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Hungary's and Rumania's Nazis-in-Red:
Hitler's Graduates Staff Stalin's New Order
Béla Fabian here presents us with a brief "Who's Who" of former fascists who have been rewarded for their past behavior with positions of authority in the Communist regimes of Hungary and Rumania; the list could be substantially expanded—apparently an impeccable fascist background is a special guarantee of fitness to rule in the "new proletarian democracies."
Two correspondents of an American news agency called on Deputy Prime Minister Rákosi of Hungary in the summer of 1945. At the very outset of their conversation, one of the newspapermen produced a printed form whose text ran somewhat like this: "I admit that I was formerly a member of the Arrow Cross party. I pledge myself, from now on, to be faithful to the Communist party." "Is this declaration authentic?" the journalist asked. "Of course it is authentic," replied Rákosi. "How else could one build up a Communist party in this country? How else could I find enough Communists?"
By this and similar routes former Nazis have been swarming into the East European Communist parties, where they are received with open arms. The Nazis need a place of refuge, the Communist party badly needs members: here, obviously, is cause for a marriage of convenience. But so vast is the scale of this recruitment that it would seem to be just as much a marriage of true minds. Indeed, by now the people of these satellite countries—liberals, intellectuals, trade-union leaders, socialists, businessmen, workers, peasants, Jews—see little to distinguish the red dictatorship from the previous brown one.
This state of affairs is not secret; and it would seem to have a rather important bearing on current political discussions. Yet it has received only the most fleeting attention in the American press or on the part of "progressive" opinion generally, so ingrained is some American liberals' habit of self-suspicion, and of focusing criticism on their own government as the real enemy. Foreign correspondents continue to devote themselves to long and indignant accounts of the elections of minor Nazis to some town council in one of the states of West Germany, while neglecting the emergence of the Communist-fascist regimes in the satellite countries. The term Communist-fascist is used advisedly. For in Eastern Europe it is not a case of the government making concessions to an unregenerate public opinion, or of a misguided generosity on the part of some leading officials, or of just plain stupidity or ill will. Rather is it a systematic policy of building a regime called Communist with personnel whose chief qualification for leadership in this enterprise is their demonstrated prowess in fascist movements.
Does this sound exaggerated? Well, here is a specimen "Who's Who" of some of the former Nazis in Hungary and Rumania who have blandly swapped the swastika for the hammer and sickle.
Kálmán Zolnay, one of the more vicious Nazi officials of the Pest county court, is remembered for having publicly urged that not only all persons of Jewish descent be deported, but also anyone having any Jewish friends or relatives, because "the Jews would certainly have contaminated his way of thinking." Under the Communist regime, Zolnay has become chief of the department that handles prison matters in the Ministry of Justice. He has there gained some further notoriety by proposing legislation to make it the legal duty of every citizen to maintain close watch over a specified section in the vicinity of his own residence, and keep a written account of the activities of his neighbors.
Márton Bodonyi, who conducted the investigations against Cardinal Mindszenty, had served, under the name of Schweitzer, as a military prosecutor during the Nazi occupation. In that post he had demanded—and usually obtained—the penalty of hanging for every officer and soldier who deserted the Hungarian army, then fighting side by side with the Wehrmacht After the Russians occupied Hungary, Bodonyi was appointed chief state prosecutor in Nyiregyhaza.
Gyula Alapi, the infamous state prosecutor of the Mindszenty trial (who received a reward of 100,000 florins for successfully carrying out this special assignment), became a Communist party member in 1945. Previously, he had played an important role in Christian – Conservative student organizations: "The principles of Christian politics are a Christian order of the world, papal encyclicals, and the fight against the barbarism of Moscow," he once announced in a speech. In 1944, Alapi, then state prosecutor of Gyor, joined the fascistic Arrow Cross party. ". . . A reliable party member and a zealous Nazi," is the notation on the margin of his party record. Today, Alapi represents the government at all major trials.
Vilmos Olty, president of the court in the Mindszenty trial, as well as in the Vogeler and Rajk trials, is today perhaps the most widely known personage in Hungarian "jurisprudence." Previous to the German invasion, Vilmos Olty had been an extreme rightist. As an official in the Ministry of Justice in 1941, he was charged with the confiscation of mills belonging to Jewish persons, and executed the law with a zeal that went far beyond regulations. Subsequently, Olty joined the staff of the Hungarian-German Society and was sent to Munich on a scholarship for basic indoctrination in Nazi ideology. After the Russians marched in, Olty joined the Communist party. In his capacity as a judge of the "People's Court" of Budapest in 1945, he was noted for the severity of his sentences. It was Olty who invented and introduced the practice of having "dress rehearsals" of trials, a technique put to efficient use in the Mindszenty and Vogeler cases. In the Rajk trial, Olty emphatically played upon anti-Semitic sentiment. He never failed to ask each defendant his father's and grandfather's names—in order to disclose a possible Jewish origin. He even forced Rajk, not a Jew, and in fact a known anti-Semite, whose brother had been an important official under the Nazis, to confess that he had conspired with the Zionists.
Sándor Zöld, Hungary's present Minister of the Interior, first came to public notice on November 14, 1934, in the town of Debrecen. That day began a series of attacks on Jews at the medical college of Debrecen, in the course of which the Jewish students were driven out by the Nazis. Among the twenty-four anti-Semitic rioters arrested were Sándor Zöld and his close friend Szilard Ujhelyi, until recently an undersecretary (the third ranking official in each ministry) of the Communist government (he was swept away in a purge early in April). Sándor Zöld went on to practice medicine in Nagyvarad, a town he had to leave after performing an illegal abortion. He then moved up to the position of gynecologist at the hospital of Berettyóujfalu, where he is remembered as having been a Nazi. He became a councilor in the Ministry of the Interior as early as 1945 and soon advanced to undersecretary. On the demand of Rajk, Zöld was removed from his position by the Communist party because of involvement in a scandal—he was charged with receiving a share of the money extorted from the inmates of concentration camps by a fraudulent lawyer. After Rajk's fall, his successor had Zöld reappointed to his former position. In the summer of 1950, he became Minister of the Interior.
János Vikol began his medical career as a leading member of the Csaba Fraternal Society, a right-wing medical students' organization. The Csaba repeatedly started riots against Jewish students, demanding that they not be allowed to sit on the same benches with the Christian students. After his graduation, Dr. Vikol became an active member of the rightist medical association, the M.O.N.E., which raised the demand that Jewish doctors should be limited to treating Jewish patients only. Captured on the Russian front in World War II, Vikol asked to be sent to Moscow, to the Communist party school there. Ernö Gerö, Rákosi's present Number Two, was one of his teachers at that school. Today, Dr. Vikol is undersecretary in the Ministry of Public Welfare.
Alajos Rottenbiller was formerly a district medical health officer in Budapest, and a card-bearing member of the Arrow Cross party. Dr. Rottenbiller joined the Communists in 194; and at the present time enjoys the special protection of a top-ranking Communist minister's wife. He is chief inspector of public health in Hungary.
Antal Babits, the present chairman of the Doctors' Union, in his former capacity of professor at the Szeged medical college, participated in the expulsion of Jewish students. A former card-bearing member of the Arrow Cross, he became a Communist party member in 1945. His wife, the actress Ilona Eszterházi, who joined the National Socialist party in Berlin, when acting in German films, is now a member of the National Theater in Budapest.
Ivan Boldizsár, Hungary's most prominent journalist, was until recently undersecretary in the Ministry of Foreign Affairs. (He has been demoted, since then, to editor in chief of the Budapest daily Magyar Nemzet.) He was born the son of Jewish parents, but changed his religion. One of his articles, published in the Nemzeti Ujsag in the pre-Nazi era, and entitled "Thomas Mann and His Jews," is a prize specimen of anti-Semitic journalism. At the time of the deportations of Jews in 1944, Boldizsár continued to perform confidential assignments for the Ministry of Foreign Affairs and to receive his salary. In the fall of 1944, an anti-Nazi conspiracy by high-ranking Hungarian army officers was betrayed to the authorities. Boldizsár, who was arrested along with the conspirators, was released after several days. Presumably, he played a dubious role in the betrayal—a fact which permitted the Communists to blackmail him mercilessly.
László Piros, an unknown former cobbler, has achieved the most staggeringly successful military career under the present regime. Piros was one of the perpetrators of the infamous Miskolc pogrom in 1946, in the course of which several Jews were murdered by a Communist mob. When the government embarked on its new anti-Semitic trend in 1948, the hitherto unknown Piros came into prominence. Today, as commander of the Frontier Guards with the rank of a major general, his chief assignments are preventing Jews from escaping the country and the carrying out of deportations to the Soviet Union. He is also an alternate member of the Politburo.
Béla Csikós Nagy can boast a long Nazi heritage. His parents were close friends and staunch supporters of the fascist murderer Szalasi, supplying him with food and other comforts during his three years' prison sentence in Szeged from 1938 to 1941. Csikós Nagy was secretary to the rightist Minister of Finance, Lajos Reményi Schneller (hanged in 1946), and wrote a pro-Nazi pamphlet proving the inevitability of Nazi victory in the war. Today he is the virtual head of Hungary's totalitarian economic system, and is known as the real power behind Zoltán Vas, head of the Planning Bureau.
Tibor Fajth, section chief of the Planning Bureau, was recently decorated with the Gold Medal of Labor by the Communist government. Fajth, a Communist party member since 1945, was in pre-Nazi times an official on the mayor's staff at the Budapest City Hall. As a member of the Arrow Cross, he conspicuously wore the party's insignia on the sleeve of his coat. Fajth bears a significant share of the responsibility for the death of his thirty-six Jewish colleagues at City Hall who were sent to the Russian front by the Nazis. After the Nazi defeat, a delegation of Jewish survivors called on the mayor of Budapest and made a complaint against Fajth's holding office. "It doesn't matter who pushes the cart, so long as it keeps rolling," was the reply they received.
Joseph Takács, another section chief in the Planning Bureau and a close friend of Fajth, is also a former member of the Arrow Cross, and also shares responsibility for the death of the thirty-six Jewish City Hall officials. During the German occupation Takács was the mayor of Szolnok, where he supervised the rounding up and deportation of the town's Jews. After the Nazi defeat, he was appointed to a post in the Ministry of the Interior, being promoted after a few months to the rank of councilor.
Sándor Scmidt, long-time Nazi, has been for years the omnipotent general manager of Hungary's greatest coal-mining company, which worked the mines of Dorog and of Salgótarján. His brutality to members of the Jewish labor battalions assigned to the coal mines under his direction in 1943 was a special scandal. The Communists have kept him at his post.
Iván Bakócz joined the Communist party while still under investigation for his activities as a member of the Arrow Cross. He became deputy chief of the Communist Economic Police, and earned a quite special reputation for blackmailing, terror, and sadism. Not long ago, after his immediate superior was purged (via the gallows), Bakócz fled to the West.
Ákos Mayor had formerly been assessor of the Military Court under the Nazis, in which capacity he passed ruthless sentences against Hungarian "deserters" from the Axis armies. In recognition of loyal service, Hitler bestowed upon him two high decorations. After the arrival of the Russians, Ákos Mayor was recruited into the Communist party by his brother, Tamás Mayor, now the director of the National Theater. Ákos Mayor became a judge, then president of the N.O.T., the court of appeal of the "People's Court," where he treated opponents of the Communists as harshly as he previously had anti-Nazis. One day Ákos Mayor disappeared without a trace and is rumored to have been "invited" to Russia.
Except for the difference in names, the situation in Rumania duplicates that in Hungary. In the fall of 1944, Techari Georgescu, Rumania's Minister of the Interior, made an agreement with Grozdea, head of the extremist branch of the Iron Guard, by which the Communist party would henceforth welcome former Iron Guardists into its ranks. In a public speech in 1945, Communist boss Ana Pauker openly called upon the Iron Guard members to join the Communist party. It was in response to this call that such a man as Ion Burdycea, one of the Iron Guard leaders chiefly responsible for the Jassy pogrom in which 12,000 Jews were massacred, joined the Communist party in 1945, and until 1946 was actually—of all things—Minister of Education.
Here are a few of the outstanding ex-Nazis who, at this writing, wield Communist power:
Stefan Vojtec, a member of the Politburo, chairman of the state-owned cooperatives, Minister of Education from 1945-48, was formerly the editor of the Sentinell, the official press organ of the joint German-Rumanian general staff during the war.
General Alexandru Petrescu, the director of concentration camps under the Antonescu regime, recently presided over the military courts that sentenced Juliu Maniu, Ion Michalache, and other anti-Communists.
Aurel Vijoli, a legionary commander of the Iron Guard, is today Assistant Minister of Finance and governor of the State Bank.
Lotar Radaceanu, alias Wurzer, a former socialist of German descent, was an agent of German economic penetration of Rumanian industry during the war, and in that capacity he made numerous trips to Germany. Today he is one of the thirteen members of the Politburo of the Communist party, and, besides, a member of the seventeen-member organizational bureau of the party. In addition, he is Minister of Labor and Social Security.
Victor Vojen, former Iron Guard commander, former editor of the Iron Guard paper, and from October 1940 to January 1941 minister to Rome, has been since 1947 the private secretary and intimate of Ana Pauker.
Approximately 200,000 members of the Rumanian middle classes are at present toiling as slave laborers on the construction of the Danube-Black Sea Canal. They live in tents, are without blankets, and their rations are below subsistence level. About 40,000 of them are Jews. Their guards are almost exclusively former members of the Iron Guards. It is no wonder that the death toll of the victims exceeds a hundred a day.
A passage in a letter recently received by the writer from a friend who succeeded in escaping from behind the Iron Curtain precisely sums up the Hungarian-Rumanian situation: "The same people as under the Nazis are today locked up in prisons and concentration camps. These same people are guarded by the same persons who guarded them under the Nazis."
East may be East, and West, West; and perhaps the twain never shall meet. But in such countries as Hungary and Rumania, Left is not Left; nor is Right, Right. The twain—being totalitarian twins—have met, mingled, and merged into a regime of uniform terror.


