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Tuesday's match between Greece and the Czech Republic has huge implications poland greece betting preview goal both sides poland greece betting preview goal terms of their chances to advance to the knockout stages of the European Championship. Greece was able to come away with a draw in its opening match against Poland in Warsaw, despite playing the majority of the match a man down. Poland greece betting preview goal Polish side looked in control, and it seemed they would start the tournament off with a win, but substitute Dimitris Salpingidis found the back of the net in the 51st minute to tie the game at one. The Czech Republic was taken behind the woodshed by Russia in its first match, falling in embarrassing fashion They'll certainly need a better effort if they want to avoid being all but mathematically eliminated after their first two games. Sokratis Papastathopoulos is suspended for this one after being sent off against Poland on a controversial call, to say the least. Avraam Papadopoulos, another key defensive contributor for the Greeks, is out of the tournament with a knee injury.

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Otacon told Snake of how Ocelot, the sole terrorist survivor, had sold Metal Gear's data on the black market and many countries would now develop their own versions of Metal Gear. Otacon told Snake that he could make a difference and convinced him to join him in forming a new group, Philanthropy, an anti-Metal Gear organization officially recognized by the United Nations. For the next two years or so, Otacon and Snake destroyed dozens of rogue Metal Gears, with nothing significant taking place.

Now every group, state, and dot-com has their own version of Metal Gear, Metal Gear is no longer a secret weapon. It has become one of the most common and effective weapons on the battlefield of the day. Snake, having spent all of his days as a mercenary working on eliminating Metal Gear, decides that he has to find out more on this new Metal Gear.

Witht he help of his new accomplice, Otacon aka Hal Emmerich, a scientist from Shadow Moses , he infiltrates the transport for the new Metal Gear. The transport has been disguised as an oil tanker descending the Hudson River, on its way out to sea. Snake must discover the purpose behind this new incarnation of Metal Gear and if necessary he must destroy it. The purpose of the Big Shell is to clean up the spilled oil from the incident two years ago and restore stability to the local ecosystem.

Now, terrorists have seized the Big Shell and captured 30 hostages and the President of the United States, who was taking an inspection tour that day. Raiden must rescue the President and the other hostages and disarm the terrorists by any means necessary. Raiden must also disarm the numerous C4 bombs that the terrorists have placed around the Big Shell. Raiden successfully infiltrates the Deep Sea Dock of the Big Shell and begins his first actual mission.

The infiltration of the Big Shell is his first actual mission. Metal Gear Solid 2: Substance is a tactical espionage game. The player must single-handedly infiltrate a facility under enemy control and carry out a covert operation. If an enemy soldier spots the player, the soldier will call his comrades and attack. Since the player doesn't stand much of a chance when vastly outnumbered in battle, unnecessary conflict should be avoided whenever possible.

Metal Gear Solid 2: Substance is composed of the original game, Metal Gear Solid 2: Sons of Liberty, a Missions pack featuring new stages to play, five additional playable stories called Snake Tales and a special skateboarding mode. Crouch Square Shoot Circle Punch Triangle Action Button R Bring up weapons menu L Auto-Aim L Bring up Item Menu Start Pause game Big Shell-Map Select This makes shooting at enemies far away a lot easier.

You can shoot handguns, assault rifles, use the HF Blade, and use fist attacks with the first person view. This will cause your character to grab the guard around the waist and toss him over his shoulder. On the easier difficulties this will knock out a guard. This will allow you to see things that are behind the corner without exposing yourself to the enemy. This will allow you to clear certain gaps, or move over metal grating quickly without making a lot of noise.

Then unequip your weapon if you have one equipped and press the weapon button. This will enable you to maneuver enemies to another area that is more secluded for purposes such as getting his Dog Tag. While he is in this chokehold he will squirm to try and get out of your grip.

You can make him stop for a moment if you quickly tap the weapon button again. When your character is hanging he will not be spotted by enemy sentries unless he is hanging right above them. You can shimmy in either direction while hanging on a ledge. Use the right analog stick, the directional pad, or the L2 and R2 buttons to move. The famous cardboard box gag from MGS. You can equip any box in your inventory and your character will duck inside it.

