two seconds later. Although, if he says "You scared the d*** off me!" it stays with you for a little bit longer, which creates the impression that the game is much fouler than it really is.
We tried to solve that by adding the language filter. Originally we forced the players to make a choice before they even got to the main menu. With best intentions to make the experience more fluid we moved the option to one of the submenus.
Big mistake. No one noticed it existed. It cost us a few prestigious reviews and a sea of tweets from angry gamers.
Next time, do it like Brutal Legend: a forced choice, during the game, right before the first f-bomb. Personally, just in case, the game should ask the players once more some time later.
2) Slow beginning, debatable ending
/// I am not a fan of action-packed openings to video games. You do get players' attention, even if the attempt reeks of desperation, but the inevitable lull after the intro is over is a pacing killer. I think the only time I have seen a hi–octane opening work well was God of War, but that's just because Kratos doesn't take a break, and the opening is actually the entire game.
But, by Crom, that does not mean your game can take 45 minutes before you get to the core gameplay! Which is exactly what Bulletstorm did.
Endings are even more important. That 2–5% of the game can increase the review score if it's awesome, and kill the other 95–98% if it sucks. A bad ending is like finishing a delicious dinner only to learn there were worms in it.
Our ending is decent, but we were just trying to be too clever. We wanted the players make a certain mental choice. Bad things start happening in main character Grayson Hunt's life when revenge becomes the only thing he cares about. At the end of the game, he can choose to continue walking down that path, or let go. What would YOU do? Do you think Grayson has learned anything?
But that was a bit fuzzy, and many people treated the lack of a clear resolution and closure—even though we destroyed an entire planet—as a blatant attempt to trick them into buying the sequel.
Guess what? Throughout the entire development time we were perfectly aware how crucial the beginning and ending of a game were, and still we got both of them wrong.
Be triple careful and assign an overkill amount of time and resources to making sure that the first and last page of your book are perfect.
3) Wrong choice for the online mode
/// I'm going to tell you a secret. We had a working, playable player-versusplayer mode in Bulletstorm. It was only a basic deathmatch, and it was a mere prototype, but it was playable—and it was tons of fun.
But we felt that the PvP space was too crowded. Ignoring the fact that our PvP was unlike anything else out there (thanks to the Skillshot system you could win with fewer kills than the opposite team), we felt we needed something different. That's how the Anarchy co-op mode was born.
That, in itself, was not a bad choice. The problem was that we decided on Anarchy too late in the development process. We managed to make the mode really fun for the hardcore, advanced Bulletstorm players, but everyone else struggled. Anarchy requires tight cooperation, and is not bulletproof. If you don't work together, the mode is just not fun. That's quite unlike PvP, where most of the time both teams and individuals can enjoy the carnage.
You only have one chance to make the first impression. It's better to release a fun and polished bare minimum effort than an unfinished experiment, no matter how unique.
4) Lack of context in the demo
/// In the game industry we often debate whether releasing a demo makes sense. It's a controversial topic. But you know what's not controversial? Bad demos.
I wouldn't say our demo was textbook "bad," but it just wasn't the right demo. Demos should be about emotions, about the atmosphere, about the vibe of the game. Remember the Batman: Arkham Asylum demo? Perfection.
With the Bulletstorm demo, we wanted to show people how fun the Skillshot system was. We wanted the players to fight for the best score, and thus replay the demo over and over again. So we took a very short, storyless fragment of one of the levels, stripped it of any story-related dialogue, and focused the gameplay flow on core mechanics. Also, because our mechanics were new in the FPS world, we added a tutorial movie.
EEEEK. Wrong.
In the full game, we take it slowly. We still have elements of the tutorial two hours into the adventure. Why did we expect that people would learn all that from a three-minute movie? Why did we expect people to watch these movies anyway?
Gamers were confused. So is this a time-attack, arcade-style kind of game? There's no story, right? How can such a shamelessly short demo be so boring? All I had to do to finish it was kick, kick, kick!
We should have chosen the opposite: a story-driven "blockbuster" fragment of the game, focused on mystery and visual appeal. The Bulletstorm demo did get two million downloads across Xbox 360 and PS3 in two weeks' time, which is a good success metric, but we needed to make sure that the demo really communicated what we wanted it to.
5) Echo mode: too little, too late
/// At PCF, we keep saying that Echo mode—a sort of trials and challenges spin on the Skillshot system that ranks players based on creativity and clearing stages—was created by gamers and journalists.
Our E3 demo was focused on the gameplay and core mechanics: we knew there was no way anyone would be able to appreciate the story with a thousand other games around fighting for attention with speakers larger than a fridge.
The one silly thing we did in the E3 demo—displaying the amount of earned skill points—turned out to be a big deal. People stayed in the demo room longer just to see how others performed. They were taking photos of their score. Even the crew held an internal competition between waves of visitors. Then, when we introduced Echoes via the demo, it spawned thousands of homemade YouTube videos.