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Keep the colors bright and vibrant! Refresh faded fabrics with permanent Simply Spray Brite Red Fabric Paint for Upholstery. Color your world and keep your fabric looking cheerful!
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4 out of 4 found this review helpful.
Recommend: Yes
Our hot TX summers took a toll on our patio cushions, I too checked into replacing them or have them upholstered, which was very expensive. Since the foundation of my furniture was in excel. condition, I decided to paint the cushions...it was so easy and they look beautiful!! Like New!! Wish I knew about this produce a lot sooner!! (i agree about the 1/2 filled cans....should have more product in them, but was still cheaper route for me)
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5 out of 5 found this review helpful.
I saw this product some time back on a Today Show segment and forgot about it until I started looking into replacing my patio seating set cushions which were badly faded from the sun. I was shocked at the replacement cushion cost which was about twice what I paid for the set; my seat cushions are not universal. The spray work great but you need a lot of it. There are 8 cushions in total, two side chairs and the love seat. These BIG 8 ounce cans are not full....they are half full or maybe less than half. I will probably need 8-10 cans for a nice coverage. That is my only gripe.
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Lambent Gunker
this wiki
- "Woah! Better stay clear of those guys!"
- — Clayton Carmine after a Gunker exploded
Lambent Gunkers, or just Gunkers, were especially-deadly Lambent lifeforms that were discovered 18 months after the Sinking of Jacinto. Both large and powerful, Gunkers were the disgusting result of Butcher Boomers being turned Lambent.
Like Lambent Drudges, Gunkers were not observed by COG forces until the Mission to Hanover. Led by Augustus Cole, a team made of Clayton Carmine, Samantha Byrne, and Damon Baird made contact with multiple Gunkers on their way through Stranded settlements at the Hanover dock warehouse and the Hanover Thrashball Stadium.
Two Gunkers were part of a Lambent squad attacking the Warehouse Stranded, shelling the Stranded barricade from afar. Short time after, Delta was attacked by another Gunker at the docks. Finally, Delta was ambushed and cut off by Lambent at the entrance to Cougars Stadium; two Gunkers were among them. Inside the stadium, another Gunker attacked Delta. Then, when the Lambent began overrunning the stadium, a second Gunker attempted to destroy the elevators Delta were in on their way up to the stadium's cable car room. But when Delta reached their destination, the cable cars were gone, so they had to ziple down the car cables. On the way down, one more Gunker tried to eliminate the team only for the bridge it was standing on exploded via shot fuel canister. A final Gunker on Centennial Bridge engaged Delta when the group landed.
A day later, Delta Squad (now composed of Marcus Fenix, Anya Stroud, Jace Stratton, and Samantha Byrne) investigates the destroyed city of Char in search of gas to reach the Endeavor Naval Shipyard. They find the local Stranded and their leader, former Griffin Imulsion Corporation CEO Aaron Griffin. Griffin, despite his hate for the COG, explains that he needs his fuel as well, sending Delta to recover it. On the way up the fuel-tower, a Gunker slams through the wooden roof of the makeshift elevator Delta were on.
When Adam Fenix's Imulsion Countermeasure went off, Sera was wiped clean of all Lambent and Locust lifeforms.
|Locust Horde
|Berserker · Boomer (Butcher, Flame Boomer, Grinder, Mauler, Mauler Elite, Savage Boomer, Tremor) · Drone (Beast Rider, Bolter, Cleaver Theron, Cyclops, Flame Grenadier, Grappler, Grenadier, Grenadier Elite, Gunner, Hunter, Hunter Elite, Miner, Palace Guard, Rager, Savage Drone, Savage Grenadier, Savage Grenadier Elite, Savage Hunter, Savage Marauder, Savage Theron, Sniper, Spotter, Theron Elite, Theron Guard, Theron Sentinel) · Kantus (Armored Kantus, Savage Kantus)
|Hollow Creatures
|Bloodmount · Brumak · Corpser · Digger · Gas Barge · Heart Leech · Hydra · Kryll · Leviathan · Mangler · Nemacyst · Reaver · Riftworm · Rock Worm · Seeder · Serapede · Shibboleth · Shrieker · Siegebeast · Tempest · Ticker · Torture Barge · Wretch
|Former · Lambent Berserker · Lambent Brumak · Lambent Drone · Lambent Drudge · Lambent Gunker · Lambent Leviathan · Lambent Polyp · Lambent Stalk · Lambent Wretch


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Until you understand the basics of functions and algebra, the thought that a number can be predicted is a surprising one. And of course `magic' and `being surprised' are often the same thing. Rob Eastaway shows us how mathemagicians trade off the fact that you can usually predict precisely the outcome of doing something in mathematics, but only if you know the secret beforehand.
Chomp is a simple two-dimensional game, played as follows.
Steven J. Brams uses the Cuban missile crisis to illustrate the Theory of Moves, which is not just an abstract mathematical model but one that mirrors the real-life choices, and underlying thinking, of flesh-and-blood decision makers.
Arguably, the exponential function crops up more than any other when using mathematics to describe the physical world. In the first of two articles on physical phenomena which obey exponential laws, Ian Garbett discusses light attenuation - the way in which light decreases in intensity as it passes through a medium.
Why can't human beings walk as fast as they run? And why do we prefer to break into a run rather than walk above a certain speed? Using mathematical modelling, R. McNeill Alexander finds some answers.


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Be the first cleveland.com user to add photos or videos of Grande Winery & Winemaking Supplies
Grande offers a good selection semi-dry & dry wines. Food is limited unless u call ahead & talk to Paul Grande personally & make group reservations. Very nice, personable guy. Jutebox of classic American-Italian & Jazz. Bar area & small tables, seats around 25-30.