This will fool enemy sentries unless they see you move or they see you get into it. You can also alert their suspicion by using the box near boxes that are not of the same type for example using the box used for storing oranges in a pile of boxes that house electronic supplies. You can also move around in a cardboard box.

This is useful for dodging enemy fire, is a kind of ducked run. You can press the R1 button to press your face up against the small opening that serves as a handle to see what is going on outside the box. You can press the L2 and R2 buttons to look in either direction. Your character will now grab him around the neck to temporarily immobilize him. Once he is in the choke hold you can either snap his neck, knock him out, or leave him.

To snap his neck rapidly press the weapon button to snap his neck, you will hear a quite distinct, and disturbingly pleasing, "SNAP! You can use the right analog stick now to move the camera around to see more detail. This will allow you hide he hind corners and step out quickly to shoot in the First Person.

This will alert the guard and cause him to come and investigate. This will provide you with an open opportunity to sneak past him if you have utilized this correctly. This will knock him over for a moment and he will remain on the ground for a second, this will allow you to escape or attack him while he is recovering.

Lay the Book on the in or near the guards patrol route so that it will catch his attention. He will stop his patrol and start browsing through it. This will allow you to sneak past unnoticed. Quickly unequipping and then equipping will automatically reload your gun. You can use this during several boss battles and this is very useful for shootouts with attack teams. You can use this to get in a quick shot at a guard with the M9, as long as you get a hit you can wait until the drugs take effect.

And, there is no risk of him seeing you after you shoot him. You can throw grenade type weapons while peeking around the corner. If you did this correctly your character will say "Freeze! The guard will put up his arms and now he is under your control.

Press L1 to keep a bead on him while you move around to the front. Now you can go into FPV and wave your gun at his head or his groin, this will cause him to panic and he will now start to drop items for you, such as ammo, bandages, and dog tags.

Occasionally you will run into a guard that does not want to give up his dog tag. He will say something like, "If you're going to shoot then shoot. Now, this will frighten him and he will now give up items for you. On an additional note, do not unequip your weapon in front of a guard that has not been properly "disposed of".

Also, do not stand in front of him for too long without your gun pointed at him or he will do the same thing as a mentioned before. One more thing, if you try to hold up and Arsenal Tengu or an attack team guard they will mock surrender and then attack you very suddenly. Useful for escaping attack teams while at the same time killing them. You can lift and drop the body several times to obtain items and ammunition. You can also drag the body into a locker to hide it by picking up the body and dragging it somewhere.

On the tanker on the Aft Deck there are two gates that you can use to dump bodies out of. Look for an enclosed white lined box on by the edge of the ship. Simply drag the body into the box and Snake will open the gate and dump him into the water. By shaking an enemy in this manner you can also wake them up faster.

This is mainly useful in the Tanker where you have tranquilized a guard, or knocked him out in some way, and you did not get his dog tag. In the Plant you will have the coolant which does the same thing just a lot faster. When a guard has discovered you can shoot his radio while he is making the report or before he does, giving you more time to get away. You can only do this in FPV. This will cause its contents to spill out.

This can be used to temporarily blind enemies or expose those damned IR beams. Beware though. Your character will sneeze sometimes after this is done. The beams will appear as faint lines stretching in between the two terminals where the beams ocme out of.

Another way is to equip the Cigarettes, Snake or Raiden will start to smoke them and the smoke produced by he cigarette will slowly reveal the IR Beams. After they are unequipped the smoke will eventually disperse and the beams will be invisible once again. The last way you can see them is to find a fire extinguisher or a bag of flour that is near the beams and sire a shot into it. The contents of the fire extinguisher or the bag will explode and saturate the air, the beams will then become visible and stay visible for a long time after the air clears.

If you are close enough to the fire extinguisher or other source when the air becomes saturated your character will begin to sneeze at random moments, this can cause certain problems when you are trying to sneak past some guards, so use this carefully.