We had never planned on making an arcade experience for Bulletstorm, but what happened at E3 could not be ignored. We decided to embrace the score hunt and added a special gameplay mode with unlocks, stars, and leaderboards—the whole shebang.
It was very risky, adding a new, big gameplay mode six months before going gold. But we did it. We didn't get it quite right, however. There were too many Echoes (maps) to make competition meaningful, and those maps didn't offer new content because they were simply locations from the campaign.
Players could win vanity items in Echo mode, but not tangible gameplay rewards, so the persistence layer was lacking. We also didn't have time to implement leaderboards such as Best of the Week or Best in the Area. The list goes on.
Statistics on how many people finish games: [http://edition.cnn.com/2011/TECH/gaming.gadgets/08/17/finishing.videogames.snow/index.html?hpt=te_t1]
The lesson here is that the time needed to put a feature into the game does not equal the time of implementation. It's possible to strike gold and have your first iteration be the last, but most of the time that's not the case.
We addressed Echoes-related issues in the DLC packs by adding completely new maps made from scratch, but it was too little, too late. We're grateful that the mode took off in popularity so quickly, and the fact that it helped round out Bulletstorm's gameplay, but we were unable to invest enough time to flesh it out.
I B E L I E V E !
/// Bulletstorm is bittersweet for us. As a new IP, it sold well over a million units, which is amazing when you consider the craziness of 2011, but disappointing if you remember that everyone expected more. It suffers from a catch-22: it would have been very profitable if it had taken less than three and a half years to create, but it would not have been the same game if we did not use all that time to make it shine.
But, as Mike Capps, the boss of Epic, said, "It was worth it." Through hard work and passion we have become a ninja team ready for any zombie apocalypse. We helped improve the Unreal Engine, adding features and improving its multiplatform support. We made a game that got praise from the most prestigious, toughest magazines in the world, and that has become a lasting memory for many gamers.
Thank you, EA, and thank you, Epic. You kick ass, PCF. We're ready for the next round.
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Solvent reorganization around the excited state of a chromophore leads to an emission shift to longer wavelengths during the excited-state lifetime. This solvation response is absent in wild-type green fluorescent protein, and this has been attributed to rigidity in the chromophore's environment necessary to exclude nonradiative transitions to the ground state. The fluorescent protein mPlum was developed via directed evolution by selection for red emission, and we use time-resolved fluorescence to study the dynamic Stokes shift through its evolutionary history. The far-red emission of mPlum is attributed to a picosecond solvation response that is observed at all temperatures above the glass transition. This time-dependent shift in emission is not observed in its evolutionary ancestors, suggesting that selective pressure has produced a chromophore environment that allows solvent reorganization. The evolutionary pathway and structures of related fluorescent proteins suggest the role of a single residue in close proximity to the chromophore as the primary cause of the solvation response.
The Stokes shift between the absorption and emission of a chromophore reflects the displacement in potential surface between the ground and excited states and loss of vibrational energy in the excited state. For chromophores that have a large increase in dipole moment between the ground and excited state, the fluorescence emission maximum often depends strongly on solvent polarity in simple fluid solvents and the emission is observed to shift to longer wavelengths during the excited-state lifetime. Such dynamic Stokes shifts have been extensively studied as a probe of solvent polarity and dynamics (1–3). For a chromophore in a protein, the solvent is much more organized and constrained than in a simple solvent, so the capacity for solvation is expected to be quite different, yet important for function, from that in a simple solvent. There are relatively few studies of dynamic Stokes shifts in proteins: a few dye–protein complexes (4–7), antibodies bound to fluorescein (8), surface tryptophan residues as a probe of hydration (9–11), unnatural amino acids (12, 13), studies on cytochrome c (14, 15), and photosynthetic antenna complexes (16). In contrast to small-molecule solvents, it should in principle be possible to dissect the contributions of individual amino acids to the solvation response of a protein and even its evolutionary history, although such an analysis has not to our knowledge been reported.
GFPs would seem to be ideal candidates for measurements of the dynamic Stokes shift because the chromophore is intrinsic to the protein and structurally well characterized (17), and the 6- to 7-debye change in dipole moment upon excitation of the chromophore (18) is as large as for most dyes used to probe solvation dynamics in simple fluid solvents. Furthermore, recent studies confirmed that, like conventional solvation probes, the emission of synthetic GFP chromophores shifts substantially as a function of solvent polarity (19). Despite these properties, the Stokes shifts in most GFPs are small, suggesting that the environment in the region around the chromophore is quite rigid. Both the synthetic GFP chromophore (20) and the chromophore in denatured protein (21) exhibit a very low fluorescence quantum yield, likely because the chromophore undergoes isomerization and nonfluorescent relaxation when the solvent is flexible. Consistent with this theory, the isolated chromophore fluorescence quantum yield increases dramatically if the solvent is frozen (22), and low temperature and/or high pressure can increase the quantum yield for GFP variants with intrinsically low quantum yields, such as blue fluorescent protein (23). Thus, a high fluorescence quantum yield, which is desirable for native function (and biotechnology), and the conditions of protein flexibility needed to observe a dynamic Stokes shift appear to be mutually exclusive for GFP variants.