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Bryan Guinn
Bryan Guinn, the president of Prism Technologies Group, has been working as a computer and information systems consultant with San Antonio and South Texas businesses. A Microsoft Gold Certified Partner, he serves the technology community as the vice president of the San Antonio International Association of Microsoft Certified Partners. He also is on the advisory board for the Technology Connexus, a non-profit organization that promotes technology within South Texas.
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Managing Mobile Devices as Enterprise Assets
I was recently talking about mobile devices with a colleague and we began discussing how companies can manage their mobile devices that contain corporate e-mail and data as an Enterprise Asset. Companies are going to need a strategy to manage and support multiple types of devices in the workplace. It will require a management plan […] [Read More]


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MOOCs and OERs in Moncton
Jun 06, 2013
Seminar presentation delivered to GTA Seminar, Universite de Moncton, Moncton, New Brunswick.
Long long loooong presentation (almost four hours, though there's 15 minutes of set-up at the start and a 15-minute break in the middle) about the thinking behind MOOCs and the tools I've developed to build them. The first half looks first at the idea of creating and sharing content as a matter of course. Then I look at how people learn, and talk about learning as a process of recognition. Then we go through the animation Connectivism link in today's newsletter. After the break, I take a step-by-step walk through gRSShopper, the software that runs this site and all of the MOOCs I've run, showing how the software and the theory both developed iteratively through use over a period of fifteen years or so.


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Geometric Constraint System Stories: Evariste Galois, CAD and Crystallography
Professor Stephen Power, Department of Mathematics and Statistics
Tuesday 15 December 2009, 1005-1030
Lecture Theatre 1, Management School Building
Geometric constraint systems are large systems of equations that arise from a configuration of geometrical objects and which feature in many structural models in applied science. I will outline three stories which have at their heart the analysis of the flexibility and rigidity of bar-joint structures. A bar (= strong bond) is inextensible while a joint (= weak angular bond) can swivel. For a 3D example imagine a telegraph pylon with all the joints loosened (and whether it will fall down).
James Clerk-Maxwell noted simple counting rules that hold when typical bar-joint systems in 2D or 3D are rigid structures. Nevertheless it is still an open problem to determine what exactly determines rigidity and in what manner bar-joint structures can flex or deform.
Evariste Galois, "revolutionary and geometer", died in a dual in 1832 before at the age of 21. The night before, the story goes, he wrote down his profound theory on the solvability of polynomial equations. In my second story I will explain how "Galois' Obstacle", as I call it, is relevant to the geometric constraint systems within core CAD software.
Max Born and Theodore von Karman developed the classical theory of crystallography. This theory incorporates infinite periodic bar-joint structures as mathematical models in the analysis of the low energy spectrum. Can the mathematics of such models shed light on the physics of crystals or, for that matter, more amorphous materials?
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Kent Anderson
Kent AndersonMusic Director & Afternoon Host
Kent Anderson began his association with KCSC/KUCO as a student employee in 1981. After graduating from then-Central State University with a B.A. in Oral Communications, he worked at other radio stations in Oklahoma City and Columbia,
MO, before returning to KCSC as a full-time staff member in 1991. He served as Program Director from 1992-2005, as part-time off-air music programmer from 2006-2013, and returned as full-time Music Director and afternoon drive
announcer in September 2013. Kent is also an award-winning magazine journalist and author of six published novels, including the 2006 winner of the Oklahoma Book Award for Fiction.
Kent lists his favorite composers as Beethoven, Chopin, Dvorak, Haydn, Holst, Mark O'Connor, and John Williams. He has a special passion for the intersection of traditional folk and classical music. Kent has three sons, and lives in Oklahoma City.


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LeMahieu flies out to center. 0-0 mid 2nd.
LeMahieu flies out to center. 0-0 mid 2nd.
It's my birthday, so I hope the Reds and Pacers can both give me a win for a present.
...the 2-2 to Woodsen and here it comes...and it is swung on and missed! And Tom Browning has pitched a perfect game! Twenty-seven outs in a row, and he is being mobbed by his teammates, just to the thirdbase side of the mound.
Bruce lines a single to right
Jay Bruce gets the inning started with a hard-hit single.
Frazier grounds into a 6-4-3 DP on the first pitch
The name DJ LeMahieu still cracks me up.
"Since I've been with the Reds in 1989, we've never had a farm system this loaded," Bowden said. "If we were the New York Yankees and had unlimited dollars, we could have traded for Colon, (Jeff) Weaver, Rolen, (Cliff) Floyd, (Kenny) Rogers and Finley and gotten them all -- and still held onto our top five prospects. That's an amazing statement."
NebraskaRed (06-03-2013)
And Paul grounds out, and that's the end of the second.
Did I hear them say Molina got a one game suspension?
Home plate is having to come out of the game.
Extended delay here as they work out the umpiring situation.
I hope they give the starters some extra time to RE-warmup. Both of them.
Wow...that's a HORRIBLE picture of Dusty. Yikes. (side-by-side with Weiss)
Roller up the center, one out.
Board Moderators may, at their discretion and judgment, delete and/or edit any messages that violate any of the following guidelines: 1. Explicit references to alleged illegal or unlawful acts. 2. Graphic sexual descriptions. 3. Racial or ethnic slurs. 4. Use of edgy language (including masked profanity). 5. Direct personal attacks, flames, fights, trolling, baiting, name-calling, general nuisance, excessive player criticism or anything along those lines. 6. Posting spam. 7. Each person may have only one user account. It is fine to be critical here - that's what this board is for. But let's not beat a subject or a player to death, please.


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The Tykes defender, who has made the first-team breakthrough at Oakwell this season, played the first 45 minutes on Tuesday night for Noel Blake's side at Telford's New Bucks Head Stadium, where Everton's John Lundstram converted a stoppage-time penalty to seal victory.
"When a lad in any academy starts at a young age and goes right through, especially with his hometown club, it's credit to him and everybody who has coached him," Barnsley's academy manager Ronnie Branson told the club's official website.
"It says a lot for the standard of coaching, he's been here since he was eight and he's had contact with a lot of good people.
"From an academy point of view it shows it can be done. Some academies want to buy in the best talent and progress them from a later age, it's quite unique now with someone like John that he's stayed with one academy for all that time.
"It goes without saying he's got a bright future in front of him. We're proud and pleased for him. He's a smashing kid off the field as well. It's not just about being a footballer, it's about developing you as a person as well."


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Who We Are
Founded by families, FBN-North America has been established to connect, support and serve families in business. We are a trusted resource, dedicated to promoting the success and sustainability of families in business locally, nationally and globally. Our primary values are confidentiality, trust, openness, honesty and mutual respect in an environment free of solicitation.
The Family Business Network North America (FBN-NA) is one of 30 chapters of the Family Business Network International (FBN-I) around the world. Each chapter organizes its own programs and events, for networking, education and training, under the motto "By Families, For Families." For more information on FBN-I and its chapters, click HERE, or paste the following link into your web browser: http://fbn-i.org/.