To climb onto it simply push the action button while in front of it and your character will climb onto it. Omnce at the top your character will climb off. To call up an elevator or to open the doors stand in front of the control panel of the elevator and push the action button. One each from the front-right, front-left, and center. You also need a close-up of the 'Marines' marking on the side of RAY. To take a picture equip the Camera through the Items menu and aim at the target. The faint green lines represent the border of the picture, get your photo in between those lines.

The workstation that will be used to transmit the photos is in the south east corner of the hold 3 where Metal Gear is housed. From there Otacon's special software will take over to confirm the quality of the photos. Equip Sensor A and go through each strut. When you see yellow stuff on the radar that means that the bomb is within that area.

The yellow stuff is the visual representation of the odor molecules given off by "Fatman's signature smell" that is on each C4 bomb. When the bomb is neutralized the yellow stuff will slowly fade away. To disarm the bomb, after you have found it, equip the coolant spray that Stillman gave you and aim at the bomb. The spray will freeze the detonator and the bomb will now be unable to explode. You will have to wait for her to get up if she has decided to sit down.

When she is holding on you will now be able to move around but only at a slow walk. Make sure that the area is clear of enemies before guiding her anywhere, she is significantly weaker than you and she will die quite soon if attacked. The Oxygen gauge represents how much oxygen is in his lungs. When it reaches zero his life bar will begin to decrease. When the life bar reaches zero you will die and it will be game over.

Unless the map has been downloaded from a node only the yellow areas will appear on the radar. Entering the yellow cone-shaped area while standing will cause the mine to explode. While in normal mode, enemy soldiers will follow set patrol routes. While on patrol enemies will use their eyes and ears to check for anything unusual on their patrol route. Some soldiers might stand guard in a single spot, while others might fall asleep on duty. When a soldier in normal mode hears a noise or spot something suspicious , such as footprints or bloodstains, he will leave his patrol route and search the surrounding area.

If he finds nothing out of the ordinary he will return to his patrol route. Creating a noise to distract an enemy is an essential technique to master. Alert Mode : The radar cannot be displayed. Occurs when the player is discovered by an enemy soldier, camera, or cypher. You will enter Evasion Mode when the gauge has reached zero. Evasion Mode : The radar cannot be displayed. You will enter Caution mode after the gauge reaches zero.

Caution Mode : Occurs when enemy soldiers are more vigilant than usual in their patrols. You will enter Normal Mode when the gauge is down to zero. Radar Jammed : Radar cannot be displayed when the radar signal is being jammed. Also, any equipment containing electronic components such as surveillance camera's and remote operated missiles will not function properly.

Furthermore, enemy soldiers will not be able to use their radios to call for backup. When you access them there will be a small loading screen while it loads the data to your radar network, and then it will show a screen in which you can change various options. You may recognize this menu, it is the Options menu that appears on the main menu of the game.

The nodes also act as in game settings changers. So , if there is a particular cutscene that you wish to view without blood you can access one of these before it happens and then proceed onwards. Now this is, obviously, just a list of the nodes and their locations.

I have indicated for each node where it is and in which part it first appears. This is just in case you are having trouble finding the nodes, which is unlikely, but then again I'm here to help you with just about everything about the game. Also, you don't have to look through the guide to find it, instead you can just come here for quick reference. In very easy you will not have to access the nodes, the radar is there no matter what, except when you are caught or it is jammed.

I guess that they just assumed that if you play on Very Easy you suck at the game. On all the other difficulties you must access them in order to get the radar and locate all of your enemies before you run around a corner right into one of them. You don't have to have radar of course, if you prefer the old fashioned sneaking around style, peeking around corners, using distractions and such.

If the life gauge drops to zero the game ends. When there is little life left the character will begin to bleed, at which point the gauge will turn orange. Crouch to stop bleeding. Radar : Appears in top-right. Displays the position of enemy soldiers and their FOV, and the players surroundings. Weapon Equipped : Appears in bottom-right. Displays the weapon that is currently equipped. Not displayed if there is no weapon equipped. Item Equipped : Appears in the lower-left.

