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Land is scarce but sky has no limits!
You might think that the two fields have nothing in common. One deals with the spaces and activities on ground whereas the latter deals with the never-ending air space as sky has no limits! This is the general perception and true superficially. If one study town planning and familiarizes himself or herself with commercial aviation sector, they will come out to be very much similar in their functionality. The only and stark difference remains in the implementation of the rules ultimately getting the things done at ground level. This is where planning is nowhere to be seen.
Similarity in the needs Urban Planning & Civil Aviation
Both the sectors needs high level of sophistication and need to make use of best available technology. Highest level of coordination and real-time data and information sharing is another necessity. The differences and similarities will be explained with use of examples to make them easy to understand for people associated with both domains.
Irrespective of the scale of work (layout plan, master plan, regional plan, economic plan) the data requirements are huge. This data and required information needs to be collected from different authorities and agencies which might be working together or independently. They may or may not be controlled by government and thus acquiring information from them will result in substantial expenditure. Since planning is not a new-born profession and its been some decades it has gained importance in some parts of World, there now exist specialized and dedicated departments working to meet the requirements of planning. These include town planning or urban planning department, data collection and assimilation units, making use of census data, need based data collection and sharing with transport authorities, municipalities and other civic bodies etc. Inspite of this planning remains distant from achieving the desired goals. There is lack of coordination among departments, lack of data, outdated data, unavailability of information, lack of expertise, lack of political will and influence (reasons are not a concern here). The direct and indirect cost incurred is paid by everyone which most of the time Is not reflected immediately.
In Aviation, a similar need of data and information exist which needs to be collected from number of concerned agencies working independently and under government. The difference remains in terms of data collected. This includes the information about the lengths of runways, weather data like wind velocity, mean direction, turbulence, types and location of clouds, routes with bad weather, distance between origin and destinations, fuel requirements and availability, access to medical and emergency facilities, real-time information about other traffic etc. Without having the sufficient data it is impossible for an aircraft to take-off. In case of any error/ mistake the results are seen immediately unlike planning. The news of crash or aborted landing starts flashing on all the news channel and a team is formed immediately to inspect the incident/accident.
Flexibility at different stages
Both the sectors have provision of corrections or adjustments in case it is evident that the desired result is not achievable or continuing the ongoing work will have negative or catastrophic results. In planning feedback and monitoring was added to the planning process under rational planning model. This provided some flexibility to the otherwise rigid planning process whose outcomes were visible only after things went wrong at end. These outcomes were at times disastrous as millions of people by this time would have suffered because of faulty planning. With the advancement and adoption of technology this improved and the time required to assess and rectify the mistake became possible. Now the corrections can be made at a much early stage to minimize the negative outcomes.
In aviation also things do go wrong. This can be because of both human error or technical glitches. Both of these are unpredictable but having a mechanism in place to minimize the damage remains to be of utmost importance. Safety in aviation is given utmost priority unlike planning. The operational standards for commercial airline transport aircraft include machine failure of order 1 in a million and 1 in a billion. No such standards or any stringent provision exist in case of Planning. In case things go wrong at starting the skills of crew, presence of safety equipment, support provided by ground staff and other nearby aircraft, a dedicated search and rescue team ensures that damage is minimized. No deliberate attempt to delay any procedure is acceptable. All the concerned people and agencies swiftly takes action with minimalist unavoidable delay.
International umbrella body and national divisions
No international umbrella body exist for urban planning. The standards adopted and functions performed varies from country to country and even within a country. No provision or need exists to detail out the practices being followed in a region. A person willing to work in a new place needs to get well versed with the system being followed by actually working and learning the practice being followed. This makes it problematic and time-consuming for a person to work in new place. No international body exist to unify the practices however in some countries there are governing bodies which work at different level but their success varies largely. Some of the founding and old bodies include American Planning Association (APA), Royal Town Planning Institute (RTPI), Indian Institute of Town Planners (ITPI), Nigerian Institute of Town Planners (NITP) etc. All of these governing bodies enjoy different rights where they work. Commonwealth Association of Planners can be considered as a pseudo international organisations but on looking at the work and influence it fails to fulfill the criteria required to qualify itself as a true international umbrella body.
International Civil Aviation Organisation (ICAO) is the apex umbrella body for civil aviation. It is a UN specialized agency, established by States in 1944 to manage the administration and governance of the Convention on International Civil Aviation (Chicago Convention). The rules framed by ICAO are binding for all members although there exists the provision to adopt rules complete or partially but in case of partial adoption, the changes from international practice needs to be informed so that all the countries have complete inform about the standards and procedures being followed in other countries.
Apart from a single governing body, all countries have their own organisations and concerned authorities dealing with civil aviation. Unlike planning, its mandatory to have a competent authority which will represent the country in conventions of ICAO and pass on the required information from time to time. These include Federal Aviation Administration (FAA), Ghana Civil Aviation Authority, Director General of Civil Aviation etc. These further have their sub divisions and coordinate among Number of other bodies to make sure the all the civil aviation operations go smoothly.
After things go wrong
In Aviation, reporting to competent authority is compulsory within a said time-frame for anything which went wrong. A well-defined classification as "incident" for relatively less serious mishaps and "accident" for serious mishaps ensures that nothing goes unreported. As soon as the information about mishap is received, investigation team starts its work. Person responsible or even merely present at time are interrogated. Pilot's licence gets suspended and the crew is grounded till the investigation is complete. The person/s responsible is prosecuted.
In case of planning, the problem starts with the identifying "what went wrong" but the final report is generally never finished. The lack of standards and reference makes it unrealistic to call the outcome as undesired and to quantify it. Unless a proper assessment is possible, it makes no sense to identify the person or agency responsible for it The margins of errors are absent in most of the cases and similarly the acceptance level of deviation from the set goals. In case by any method some people or a department can be identified, there exist the problem of overlapping or lack of detailed function or some other loophole which will prevent from any strict action being taken. This further gets complicated as the outcome is visible only after few years and planning itself is prone to drastic changes due to external factors. Accountability in true sense can't be established without very refined procedures and rules.
Accountability and actions taken
Having a clear and well-defined structure is essential for any task which requires input from multiple sources. Avoiding overlapping functions and identification of gaps is of utmost importance in efficient working. Accountability for delays and wrong information needs to be taken seriously to do quality work and this requires continuous and timely evaluation. Having standards and fix procedures helps in ensuring that minimalist deviations occur from the set rules. This is what makes Aviation a rapidly growing sector.
Urban planning on the other hand is dominated by "Guidelines" instead of rules. Number of overlapping functions, conflicts in administrative work and boundaries, lack of penalties. No well-defined standards or complete lack of standards makes assessment next to impossible. This results to serious issues and questions over the quality of work. Further, because of the scattered authorities it is difficult to find out the actual person/department who is responsible for the loss. Even if the concerned person is found out than the overlapping functions or the lack of clear procedure of what to do next will be at disposal to save the person/department from facing and serious actions. As a result no one gets punished except people for whom planning was being done. In such cases people become victims of wrong or ill-defined process and part of experimentation.
Data sharing and updating
The data requirements in planning is largely dependent on the secondary data collection. Plans being prepared get most of the required information by indirect means and thus at mercy of other departments. This is unfortunate but true in most parts of the world. Very few dedicated data collection takes place which is meant solely for planning because of the fact that there is very little need to conduct surveys again. The problem lies in making the existing data available for the concerned planning agency. The data thus gets updated at discretion of "other" fields for which data was originally collected. This results in delay or highly unreliable data for planning purposes. But this remains to be the only option available, sort of best out of trash in some cases. The frequency of updating this data ranges from days, weeks, months, years and even a decade in some cases. To list some examples, traffic volume count data is collected several times a month/year depending on the need and place. This collection is relatively easy and less tedious. But census data is updated at interval of 5 years in some countries, in 10 years in most of the countries and even more in some cases. This very basic demographic information is too often outdated if used as it is without care. Information about pollution and environmental changes might be unavailable or last updated decades ago.
In case of Aviation no delay can take place in obtaining the data. All the data collection needs to be done before hand, no take-off is possible without the crucial information. Thus for a successful and uninterrupted operations data needs to be collected and disseminated in almost real-time. This collection and sharing in done within and outside the geographical boundaries of a region. Some of the data is updated periodically and needs to be checked before starting of flights. In case of any unexpected or sudden change notices (NOTAM) is issued which needs to be checked. This ensures highest level of safety and awareness about the complete work. The collection and distribution of data also needs to be free from errors and negligible error is acceptable. All this comes at a good monetary cost but it is available and can be accessed by paying the required amount. The cost is high because of the perfection in the data collection and the responsibility of the data provider.


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About Me
CurrentUniversity of Evansville
MajorExercise Science
Graduating Class2016
HometownNewburgh, IN
Follow Benjamin's Progress
Established in 1854, the University of Evansville, located in southwestern Indiana, has a rich history that spans more than 160 years. UE's 2,486 students from 42 states and 55 countries experience a distinctive curriculum built on exposure to grea...