Recently, Wang et al. (24) reported a very red-emitting fluorescent protein named mPlum that was created by a directed evolution strategy. mPlum is derived from DsRed, which was isolated from Discosoma coral. DsRed displays bright, red fluorescence, but it is a strongly associated tetramer, so its use as a genetic marker is limited. Through a combination of rational design and stepwise random mutagenesis, Tsien and colleagues (10) generated a monomeric version of DsRed known as monomeric red fluorescent protein (mRFP). Wang et al. then used somatic hypermutation to generate a huge and diverse library of mRFP mutants, which were screened for brightness and the reddest possible emission maximum by using a FACS; the reddest populations were subjected to further rounds of directed evolution. In this selection process, the FACS excitation wavelength was fixed at 568 nm, so the selection was not just for a red emitter but also for a large Stokes shift. Interestingly, both mRFP and mPlum absorb maximally at ≈588 nm, but they emit at 607 and 649 nm, respectively. Thus, mPlum has a much larger Stokes shift than is typical for GFPs, and here we investigate the nature of this Stokes shift.
The absorption spectra of the isolated and purified mPlum and immature mPlum are shown in Fig. 1. The extra conjugated double bond of the mature form of chromophore leads to the large red shift in the spectrum relative to the immature form. An excitation spectrum of the mature protein confirmed that the minor peaks at 375 and 345 nm in the absorption spectrum of the mature form (Fig. 1 C) are the higher energy states of the mature form and not contamination from the immature form. Isolating the mature form was important to ensure that the spectroscopic and crystallographic data were of a pure protein and not a mixture of the mature and immature forms. The structures of the chromophores for mature and immature mPlum are shown in Fig. 1 B and D (S. J. Remington and X. Shu, personal communication). The extension of the chromophore is confirmed in the crystal structure by a change in the hybridization state of the exocyclic carbon (indicated by an arrow in Fig. 1) from sp3 to sp2. Neighboring residues to the chromophore, E16 and E215, also are shown.
Chromophore characterization. (A) Absorption spectra of isolated immature mPlum with corresponding chromophore chemical structure. (B) Crystal structure of immature mPlum chromophore including neighboring residues E16 and E215 (S. J. Remington and X. Shu, personal communication). The arrow points to the sp3 hybridized carbon of immature mPlum. The dotted line indicates the hydrogen bond between the glutamic acid and carbonyl of residue 65. (C) Same as A for mature mPlum. (D) Same as B for mature mPlum. The arrow points to the sp2 hybridized carbon of mature mPlum.
Fig. 2 shows the fluorescence excitation and emission spectra of mRFP, mRaspberry, and mPlum at pH 7 and pH 11. The pKa of the mPlum chromophore is <4.5 (25); therefore, the chromophore is almost exclusively in the anionic form at all pH values described here. mRFP has an excitation maximum at 588 nm and an emission maximum at 612 nm. mRaspberry is an intermediate in the evolution process from mRFP to mPlum and has excitation and emission maxima of 596 and 624 nm, respectively. The mPlum spectra are characterized by an excitation maximum at 588 nm and an emission maximum at 649 nm in pH 7 buffer. At higher pH, the excitation shifts to the blue and the emission shifts to the red, so that in pH 11 buffer, the Stokes shift is 21 nm larger with peak wavelengths of 576 nm and 658 nm for excitation and emission, respectively. Treating the pH dependence as a titration, the apparent pKa of the titratable group in mPlum is 10.7. Similar pH-dependent shifts in the excitation peak were observed for other proteins evolved from mRFP, such as mCherry (apparent pKa of 10.29) and mStrawberry (apparent pKa of 9.75) (26); however, for these proteins, the emission shift is to the blue rather than to the red, as observed for mPlum. The fluorescence quantum yield of mPlum is 0.10, substantially lower than mRFP, which is 0.26 (24).
Steady-state fluorescence spectra. Normalized steady-state excitation (dotted line) and fluorescence spectra (solid line) of mRFP at pH 7 (red), mRaspberry at pH 7 (green), and mPlum at pH 7 (black) and pH 11 (blue). The excitation wavelength for the emission spectrum and time-resolved experiments of mPlum is 580 nm (540 nm for mRFP and mRaspberry) and is indicated by an arrow. All excitation spectra were obtained at a wavelength of 650 nm.