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Shows how the upgrade of this free program will help photographers better organize, share, and edit digital photos more easily than before.
"I took several Photoshop classes at local community colleges and couldn't seem to master it—until lynda.com. It was exponentially easier with video learning, and now I create beautiful digital art that gets rave reviews." —Cathy N.
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The present invention provides fusion proteins for the detection and treatment of neuroectodermal tumors. Previous work demonstrated that chlorotoxin is specific for glial-derived or meningioma-derived tumor cells. The current invention has extended the use of chlorotoxin-cytotoxin fusion proteins to treat the whole class neuroectodermal tumors such as gliomas, meningiomas, ependymonas, medulloblastomas, neuroblastomas, gangliomas, pheochromocytomas, melanomas, PPNET's, small cell carcinoma of the lung, Ewing's sarcoma, and metastatic tumors in the brain. Also, diagnostic methods are provided for screening neoplastic neuroectodermal tumors.
1. A method of detecting a neuroectodermal tumor in a patient comprising:
(a) adding chlorotoxin to a patient tissue sample; and
(b) detecting the binding of chlorotoxin to the tissue sample wherein an elevated level of binding relative to normal tissue is indicative of the presence of the neuroectodermal tumor.
2. The method of
3. The method of
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5. The method of
6. The method of
7. The method of any one of claims 5 or 6 wherein the radiolabel is detected by positron emission tomography scanning.
8. The method of
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10. The method according to
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16. The method of
17. The method of any one of claims 15 or 16 wherein the tissue sample is counterstained.
18. The method of
This application is a continuation-in-part of U.S. application Ser. No. 08/774,154 filed Dec. 26, 1996) now U.S. Pat. No. 5,905,027 which claims the benefit of U.S. Provisional Application No. 60/009,283 (filed Dec. 27, 1995).
This invention was produced in part using funds from the Federal government under NIH grant no. R01 NS 36692. Accordingly, the Federal government has certain rights in this invention.
1. Field of the Invention
The present invention relates generally to the fields of cell physiology, neurology, developmental biology, and oncology. More specifically, the present invention relates to novel methods of using a chlorotoxin sensitive cytoplasmic protein for the diagnosis and treatment of primitive neuroectodermal tumors (PNET).
2. Description of the Related Art
During embryonic development, the future nervous system forms from a specialized layer of ectodermal cells called the neuroectoderm. This layer extends longitudinally along the body axis congruent with the future spinal column. Invagination of the neuroectoderm gives rise to the neural tube from which essentially all central nervous system (CNS) components including the spinal cord develop. Specialized cell clusters along the rim of the invaginating neural tube stay separate from the tube and from the neural crest. These highly migratory neuroectodermal cells give rise to specialized cells throughout the body including Schwann cells, neuronal cells of the peripheral nervous system (PNS) (enteric, parasympathetic, sympathoadrenal, and sensory neurons), pigment cells (melanocytes), endocrine cells and cells forming connective tissue of the face and neck. Since these cells share a common embryonic origin with central nervous system cells, it is not surprising that these cells, or the tumors developing from these cells, share some genetic and antigenic phenotypes with central nervous system cells.
For example, melanomas and glioblastomas share a common mutation in the gene encoding for the epidermal growth factor receptor (EGFR) (1). Malignant astrocytomas and neurofibromas not only express high levels of the epidermal growth factor receptors but also vascular endothelial growth factor receptor (VEGF-R) and platelet-derived growth factor receptor (PDGF-R) (2). A high expression of the mutant variant, EGFRvIII, has been demonstrated in glial tumors as well as the extracellular matrix proteins, GP 240 and tenascin (3). Tenascin and the ganglioside-3′,6′-isoLD1 have been found in both gliomas and primitive neuroectodermal tumor tissues (3). In another study, antibodies to tenascin bound extensively to CNS gliomas and also to melanomas, breast, lung and squamous cell carcinomas (4). Another ganglioside, GD2, has been shown to be a common antigen marker in both gliomas and primitive neuroectodermal tumor tissues (5). Other common antigens between melanomas and gliomas were demonstrated by showing that Tyr, TRP-1, TRP-2 and gp100 gene products are commonly found in both melanoma and glioma tumors (6). Common cytokines or their receptors linking tumors of astrocytomas, ependymomas and primitive neuroectodermal tumors have been identified as: interleukin (IL) IL-1 alpha, IL-1, IL-1R1, IL-1R antagonist and transforming growth factor (TGF) TGF-beta 1 (11).
Another class of proteins used as markers for gliomas and primitive neuroectodermal tumors are the cytoskeletal proteins, neurofilament (NF), glial fibrillary acidic protein (GFAP), intermediate filaments (IF), intermediate associated protein filament (IFAP), vimentin, nestin and keratins. These markers have been used to determine stages of differentiation along the various cell lineages (12). New evidence linking astrocytomas with certain primitive neuroectodermal tumor tumors is the cytoskeleton marker of IFAP-300 kDa, a marker of immature glia (13).
Further arguments for a tight linkage of neuroectodermally derived cells in the central nervous system and periphery can be made based on their similar dependence on epigenetic influences. For example, sympathoadrenal precursor neurons require basic fibroblast growth factor (bFGF) to proliferate and differentiate, but survival of these cells depends on nerve growth factor (NGF) responsiveness and nerve growth factor availability (14). A similar scenario is required for each of the other cell types. Not only are growth and trophic factors necessary but cytokines and hormones are needed for which links remain to be elucidated between primitive neuroectodermal tumors and gliomas.
However, despite this list of similarities shared between neuroectodermally derived cells, these cells are distinct entities with unique cytological, biochemical and functional features. Indeed, the list of unique features not shared with other neuroectodermally derived cells by far exceeds the above mentioned shared phenotypes. Thus, one can not assume a priori that expression of a certain antigen or phenotype is to be expected in a given cell type based on expression by any other member of the neuroectodermally derived cell types.
Neuroblastomas generally express a selective increase in the gene copy number of the MYCN gene found in fetal stages of brain development suggesting links between the origin of the cells and the ability of neoplastic cells to dedifferentiate (7). However, this gene has yet to be demonstrated in the glioma cells. Other proteins that are not common to both glioma and primitive neuroectodermal tumors have been demonstrated. CD99 immunoreactivity is used as a tool in identifying primitive neuroectodermal tumors (8) and has been shown in Ewing's sarcoma tumors although not in gliomas (9). Another factor, stem cell factor and its receptor, c-kit, are also expressed in both primitive neuroectodermal tumor and Ewing's Sarcoma tumors (10).
The common origin and ability to respond to internal and external signals during the normal developmental processes suggests that central nervous system cells and peripheral neuroectodermally derived cells may also share common mechanisms during pathological developments as for example, during neoplasia. Such neoplastic tissues include CNS gliomas that are glial-derived tumor cells specific to the CNS. They metastasize only within the CNS including the spinal column. They are believed to originate from at least three separate lineages either from undifferentiated precursor cells or by dedifferentiation of astrocytes, oligodendrocytes or ependymal cells.
Primitive neuroectodermal tumors (PNET) are found both in the CNS and PNS. Primitive neuroectodermal tumors found only in the PNS are referred to as peripheral primitive neuroectodermal tumors (PPNET). Primitive neuroectodermal tumors manifest preferentially in children and have capacity for developing into a variety of neuronal, astrocytic, ependymal, muscular and melanotic lines. The conceptual basis of grouping these tumors together is based upon sharing common progenitor cells as well as sharing similar neoplastic transformations leading to tumors of similar morphological features and biological behavior. However, there remains controversy in placing all primitive neuroectodermal tumors into the same categories. The following paragraphs demonstrate examples of the overlap of common antigens between the various types of CNS and PNS tumors.
Supratentorial primitive neuroectodermal tumors include cerebral medulloblastomas, cerebral neuroblastomas, 'blue' tumors, ependymoblastoma and other primitive neuroectodermal tumors, such as pineoblastomas (WHO grade IV). The most useful markers for these tumors include GFAP, NFP, desmin and melanin. Others antigens found in these tumors are vimentin, nestin, keratin but are not useful for diagnostic purposes.
Peripheral neuroblastic tumors of the adrenal gland (medulla) and sympathetic nervous system are the most common type of childhood tumor outside of the CNS. Primary sites for these primitive neuroectodermal tumors are in the adrenals, abdominal, thoracic, cervical and pelvic sympathetic ganglia but include other primary sites as orbit, kidney, lung, skin, ovary, spermatic cord, and urinary bladder. Specific names of these related tumors are pheochromocytomas, paraganglioma, neuroblastomas, ganglioneuromas, ganglioneuroblastomas, neurofibromas, schwannomas, and malignant peripheral nerve sheath tumors. These all share common origin in the neural crest. Neuroblastomas all share high TRK-A (NGFR) and CD44 expressions. Neuronal specific enolase (NSE), synaptophysin, neural filament (NF) protein, GD2, tyrosine hydroxylase (TH) and chromogranin are used as diagnostic markers also found in medulloblastomas. Neuroblastomas generally express a selective increase in the gene copy number of the MYCN gene found in fetal stages of brain development (7).
Medulloblastomas are members of the primitive neuroectodermal tumors that are described as highly malignant embryonal tumors of the CNS found in the cerebellum (WHO grade IV). A common antigen of these medulloblastoma and other neuronal lineage tumors is synaptophysin (not found in glial or mesenchymal brain tumors). Nestin (IF protein) is found in developing CNS precursor cells and in medulloblastomas and in some peripheral neuroectodermal origin cells. Nestin (and vimentin) are found in medulloblastomas, astrocytomas, glioblastomas, ependymomas, gangliogliomas and meningiomas (only GFAP is found in the astrocytic-derived cells, which are occasionally 'trapped' in medulloblastomas). Increased levels of neural-cellular adhesion molecule (N-CAM) found in these tumors, may reflect levels of differentiation in the development of tumors (15). While varying levels of nerve growth factor (NGF), are found in nearly all tumors, medulloblastomas exhibited substantial reactivity to the NGF receptor and related proteins, neurotrophin (NT) NT-3, TRK-C and brain derived neurotrophic factor (BDNF) (16).
Melanomas, arising from melanocytes follow a graded development from diffuse melanocytosis, to melanocytoma to malignant melanomas. S100 protein is a marker for these tumors, as vimentin and NSE reactivity are variable.
Small cell neuroendocrine carcinomas of the lung are highly invasive and typically found in adult smokers. They have been shown to exhibit reactivity to many of the neural and neuroendocrine markers (some of them similar to N-CAMs) for tumor differentiation as peripheral primitive neuroectodermal tumors, gliomas, and ependymomas. These markers include neural specific enolase and extremely high c-src expression (17).
A feature conspicuously shared between developing CNS cells and neural crest derived cells is their propensity to migrate either towards a target or target area. It is believed that this ability is lost after cell differentiation and maturation. However, tumors of the CNS show significant cell migration and invasion into healthy brain, suggesting that cell have maintained or regained this enhanced migratory ability. It is, thus, not surprising that neoplastic transformation of neuroectodermally derived cells outside the CNS would have similar migratory abilities. At the intended destination, these cells differentiate into their final phenotype, similar to normal development, influenced by several trophic factors crucial for the proliferation and differentiation of various cell types.
The prior art is deficient in the lack of an diagnostic and therapeutic agents specifically targeted to primitive neuroectodermal tumors. The present invention fulfills this longstanding need and desire in the art.
In one embodiment of the current invention, a method is described for the treatment of tumors of neuroectodermal origin by administering a ligand specific for this class of tumors fused to a cytotoxic moiety. Specific neuroectodermal tumor tumors which can be treated in this manner include gliomas, meningiomas, ependymomas, medulloblastomas, neuroblastomas, gangliomas, pheochromocytomas, melanomas, peripheral primitive neuroectodermal tumors, small cell carcinoma of the lung, Ewing's sarcoma, and metastatic tumors of neuroectodermal origin in the brain.
In the preferred embodiment, the neuroectodermal tumor specific ligand is chlorotoxin in the form of a fusion protein of chlorotoxin with cytotoxic moiety. The chlorotoxin may be native, synthetic or recombinant chlorotoxin. Possible cytotoxic moieties include gelonin, ricin, saponin, pseudonomas exotoxin, pokeweed antiviral protein, diphtheria toxin, and complement proteins.
In another embodiment of the current invention, the neuroectodermal tumor specific ligand is an antibody against the chlorotoxin receptor, presumably a 72 kDa chloride channel. The antibody may be fused to gelonin, ricin, saponin, pseudonomas exotoxin, pokeweed antiviral protein, diphtheria toxin, and complement proteins.
In yet another embodiment of the current invention, a method of differentiating neuroectodermal tumor-derived neoplastic tumor tissue from non-neoplastic tissue is presented. This is accomplished by exposing the tissue with labeled chlorotoxin and measuring the binding of the labeled chlorotoxin. An elevated level of binding relative to normal tissue is indicative that the tissue is neoplastic. In one embodiment, the label is a fluorescent moiety which is detected by fluorescent microscopy, fluorescent activated cell sorting or a fluorescent plate reader. Alternatively, the chlorotoxin may be radiolabeled (eg. 131I-chlorotoxin or 125I-chlorotoxin; a person having ordinary skill in this art would readily recognize other useful radiolabels) and detected by positron emission tomography scanning. Alternatively, the chlorotoxin may be conjugated to a non-fluorescent detection moiety such as biotin and detected immunohistochemically or by use of a calorimetric assay.
So that the matter in which the above-recited features, advantages and-objects of the invention, as well as others which will become clear, are attained and can be understood in detail, more particular descriptions of the invention briefly summarized above may be had by reference to certain embodiments thereof which are illustrated in the appended drawings. These drawings form a part of the specification. It is to be noted, however, that the appended drawings illustrate preferred embodiments of the invention and therefore are not to be considered limiting in their scope.
FIG. 1 shows the positive immunohistochemical staining of a glioblastoma multiform (GBM) tumor with chlorotoxin. The brown reaction product of DAB 3′3′-diaminobenzidine with biotinylated chlorotoxin is clearly visible in the TM-601 stained section. TM-601: biotinylated chlorotoxin stained, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 2 demonstrates that normal brain is not immunohistochemically stained by biotinylated chlorotoxin. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin. Normal brain was also stained with biotinylated antibodies against GFAP (Glial Fibrillary Acidic Protein) which positively stains astrocytes in normal brain tissue.
FIG. 3 shows chlorotoxin staining of an adrenal mass neuroblastoma tumor. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 4 illustrates that biotinylated chlorotoxin immunohistochemically stains pheochromocytomas. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 5 illustrates that normal adrenal tissue is not immunohistochemically stained by biotinylated chlorotoxin. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 6 shows the immunohistochemical staining with biotinylated chlorotoxin of melanoma tumor cells metastasized to the brain TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 7 illustrates biotinylated chlorotoxin immunohistochemical staining of melanoma tumor cells metastasized to the lung. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 8 shows immunohistochemical staining of normal skin with biotinylated chlorotoxin. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 9 shows immunohistochemical staining of small cell lung carcinomas with biotinylated chlorotoxin. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 10 shows that biotinylated chlorotoxin does not immunohistochemically stain normal lung tissue. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 11 shows the immunohistochemical staining of a medulloblastoma tumor with biotinylated chlorotoxin. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 12 shows that the rare, neuroectodermally derived bone cancer, Ewing's sarcoma, also exhibits positive immunohistochemical staining with biotinylated chlorotoxin. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 13 shows the negative results obtained in the immunohistochemical staining of normal stomach tissue with biotinylated chlorotoxin. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 14 shows the lack of immunohistochemical staining of normal liver tissue with biotinylated chlorotoxin. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
FIG. 15 demonstrates that normal spleen tissue is not immunohistochemically stained by with biotinylated chlorotoxin. TM-601: biotinylated chlorotoxin, counter-stained with methyl green; Control: methyl green only; and, H&E stain: hematoxylin and eosin.
In accordance with the present invention there may be employed conventional molecular biology, microbiology, and recombinant DNA techniques within the skill of the art. Such techniques are explained fully in the literature. See, e.g., Maniatis, Fritsch & Sambrook, "Molecular Cloning: A Laboratory Manual (1982); "DNA Cloning: A Practical Approach," Volumes I and II (D. N. Glover ed. 1985); "Oligonucleotide Synthesis" (M. J. Gait ed. 1984); "Nucleic Acid Hybridization" [B. D. Hames & S. J. Higgins eds. (1985)]; "Transcription and Translation" [B. D. Hames & S. J. Higgins eds. (1984)]; "Animal Cell Culture" [R. I. Freshney, ed. (1986)]; "Immobilized Cells And Enzymes" [IRL Press, (1986)]; B. Perbal, "A Practical Guide To Molecular Cloning" (1984).
Therefore, if appearing herein, the following terms shall have the definitions set out below.
As used herein, the term "cDNA" shall refer to the DNA copy of the mRNA transcript of a gene.
As used herein, the term "derived amino acid sequence" shall mean the amino acid sequence determined by reading the triplet sequence of nucleotide bases in the cDNA.
As used herein the term "screening a library" shall refer to the process of using a labeled probe to check whether, under the appropriate conditions, there is a sequence complementary to the probe present in a particular DNA library. In addition, "screening a library" could be performed by PCR.
As used herein, the term "PCR" refers to the polymerase chain reaction that is the subject of U.S. Pat. Nos. 4,683,195 and 4,683,202 to Mullis, as well as other improvements now known in the art.
The amino acid described herein are preferred to be in the "L" isomeric form. However, residues in the "D" isomeric form can be substituted for any L-amino acid residue, as long as the desired functional property of immunoglobulin-binding is retained by the polypeptide. NH2 refers to the free amino group present at the amino terminus of a polypeptide. COOH refers to the free carboxy group present at the carboxy terminus of a polypeptide. In keeping with standard polypeptide nomenclature, J Biol. Chem., 243:3552-59 (1969), abbreviations for amino acid residues are known in the art.
It should be noted that all amino-acid residue sequences are represented herein by formulae whose left and right orientation is in the conventional direction of amino-terminus to carboxy-terminus. Furthermore, it should be noted that a dash at the beginning or end of an amino acid residue sequence indicates a peptide bond to a further sequence of one or more amino-acid residues.
A "replicon" is any genetic element (e.g., plasmid, chromosome, virus) that functions as an autonomous unit of DNA replication in vivo; i.e., capable of replication under its own control.
A "vector" is a replicon, such as plasmid, phage or cosmid, to which another DNA segment may be attached so as to bring about the replication of the attached segment.
A "DNA molecule" refers to the polymeric form of deoxyribonucleotides (adenine, guanine, thymine, or cytosine) in its either single stranded form, or a double-stranded helix. This term refers only to the primary and secondary structure of the molecule, and does not limit it to any particular tertiary forms. Thus, this term includes double-stranded DNA found, inter alia, in linear DNA molecules (e.g., restriction fragments), viruses, plasmids, and chromosomes. In discussing the structure herein according to the normal convention of giving only the sequence in the 5′ to 3′ direction along the nontranscribed strand of DNA (i.e., the strand having a sequence homologous to the mRNA).
An "origin of replication" refers to those DNA sequences that participate in DNA synthesis.
A DNA "coding sequence" is a double-stranded DNA sequence which is transcribed and translated into a polypeptide in vivo when placed under the control of appropriate regulatory sequences. The boundaries of the coding sequence are determined by a start codon at the 5′ (amino) terminus and a translation stop codon at the 3′ (carboxyl) terminus. A coding sequence can include, but is not limited to, prokaryotic sequences, cDNA from eukaryotic mRNA, genomic DNA sequences from eukaryotic (e.g., mammalian) DNA, and even synthetic DNA sequences. A polyadenylation signal and transcription termination sequence will usually be located 3′ to the coding sequence.
Transcriptional and translational control sequences are DNA regulatory sequences, such as promoters, enhancers, polyadenylation signals, terminators, and the like, that provide for the expression of a coding sequence in a host cell.
A "promoter sequence" is a DNA regulatory region capable of binding RNA polymerase in a cell and initiating transcription of a downstream (3′ direction) coding sequence. For purposes of defining the present invention, the promoter sequence is bound at its 3′ terminus by the transcription initiation site and extends upstream (5′ direction) to include the minimum number of bases or elements necessary to initiate transcription at levels detectable above background. Within the promoter sequence will be found a transcription initiation site, as well as protein binding domains (consensus sequences) responsible for the binding of RNA polymerase. Eukaryotic promoters often, but not always, contain "TATA" boxes and "CAT" boxes. Prokaryotic promoters contain Shine-Dalgarno sequences in addition to the −10 and −35 consensus sequences.
An "expression control sequence" is a DNA sequence that controls and regulates the transcription and translation of another DNA sequence. A coding sequence is "under the control" of transcriptional and translational control sequences in a cell when RNA polymerase transcribes the coding sequence into mRNA, which is then translated into the protein encoded by the coding sequence.
A "signal sequence" can be included near the coding sequence. This sequence encodes a signal peptide, N-terminal to the polypeptide, that communicates to the host cell to direct the polypeptide to the cell surface or secrete the polypeptide into the media, and this signal peptide is clipped off by the host cell before the protein leaves the cell. Signal sequences can be found associated with a variety of proteins native to prokaryotes and eukaryotes.
The term "oligonucleotide", as used herein in referring to the probe of the present invention, is defined as a molecule comprised of two or more ribonucleotides, preferably more than three. Its exact size will depend upon many factors which, in turn, depend upon the ultimate function and use of the oligonucleotide.
The term "primer" as used herein refers to an oligonucleotide, whether occurring naturally as in a purified restriction digest or produced synthetically, which is capable of acting as a point of initiation of synthesis when placed under conditions in which synthesis of a primer extension product, which is complementary to a nucleic acid strand, is induced, i.e., in the presence of nucleotides and an inducing agent such as a DNA polymerase and at a suitable temperature and pH. The primer may be either single-stranded or double-stranded and must be sufficiently long to prime the synthesis of the desired extension product in the presence of the inducing agent. The exact length of the primer will depend upon many factors, including temperature, source of primer and use the method. For example, for diagnostic applications, depending on the complexity of the target sequence, the oligonucleotide primer typically contains 15-25 or more nucleotides, although it may contain fewer nucleotides.