CollegeUniversity of Evansville
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Benjamin Johnson Graduates from University of EvansvilleBenjamin Johnson of Newburgh, IN, 47630, has graduated from the University of Evansville with the degree of Bachelor of Science. Johnson, majoring in Exercise Science, graduated during UE's 158th ...Spring 2016
Benjamin Johnson of Newburgh Named to University of Evansville Dean's ListBenjamin Johnson of Newburgh, IN, has achieved a place on the University of Evansville Dean's List for academic achievements during the Spring Semester 2013. Johnson is majoring in Exercise Scienc...Spring 2013
Benjamin Johnson of Newburgh Named to University of Evansville Dean's ListBenjamin Johnson of Newburgh, IN, has achieved a place on the University of Evansville Dean's List for academic achievements during the Fall Semester 2012. Johnson is majoring in Exercise Science....Fall 2012
Benjamin Johnson of Newburgh Enrolls at University of EvansvilleBenjamin Johnson of Newburgh, IN, has enrolled at the University of Evansville. Johnson, majoring in Exercise Science, joins an entering class defined by its strong academic ability and diversity. ...Fall 2012
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High SchoolCastle High School
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Benjamin Johnson of Newburgh Participates in University of Evansville Freshman Service ProjectThe University of Evansville is proud to announce that Benjamin Johnson of Newburgh, IN, participated in the Freshman Service Project on Monday, August 20. The project sent new UE students to 20 lo...Summer 2012
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Three Named All-American at NCAA Indoor Track and Field Championships
March 13, 2010
FAYETTEVILLE, ARK. - The University of Kentucky Track and Field team concludes NCAA Indoor Track and Field Championship competition earning three All-America titles in front of a crowd of 5,475 at the Randal Tyson Track Center in Fayetteville, Ark.
The men and women's track and field team had a total of eight athlete's travel to the NCAA Indoor Track and Field Championships to represent the University of Kentucky in national competition.
"It is a huge accomplishment for all of the athletes to qualify for the championship meet," Coach Don Weber said.
Senior thrower Ashley Muffet (N. Canton, Oh.) won the second All-America honor of her career and the second of the meet for the Wildcats with a second-place finish in the women's shot put. Muffet tossed her winning mark of 17.31m (56 feet, 9.5 inches) on her final attempt in the preliminary round. Scoring the largest amount of points for the women, Muffet earned eight with her second-place mark.
"I am very happy for Ashley Muffet," Weber said. "She has struggled this season and it wasn't until the qualifying meets and the championships that she's finally performed to the best of her abilities."
Muffet's second-place finish and eight-point contribution moved the women's team, which was in 39th place at the time, into a much higher 12th place. Muffet finished second only to the national leader, Mariam Kevkhishviilli from the University of Florida.
Honored with the All-American title for the sixth time in his UK career, senior Rondel Sorrillo (Vessigny, Trinidad and Tobago) took fifth place in the men's 60m dash. Running a time of 6.66, Sorrillo put the UK men's team on the scoreboard with four points, which put the Cats into 37th place at that time. He is also ranked fifth nationally in the men's 200m dash but just missed qualifying for the finals by one-hundredth of a second with a ninth-place time of 20.98.
"Rondel has been battling a leg injury and for him to come through in this meet is big for him and big for us as a team," Weber said.
Multi-event athlete Precious Nwokey (Houston, Texas) was the Cats first competitor on Saturday competition with the women's pentathlon, in which she set a new personal record of 3,985 points. Nwokey missed point contention and a spot on the podium by a small margin of 70 points, finishing in ninth place.
In each of her meets this season, Nwokey has set a new personal best, consistently increasing her point total while remaining nationally ranked. She ranked 16th nationally coming into this weekends championship meet.
The school record holding women's 4x400m relay team of Jenna Martin (Nova Scotia), Precious Nwokey (Houston, Texas), Jazmyn Shorter (St. Petersburg, Fla.) and Brittany Cabbler (Roanoke, Va.) ran in the NCAA Indoor Championships as the first qualifying 1600-meter relay team in school history.
The women finished three-tenths of a second off of their best time with a sprint of 3.38.15. Their time gave them a meet finish of 10th placeoverall.
Senior thrower Kristin Smith (Marietta, Ga.) made her point contribution during Friday's competition in the women's weight throw where she finished in eighth place. Earning one point for the Wildcats with a throw of 19.86m (65 feet, 2 inches), Smith was named top honor of All-America for the first time in her career.
Men's 800-meter runner, Sharif Webb (Paramus, N.J.) also competed for the Wildcats on Friday. The junior barely missed the 800 final by five-hundredths of a second during the preliminary race. His time of 1:49.78 put him just slightly behind the eighth-place finisher's qualifying time of 1:49.73.
The UK Women's team scored a total of nine points, giving the Cats a 26th-place team finish in the meet. The men's team finished the championships in 40th place with a point total of four.
To view all NCAA Indoor Track and Field Championship final results go to www.NCAA.com.
The Wildcats open the 2010 outdoor track and field season next weekend at the Black and Gold Challenge, March 19-20 in Orlando, Fla.
Stay up to date on all of your latest Kentucky Track and Field information at www.UKathletics.com.
Final Kentucky Team Standings:
26. Kentucky Women (9 points) 40. Kentucky Men (4 Points)
Saturday Results:
Women's Pentathlon - 11:30 a.m. CST (AUTO: 4,075; PROV: 3,725)
9. Precious Nwokey - Final: 3,985
Women's 60m Hurdles - (AUTO: 8.14; PROV: 8.43)s 10. Time: 8. 64; Points: 987
Women's High Jump - (AUTO: 1.85m; PROV: 1.78m) 13. Jump: 1.62m; Points: 759
Women's Shot Put - (AUTO: 16.90m; PROV: 15.20m) 12. Throw: 10.93m; Points: 591
Women's Long Jump - (AUTO: 6.38m; PROV: 6.13m) 10. Jump: 5.83m; Points: 798
Women's 800 Meters - (AUTO: 2:05.00; PROV: 2:09.00) 4. Time: 2:18.09; Points: 850
Women's Shot Put - 4:30 p.m. CST (AUTO: 16.90m; PROV: 15.20m)
2. Ashley Muffet - 17.31m (56-09.50)
Men's 60m Dash Final - 7:15 p.m. CST (AUTO: 6.60; PROV: 6.71)
5. Rondel Sorrillo - 6.66
Women's 1,600-Meter Relay - 8:40 p.m. CST (AUTO: 3:33.00; PROV: 3:40.00)
10. Kentucky A - 3:38.15
Friday's Results
Women's Weight Throw - 12 p.m. CST (AUTO: 21.00; PROV: 18.75)
8. Kristin Smith - 19.86m (1 point)
Men's 60m Dash Prelims - 6:25 p.m. CST (AUTO: 6.60; PROV: 6.71)
6. Rondel Sorrillo - 6.66q
Men's 800m Run Prelims - 6:55 p.m. CST (AUTO: 1:48.00; PROV: 1:50.50)
9. Sharif Webb - 1:49.78
Men's 200m Dash Finals - 7.25 p.m. CST (AUTO: 20.83; PROV: 21.23)
9. Rondel Sorrillo - 20.98


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|2006-06-21, 15:20
|Link #1
Join Date: Jan 2004
Location: もう東京ではない
Age: 36
[Manga] "That jutsu"
First off, if there's a thread about this already then please delete a.s.a.p. I tried searching but "that jutsu" returns quite a few hits on the Naruto board.
Anyway, as many of you remember about 15 months ago Jiraiya warned Naruto about using that jutsu. Now there is still a lot of speculation and wishful thinking concerning the specifics of this jutsu, even though we've already probably seen it. Yes, you've probably seen me arguing in the manga threads that that jutsu is simply Naruto's ability to release kyuubi chakra forming a number of tails. I thought this finally deserved a topic for itself where we can debate this subject without interfering with other discussions.
So without further ado, let's present the arguements for
the "the tails are the jutsu" side.
The "we haven't seen that jutsu yet" side:
I realize most of you would be heavily dissapointed if that jutsu turned out to be the tails, but logically speaking, isn't it the most likely answer? Anyway, I'm turning the word over to an open debate. This is my first poll so I hope it turns out ok, I've never seen anyone but me really argue that that jutsu is the tails so I wonder what the outcome will be...
|2006-06-21, 15:51
|Link #2
Join Date: Jul 2004
Location: 9th Temple
Age: 38
Ok, I voted for the, It is not the tail.
Here is why:
A) The tail growth should not be considering a Jutsu; it is just a Chakra release. I think is very cheep that Jiraya is calling that a Jutsu.
B) Given what Jiraya said to Naruto, about not using that Jutsu, and apart of this, Naruto making a face expression as he knew the consequence of this Jutsu, makes me believe is not the tails, at least 4 tails, given that Naruto has no recollection on the Tails consequence, yet, it seems he knows about that Jutsu consequence.
C) The Author would had trown a random quote saying something like: Im going to use that Jutsu That Jutsu is dangerous, but I will use it, He used that Jutsu etc.
|2006-06-21, 16:22
|Link #4
Bubbly and super fun
Join Date: May 2003
Location: Doesn't look like Kansas
A jutsu merely means a technique, the art to do something or an ability. There are arguements to believe that growning tails can't be "That" jutsu but this one -which funny enough is the most often used- isn't one of them.
Right now the only single thing making me believe the possibility of Naruto still having this so mysterious that jutsu is the fact that Sasuke got one of them.
If by any mean Kakashi's training involve a new that jutsu it would be the end of this possibility for me.
|2006-06-21, 16:33
|Link #5
nothing at all...