Fluorescence decays were obtained by upconversion spectroscopy and time-correlated single photon counting (TCSPC) at 10 discrete energies from 13,946 cm−1 (717.0 nm) to 16,946 cm−1 (617.4 nm) in 250-cm−1 increments. Fig. 3 A shows the 627- and 692-nm fluorescence decays obtained by fluorescence upconversion spectroscopy for mPlum. The decays were well fitted by the sum of two exponentials convoluted with the instrument response. The 627-nm trace is characterized by a fast decay of 39 ps that makes up 28% of the total amplitude followed by a slower decay of 573 ps. The kinetics at redder wavelengths show a fast rise with a lifetime of 39 ps and a −27% amplitude contribution followed by a slower decay of 723 ps. The decay could not be fitted well with either a sum of exponentials or a stretched exponential without a negative amplitude contribution. The negative amplitude contribution is consistent with an increase in the excited-state population at this wavelength, the hallmark of a dynamic Stokes shift associated with the solvation of excited chromophores. mPlum's evolutionary ancestors, mRFP and mRaspberry, do not show a similar solvation response. In the case of mRFP, the fluorescence decays show no rise, and decays at both red and blue wavelengths are well described by a single exponential decay. For mRaspberry, there may be a rise at red wavelengths, but it is very small compared with mPlum with a total amplitude of <4%.
Time-resolved measurements. (A) Normalized time-resolved fluorescence decays for mPlum, pH 7, at 627 nm (blue) and 692 nm (red) as obtained by upconversion spectroscopy with excitation at 580 nm. Excitation occurs at 1 ps on the time axis. (B) Reconstructed time-resolved fluorescence spectra obtained at 1, 20, 50, 100, and 1,000 ps as obtained by upconversion spectroscopy and TCSPC. The solid lines are log normal line-shape fits to the data. (C) Fluorescence emission maximum as a function of time for mPlum at pH 7 and pH 11 as obtained from upconversion spectroscopy and TCSPC as described in the text. (Inset) Detail for the first 0.2 ns. (D) Fluorescence emission maximum as a function of time for mPlum, mRaspberry, and mRFP in pH 7 buffer as obtained from TCSPC. The excitation wavelength for these measurements was 545 nm.
Fig. 3 B shows normalized, time-resolved fluorescence spectra. The spectra are fit to log-normal line shapes and show a fast shift in time to lower energy. Peak fluorescence energy versus time for samples in pH 7 and pH 11 buffers are shown in Fig. 3 C. The dynamic Stokes shift at pH 7 is characterized by a fast shift that is largely completed by 500 ps. It can be fitted to a double exponential with amplitudes of 301 and 492 cm−1 and lifetimes of 4.3 and 71 ps, respectively. At pH 11, a similar fast component is observed with amplitudes of 268 and 357 cm−1 and lifetimes of 3.3 and 50 ps, respectively. In pH 11 buffer, the dynamic Stokes shift also exhibits a longer time component, with an amplitude of 175 cm−1 and a lifetime of 900 ps. Note that uncertainty in the emission maximum is greater at early time due to the lack of data points on the high-energy side. The lack of data points is due to light contamination from upconverted scattered excitation light. See supporting information (SI) Text for a detailed description of the data analysis. Dynamic Stokes shifts for mPlum, mRaspberry, and mRFP at pH 7 from TCSPC are shown in Fig. 3 D. Both mRaspberry and mRFP show a negligible shift in peak emission wavelength. Upconversion experiments confirmed that there is also not a significant shift on faster timescales.
The effect of temperature on the steady-state fluorescence emission spectrum of mPlum was studied from 298 to 140 K in pH 7 buffer (Fig. 4 A). As the temperature was lowered, the steady-state emission maximum decreased from 648 nm (15,432 cm−1) at 298 K to 624 nm (16,026 cm−1) at 140 K. The glass transition temperature for the 60/40 glycerol/buffer solution is ≈200 K. There is a steady shift in emission maximum above the glass transition; however, below the glass transition, there is only a small dependence of the emission maximum upon a further decrease in temperature. The peak of the excitation spectrum also shifts to lower wavelength with decreasing temperature, from 588 nm (17,007 cm−1) at 298 K to 582 nm (17,182 cm−1) at 140 K (data not shown). The temperature effect on the dynamic Stokes shift is shown in Fig. 4 B and was obtained by TCSPC at 298, 270, 230, and 140 K. At 298, 270, and 230 K, the dynamic Stokes shift is characterized by a fast component that is largely complete by 200 ps. Below the glass transition, at 140 K, no dynamic Stokes shift is observed.