The primers herein are selected to be "substantially" complementary to different strands of a particular target DNA sequence. This means that the primers must be sufficiently complementary to hybridize with their respective strands. Therefore, the primer sequence need not reflect the exact sequence of the template. For example, a non-complementary nucleotide fragment may be attached to the 5′ end of the primer, with the remainder of the primer sequence being complementary to the strand. Alternatively, non-complementary bases or longer sequences can be interspersed into the primer, provided that the primer sequence has sufficient complementary with the sequence or hybridize therewith and thereby form the template for the synthesis of the extension product.
As used herein, the terms "restriction endonucleases" and "restriction enzymes" refer to enzymes, each of which cut double-stranded DNA at or near a specific nucleotide sequence.
A cell has been "transformed" by exogenous or heterologous DNA when such DNA has been introduced inside the cell. The transforming DNA may or may not be integrated (covalently linked) into the genome of the cell. In prokaryotes, yeast, and mammalian cells for example, the transforming DNA may be maintained on an episomal element such as a plasmid. With respect to eukaryotic cells, a stably transformed cell is one in which the transforming DNA has become integrated into a chromosome so that it is inherited by daughter cells through chromosome replication. This stability is demonstrated by the ability of the eukaryotic cell to establish cell lines or clones comprised of a population of daughter cells containing the transforming DNA. A "clone" is a population of cells derived from a single cell or ancestor by mitosis. A "cell line" is a clone of a primary cell that is capable of stable growth in vitro for many generations.
A "heterologous" region of the DNA construct is an identifiable segment of DNA within a larger DNA molecule that is not found in association with the larger molecule in nature. Thus, when the heterologous region encodes a mammalian gene, the gene will usually be flanked by DNA that does not flank the mammalian genomic DNA in the genome of the source organism. In another example, coding sequence is a construct where the coding sequence itself is not found in nature (e.g., a cDNA where the genomic coding sequence contains introns, or synthetic sequences having codons different than the native gene). Allelic variations or naturally-occurring mutational events do not give rise to a heterologous region of DNA as defined herein.
The labels most commonly employed for these studies are radioactive elements, enzymes, chemicals which fluoresce when exposed to ultraviolet light, and others. A number of fluorescent materials are known and can be utilized as labels. These include, for example, fluorescein, rhodamine, auramine, Texas Red, AMCA blue and Lucifer Yellow. A particular detecting material is anti-rabbit antibody prepared in goats and conjugated with fluorescein through an isothiocyanate link.
Proteins can also be labeled with a radioactive element or with an enzyme. The radioactive label can be detected by any of the currently available counting procedures. The preferred isotope may be selected from 3H, 14C, 32P, 35S, 36Cl, 51Cr, 57Co, 58Co, 59Fe, 90Y, 125I, 131I, and 186Re.
Enzyme labels are likewise useful, and can be detected by any of the presently utilized colorimetric, spectrophotometric, fluorospectrophotometric, amperometric or gasometric techniques. The enzyme is conjugated to the selected particle by reaction with bridging molecules such as carbodiimides, diisocyanates, glutaraldehyde and the like. Many enzymes which can be used in these procedures are known and can be utilized. The preferred are peroxidase, β-glucuronidase, β-D-glucosidase, β-D-galactosidase, urease, glucose oxidase plus peroxidase and alkaline phosphatase. U.S. Pat. Nos. 3,654,090, 3,850,752, and 4,016,043 are referred to by way of example for their disclosure of alternate labeling material and methods.
A particular assay system developed and utilized in the art is known as a receptor assay. In a receptor assay, the material to be assayed is appropriately labeled and then certain cellular test colonies are inoculated with a quantity of both the label after which binding studies are conducted to determine the extent to which the labeled material binds to the cell receptors. In this way, differences in affinity between materials can be ascertained.
An assay useful in the art is known as a "cis/trans" assay. Briefly, this assay employs two genetic constructs, one of which is typically a plasmid that continually expresses a particular receptor of interest when transfected into an appropriate cell line, and the second of which is a plasmid that expresses a reporter such as luciferase, under the control of a receptor/ligand complex. Thus, for example, if it is desired to evaluate a compound as a ligand for a particular receptor, one of the plasmids would be a construct that results in expression of the receptor in the chosen cell line, while the second plasmid would possess a promoter linked to the luciferase gene in which the response element to the particular receptor is inserted. If the compound under test is an agonist for the receptor, the ligand will complex with the receptor, and the resulting complex will bind the response element and initiate transcription of the luciferase gene. The resulting chemiluminescence is then measured photometrically, and dose response curves are obtained and compared to those of known ligands. The foregoing protocol is described in detail in U.S. Pat. No. 4,981,784.
The current invention is directed to a method of treating neuroectodermal tumors with a neuroectodermal tumorspecific ligand fused to a cytotoxic moiety. Possible neuroectodermal tumor targets include gliomas, meningiomas, ependymonas, medulloblastomas, neuroblastomas, gangliomas, pheochromocytomas, melanomas, PPNET's, small cell carcinoma of the lung, Ewing's sarcoma, and metastatic tumors in the brain. Preferably, the neuroectodermal tumor specific ligand is chlorotoxin fused to a cytotoxic moiety. Examples of possible cytotoxic moieties include gelonin, ricin, saponin, pseudonomas exotoxin, pokeweed antiviral protein, diphtheria toxin, and complement protein.
The current invention is also directed to a neuroectodermal tumor specific therapeutic agent in which the neuroectodermal tumor specific ligand is an antibody against the chlorotoxin receptor believed to be a 72 kDa chloride channel. The antibody may be fused to gelonin, ricin, saponin, pseudonomas exotoxin, pokeweed antiviral protein, diphtheria toxin, or complement proteins.
The current invention is also directed to a method of differentiating neuroectodermal tumor-derived neoplastic tumor tissue from non-neoplastic tissue by incubating the tissue of interest with labeled chlorotoxin and measuring the binding of the labeled chlorotoxin, relative to normal tissue where available. The chlorotoxin may be labeled with either a fluorescent moiety or may be radiolabeled with radiolabels such as 131I or 125I. Fluorescent moieties can be used for detection by fluorescent microscopy or fluorescent activated cell sorting. Radiolabeled chlorotoxin can be detected by positron emission tomography scanning.
The following examples are given for the purpose of illustrating various embodiments of the invention and are not meant to limit the present invention in any fashion.
Summary of Chlorotoxin Results from Glioma Experiments
Recent studies demonstrated a common antigen is expressed by the vast majority of glioma cells. This antigen is targeted by chlorotoxin (Ctx or TM-601), a 36 amino acid peptide originally isolated from Leiurus quinquestriatus scorpion venom. Chlorotoxin selectively binds to the membrane of glioma cells allowing selective targeting of these cells within the CNS (18). The antigen targeted by this peptide appears to be a chloride ion channel although the antigen has not yet been unequivocally identified at the molecular level. Thus far, the data indicates that chlorotoxin binds to a membrane protein of 72 kDa molecular weight that is preferentially expressed in the cytoplasmic membrane of glioma cell. Binding of the peptide enhances glioma cell proliferation (19) and inhibits the ability of glioma cells to migrate in Transwell assays, an in vitro assay to evaluate tumor invasiveness (20). Chlorotoxin appears to exert these effects by reducing the membrane permeability to Cl− ions thereby preventing cell volume changes that are required to allow cells to invade healthy tissue (20). Thus, the most likely action of chlorotoxin is on a glioma chloride channel previously extensively characterized (19).
Immunohistochemical Staining of Gliomas with Chlorotoxin
Over 250 frozen or paraffin sections of human biopsy tissues were histochemically stained with a chemically synthesized form of chlorotoxin containing a detectable biotin group chemically attached to the N terminus (TM-601). Binding of the TM-601 molecule was observed on selective cells associated with the essentially all glioma tumors with up to 95% positive cells per tumor. Based on these studies, it has been proposed to utilize chlorotoxin as a glioma specific marker and as a potential therapeutic tool for targeting glioma tumors. For such purposes, chlorotoxin linkage of radioactive molecules or cytotoxic moieties such as saporin could be employed.
Recombinant DNA Manipulation of Chlorotoxin
Using techniques well known in the art, one may prepare recombinant proteins specifically engineered to mimic the binding and action of the native toxin. The biological activity of the synthetic chlorotoxin is as effective for chloride ion channel blockade as the native venom toxin. Recombinant techniques are used to synthesize chlorotoxin in E coli using a modified PGEX vector system and the toxin may be linked to various fusion proteins using common restriction sites. After synthesis of recombinant chlorotoxin, it may be linked to various cytotoxic fusion proteins including glutathione-S-transferase (GST), gelonin, ricin, diptheria toxin, complement proteins and radioligands and other such proteins as are well known in the immunotoxin art.
Antibodies Against the Chlorotoxin-Binding Chloride Ion Channel
Antibodies to the chloride ion channels in glial-derived tumors may be prepared as follows. Polyclonal antisera are generated by injecting fusion proteins created between the glutathione-S-transferase and the chlorotoxin insert into mice or rabbits. Mice are immunized with 0.5 ml of a 1:1 emulsion of 1 mg/ml purified fusion protein in Freund's complete adjuvant and subsequently with two additional injections after 14 and 28 days in Freund's incomplete adjuvant. The mouse and rabbit antibodies are purified from the antisera using the GST fusion protein immobilized on nitrocellulose filters. The antibodies are then examined for binding specificity in various tissues.
Rationale for the Examination of Neuroectodermally Derived Tumors for Chlorotoxin Binding
Given the similarities that can be shared between gliomas and other neuroectodermally-derived cells, and the arguments developed that propose similar propensities of neuroectodermal cells to migrate, a thorough investigation was undertaken to examine neuroectodermally derived tumors for the expression of chlorotoxin binding sites.
Preparation of Sections from Frozen or Paraffin-Embedded Human Biopsies
Most of the samples of human tissuel, from both sexes, all ages and race were obtained through the Cooperative Human Tissue Network, Tissue Procurement at UAB, UAB hospitals and the Human Brain Tissue Bank in London, Canada. Snap frozen tissue and fresh tissue embedded in a freezing gel were sliced at 8 microns and picked up onto positively charged glass slides. The sections were then fixed in 4% paraformaldehyde or milloniqs according to the staining protocol. Paraffin blocks were sectioned and prepared according to standard procedures.
Examination of Biopsy Samples for Chlorotoxin Binding
Biopsy sections were blocked for 1 hour in 10% normal goat serum in PBS and treated with a dilution of biotinylated chlorotoxin overnight at 4� C. After thorough rinsings, the stainings were developed by avidin-biotin complex (ABC) technique (Vectastain Elite ABC Kit from Vector Laboratories, Burlington, Calif.) and visualized by the colorimetric reaction of DAB (3′3′-diaminobenzidine; Vector Laboratories) with the ABC complex.
The biopsy sections were counterstained with methyl green, a nuclear dye, to more easily visualize the unstained cells. Non-specific background label can vary from experiment to experiment due to changes in the effective concentrations of the label, condition of the tissue or the duration of the reaction. Therefore, a control section was identically stained with methyl green but without the biotinylated chlorotoxin. Positive cell staining is identified by chlorotoxin-labeling above background when compared to its individual control of a successive slice. Cells containing high amounts of endogenous peroxidase exhibit dark background staining in the controls due to the reaction of DAB with the peroxidases.
Finally, a third adjacent section was stained with both hematoxylin, a cell nuclei specific stain, and eosin, a cytoplasmic stain. Therefore, for each tissue analyzed, three adjacent section were stained. These are shown in photomicrographs providing evidence of the specificity of TM-601 chlorotoxin binding to tumors of neuroectodermal derivation in comparison to controls. In the photomicrographs, adjacent sections are identified as follows: TM-601: biotinylated chlorotoxin detected by a brown reaction product of DAB with the biotin and further counterstained with methyl green; Control: the control section stained with only methyl green; and, H&E: the hematoxylin and eosin stained section.
Glioblastoma Multiforme (GBM) Tumors
Glioblastoma multiforme (GBM) were stained with the biotinylated chlorotoxin (TM-601). These tumors are extremely reactive to biotinylated chlorotoxin as 25 out of 25 patient samples tested positive as seen in FIG. 1. This glioblastoma multiforme can be compared to the staining of the normal human brain tissue with biotinylated chlorotoxin (18/23 negative). FIG. 2 shows a representative staining. Normal brain tissue demonstrates a lack of TM-601 staining. This is consistent with earlier evidence of specificity of chlorotoxin binding to gliomas.
GFAP Staining in the Normal Brain Tissue
The biopsy section was blocked for 1 hour in 10% normal goat serum in PBS and then stained with antibodies against glial fibrillary acidic protein (GFAP; DAKO corporation, Carpinteria Calif.) overnight. The secondary antibody conjuaged to peroxidase was applied to the rinsed tissue for 2 hours, rinsed again before the stain was visualized with DAB. A typical glial fibrillary acidic protein stain of normal brain is shown in FIG. 2. Normal brain was positive for glial fibrillary acidic protein staining where it stained the astrocytes typically present in normal tissue.
Neuroblastomas are a tumor primarily found in children with a high incidence in the adrenals. Neuroblastoma show TM-601 reactivity above the control staining as seen in FIG. 3. Six out of seven neuroblastomas were positive for chlorotoxin binding.
Pheochromocytomas are neoplastic chromaffin cells of the adrenal glands. This tumor also show a high degree of staining as seen in FIG. 4. Five out of six pheochromocytomas were positive for staining with biotinylated chlorotoxin, especially in comparison to TM-601 staining of the normal adrenals (3/3 negative) seen in FIG. 5.
FIG. 6 shows the biotinylated chlorotoxin staining of a melanoma metastasized to the brain. Seven out of seven melanoma brain metastasis were positive for TM-601. In addition, melanoma metastasized to the lung were analyzed as seen in FIG. 7. Normal skin, however, is unreactive to TM-601 (6/6 negative) (FIG. 8) although there is some background staining in the melanocytes even in the controls.
Small Cell Lung Carcinomas
Small cell lung carcinomas are reactive to TM-601. There is good contrast between the cells that stain and those that do not (FIG. 9). The cells positive for TM-601 in the control (middle panel) are red blood cells which present high levels of background peroxidase stain. This TM-601 specificity can be further demonstrated by comparing the TM-601 staining of the small cell carcinoma (2/3 positive) and the normal lung (3/3 negative) (FIG. 10).
Another neuroectodermally derived tumor type are the medulloblastomas. They exhibit specific reactivity to TM-601 as seen in FIG. 11 (4/4 positive).
Ewing's Sarcoma
Ewing's sarcoma, a rare bone cancer sometimes found in soft tissue, is TM-601 positive (2/2) (FIG. 12).
Testing of Potential Sites of Chlorotoxin Administration for Side Effects
To aid in the design of drug therapy with this product, various normal tissues were stained with TM-601 to determine possible sites where side effects may occur. Preliminary evidence indicates that some of the most common targets for side effects such as the stomach and liver, are TM-601 negative (2/2 negative samples for both tissues, thus far) (FIGS. 13 and 14 respectively). The staining of spleen tissue is also shown in FIG. 15 (3/3 negative). The TM-601 staining of other normal human tissues is summarized in Table 1.
Summary of Tested Tumors and Tissues
As summarized in Table 1, the vast majority of neuroectodermally derived tumors bind chlorotoxin, indicating that chlorotoxin has a more widespread utility to target tumors of neuroectodermal origin. Specifically, primitive neuroectodermal tumor tumors have been tested from 34 patients, 31 of which showed chlorotoxin specificity in the tumor material as seen in Table 1. This staining was compared with the chlorotoxin staining of other types of CNS and PNS tumors as well as the comparison to various normal human tissues.
TM-601 specifically associates with neuroectodermally-derived tumors including medulloblastomas, neuroblastomas, ganglioneuromas, melanomas, pheochromocytomas, small cell lung carcinomas and Ewing's sarcomas. Thus, chlorotoxin-derived molecules can be utilized to target specifically for therapeutic or diagnostic purposes the above identified neuroectodermally-derived tumors. Likewise, these tumors can also be targeted by other molecules such as antibodies that bind to the chlorotoxin receptor, presumed to be the 72 kD Cl− ion channel.
The following references were cited herein:
1. Hemizygous or homozygous deletion of the chromosomal region containing the pl6INK4a gene is associated with amplification of the EGF receptor gene in glioblastomas. Hegi M E, Hausen A Z, Ruedi D, Malin G and Kleihues P. (1997) Int. J. Cancer 73:57-63.
2. Ras activation in astrocytomas and neurofibromas. Guha A. (1998) Can J. Neurol. Sci. 25:267-281.
3. Tumor antigens in astrocytic gliomas. Kurpad S N, Zhao X G, Wikstrand C J, Batra S K, McLendon R E, and Bigner D D. (1995) Glia 15:244-256.
4. Iodine-131-labeled anti-tenascin monoclonal antibody 81C6 treatment of patients with recurrent malignant gliomas: phase 1 trial results. Bigner D D, Brown M T, Friedman A H, Coleman R E, Akabani G, Friedman H S, Thorstad W L, McLendon R E, Bigner S H, Zhao X-G, Pegram C N, Wikstrand C J, Herndon J E, Vick N A, Paleologos N, Cokgor I, Provenzale J M and Zalutsky M R. (1998) J. Clin. Onco. 16:2202-2212.
5. Trilateral tumors in four different lines of transgenic mice expressing SV40 T-antigen. (1996) Marcus D M, Lasudry J G, Windle J, Howes K A, al Ubaidi M R, Baehr W, Overbeek P
This invention was produced in part using funds from the Federal government under NIH grant no. R01 NS 36692. Accordingly, the Federal government has certain rights in this invention. A, Font R L, and Albert D M. Invest. Ophthalmol. Vis Sci 37:392-396.
6. Molecular detection of tumor-associated antigens shared by human cutaneous melanomas and gliomas. Chi D D J, Merchant R E, Rand R, Conrad A J, Garrison D, Turner R, Morton D L, and Hoon D S B. (1997) Am. J. Pathol. 150:2143-2152.
7. Pathology and Genetics of Tumors of the Nervous System. Eds. Paul Kleihues and Webster K. Cavenee, International Agency for Research on Cancer, Lyon, 1997.
8. Peripheral primitive neuroectodermal tumor of the ovary confirmed by CD99 immunostaining, karyotypic analysis, and RT-PCR for EWS/FLI-1 chimeric mRNA. Kawauchi S, Fukuda T, Miyamoto S, Yoshioka J, Shirahama S, Saito T, and Sukamoto N. (1998) Am J Surg. Pathol. 11:1417-1422.
9. Cytology of typical and atypical Ewing's sarcoma/PNET. Renshaw A A, Perez-Atayde A R, Gletcher J A, and Granter S R. (1996) Am J. Clin Pathol 106:620-624.
10. C-kit is expressed in soft tissue sarcoma of neuroectodermic origin and its ligand prevents apoptosis of neoplastic cells. Ricotti E, Fagioli F, Garelli E, Linari C, Crescenzio N, Horenstein A L, Pistamiglio P, Vai S, Berger M, Cordero di Montezemolo L, Madon E, and Basso G. (1998) Blood 91:2397-2405.
11. Interleukin-1 alpha, IL-1 beta, IL-1R type1, IL-1 R antagonist, and TGF-beta 1 mRNAs in pediatric astrocytomas, ependymomas, and primitive neuroectodermal tumors. Ilyin S E, Gonzalez-Gomez I, Gilles F H, and Plata-Salaman C R. (1998) Mol. Chem. Neuropathol. 33:125-137.
12. Immunohistochemical characterization of primitive neuroectodermal tumors and their possible relationship to the stepwise ontogenetic development of the CNS. 2. Tumor studies. Kleinert R. (1991) Acta Neuropathol 82:508-15.
13. Proteins of the intermediate filament cytoskeleton as markers for astrocytes and human astrocytomas. Yang H Y, Lieska N, Shao D, Kriho V, and Pappas G D. (1994) Mol.Chem. Neuropathol 21:155-176.
14. Human primitive neuroectodermal tumour cells behave as multipotent neural precursors in response to FGF2. Derrington E A, Dufay N, Rudkin B B, and Belin M-F. (1998) Oncogene 17:1663-1672.
15. Neuroectodermal tumors of the peripheral and the central nervous system share neuroendocrine N-CAM-related antigens with small cell lung carcinomas. Molenaar W M, de Leij L, and Trojanowski J Q. (1991) Acta Neuropathol. 83:46-54.
16. Neurotrophins and neuronal versus glial differentiation in medulloblastomas and other pediatric brain tumors. Tajima Y, Molina R P Jr, Rorke L B, Kaplan D R, Radeke M, Feinstein S C, Lee V M, and Trojanowski J Q. (1998) Acta Neuropathol. 95:325-332.
17. Expression of c-src in cultured human neuroblastoma and small-cell lung carcinoma cell lines correlates with neurocrine differentiation. Mellstrom K, Bjelfman C, Hammerling U, and Pahlam S. (1987) Mol Cell Biol 7:4178-4184.
18. Use of chlorotoxin for targeting of primary brain tumors. Soroceanu L, Gillespie Y, Khazaeli M B and Sontheimer H W. (1998) Cancer Res. 58:4871-4879.
19. Cell cycle-dependent expression of a glioma-specific chloride current: proposed link to cytoskeletal changes. Ullrich N and Sontheimer H. (1997) Am J. Physiol. 273:C1290-1297.
20. Modulation of glioma cell migration and invasion using Cl− and K+ ion channel blockers Soroceanu L, Manning T J Jr., and Sontheimer H. (1999) J. Neuroscience. Submitted.
Any patents or publications mentioned in this specification are indicative of the levels of those skilled in the art to which the invention pertains. These patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.
One skilled in the art will readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The present examples along with the methods, procedures, treatments, molecules, and specific compounds described herein are presently representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art which are encompassed within the spirit of the invention as defined by the scope of the claims.
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County girl wins Miss Maine U.S. International Crown
Local pageants in the County have been a part of one girls life for years; and now that same girl that was born and raised in Presque Isle gets to represent not just the County, but the entire state on the National level, as well as the chance to compete for the national title of one of the largest pageant systems in the world.
Brittany Bearden is an Assistant Manager at Panscofar's Boutique, and a recent graduate of UMPI. But what you might not know about her is she's a pageant queen; and she just won the state title of the 3rd largest pageant system in the world.
Brittany Bearden, Miss Maine U.S. International 2014 says, "It's an honor really being born and raised here being able to represent your state."
Brittany shined in evening gown, on stage question, and the swimsuit phases of competition that won her the Miss Maine U.S. International Crown in September.
Brittany Bearden, Miss Maine U.S. International 2014 says, "I can get up in front of a crowd and not be scared, my speaking ability my interview skills have really improved from that."
Brittany says one of her favorite things about being a pageant queen is getting to wear the rhinestone embellished crown to appearances- but there's a lot more to being a pageant titleholder than meets the eye.
Brittany Bearden, Miss Maine U.S. International 2014 says, "If you've never been on stage before getting up on stage is something to overcome for girls. Getting on stage in swimsuit is not the easiest thing either when you have a lot of people all eyes are on you."
She has about 9 months to prepare for nationals and during that time she'll be traveling the state making appearances, hitting the gym and of course getting a glamorous wardrobe in order! But with the former title of Miss Maine Collegiate America and several local pageants under her belt, she's feeling pretty confident.
Brittany Bearden, Miss Maine U.S. International 2014 says, " I'm still working I want the national title I'm not giving up yet."
Only one person gets to walk away with the crown; and Brittany says she's in it to win it.
Brittany Bearden will be in Orlando, Florida from June 23rd to the 29th to compete in the national Miss U.S. International Pageant; and if she wins the national title, she'll get to compete in the Miss International pageant which will be held in Asia.
Category: News