Join Date: Jul 2004
Age: 31
I think that "that jutsu" is the tails... Remember that i'm not talking about 4-tails, i'm talking about the ability of creating tails only. Maybe that's what Jiraiya taught Naruto, how to make use of kyubi's chakra... of course, when naruto reached the 4-tails and went berserk, Jiraiya noticed that was a bad thing and told naruto not to use it.
And... what else could it be? And why naruto didn't use it until now?
|2006-06-21, 16:36
|Link #6
Bubbly and super fun
Join Date: May 2003
Location: Doesn't look like Kansas
Yeah, I have a question for people thinking this jutsu being yet to be disclosed : what do you expect this jutsu to be?
I mean obviously Jiraiya was serious about it, solemn even. Yet we saw him talking about the danger of the tails to Kakashi and how to seal if that begins. Then we saw the same thing with Tsunade and Yamato and how he was chosen for this reason.
So the question is : what could be worse than something leading to MiniKyubi killing anyone at sight and destroying Naruto's body?
|2006-06-21, 16:57
|Link #7
Join Date: May 2004
Age: 30
I agree with all the points made thus far; the four tails were not treated like a "that jutsu" traditionally is in this series. However, they definitely fit the bill of what Jiraiya was talking about given the context, and the more time goes on the more I think that it was the tails. It has been a really long time since Jiraiya said that. We've been through two full arcs, and Naruto is about to get another big training session. It doesn't make sense that there's a big important jutsu he's known all this time and still hasn't used.
Then again, it's still very likely that he hasn't really fought to his own full capacity yet. Up until the 4 Tails incident, Naruto was too emotional about all his major fights and relied heavily on Kyuubi every time. He didn't have any opportunities to show such a powerful new jutsu because Kyuubi kept taking over. After Yamato had that talk with him and helped him hold Kyuubi back, he was still in Kyuubi hangover mode and couldn't fight properly, not that there's been a real fight since then anyway. In other words, not once in Part 2 has he fought at full power without going Kyuubi. There is technically room in his repertoire for another jutsu.
|2006-06-21, 17:07
|Link #8
The Northern Guy
A jutsu he c<hasnt yet fully mastered.
Like if he knows how to use yellow flash but hasnt mastered it yet and cant fully control it. It would be prety disaterous if he teleported into solid mater.
It can't be Naruto growing tails since Naruto did know about it consequences. Were when Jiraya warned Naruto, Naruto knew what they were talking about.
As for warning Kakashi and Yamato about the tails its only logical since at 4 Naruto loses control and doesnt remember a thing. And the reason for that is because tails seem to be released when Naruto loses self control over his anger. And Naruto would realy need some anger management clases. But dunno if that would help since he is just and emotional guy.
An other possibilaty but not as likely i thought up is: A way for Naruto to use the Kyubys chakra by not having to go into tails and losing control. Also since Jiraya knew he would be facing Uchihas in the future, maybe a way to mix Kyubys chakra with his own so the chakra couldnt be surpresed.
Because we know Jiraya wante dto teach Naruto how to better use the Kyuby. But obviously it isnt the only thing he would be teaching him.
|2006-06-22, 13:25
|Link #9
Join Date: Jul 2004
Location: 9th Temple
Age: 38
Now, What this Jutsu would be? Quite simply, Im expecting something that It is Dangerous to both Naruto and his opponent, and even maybe someone close to him (Yeah, Redundancy as I expect it to be almost the same as His Kyuby Manifestation), I go far as to say it could be A Kinjutsu, something that when Sasuke and Naruto go at it, instead of Having a Clash of Rasengan and Chidory, we would have a clash of Both Sasukes and Narutos That Jutsu.
I would not rule out the possibility that a the end that Jutsu turn out to be Kyuby Tails, but after Seen how Naruto reverted back to his normal from was in like Ok Lets get Orochimaru- He was unaware, and did not even felt any sort of guilt towards what happened, only after Yamato told him what Happened.
|2006-06-22, 13:40
|Link #10
Bubbly and super fun
Join Date: May 2003
Location: Doesn't look like Kansas
So you think it will be something with the same kind of impact than the tails but different in its application. Do you have something more precise in mind?
And since the chapter 312 where we get to know for sure that Naruto will develop his own "ultimate Ninjutsu" you don't think that in a future jutsu clash between Sasuke & Naruto he would rather use this one?
I have two others questions for people believing in this that jutsu:
How many time would you still believe in the existance of this jutsu if there is no mention of it in the story?
What kind of event would it take for you to stop believing into it?
Last edited by Hunter; 2006-06-22 at 13:55.
|2006-06-22, 14:50
|Link #12
Junior Member
Join Date: Jun 2006
Im gunna go against the wholy kyuubi thing, just because yamato tells naruto NOT to use the kyuubi chakra, i know yamato is the man, so i dont think he would advise naruto against using what jiraiya has called "that jutsu". Also the times he does go into his tailed form its usually through severe distress. Against itachi he does go kyuubi (the face marks and what not) But he doesnt go tailed form, and still manages to pull of some powerful moves (oodoma rasengan). It doesnt seem to fit that going berzerk is a jutsu :P Maybe what jiraiya meant, is that the jutsu he taught naruto is too powerful to just be casually used, and if naruto were to get hot headed or pissed off he would use it on a whim
|2006-06-22, 15:08
|Link #13
Join Date: Jun 2006
Location: Lyon (France)
Age: 32
Considering that Kakashi is going to train Naruto in order for him to create his own über jutsu that will be more powerfull than the rasengan etc... it's seems that the infamous "that jutsu" who Jiraiya was talking about is just the tails coz obviouly Naruto new super attack will be the one that he'll learn with Kakashi.
If there was a "that jutsu" then why bother learning a new one with Kakashi ?
Last edited by xyz; 2006-06-22 at 16:41.
|2006-06-22, 16:07
|Link #14
Senior Member
Join Date: Oct 2004
Location: Hidden Village of Sake
My ultimate answer is that it would totally suck if in 2,5 years with Jiraiya Naruto did not TRY to learn any "that"-level jutsu. I mean he did not even learn to use it safely, so how could i believe that the main character did not half-learn only 1 (one) high level jutsu.
|2006-06-22, 16:19
|Link #15
Join Date: Feb 2006
Location: Location is all relative.
-Why bother learning a new one you ask? Well why not? It is always good to up the arsenal of your attacks so I don't see a problem with Naruto learning his own ''ultimate'' jutsu. If 1 jutsu of your techniques is forbidden for you to use then why not learn a new one to make sure you don't use the forbidden one? In conclusion I would just like to say just because we haven't seen Naruto use "that" jutsu in the last 2 Arcs doesn't mean that it is simply the releasing of more tails. Maybe Kishimoto-sensei is saving it for another occassion or he himself hasn't really thought of how it works yet.
|2006-06-22, 16:23
|Link #16
Join Date: Jul 2004
Location: 9th Temple
Age: 38
In other hand, we can say that Naruto consciously decided to convert in to 4 tails, but then you have to convince me that he knew exactly what happened when he almost killed jiraiya.
And the new Jutsu that Naruto is going to learn is what have open my doubt of this Jutsu been something not shown yet, Why learn another Bigger than life Jutsu when he has already has one to show?
BTW, Regarding calling it that Jutsu, I also remembered that Gaara's Shukaku's manifestation was considered as "that Jutsu" also.
Really, I would leave the benefit on the Jutsu for at least Naruto next fight participation.
-If in this fight Naruto Throw everything including the Kitchen sink, and no sighting of this Jutsu, then I will 100% convinced that it was the Kyuby 4T manifestation.
Remember that Naruto has not fought without converting himself into the tail thingy, maybe this is the reason why he has not had the opportunity to use that Jutsu.
Last edited by Rurik; 2006-06-23 at 15:04.
|2006-06-22, 16:28
|Link #17
Join Date: Feb 2006
Location: Location is all relative.
|2006-06-22, 17:01
|Link #18
Join Date: Jun 2006
Location: Lyon (France)
Age: 32
If there was a "that jutsu" that would be Naruto's ultimate trump card, something he could not use carelessly and only to defeat a very strong opponent ...& as far as we known the jutsu he'll learn with Kakashi is going to be "that jutsu".
Of course the more you know powerfull jutsu the better it is but you cannot have a collection of "that jutsu" so unless the one he'll learn with Kakashi is going to be crap I don't think there will be another very special jutsu.
Last edited by xyz; 2006-06-22 at 18:58.
|2006-06-22, 18:39
|Link #19
Senior Member
no no no no its not the tails. We will eventually see "that jutsu" in a fight. Maybe we will see Naruto use his "That Jutsu" against Sasuke's "That Jutsu"
manga chapter 335 "The Amazing showdown of "That Jutsu" Naruto and sasuke face off using "that jutus"
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A group of men and women 12 in size were given a special task utilizing new-found weapons and tactics from the battle of hope to complete said task this battle-group was to be sent in on a mission to deeply infested territory in Africa. once there they would be airdropped roughly 600 meters from what the military designated as hives for the undead. what makes this task so unlike the others you ask ? it was the fact that this was one of the larger hives which contained some of the most mutated and vicious results of the plague creatures so deadly that very few have lived to tell of their existence. it was supposed to be a get in deploy the bomb and get out op however things went horribly wrong intel was off (as per the usual) a massive herd of over a million undead had amassed at the site. six of the team mates were killed by creatures that have been designated as "redacted" the other six tried to run to the evac point but only two made it. The whereabouts of the other four remain unkown.