Temperature dependents. (A) Normalized steady-state fluorescence for mPlum at pH 7 from 298 K (red) to 140 K (blue). The excitation wavelength was 466 nm in all cases. (B) Fluorescence emission maximum as a function of time for mPlum at pH 7 for temperatures of 298, 270, 230, and 140 K as obtained by TCSPC.
mPlum is the final product of a long series of mutagenesis steps starting with the tetramer, DsRed, proceeding through monomeric mRFP, and finally via somatic hypermutation to its final red-emitting form. Approximately 5 nm of the shift to redder emission can be attributed to a mutation that is incorporated into the chromophore itself: The Gln-Tyr-Gly sequence that forms the chromophore in DsRed was changed to Met-Tyr-Gly in mRFP (27). The remaining shift results from mutations to residues around the chromophore. mPlum exhibits a substantial dynamic Stokes shift, possible only if the region of the protein solvating the chromophore is capable of some flexibility, yet the mPlum fluorescence quantum yield is reasonably high, suggesting that the directed evolution process has selected for a compromise between flexibility and local rigidity. We consider other possible processes in this Discussion.
Solvent reorganization can be monitored by examining the spectral and kinetic components of the fluorescence decay. As shown in Fig. 3 A, a fast decay component is observed at wavelengths toward the red end of the steady-state emission maximum, and a negative amplitude exponential is required to accurately fit the data. This rise in fluorescence amplitude on the red side of the band is characteristic of solvent reorganization as the (protein) environment reorganizes to better solvate the dipolar chromophore excited state. The kinetics are not consistent with two noninterconverting species (or species that interconvert slowly compared with the excited-state lifetime) of different fluorescence lifetimes because only a population transfer, not a difference in lifetime, can produce a rise. Two or more species that interconvert on the timescale of the excited-state lifetime could potentially produce the observed kinetics. However, other data argue against this possibility. The shape of the emission spectra (Fig. 2) and the steady-state anisotropy (data not shown) are the same, irrespective of excitation wavelength. If there were multiple species, it would be likely that each could be preferentially excited at different wavelengths, leading to measurable differences in fluorescence line shape and/or anisotropy. The steady-state anisotropy is high (>0.35) and independent of wavelength; thus, there also is no evidence of a major reorientation or reaction of the chromophore to a photoactive state upon excitation. The time-resolved anisotropy obtained from TCSPC that decays with a single exponential lifetime of ≈14 ns (data not shown) is consistent with the overall rotational correlation time of the protein of 16 ns, which was previously reported for WT GFP and S65T GFP (28), and discounts faster rotation of the entire chromophore with respect to the molecular frame of the protein.
The nature of the fluorescence shift is made clearer by the time-resolved spectra in Fig. 3 B. The data are consistent with a chromophore excited state that progresses to lower energy as it becomes better solvated. Although the shape of the emission spectrum is largely unchanged as it shifts in energy over time, the FWHM [Γ(t) in Eq. 7 of SI Text ] at pH 7 becomes narrower [from 2,350 cm−1 at 10 ps to its final value of ≈2,000 cm−1 after 500 ps; before 10 ps, the lack of data points on the high energy side of the band leads to poorly defined Γ(t) and inconsistent results between data sets]. Some narrowing on a similar time scale and magnitude also has been observed for a dye in simple solvent (2). This narrowing has been interpreted as an environment that is becoming more organized and ordered over time, leading to slightly less inhomogeneous broadening (2). Apparently, a similar process is possible in the interior of a protein.
The dynamic Stokes shift of mPlum at pH 7 occurs on a timescale of tens of picoseconds, with no evidence for further solvation on the nanosecond time scale. Fitting the peak energy as a function of time with two exponentials (4.3 ps and 71 ps) (Fig. 3 C) yields results that are slower than observed for dyes in simple, fluid, protic solvents (2) and closer to that observed for tryptophan residues on the surface of a protein (10, 29). The fastest component is likely part of the initial response observed for dyes in protic solvents (30), a fast reorganization and rotation of the relatively unrestrained solvent and side chains around the chromophore. The magnitude of the initial response is likely underestimated given the time resolution of the experiment (instrument response of 400 fs). The solvation response lacks the very slow components (hundreds of picoseconds to nanoseconds) reported for some probes buried or partly buried in proteins, such as dye–protein complexes (5, 7, 31) and unnatural fluorescent amino acids inserted site-specifically at several locations in the sequence of the small protein GB1 (12). However, direct comparison is difficult because, unlike these other systems, mPlum has an intrinsic chromophore that is part of the backbone and protected deep within the protein's β-barrel. Therefore, motions that may be possible within a binding pocket or small protein may be inhibited in mPlum at neutral pH. In contrast, mPlum in pH 11 buffer shows a slow and fast component. The slow component has a lifetime of ≈900 ps compared with the fast components of 3 and 50 ps. The slower component at high pH could be caused by the chromophore environment becoming less rigid as the pH increases. Interestingly, this added flexibility does not come with a major loss in fluorescence because the quantum yield at pH 11 is only slightly lower than at pH 7.