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Community Support
Steps to shut down a wordpress installation
Category: Cpanel
What are the steps to properly shut down (delete) an existing wordpress installation from cPanel? Delete directory from file manager? Run a script? Use MySQL manager?
If I need to delete a database, how do I know which database is associated with which site (assuming I have more than one running)?
If I need to delete a database, how do I know which database is associated with which site (assuming I have more than one running)?
You can remove the WordPress installation through Softaculous. Softaculous will delete the files and the database for you. See the following link.
How to uninstall programs using Softaculous
To find your database for your WordPress, you can look it up in your wp-config.php file in your WordPress files.
1. Look in your wp-config.php file found in the installation directory of your WordPress site.
2. Your database name associated with this WordPress install will be labeled as:
/** The name of the database for WordPress */
define('DB_NAME', 'putyourdbnamehere');
Best Regards,
James R
10 Points
2013-07-02 3:50 am EST
1) Delete all WordPress files. If you installed into your root directory, delete all the files and directories installed by WordPress (wp-config.php, /wp-content/, /wp-admin/ etc). Be careful to not delete any other files you might have had there before WordPress. If WordPress is in a subdirectory, simply delete the subdirectory and all its contents.
2) Go into PHPMyadmin or whatever else you have on your server to edit MYSQL databases. Select the database used by your WordPress install and drop all the WordPress tables. Again, if this was a fresh database only used by WordPress, you can drop all tables. If the database had existing tables before, make sure you don't lose them.
WordPress is now un-installed and you can use the same blank database with the old settings to install it again.
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PANAMA — While a lot of high schoolers were enjoying a relaxing, albeit hot, July on their summer break, Panama Lady Razorback softball player Taylor Eaves did anything but relax. The Lady 'Back junior played in two national tournaments — one in Denver and one in New York City.
The Sparkler Tournament was first up in Denver.
And there wasn't much rest for Eaves before her New York adventure.
"I went to Colorado for a week," the Panama Lady 'Back softballer said. "Then, I came home for five days, then went to New York."
Three LeFlore County high school football teams, the Pocola Indians, the Talihina Golden Tigers and the Heavener Wolves will get their 2011 practices under way with "Midnight Madness" practices — like they did a season ago.
However, Heavener's will be at 12:01 a.m. Friday — the first day in which players can put pads on after going three first three days of practice in just helmets and shorts.
If you're a high school football player or coach, Tuesday, Aug. 9, 2011 has been circled on the calendar for quite some time. That is the day which high school football teams can begin practicing for their 2011 seasons, which officially begin in September.
There's always some kind of heat issues high school football teams will deal with when practice begins every August. But this year, when practice begins Tuesday for the 2011 season, this summer's heatwave may be the worst ever. Thus, it's making the teams plan accordingly.
"I've never seen heat like this," Spiro Coach Chris Bunch said. "We've got to be real careful because of it. We can't take a chance of the kids getting hurt. One thing, we're going to give them as much water as possible, and we probably won't go as long as we normally would."
While at Briggs Middle School in Tahlequah, Tyler Massey was only coaching football. When the chance presented itself to be able to coach even more, he took the opportunity by the horns.
Massey is the new Bokoshe football coach, and will also help out in basketball and baseball.