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2014-Uav drones patrolling the site of the hive notice gunfire appearing from four locations within the herd
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Click to Watch in HD > Katakuti Khela | Acoustic Version | Full Audio Song | Srijit | Anupam Roy | 2016
Watch There is a different emotion attached to the acoustic version of the song Katakuti Khela from the film Zulfiqar. One can never get over this soulful version sung by the ever so tuneful Anupam Roy. Song Name: Katakututi Khela Singer: Anupam Roy Music: Anupam Roy Film : Zulfiqar Starring : Prosenjit Chatterjee, Dev, Parambrata Chatterjee, Jisshu Sengupta, Ankush Hazra, Koushik Sen, Rahul, Arunoday Banerjee, Kanchan Mullick, Nusrat Jahan, Paoli Dam, Kyra Dutt, June Malia. Presenter : Shrikant Mohta and Mahendra Soni. Produced by : Shree Venkatesh Films. Screenplay, Dialogue and Direction: Srijit Mukherji. D.O.P : Soumik Haldar. Music and Lyrics: Anupam Roy. Singer: Anupam Roy Background Music - Indraadip Dasgupta. Edit - Srijit Mukherji, Anindya Chattopadhyay. Action Director - Javed-Aejaz. ---------------------------------------------------------------------------- Set it as your Caller Tune SMS ZULFI4 to 54646 ---------------------------------------------------------------------------- Enjoy and stay connected with us!! Subscribe VenkateshFilms channel for unlimited entertainment https://www.youtube.com/venkateshfilms Like us on Facebook http://www.facebook.com/venkateshfilms Follow us on http://www.twitter.com/venkateshfilms Follow us on http://www.instagram.com/venkateshfilms
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Police say four people face felony charges after a vandalism spree involving paintball guns and marbles in the Savannah area that damaged dozens of businesses, houses, vehicles and a church over two weekends.
The four were arrested by officers from the Savannah-Chatham and Thunderbolt police departments early Monday morning.
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All four: Lavontay Frank Gatson, 20, of the 300 block of Young Avenue; April Ann Mizer, 18, of the 100 block of Tabby Lane; and Renee Kelsey Raker, 17, and William Cody Smith, 21, both of the 9900 block of Ferguson Avenue; were charged with two counts of criminal damage to property and nine counts of criminal trespass. Mizer and Smith also were charged with vandalism to a place of worship.
All four admitted to shooting marbles at multiple vehicles and businesses causing the glass and windows to shatter and spray painting multiple vehicles and the church on Lehigh Avenue, said Julian Miller, metro police spokesman.
Police identified dozens of cars and buildings with windows and other damage from marbles that had been fired from a paintball gun, he said.
The damage was similar to reports from Jan. 29 and 30 when numerous incidents of vandalism were reported.
Damage included broken windows at Food Lion on Skidaway Road, K-Mart on Montgomery Crossroad, Farmers Furniture, numerous car windows, vehicles spray painted, and a church spray painted in Pin Point.
Reports of damage continue to be received by metro police, Miller said. Islands and Southside Precinct detectives continue to investigate.
Pooler man indicted on robbery charges
A Pooler man was indicted last week by a federal grand jury in Savannah on robbery charges.
Bobby Joe White Jr., 42, is accused of robbing the Sea Island Bank and the Biomat USA Savannah Blood Plasma Donation Center, said James Durham, first assistant U.S. attorney for the Southern District of Georgia.
According to the indictment, White allegedly robbed the Sea Island Bank on Nov. 1 by "force, violence and intimidation." Then, on Nov. 22, he allegedly robbed the plasma donation center of about $8,000 in cash.
White faces a maximum sentence of 20 years in prison, a $250,000 fine and three years of supervised release for each robbery charge, Durham said.
The charges originated from an investigation by the FBI's Safe Streets Violent Crime Task Force of Southeast Georgia, which was assisted by officers from the Pooler and Savannah-Chatham police departments.
Assistant U.S. Attorney Charlie Bourne is prosecuting the case.
Cats quarantined at county animal shelter
Officers with the Savannah-Chatham police department's Animal Control division have quarantined cats at their own shelter after identifying a serious illness in the feline areas.
Shelter employees noticed some of the cats had become ill and sent cultures to be analyzed, said Julian Miller, police spokesman. Results received this morning indicated the presence of feline herpes virus infection, microplasma and bordetella, he said.
Officers reported the situation the Georgia Department of Agriculture and initiated a self-imposed quarantine for cats over the next three weeks. Cats will not be accepted or adopted out during this period but they will be taken in cases of injured animals, bite cases, feral and obviously ill cats, Miller said.
Medical staff have advised that cats displaying signs of the illness be euthanized upon arrival to prevent the spread of the illnesses, Miller said.
Police: Probationer tied to southside robbery
Savannah-Chatham police are asking the public's help locating a repeat offender with possible ties to a southside Savannah robbery.
Rashard Mosley, 18, of the 1600 block of Staley Avenue, is wanted for questioning by detectives in for a Dec. 14 armed robbery on the 10,000 block of Abercorn Street, where two gunmen stole cash and several personal items from the victim, said Eunicia Baker, metro police spokeswoman.
Mosley, a probationer since 2009, also is wanted by Savannah Impact Program after removing his electronic monitoring device on Jan. 29. This is the second leg monitor removed by Mosley in less than six months. Mosley is on Savannah Impact Program's top 10 list of prolific juvenile offenders. He is charged with removal of electronic monitoring device.
Mosley is a black male, standing 5-8 and weighing 155 pounds. He has a tear drop tattoo under his right eye and a cross under his left eye. There are several local gang-related tattoos on his hands and arms. He is known to frequent the 8500 block of Lyn Avenue and hotels throughout Chatham County. Mosley is believed to be accompanied by a female.
Detectives ask individuals with knowledge of Mosley's location to call 911 immediately. Do not approach him.Information can also be directed to CrimeStoppers by calling 912-234-2020 or texting CRIMES (274637) using the keyword CSTOP2020. Tipsters remain anonymous and may qualify for a cash reward.
A confidential tip line also is open directly to investigators at 912-525-3124.
Sewage spilled on southside
City of Savannah crews responded Monday to a minor sewage spill at a ditch at 12328 Largo Drive. A grease blockage in an 8-inch diameter gravity sewer main forced sewage to spill from a nearby private lateral line that had its cleanout cap removed. The total volume discharged was 385 gallons, which is classified as a minor spill under Georgia Environmental Protection Division guidelines.
Savannah Water and Sewer crews were notified of the spill at 2:58 p.m. and the spill was corrected at 4:15 p.m.
The blockage was cleared and the mix of contaminated and drainage water was pumped from the canal and back into the sewer collection system. The ditch was flushed and lime was placed on the affected area. Sampling will be done. Spill notifications were put at the spill site and Wilshire Canal at White Bluff, city officials report.
All agencies required by the EPD were notified.
Compiled by Dash Coleman and Mary Landers
To see pictures and details for individuals serving their initial incarceration in Chatham County, as reported by the Chatham County Sheriff's Office, go to booked.savannahnow.com.


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And Now Back To Celebrity Boobs, Mainly Jessica Simpson's
Since being predictable as fuck is my super power, after every single soapbox post, I go "Hey, guys, sorry about that. Giant boobs in your face?" because it works. So to make up for 18,000 words on pussy cops being afraid of Beyoncé, here are Jessica Simpson's breasts fresh off of having sex with Eric Johnson's penis on Valentine's Day like she told everyone for some reason. And if it wasn't to get him murdered in a violent crime of passion, then I should probably get out of this closet. And put clothes on. *tries to turn knob but hands keep slipping* Yup, I used too much Crisco.
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Enabling SDEE on AIP-SSM
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I have an AIP-SSM on a 5510. I'd like to pull events via SDEE. How do I go about setting this up? I have read some docs to just add host to the allowed hosts list, and setup a readonly account.
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Cytostasis and morphological changes induced by mifepristone in human metastatic cancer cells involve cytoskeletal filamentous actin reorganization and impairment of cell adhesion dynamics
Experimental therapeutics and drug development
Changes in cell shape and plasticity in cytoskeletal dynamics are critically involved in cell adhesion, migration, invasion and the overall process of metastasis. Previous work in our laboratory demonstrated that the synthetic steroid mifepristone inhibited the growth of highly metastatic cancer cells, while simultaneously causing striking changes in cellular morphology. Here we assessed whether such morphological alterations developed in response to cytostatic concentrations of mifepristone are reversible or permanent, involve rearrangement of cytoskeletal proteins, and/or affect the adhesive capacity of the cells.
Cancer cell lines of the ovary (SKOV-3), breast (MDA-MB-231), prostate (LNCaP), and nervous system (U87MG) were exposed to cytostatic concentrations of mifepristone and studied by phase-contrast microscopy. The transient or permanent nature of the cytostasis and morphological changes caused by mifepristone was assessed, as well as the rearrangement of cytoskeletal proteins. De-adhesion and adhesion assays were utilized to determine if mifepristone-arrested and morphologically dysregulated cells had abnormal de-adhesion/adhesion dynamics when compared to vehicle-treated controls.