The solvent response in mPlum that produces its far-red emission is not observed in mRFP, its evolutionary ancestor (Fig. 3 D). The changes in dipole moment between the ground and excited state of mRFP and mPlum were determined to be 6 ± 1 and 5 ± 1 debye, respectively, by electronic Stark spectroscopy (data not shown), similar to GFP (18). Therefore, from the perspective of the electrostatic perturbation associated with excitation, these proteins should be comparable, and the difference in Stokes shift is likely due to the difference in protein solvent.
A blue shift in steady-state fluorescence spectra with decreasing temperature has been attributed to solvation in both small, fluorescent dyes in simple solvents and fluorescent probes in proteins. For dyes in simple solvents, when the temperature is decreased, solvation is slowed due to an increase in viscosity. When the rate of solvation approaches the inverse of the lifetime of the dye, there is a significant contribution to the steady-state spectrum from higher-energy, partially solvated configurations leading to a blue shift in the steady-state spectra at low temperatures. In the few studies of fluorescent probes located in the protein interior, the steady-state spectra also have been shown to shift to higher energy at lower temperatures (5, 32); however, in contrast to simple solvents, studies on a dye–apomyglobin complex showed that, within the studied temperature range of 243–298 K, the rate and magnitude of the solvation response was largely independent of temperature (5). Instead, it was the initial and final emission energies that appear to change with temperature.
As the temperature is lowered, the mPlum steady-state fluorescence emission maximum shifts to higher energy (Fig. 4 A). This observation can be explained by one of two models. The initial and long-time emission energy may be temperature-dependent. In this case, the protein structure would change with temperature, and we would observe the solvation response between different protein conformations at different temperatures. Alternatively, the initial emission energy may be temperature-independent, but the magnitude of the fast dielectric response that occurs on the timescale of our instrument response may be temperature-dependent. In this case, we may not observe the very fast solvation component, and it would appear as if the solvation response originated from a lower energy state. The time-zero and long-time emission energies increase with decreasing temperature (Fig. 4 B). However, the rate of the observed solvent reorganization is largely temperature-independent, and a fast solvation component is observed for all temperatures above the glass transition. At 140 K, below the glass transition, no time-dependent Stokes shift is observed. The final emission energies can be confidently determined as all solvation occurring in the first few hundreds of picoseconds, well within the 12-ns measurement window. The lack of appreciable change in the rate of solvation is consistent with a buried chromophore that is largely isolated from the large change in bulk solvent viscosity. The change in steady-state emission spectra can be understood as a change in the final emitting state that is the result of either a temperature-dependent protein structure or a temperature-dependent magnitude of the initial solvation response.
Because mPlum was created through a combination of design and in situ evolution and we have several of its ancestors in hand, this system offers a unique opportunity to dissect the contributions to spectral shifts and the dynamic Stokes shift. mRaspberry is particularly useful because it emerged during the evolution from mRFP to mPlum. Because mRaspberry lacks a time-dependent Stokes shift (Fig. 3 D), the solvation response must be a consequence of mutations that occurred after its development. The mutations that differentiate mRaspberry and mPlum are V16E, R17H, K45R, C65I, G71A, L124V, and K166R. The mutation L124V is associated with a narrowing of the high-energy side of the emission peak but does not change the peak wavelength. Most of these mutations were introduced late in the directed evolution and lead to moderate changes in the extinction coefficient, with little effect on the peak emission maximum that is most indicative of the solvent response. Two mutations however are essential to the far-red emission of mPlum, V16E, and C65I. Saturation mutagenesis showed that variation at either site to any other amino acid led to significant blue-shifted emission or loss of fluorescence (see figure 4 in ref. 24). Therefore, these residues are the focus of discussion.
Residue 65 directly precedes the heterocyclic ring of the chromophore, and the oxidation of its peptide bond extends the chromophore, producing the characteristic red emission of DsRed-derived variants such as mPlum (Fig. 1 D). The sidechain of E16 is positioned nearby and forms a 2.6-Å-long hydrogen bond with the backbone carbonyl of residue 65, which is part of the extended chromophore. The side chain of residue 65 points away from the chromophore (Fig. 1 D). The size and shape of the side chain is crucial because mutation to another hydrophobic residue, such as leucine or alanine, leads to a blue shift of the emission by 25 nm. Mutation back to a cysteine, found at residue 65 in mRasberry, leads to a similar blue shift. Because the side chain is hydrophobic and has no direct contact with the chromophore, its contribution to the red emission of mPlum is likely due to packing, which affects the specific interactions of the chromophore within the β-barrel, probably with residue 16.