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This 20 tray cubbie unit is made of heavy duty construction-built to last. It has bright 3/4" thermofused laminate. It has durable rounded corners which eliminates sharp edges. It will not fade or discolor and is easy wipe clean. It has specially designed, smooth wraparound T-mold accents. Its safe, non-toxic and stain resistant. Cubbie unit comes with casters but cubbie trays are sold separately. Please select the color of your choice for the trim.
(Cubbie does not come with tray)


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Box Score
SAN LUIS OBISPO, Calif. – A pair of late goals proved to be the difference as the Lamar Lady Cardinals dropped a 2-1 decision at Cal Poly in a non-conference women's soccer game Sunday afternoon.
Lamar (1-1) jumped into the lead 11:05 into the second half when Kimmy Albeno converted a pass from Jannet Hernandez. It was Albeno's first of the season and 26th of her career as she extended her school record.
Lamar goalkeeper Bailey Fontenot kept the score at 1-0 minutes later when she stoned Elise Krieghoff on a penalty kick. The score stayed at 1-0 until Cal Poly's Kendra Bonsall evened things at 1 with 11:16 remaining in regulation.
Krieghoff got revenge for her missed penalty kick when she scored the game-winner with 7:00 remaining. Krieghoff took a pass from Megan Abutin and beat Fontenot from about 20 feet to give the Mustangs (1-1) a 2-1 lead.
"I thought we played well," Lamar coach Orlando Cervantes said. "They took advantage of some opportunities late in the game, but I was very happy with our performance."
Fontenot made three saves for Lamar.
The Lady Cardinals return to Beaumont to open a five-game homestand when they meet Rice at 7 p.m. Friday at the Lamar Soccer Complex. Season tickets and individual-game tickets are available at the Montagne Center ticket office from 8 a.m. to 5 p.m. weekdays. Tickets are also available by calling (409) 880-1715, or online at www.LamarCardinals.com.