Mifepristone-treated cells displayed a long, thin, spindle-like shape with boundaries resembling those of loosely adhered cells. Growth arrest and morphology changes caused by mifepristone were reversible in SKOV-3, MDA-MB-231 and U87MG, but not in LNCaP cells that instead became senescent. All cancer cell types exposed to mifepristone displayed greatly increased actin ruffling in association with accelerated de-adhesion from the culture plate, and delayed adhesion capacity to various extracellular matrix components.
Cytostatic concentrations of mifepristone induced alterations in the cellular structure of a panel of aggressive, highly metastatic cancer cells of different tissues of origin. Such changes were associated with re-distribution of actin fibers that mainly form non-adhesive membrane ruffles, leading to dysregulated cellular adhesion capacity.
Originally developed as an anti-glucocorticoid agent in the 1980s, the synthetic steroid mifepristone was also found to modulate the progesterone receptor. This unexpected finding led mifepristone to be rapidly repurposed for its use for early termination of pregnancy. However, aside from this most common usage, mifepristone has been proven effective as a growth inhibitor in endometriosis
Previous work in our laboratory demonstrated that mifepristone: i) arrests the growth of ovarian cancer cells by inhibiting DNA synthesis and halting progression of the cell cycle at the G1-S transition
Changes in cellular structure are a consequence of the rearrangement of cytoskeletal proteins, and are critically involved in adhesion turnover and polarized cell migration required for the success of the metastatic process
Cell culture and
The human ovarian carcinoma cell line SKOV-3, the human breast carcinoma cell line MDA-MB-231, the human glioblastoma cell line U87MG, and the human prostate carcinoma cell line LNCaP were obtained from the American Type Culture Collection (ATCC, Manassas, VA) and cultured as previously detailed
Time-course of morphology
Cells were seeded in 6-well plates at a density of 100,000 cells per well and allowed 24 h for attachment. Using previously established cytostatic doses of mifepristone (SKOV-3: 23.5 μM, MDA-MB-231: 30 μM, U87MG: 20 μM, LNCaP: 20 μM)
Reversal of cell proliferation and morphology
In order to determine the long-term effect of mifepristone treatment, cell morphology was assessed after removal of treatment media. After 72 h of treatment, mifepristone-containing media was removed, and media for all cells was replaced with control media. Phase contrast images were taken at 0 h, 12 h, 24 h, 48 h, and 72 h after mifepristone withdrawal to observe cell morphology. In addition to images taken at each time-point, cell number and viability were determined using the Guava EasyCyte Mini microcapillary cytometer (Guava Technologies, Hayward, CA). Samples were collected at the beginning of the experiment, for vehicle and mifepristone-treated cells after 72h of treatment, and at each time-point after treatment withdrawal. Triplicate wells were trypsinized, the cells pelleted by centrifugation at 500
Senescence Associated (SA)-β-galactosidase staining
Using 6-well plates, LNCaP cells were plated at a density of 50,000 cells per well. Cells were treated with mifepristone for 72 h followed by vehicle for 5 days, or vehicle, 5% charcoal-stripped (CS)-FBS, or 10% CS-FBS media for 8 days prior to SA-β-galactosidase staining. Cells were fixed in 0.5% glutaraldehyde at 4°C for 10 min and then washed three times with PBS. Cells were then incubated for 3 h with 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining solution consisting of 1 mg/ml X-gal, 40 mM citric acid/sodium phosphate (pH 6.0), 5 mM potassium ferricyanide, 5 mM potassium ferrocyanide, 150 mM NaCl, and 2 mM MgCl2. Following incubation, cells were washed briefly with PBS and stored in 100% methanol for analysis and imaging. Cells that expressed SA-β-galactosidase were stained blue when viewed using a Zeiss Axiovert M1-Imager (Carl Zeiss) microscope. Senescence was quantified as the number of blue-stained cells per field and expressed as a percent of total number of cells per field and corrected against a negative control. The average of 9 fields per well was calculated, with 3 wells per treatment group. Positive controls for senescence staining were LNCaP cells that had been depleted of steroids by culture in 5% or 10% CS-FBS media as reported
All cell lines were plated in 8-well chamber slides at a density of 5,000 cells per well. Cells were allowed to attach for 24 h before treatment began. Wells were treated with vehicle or media containing mifepristone for 72 h at doses tailored to induce cytostasis and morphological changes to individual cell lines. Following treatment cells were fixed according to Waterman-Storer et al.
SDS-PAGE and western blotting
Cells were treated with vehicle or mifepristone for 72 h, after which cultures were trypsinized, stained with trypan blue, and counted using a hemacytometer. Equal numbers of viable vehicle and mifepristone-treated cells were then pelleted, washed twice with PBS, and snap frozen followed by storage at −80°C. Whole cell extracts were obtained, protein quantitated, separated by SDS-PAGE, electro-transferred to PVDF membranes, and then probed for 1 h at room temperature using primary antibodies against α-tubulin (A-11126; 1:1,000; Molecular Probes), β-actin (clone AC-15; 1:10,000; Sigma), or GAPDH (ab94985; 1:8,000; Abcam Inc., Cambridge, MA).
De-adhesion assay
Cells were first grown to 50% confluence in 6-well plates, and then treated with vehicle or mifepristone-containing media for 72 h. Following treatment, cells were exposed to 0.025% trypsin/0.265 mM EDTA for 30 sec, 2 min or 4 min. Detached cells were removed with a washing of PBS. Cells remaining adhered were fixed with 100% methanol, stained with crystal violet, and quantified using bright field microscopy. Cell adhesion was expressed as percent of adherent cells for each of the times of exposure relative to the adhesion measured in a culture not exposed to trypsin (considered to be 100%).
Adhesion assays
The first adhesion assay was performed under sterile conditions using the CytoSelect 48-Well Cell Adhesion Assay (CBA-070, Cell Biolabs Inc., San Diego, CA). Briefly, the adhesion plates containing various extracellular matrix components (fibronectin, collagen I, collagen IV, fibrinogen, or laminin) were allowed to warm up at room temperature for 10 min. Cell suspensions were then prepared containing 1 × 106 cells per ml in serum-free media with or without mifepristone; 150 μl of each cell suspension was added to the appropriate wells and the plates were incubated for 60 min at 37°C in a humidified atmosphere in the presence of 5% CO2. The media was discarded from each well and all wells were washed 4–5 times with 250 μl PBS. PBS was removed and 200 μl of the provided cell stain solution was added to each well. The plates were incubated for 10 min at room temperature. After incubation, the cell stain solution was removed and each well was washed 4–5 times with 500 μl of deionized water. The final wash was discarded and wells were allowed to air dry. Next, 200 μl of provided extraction solution was added to each well and the plates were incubated for 10 min on an orbital shaker at room temperature. Finally, 150 μl of each sample was transferred to 96-well microtiter plates and the optical density at 540 nm was measured using a Titertek Multiskan MCC/340 Microplate Reader II (Dupont, Labsystems, Finland).
When adhesion to fibronectin was further studied, cells were cultured in the presence of 20 μM mifepristone for 72 h or left untreated in controls. The cells were trypsinized and incubated in suspension for 20 min to allow recovery from trypsinization. Thereafter, 100,000 cells were placed per 35 mm diameter plates that had been pre-coated with 0.1% fibronectin (Sigma), and were incubated for various times. Cells were fixed with methanol and stained with crystal violet. Counting was achieved under a microscope by recording number of adherent cells per 20 X microscopic field.
Statistical analysis
Data processing and statistical analysis were performed using GraphPad Prism (GraphPad Prism Software, Inc., San Diego, CA). All data are represented as means ± SEM, and statistical significance was defined as
Cytostatic concentrations of mifepristone cause morphological changes in cancer cell lines of the ovary, breast, prostate, and nervous system
In a previous study we made the serendipitous observation that when cancer cells of various tissues of origin were exposed to concentrations of mifepristone that inhibited their growth
Additional file 1: Figure S1
Time-course effect of mifepristone on cancer cells of the ovary (A), breast (B), prostate (C), and nervous system (D). Cells were seeded at a density appropriate for exponential growth for each cell line, allowed to adhere for 24 h, and then exposed to a previously determined cytostatic concentration of mifepristone (MF) for 72 h. Cells that received vehicle for 72 h were used as positive control of growth (right panel, VEH). Images were taken using phase contrast microscopy every 12 h and examined for morphological changes. Scale bar = 50 μm.
Click here for file
Additional file 2: Figure S2
Treatment of SKOV-3 cells with mifepristone induced inhibition of growth associated with changes in cell morphology. Cells were cultured in 8-well chamber slides in the presence of vehicle (VEH) or 20 μM mifepristone (MF) for 4 days. At the end of the incubation cells were fixed with 4% paraformaldehyde and stained with hematoxylin. X 400.