The hydrogen bond between E16 and the chromophore is believed to be the major cause of mPlum's far-red emission. Removal of this hydrogen bond by mutation to glutamine did not change the position of the sidechain, as confirmed by crystallography (26), but led to a 19-nm shift of emission maximum to the blue. Mutations to other amino acids, such as leucine and histidine, lead to even larger shifts of 30 nm. The solvation response in mPlum can then be explained as a time-dependent interaction between E16 and the excited state of the chromophore. The simplest explanation for the time dependence is a rotation of the glutamic acid in response to the new charge distribution of the excited state.
The link between E16 and the red shift suggests that the dynamic Stokes shift is the result of a specific excited-state rearrangement between the chromophore and a strongly interacting sidechain. This interpretation differs from canonical solvation theory, where a collection of solvent dipoles stochastically rearranges around the excited-state dipole of a chromophore. However, the possibility that the mutation causes other changes or increases the general flexibility of the chromophore environment, including water inside the β-barrel, cannot be dismissed. Flexibility of the chromophore environment would be expected to lead to a lower quantum yield. It is unclear whether increased flexibility around the chromophore is the cause of the lower quantum yield for mPlum compared with mRFP and mRaspberry.
Separation of immature and mature mPlum was achieved by anion exchange chromatography: a P900 (Amersham Biosciences) FPLC provided a constant flow rate of 2.00 ml/min through four 5-ml HiTrap SP HP columns (Amersham Biosciences) connected in tandem. Elution occurred with a pH gradient in 50 mM citrate buffer from pH 5 to pH 6 over 150 ml. After elution, protein was exchanged into pH 7, 50 mM Mops, 100 mM NaCl buffer and stored at 4°C. mRFP (10) and mRaspberry (24) were grown and purified by the same procedure. The mRFP1.2 variant (27) was used exclusively in this study and is referred to as mRFP for simplicity.
Absorption and Steady-State Fluorescence Spectroscopy.
Absorption and steady-state fluorescence spectra were obtained with a Cary 6000i (Varian) and a FluoroLog-3 (Horiba Jobin Yvon) spectrometer, respectively. Variable temperature measurements from 298 to 140 K were performed on mPlum in 60/40 glycerol/buffer solution using a miniature Joule–Thomson refrigerator (MMR Technologies) and a 1-mm sample geometry, with temperature calibration as described in detail elsewhere (8). Electronic Stark spectra were obtained and analyzed as previously described (34).
With the exception of the excitation source described below, fluorescence upconversion spectroscopy (34, 35) and TCSPC (36) were performed largely as described previously. Excitation light at 580 nm (or 540 nm for some experiments) was obtained from the pulsed, supercontinuum output from a photonic crystal fiber (Femtowhite). Upconversion with photonic crystal fiber excitation produced a Gaussian instrument response function of ≈400 fs at FWHM, for TCSPC it is a series of Gaussians of ≈30 ps at FWHM. The photonic crystal fiber was pumped with 800 nm light at 82 MHz with a Ti:Sapphire oscillator (average pump power of the 100-fs pulses was 400 mW). A Faraday isolator (OFR, Inc.) was inserted before the photonic crystal fiber because back reflection from the fiber was sufficient to interrupt the mode locking of the laser. A half waveplate (Newport) was used to control the polarization of the input beam, and a ×60 microscope lens (Thorlabs) was used to couple the light to the fiber. The fiber generated a continuum of light from ≈500 to 700 nm, and a 20-nm band pass filter centered at 580 nm (Chroma Technology) was used to select the appropriate light for mPlum excitation with an average excitation power between 2 and 3 mW. For some experiments, excitation light centered at 540 nm with a 10-nm bandpass was used. Collimation after the photonic crystal fiber was achieved with a ×60 microscope lens identical to the input lens, and a broadband half waveplate (Newport) was used to control excitation polarization.
We thank Profs. Roger Tsien and Lei Wang (now at the Salk Institute, La Jolla, CA) for providing the clone for mPlum and for many useful discussions; Prof. S. James Remington and Dr. Xiaokun Shu (both at University of Oregon, Eugene, OR) for crystallizing mature and immature mPlum and providing the coordinates for the chromophore and nearby amino acids (Fig. 1); Timothy B. McAnaney for helpful discussion; and Tony Kanchanawong for the determination of the change in dipole moment for mRFP and mPlum by electronic Stark spectra. This work was supported in part by National Institutes of Health Grant GM27738 (to S.G.B.). The fluorescence upconversion facilities were supported by the Medical Free Electron Laser Program of the Air Force Office of Scientific Research through Grant F49620-00-1-0349. X.S. was supported by a William R. and Sara Hart Kimball Stanford Graduate Fellowship. P.A. was supported by the Fonds de Recherche sur la Nature et les Technologies and the National Science and Engineering Research Council of Canada.
Stanford University, Keck Science Building, 380 Roth Way, Stanford, CA 94305.