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Home Dartford Sport Article
Unbeaten Charlton claimed an impressive 3-2 win over Championship favourites Derby at the Valley on Tuesday night.
George Tucudean finished Jordan Cousins' cross to put the Addicks in front after 11 minutes.
Steve McClaren's side levelled just after the half-hour when Jamie Ward found the top corner from the edge of the box, giving debutant keeper Nick Pope no chance.
Charlton regained the lead seconds before half-time with Yoni Buyens scoring a penalty following a foul on Tucudean by Richard Keogh.
The Addicks went 3-1 up in the 78th minute when Igor Vetokele headed home a Lawrie Wilson cross.
A scruffy goal from Ward gave Derby hope with five minutes left but Bob Peeters' men held on and sit fourth in the early Championship table.
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text: Prev | Next West Virginia Gov. Earl Ray Tombin, center, and first lady Joanne Jeager Tomblin, right, greet supporters during an inauguration ball at the Clay Center on Monday, Jan. 14, 2013, in Charleston, W.Va. Gov. Tomblin, a Democrat, was celebrating his second term in office. (AP Photo/Randy Snyder) Gallery: Tomblin inauguration at Capitol Jan. 15, 2013 @ 09:53 AM West Virginia Gov. Earl Ray Tomblin vowed to continue seeking improvements for the state economy as he took the oath for his first full four-year term. (u'nobuy',)


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Riverside heads into state playoffs unbeaten in PAC-6
Riverside 3, Northern 0
DURHAM — Riverside coach Mike Hodge navigated the Pirates to an undefeated 4-A PAC-6 season with a 3-0 win at Northern on Monday.
Riverside got 12 kills apiece from Rachel Kallianos and Zariya Thomasville.
Victoria Pearce had eight digs for the Pirates and teammate Casey Speer lofted 23 assists.
The Pirates didn't lose a single game in the PAC-6, going 10-0.
Three of the last four PAC-6 titles ended up at Riverside.
The state playoffs begin on Saturday, and Riverside will be a No. 1 seed hosting the No. 4 seed from the Cap-8 Conference.
East Chapel Hill 3, Jordan 0
EAST CHAPEL HILL — East Chapel Hill shutout Jordan 3-0 Monday.
Jordan got eight kills from Margaret Dunn and 18 assists from Rose Gunn, but the Wildcats were too much for the Falcons.
Franklinton 3, NCSSM 2
FRANKLINTON — Franklinton's 3-2 win over N.C. School of Science and Mathematics in the 2-A Northern Carolina Conference quarterfinals ended the Unicorns' season on Monday.
Sheridan Earnhardt had 24 digs and three aces for NCSSM, and Sam Zarnick was setting up her teammates with 23 assists to go with five digs.
"For the third time this season we went five games with Franklinton," NCSSM coach Davene Mainwaring said. "We stuck to our game plan in the first two games, but once we deviated from it, we obviously didn't get the same results. So our season ends."
South Granville gets a crack at Franklinton today in the semifinals.
NCSSM 8, Bunn 0
BUNN — N.C. School of Science and Mathematics shutout Bunn 8-0 on Monday.
Jarod Glover led the Unicorns with three goals, with Cameron Pharr, Jacob Einstein, Thomas Winslow, Henry Bristol and Thomas Baraldi all getting in on the scoring with a goal apiece.
Pharr, Einstein and Baraldi each had an assist.