Click here for file
Figure 1
Cells lines representing cancers of the ovary (SKOV-3), breast (MDA-MB-231), nervous system (U87MG), and prostate (LNCaP) display distinct morphological changes in response to mifepristone treatment
Cells lines representing cancers of the ovary (SKOV-3), breast (MDA-MB-231), nervous system (U87MG), and prostate (LNCaP) display distinct morphological changes in response to mifepristone treatment. Equal number of cells were plated and allowed to attach for 24 h. Cells were then exposed to vehicle (VEH)-containing media, or media containing a previously tailored cytostatic dose of mifepristone (MF) for a period of 72 h. At the end of the experiment the total number of cells was recorded by microcytometric analysis (A) and images were taken using phase contrast microscopy (B). Cells were exposed to the following cytostatic concentrations of MF: 23.5 μM for SKOV-3, 30 μM for MDA-MB-231, and 20 μM for U87MG and LNCaP. *p < 0.01 and **p < 0.005 vs. VEH (Student's
Cellular proliferation and morphological changes are reversible upon removal of mifepristone in all cell lines except for LNCaP prostate cancer cells that undergo cellular senescence
To determine the long-term effect of mifepristone on cell proliferation and morphology, each cell line was cultured in the presence of vehicle or mifepristone for 72 h. Thereafter, mifepristone was removed from the cultures and replaced with media lacking the drug for 5 days. Subsequently, cultures were imaged and cell number was determined every 24 h to monitor for reversal of morphology and proliferation. While the proliferation of SKOV-3, MDA-MB-231, and U87MG cells remained relatively slow 1–2 days post-treatment, by day 3 after mifepristone withdrawal, cultures were proliferating at the same rate as cultures never treated with the synthetic steroid (Figure
Additional file 3: Figure S3
Long-term effect of mifepristone on LNCaP cells. Cells were seeded, allowed to adhere for 24 h, and then exposed to a cytostatic concentration of mifepristone (MF) for 72 h. Thereafter, triplicate wells were harvested by trypsinization and counted by microcytometry. Remaining wells were returned to vehicle treatment and monitored after 1, 3, 7, or 9 days for their growth in the absence of MF and compared against the growth of similar number of cells never exposed to the steroid.
Click here for file
Figure 2
Time-course effect of mifepristone withdrawal on cancer cells of the ovary (A), breast (B), prostate (C), and nervous system (D)
Time-course effect of mifepristone withdrawal on cancer cells of the ovary (A), breast (B), prostate (C), and nervous system (D). Cells were seeded at a density appropriate for exponential growth for each cell line, allowed to adhere for 24 h, and then exposed to the previously determined cytostatic concentrations of mifepristone for 72 h. Thereafter mifepristone-containing media was replaced with normal growth media and images were taken using phase contrast microscopy after 0, 24, 48 or 72 h (A-D). Scale bar = 50 μm.
Figure 3
Long-term effect of mifepristone on tumor cell lines of the ovary (A), breast (B), nervous system (C), and prostate (D, E)
Long-term effect of mifepristone on tumor cell lines of the ovary (A), breast (B), nervous system (C), and prostate (D, E). Cells were seeded at a density appropriate for each cell line, allowed to adhere for 24 h, and then exposed to concentrations of mifepristone (MF) specific for each cell line for 72 h. Thereafter, triplicate wells were harvested by trypsinization and counted by microcytometry. Remaining wells were returned to vehicle treatment and monitored for 5 days, during which time triplicate wells were harvested and counted every 24 h. Growth expressed as number of cells per well are shown for each cell line (A-D). Data points represent the mean ± s.e.m of 3 independent experiments completed in triplicate. Viability of LNCaP cells was also determined by microcytometric analysis (E).
Figure 4
Mifepristone treatment induces senescence in LNCaP cells
Mifepristone treatment induces senescence in LNCaP cells. LNCaP cells were seeded at 50,000 cells per well and treated with mifepristone (MF) for 72 h followed by vehicle (VEH) for 5 days; or vehicle, 5% or 10% charcoal-stripped (CS)-FBS-containing media for 8 days. SA-β-galactosidase staining was performed as surrogate marker of senescent cells. Relative senescence was quantified as the number of cells with blue cytoplasm per field and expressed as the percent of total number of cells per field (A, B). Nine fields per well were quantified and completed in triplicate for each treatment group. All treatment groups were corrected against the background provided by the negative control, WI-38 cells that do not undergo senescence (data not shown). *** p < 0.0001 vs. VEH (one-way ANOVA followed by Tukey's post-test). Scale bar = 50 μm.
Cytostatic doses of mifepristone dysregulate the cytoskeletal architecture of cancer cells
To further characterize the morphological changes caused by cytostatic, non-lethal concentrations of mifepristone, the various cancer cell types were cultured in the presence of vehicle or mifepristone for 72 h, and the arrangement of filamentous actin (F-actin) and tubulin filaments contributing to cytoskeletal structure and overall cell morphology were assessed. Figure
Additional file 4: Figure S4
Effect of mifepristone on the cellular distribution of filamentous actin (F-actin) and tubulin. SKOV-3 cells, U87MG, MDA-MB-231 and LNCaP were cultured in the presence of vehicle (VEH) or mifepristone (MF) for 72 h, following which immunocytochemistry was used to visualize the cytoskeletal protein α-tubulin, AlexaFluor 594® phalloidin was used to visualize filamentous actin (F-actin), and DAPI to label cell nuclei. Images were taken using confocal microscopy. Scale bar = 50 μm.
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Figure 5
Effect of mifepristone on cytoskeletal actin fibers and tubulin filaments
Effect of mifepristone on cytoskeletal actin fibers and tubulin filaments. SKOV-3 (A), U87MG (B), MDA-MB-231 (C) or LNCaP (D) cells were cultured in the presence of vehicle (VEH) or a cytostatic concentration of mifepristone (MF) for 72 h, following which immunofluorescence was used to visualize the cytoskeletal protein α-tubulin. AlexaFluor® 594 phalloidin was utilized to visualize F-actin and DAPI to label cell nuclei. Images were taking using confocal microscopy. Scale bar = 50 μm. The inset in panel D represents a cell that was in a different field within the same image and that denotes the characteristic increase in membrane ruffles induced by MF (for the complete image see Additional file
To evaluate whether mifepristone was merely dysregulating the distribution of actin and tubulin or changing their abundance, we assessed the expression levels of one of the isotypes of actin, β-actin, and one of the isotypes of tubulin, α-tubulin. Given that actin and tubulin are both commonly used as loading controls in Western blot studies, we sought to evaluate any differences in their expression by loading lysates obtained from an equal number of vehicle or mifepristone-treated cells. Membranes were immunoblotted for β-actin, α-tubulin, and GAPDH, which was used as protein loading control. Densitometry analysis was performed and protein levels expressed as the ratio of β-Actin/GAPDH (Additional file
Additional file 5: Figure S5
Expression of β-actin and α-tubulin in mifepristone-treated cells. Cells were plated and exposed to either vehicle (VEH) or the cytostatic concentration of mifepristone (MF) optimized previously for each cell line for 72 h. Following treatment, cells were subsequently harvested, lysed, and whole-protein extracts, representing equal numbers of VEH or MF-treated cells were separated by electrophoresis. Immunoblots were then probed for β-actin and α-tubulin. GAPDH was included as a loading control. Densitometry analysis was performed from three different experiments and protein levels expressed as the ratio of β-actin/GAPDH (A), or α-tubulin/GAPDH (B). Densitometry graphs represent the mean ± s.e.m.
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Effect of mifepristone on cellular de-adhesion and adhesion dynamics
One commonality in the cancer cells that were treated with cytostatic doses of mifepristone was the increased density of membrane actin ruffles along the surface of the cells (Figure
Figure 6
Exposure to mifepristone impairs the ability of cancer cells to remain adherent
Exposure to mifepristone impairs the ability of cancer cells to remain adherent. SKOV-3 (A), LNCaP (B), MDA-MB-231 (C) or U87MG (D) cells were plated at equal densities and treatment with vehicle (VEH) or mifepristone (MF)-containing media was administered for 72 h. Cells were then exposed to 0.025% trypsin/0.265 mM EDTA for 0.5, 2, or 4 min. The detached cells were removed, while those remaining adherent were fixed with 100% methanol and stained with 0.25% crystal violet (as shown in panel E for U87MG cells; 4 min trypsin exposure; 200 X). Stained cells were then imaged and counted via bright field microscopy. Adherent cells were quantified as a percent of control (0 min trypsin exposure). Data shown represent the mean ± s.e.m. of 3 independent experiments completed in triplicate. * p < 0.01 vs. VEH; ** p < 0.001 vs. VEH (A-D). Statistical analysis was done using two-way ANOVA followed by Bonferroni's post-tests.
To assess whether mifepristone impairs the capacity of cancer cells to adhere to extracellular matrix, cells were pre-treated for 72 h with or without a cytostatic concentration of mifepristone, trypsinized, and re-plated in commercially available plates that had been pre-coated with an array of extracellular matrix proteins including fibronectin, collagen I, collagen IV, laminin or fibrinogen. Cells were allowed 1 h to adhere, except for LNCaP cells that underwent adhesion for 24 h. Thereafter, the cells were stained and optical density (OD) was measured. All cell lines showed different kinetics of adhesion to the substrates offered, as observed by the different ranges of OD detected (Figure
Additional file 6: Table S1
Semi-quantitative representation of the effect of mifepristone on the adhesion of cells to individual extracellular matrix proteins.