Author contributions: P.A., W.C., X.S., and S.G.B. designed research; and P.A., W.C., and X.S. performed research.
This article contains supporting information online at www.pnas.org/cgi/content/full/0706185104/DC1.
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In this lesson students, will learn about common law, statute law, and constitutional law. They will begin by learning about the three branches of government. Students will have the opportunity to understand the process used to pass legislation in our Parliament. This will prepare them for a mock parliament. Students will also learn about Canada's constitutional framework, what it is and how it came to be. Students will learn about the areas of jurisdiction carved out under a federalist state. Students will use their learning to explore disaster scenarios and to write and present a disaster relief plan using emergency services provided by the different levels of government – Federal, Provincial, and Municipal.
What is the difference between common law and statute law?
What is the function of the three branches of government (legislative, executive, and judicial)?
How does a bill become a law in Canada?
Why does our government divide its powers between the federal and provincial governments?
The information on this topic is brief. Students will read Handout 1: Common Law for homework.
This topic is covered in Activities 1-5. Before starting the lesson, read the content for this topic.
For Activity 1, students will begin by reading Handout 2: The Branches of Government and will use this information to complete the chart in Handout 3: The Three Branches of Government. In Activity 2, students will complete a vocabulary list on Handout 4: Parliamentary Vocabulary. Students will then learn how a bill becomes a law by watching a short video. Next, students will complete a short writing assignment on Handout 5: A Day in the Life of a Bill and fill out a chart on Handout 6: Bills! Bills! Bills! for Activities 3 and 4. Activity 5 is a role play exercise, and will require Handout 7: Canada's Parliamentary System, Handout 8: Parliamentary Roles, Handout 9: Model Parliament and Handout 10: Legislative Bills for Model Parliament.
Activities 6 and 7 discuss constitutional law. Before introducing students to this topic, review the content. Students will read Handout 12: Constitutional Framework for homework and use it to complete five questions on Handout 13: What Did I Learn?
For Activity 6, discuss Handout 14: Levels of Government Responsibilities/Services to use here. Students will use this to answer the questions on Handout 15: Who is Responsible?
Activity 7 is a group exercise where students choose a natural disaster and create a disaster relief plan. They will identify which level of government will be responsible for what. They will need Handout 17: Disaster Plan Outline and can use Handout 18: How the Government Helps in a Disaster to test their knowledge.


text:
Q: Eclipse open declaration (F3) for resources as you know :
In Eclipse when F3 is pressed on a class, its declaration appears.
But if we press F3 on a resource, it opens the R.java and shows resource's ID.
My questions is :
Is there a way to open the resource declaration in the xml file instead of R.java ?
(As on earlier versions of ADT plugin ?)

A: Press ctrl and then click on the name of resource it will prompt you to open that resource in related.xml file.
Other think could be as @Roman mentioned but that would not be good if you have same xml file in you project.

A: I doubt that you can open android resources this way. It is not quite that, but for workaround you can use Ctrl+Shift+R to open resource files by name. Very convenient.


Obama's Katrina? How About Obama's Medicare Part D?
Ezra Klein writes today that reporters should stop asking if Obamacare is "Obama's Katrina." After all, Katrina killed 1,833 people. The Obamacare rollout has killed zero people. So knock it off.
However, Klein raises another point that's interesting for a different reason: nobody really needs to compare Obamacare to Katrina because there's a much more apt comparison at hand: the rollout of Medicare Part D in 2006. It was a disaster! And it was a health care plan! What better comparison do you need?
didn't complain loudly. They did. And yet, despite all that, no one uses it. Why?
Here's my guess: It's because in 2006 there was no liberal equivalent of Drudge and Limbaugh and Fox News on the left. That's changed a bit since, but MSNBC is still a shadow of the Drudge/Fox/Limbaugh axis. These guys are simply way better at milking a narrative and getting the traditional media to play along. And the Obamacare narrative is tailor-made for them. Bureaucratic failure. Broken promises. Rising costs. Their outrage is taken as entirely sincere, and for that reason it gets amplified into a feeding frenzy in the media that makes the Obamacare rollout seem not just modestly worse than the Medicare Part D rollout, but an epic disaster unparalleled in the history of social welfare.
In fairness, there's a second reason: The Medicare Part D rollout might have been a debacle, but it didn't cost anyone anything. It was literally something for nothing, so nobody ended up with higher monthly bills to complain about. That's the miracle of being a fiscally irresponsible party that funds new programs without bothering to pay for them. Democrats could have done the same thing with Obamacare and avoided a lot of its rollout problems, but they mostly decided to be responsible and pay for things honestly. As usual, it turns out that Americans don't appreciate that much.
1This is not a license to be an idiot. If you start calling Obamacare "Obama's Holocaust," then you're an idiot.