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Figure 7
Mifepristone-pretreated MDA-MB-231 (A), U87MG (B), LNCaP (C), and SKOV-3 (D) cells have delayed adhesion to extracellular matrix proteins
Mifepristone-pretreated MDA-MB-231 (A), U87MG (B), LNCaP (C), and SKOV-3 (D) cells have delayed adhesion to extracellular matrix proteins. For all cell lines, treatment with cytostatic concentrations of mifepristone was administered for 72 h prior to plating. After treatment, cells were trypsinized and re-plated in a pre-coated plate at a density of 100,000 cells per well. FN: fibronectin, Col I: collagen I, Col IV: collagen IV, LN: Laminin, FG: Fibrinogen. Data shown in A-D represent one experiment that was repeated three times with a similar outcome. In E, SKOV-3 cells were cultured in the presence of vehicle (VEH) or mifepristone (MF) for 72 h. The cells were trypsinized and placed in plates that had been pre-coated with 0.1% fibronectin or bovine serum albumin (BSA) used as a negative control of adhesion. Adhesion was quantified as the number of cells attached per 20 X microscopic field. * p < 0.05 vs. VEH; ** p < 0.01 vs. VEH. Statistical analysis was done using two-way ANOVA followed by Bonferroni's post-test. Bright field Images in F are representative of vehicle (VEH) or mifepristone (MF)-pretreated cells observed after 60 min of incubation (200 X).
To confirm the impairment of mifepristone-pretreated cells to adhere to an extracellular matrix-related protein, an equal number of either vehicle-pretreated or mifepristone-pre-treated SKOV-3 cells were incubated at 37°C for various times (0.5, 1, or 2 h) on plates that had been pre-coated with fibronectin. Thereafter, the cultures were washed to remove the non-adherent cells, whereas the cells that had already attached to the plate at each time point were fixed with methanol, stained with crystal violet, and their number was quantified per microscopic field. The results shown in Figure
In this study we observed that, at known cytostatic doses, mifepristone induced a strikingly similar change in shape of four highly metastatic and aggressive cancer cell lines. This included shrinkage of the cell body with long, thin, neurite-like cellular extensions. Morphology changes were found to be dependent on time of exposure to the drug, with confirmed and observable phenotypic changes occurring after 48 h. These alterations were not influenced by the cellular density of culture, suggesting that mifepristone has an intrinsic effect upon cellular structure, and that this change in morphology is not a result of lack of perceived cell number. In addition, treatment with mifepristone was not associated with changes in total expression of β-actin and α-tubulin.
Mifepristone is known to exert anti-progestin activity by blocking the action of progesterone on progesterone receptors (PR)
Potential mediators of the effect of mifepristone are the glucocorticoid receptors (GR). It was reported that mifepristone binds GR when used at concentrations higher than those needed to bind PR
When operating at micromolar concentrations, mifepristone has a distinctive effect not shared by other natural and synthetic steroids. For instance, when used at equimolar concentrations, mifepristone was a much potent inhibitor of ovarian cancer cell growth than progesterone, medroxyprogesterone acetate or levonorgestrel
Important in studying mifepristone as a possible treatment option in oncology was to address its long-term effect on the cancer cells. Interestingly, upon removal of mifepristone following exposure for 3 days, cell morphology and proliferation returned to that of untreated cells in all cell lines except for the LNCaP prostate cancer cells. In these cells, the non-proliferative effect of mifepristone remained as long as 9 days following treatment removal, suggesting an irreversible growth arrest. Previous research has shown the propensity of LNCaP cells to enter irreversible growth arrest and senescence in response, for instance, to treatment with doxorubicin or to culture in androgen-free media
An alternative explanation for the senescence induced by mifepristone in LNCaP cells is the possible mediation by androgen receptors (ARs). LNCaP is the only cell line in the studied cohort that expresses ARs
Mifepristone likely altered cellular morphology as a consequence of the dysregulation of the cytoskeletal structure, which was observed via fluorescent staining of actin fibers (F-actin) and tubulin filaments; these were both found to rearrange in response to mifepristone. Actin fibers were found reorganized to sites located at the ends of tubulin-rich extensions. As this was seen multiple times in different directions within individual cells, there appears to be a loss of cell polarity following mifepristone treatment. Upon mifepristone treatment, tubulin filaments were mainly located in the neurite-like extensions, in contrast to their original localization throughout the cell body with particular intensity around the nucleus observed in controls.
F-actin is formed by polymerization of actin molecules that assemble at different times and locations, depending upon the extracellular environment
We observed that cells under the stress of mifepristone are easily de-adhered from the extracellular surface than untreated cells when exposed to a sub-optimal concentration of trypsin. Because an inverse relationship between de-adhesion time and cell contractility assessed by trypsin-induced de-adhesion has been demonstrated
Since the survival, movement and invasiveness of cancer cells in the organism require great plasticity in the distribution of F-actin, our data suggest the mifepristone may interfere with such actin polymerization dynamics, disturbing the metastatic process. Blocking actin plasticity with mifepristone can be therapeutically beneficial to reduce the seeding at secondary sites by cells that had detached from a primary tumor.
Microtubules are important to maintain cell shape, play a key role in the polarized distribution of signals within a cell, and have been implicated in the asymmetric regulation of adhesion dynamics; in particular, they promote adhesion disassembly triggering the destabilization first, and then the detachment of adhesion components. At the same time, adhesions can be pulled off the substrate by stress fibers, which contract in response to microtubule depolymerization
The anti-cancer effect of mifepristone manifested in the inhibition of cellular growth is related to drastic alternations in cellular morphology with the formation of neurite-like protrusions and altered cytoskeletal architecture characterized by an increase in membrane F-actin ruffling and concentration of tubulin filaments at the neurite-like cellular extensions. Such effect of mifepristone is associated with dysregulated cellular adhesion; it is reversible in most cell lines, except for prostate cancer cells that instead undergo senescence. Whether or not mifepristone-induced cytostasis, and alterations in cell shape, cytoskeletal structure, and adhesion capacity affect the migratory and invasive properties of cancer cells, warrant further investigations.
Competing interests
The authors declare that there is no conflict of interest that could influence the impartiality of the research reported.
Authors' contributions
Conceived and designed experiments: BNB AAG CMT. Performed experiments BNB CRT TMU MST AAG. Analyzed the data: BNB AAG CMT. Contributed reagents/materials/analysis tools: CMT. Wrote the paper: BNB CMT. All authors read and approved the final manuscript.
This study was supported by National Cancer Institute Grant K22CA121991, ARRA Supplement K22CA121991-S1, and R15CA164622 (to CMT) and in part by a South Dakota Board of Regents competitive research grant award (SDBOR/USD 2011-10-06) (to CMT), and funds from the Division of Basic Biomedical Sciences, Sanford School of Medicine (to AAG). BNB and CRT were recipients of graduate student scholarships from the University of South Dakota. MST was a recipient of a summer medical student research scholarship from the Sanford School of Medicine.
Emerging drugs for endometriosis
Preliminary report on the treatment of endometriosis with low-dose mifepristone (RU 486)
Low-dose mifepristone for uterine leiomyomata
Regression of uterine leiomyomata to the antiprogesterone RU486: dose–response effect
Systematic review of mifepristone for the treatment of uterine leiomyomata
Antitumor effects of antiprogesterones on human meningioma cells in vitro and in vivo
Cisplatin cytotoxicity is increased by mifepristone in cervical carcinoma: an in vitro and in vivo study
Antiprogestins in breast cancer treatment: are we ready?
Endometrial effects of RU486 in primates–antiproliferative action despite signs of estrogen action and increased cyclin-B expression
Low-dose mifepristone inhibits endometrial proliferation and up-regulates androgen receptor
Inhibition of endometrial cancer cell lines by mifepristone (RU 486)
Levonorgestrel, medroxyprogesterone and progesterone cause a concentration-dependent reduction in endometrial cancer (Ishikawa) cell density, and high concentrations of progesterone and mifepristone act in synergy
Response of human ovarian carcinoma cell lines to antiprogestin mifepristone
Growth inhibition induced by antiprogestins RU-38486, ORG-31710, and CDB-2914 in ovarian cancer cells involves inhibition of cyclin dependent kinase 2
Mifepristone abrogates repopulation of ovarian cancer cells in between courses of cisplatin treatment
Resistance to cisplatin does not affect sensitivity of human ovarian cancer cell lines to mifepristone cytotoxicity
Mifepristone inhibits ovarian cancer cell growth in vitro and in vivo
Effects of mifepristone on invasive and metastatic potential of human gastric adenocarcinoma cell line MKN-45 in vitro and in vivo
Antitumor activity of mifepristone in the human LNCaP, LNCaP-C4, and LNCaP-C4-2 prostate cancer models in nude mice
Induction of apoptosis by mifepristone and tamoxifen in human LNCaP prostate cancer cells in culture
Mifepristone treatment improves length and quality of survival of mice with spontaneous leukemia
Mifepristone treatment improves length and quality of survival of mice with spontaneous lung cancer
Efficacy of the progesterone receptor antagonist mifepristone for palliative therapy of patients with a variety of advanced cancer types
Endocrine therapy in metastatic breast cancer
Mifepristone prevents repopulation of ovarian cancer cells escaping cisplatin-paclitaxel therapy
Antiprogestin mifepristone inhibits the growth of cancer cells of reproductive and non-reproductive origin regardless of progesterone receptor expression
Asymmetric focal adhesion disassembly in motile cells
Tumor metastasis: molecular insights and evolving paradigms
Androgen depletion induces senescence in prostate cancer cells through down-regulation of Skp2
Feedback interactions between cell-cell adherens junctions and cytoskeletal dynamics in newt lung epithelial cells
Regulation of microtubules in cell migration
The many faces of actin: matching assembly factors with cellular structures
Membrane ruffles in cell migration: indicators of inefficient lamellipodia adhesion and compartments of actin filament reorganization
Mechanism of action of an antiprogesterone, RU486, in the rabbit endometrium. Effects of RU486 on the progesterone receptor and on the expression of the uteroglobin gene
RU 38486: potent antiglucocorticoid activity correlated with strong binding to the cytosolic glucocorticoid receptor followed by an impaired activation
Molecular origins for the dominant negative function of human glucocorticoid receptor beta
Human glucocorticoid receptor beta binds RU-486 and is transcriptionally active
Glucocorticoid receptor (GR) beta has intrinsic, GRalpha-independent transcriptional activity
Antiprogestins in ovarian cancer
Drug-induced senescence bystander proliferation in prostate cancer cells in vitro and in vivo
Adenovirus-mediated p16 gene transfer changes the sensitivity to taxanes and Vinca alkaloids of human ovarian cancer cells
p53 disruption profoundly alters the response of human glioblastoma cells to DNA topoisomerase I inhibition
Predicting chemoresistance in human malignant glioma cells: the role of molecular genetic analyses
INK4a gene expression and methylation in primary breast cancer: overexpression of p16INK4a messenger RNA is a marker of poor prognosis
Expression of the cyclin-dependent kinase inhibitors p16INK4, p15INK4B and p21WAF1/CIP1 in human breast cancer
Molecular mechanisms of androgen-independent growth of human prostate cancer LNCaP-AI cells
Involvement of the cyclin-dependent kinase inhibitor p16 (INK4a) in replicative senescence of normal human fibroblasts
Cooperative effect of antisense-Rb and antisense-p53 oligomers on the extension of life span in human diploid fibroblasts, TIG-1
A role for both RB and p53 in the regulation of human cellular senescence
Reversal of human cellular senescence: roles of the p53 and p16 pathways
Pro-senescence therapy for cancer treatment
BRAFE600-associated senescence-like cell cycle arrest of human naevi
Crucial role of p53-dependent cellular senescence in suppression of Pten-deficient tumorigenesis
A senescence program controlled by p53 and p16INK4a contributes to the outcome of cancer therapy
Antiandrogen effects of mifepristone on coactivator and corepressor interactions with the androgen receptor
Mechanics of the F-actin cytoskeleton
Coupling membrane protrusion and cell adhesion
Adhesion-dependent cell mechanosensitivity
Cell-matrix De-adhesion dynamics reflect contractile mechanics
Role of RhoA, mDia, and ROCK in cell shape-dependent control of the Skp2–kip1 pathway and the G1/S transition
Cytoskeletal control of growth and cell fate switching
Growth control by intracellular tension and extracellular stiffness
Pre-publication history
The pre-publication history for this paper can be accessed here:


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text: 8-Ball is a character in the Grand Theft Auto series, only appearing in Grand Theft Auto III and Grand Theft Auto: Liberty City Stories. He is a shop owner who owns 8-Ball Autos, a shop that installs car bombs on vehicles. He never makes an appearance in Grand Theft Auto: Vice City and Grand Theft Auto: Vice City Stories.


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Posted: Sunday, May 14, 2017 4:43 AM
By the end of that season, he made his Ptan ebut, thrown into the deep end, sent in at No. 3 in the second innings in Aand. He failed to score enough runs there, but what he gained became the basis for all that was to come.
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MILWAUKEE (AP) The Milwaukee Bucks acquired a fan favorite when they completed a trade Wednesday for Sacramento Kings forward Jon Brockman (FSY).
Bucks general manager John Hammond said he thinks Bucks fans will love Brockman, too.
"Any fan base anywhere will appreciate a player who plays with the type of energy he does," Hammond told the Associated Press.
The Bucks completed a sign-and-trade deal for Brockman in exchange for forward Darnell Jackson (FSY) and a 2011 second-round pick.
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The 23-year-old Brockman averaged 2.8 points and 4.1 rebounds in limited minutes as a rookie with the Kings. He appeared in 52 games, including four starts, after being picked in the second round of last year's draft.
"We're excited about adding a player with his energy, his toughness and a player we think we can put on the floor and help us win games," Hammond said.
Jackson began last year with Cleveland and appeared in 28 games before moving to Milwaukee when the Cavs waived him to bring back Zydrunas Ilgauskas (FSY). Jackson, a second-year forward, played in one game with the Bucks.
"We're looking forward to greeting Darnell and moving forward with the rest of the roster over the course of the summer," Kings' president of basketball operations Geoff Petrie said.
Brockman participated in a live chat with The Seattle Times earlier Wednesday and said it was bittersweet leaving the Kings. He said he had just signed a new contract at the newspaper's office before participating in the chat. That deal is worth about $3 million over the next three years.
"It really did feel like home. It was a great place to live and play, but I'm very excited about my new opportunity in Milwaukee," Brockman wrote. "I truly am going to miss the support I received in Sacramento and am touched by all the messages, poems, calls I've gotten from people telling me I will be missed."
Brockman was known as The Brockness Monster in Sacramento and has a Twitter audience of more than 320,000 followers on his verified account. Brockman had been told not to communicate on Twitter as the deal was being negotiated, but he wrote Wednesday that he was excited about his move.
"You have no idea how pumped I am to be in Milwaukee!" he wrote. "Great people, great city, great team!"
On the floor, the Bucks believe the rugged 6-foot-7 power forward also will be able to guard centers.
Milwaukee may not be done with all of its moves, either.
The Bucks signed Keyon Dooling (FSY) to a two-year contract on Monday to be the backup point guard and Hammond said work remains to finish signing their draft picks.
Outside of Miami, Milwaukee has been one of the busiest teams this offseason, needing to fill eight roster spots following last season's 46-36 run that ended with the franchise's first playoff berth in four years.
Milwaukee traded Dan Gadzuric (FSY) and Charlie Bell (FSY) for small forward Corey Maggette (FSY) in the days leading up to the draft, then selected forward Larry Sanders (FSY), guard Darington Hobson (FSY) and forward Tiny Gallon (FSY).
The Bucks also struck two major deals in free agency, bringing back shooting guard John Salmons (FSY) and signing power forward Drew Gooden (FSY) to five-year contracts to add them to a core that includes center Andrew Bogut (FSY) and point guard Brandon Jennings (FSY).
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St Lucia: Pakistan captain, Shahid Afridi believes that the senior players should make way for younger talents for the better of Pakistan cricket.
He said this after the recently concluded ICC T20 World Cup 2010 in which his team managed to make it till the semi-final stage before they lost out against Australia.
Before the team's departure to London from St Lucia said to the PA sports network, " It is time for some senior players to go because we can't expect a good show from them any more in the future."
He added: "There is no dearth of talent in Pakistan, especially in the T20 format; the youngsters should be given a chance in the national team."
Especially, for the shortest version of the game, Afridi feels it is time to look out for talented youngsters.


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Winterball continues for the Big Red. Get all the information you need about Cornell's games at Stony Brook, Sunday, December 22, 2013, at Saint Peter's, Saturday December 28, 2013, at St. Bonaventure (TV: NBC Sports Network), Saturday January 4, 2014 and vs. Oberlin, Saturday, January 11, 2014, right here with The Cornell Basketball Blog's Game Preview Center.
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The comely, curly haired Melanie Merkosky first came to prominence with her role on the Canadian comedy series Slings and Arrows. While she is certainly an accomplished actress, noted for her roles in the Golden Globe-nominated films Away from Her and Door to Door, she's actually dipped her toe into the writing and directing waters as well, making her quite a versatile talent. She came to Mr. Skin's attention however when she appeared in the direct to video spin-off American Pie Presents The...Join Mr. Skin for the full bio >>
An Eye for Beauty (2014) - as Lindsay Walker
American Pie Presents The Naked Mile (2006) - as Natalie
Runaway - as Sam
Slings and Arrows (2003) - as Megan


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Tatum turns producer
The G.I. Joe: The Rise of Cobra star will play a young forger with multiple identities in the story based on Craig Clevenger's bestselling book, as well as taking on producing duties, reports Deadline.com.
He will helm the picture alongside Reid Carolin, Sriram Das, John Penotti, Tim Williams and Sidney Kimmel, while newcomer Robin Shushan has taken on scriptwriting duties.


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EDMC - Education Management Corporation [www.edmc.edu], among the largest providers of private post-secondary education in North America, chose Big Clic to launch and grow enrolment for their newest Canadian school, The Art Institute of Toronto.
The Art Institute of Toronto, part of The Art Institutes group of schools [www.artinstitutes.edu], entered the Toronto market in June 2003 with aggressive enrolment plans.
The principle challenges presented to us were to make the school a Top 5 school when considering an education in the crowded Toronto design school market, to grow enrolment and market share, and to make the Toronto campus profitable within 3 years. And though a leading design school for students in the United States boasting over 40,000 current students, The Art Institutes brand was a relative unknown in Canada.
The Art Institute of Toronto offers diploma courses in 2D and 3D Animation, Game Art and Design, Graphic Design, Web Design, Interior Design, Residential Design, Fashion Design, and Fashion Marketing.
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