Displays the current item equipped. When the gauge reaches zero the boss battle will end. Boss Stun Gauge : When a boss is hit with a tranquilizer or punching attack this bar will decrease, when this reaches zero the battle will end. Grip Gauge : While in hanging mode this will appear.

Appears below character's life bar. When this gauge reaches zero your character will lose grip and fall. You can increase the duration of hanging by doing pull-ups, or 10 hang-drops. There are 3 levels of grip.

Oxygen Gauge : When underwater this bar shows the amount of air left in the character's lungs. When the gauge reaches zero the life bar will begin to decrease gradually. The rate at which the oxygen gauge decreases can be slowed by pressing the action button. Partner Life Gauge : When working with a partner in certain circumstances this will appear.

When the gauge reaches zero the character will die and that will result in a game over. Just as if you had died. Sleeping : Sometimes you will come across a guard that has been dozing off on duty. You will see blue Z's circling over his head.

They will also appear when you have tranquilized a guard. The more Z's a guard has over his head the longer he will stay asleep. Unconscious : When you have knocked out an enemy with your fists he will lie on the ground with yellow start circling around his head.

The more stars there are the longer it will be until he comes around. Suspicious : When a guard has heard or seen something that has alerted his suspicion a big "? This appears when he has seen something just out of his vision cone or heard a noise such as a magazine being thrown or you knocking on the wall.

Discovery : When a guard has discovered your character a big "! This also appears when a guard has discovered dead body or an unconscious comrade. All action is paused when using the codec. Useful for contacting other characters for helpful information. To use the codec first adjust the frequency with he directional pad or the right analog stick. You can also chose a recipient of a codec call by pressing the right analog stick down or down on the directional pad.

This will display a list of all the known contacts on your codec. Press the circle button to make the call or push up on the analog stick or the directional pad. You can exit the codec by pushing select again or pressing X after the call. You can advance the conversation with several buttons.

Press Triangle, X, or Circle to stop the conversation and then press Triangle again to automatically fast forward to the end. Press Square to fast forward through the conversation or X to go one line at a time. Otacon-Save : Rose : X : Stillman : Ames : Johnson : Instructs you to find Emma Emmerich. Emma : Olga : Push the sticks in to zoom in. Push R1 and R2 to hear your characters thoughts.

This can be quite funny at times. Lockers are useful for stuffing mmmm If you want to keep other guards from finding out about guards that you riddled with bullets you can hide them, conveniently, in a locker. Drag them into a nearby open duh!

Lockers will almost always contain an item or ammo. You can kick and punch the doors off of the lockers if you feel in the mood, or find that one is locked. The hinges will break and the door will fall onto your character if you haven't moved out of the way. Don't worry this won't reduce your health.

Shooting at the hinges also works. If you are a pinch for a good hiding spot i. You will most often be safe in a locker because guards will rarely search them. Pressing L2 and R2 will allow you to look out both sides of the lockers vent. Use R1 to lean up against the vent and look out. However, do not do this quickly when a search team is in the area or a guard is patrolling nearby, your character will bang his head on the vent alerting any nearby guards.

If you need a good laugh you can enter the locker room on Deck-A of the tanker. Open the locker and look at the girly poster on the inside in FPV. Call Otacon and see what happens. If you get a message from Otacon about the correct utilization of lockers then you may be standing too close or too far. When your character has a cold he will sneeze periodically and unexpectedly. This may alert nearby guards. Which is always bad. To combat this your character must find Medicine.

Do NOT let me catch you playing this, unless you are trying to get dog tags! Anyway, the name speaks for itself. Normal : Guards are fairly intelligent; bosses take about as much damage as they do to you. Guards can't hear very well or see for that matter.

There is a good scattering of guards throughout the levels. Hard : Harder than normal, get it? You have a small chance of making it out of a shootout alive if you get caught. Extreme : If you can finish this difficulty without making a hole in your wall then good job. Bosses are 2 hit kills, if you are caught. Guards see about twice as far as in normal and can hear very well. European Extreme : curses wildly!

Guards see almost by human standards, have amazingly good hearing, and bosses are one hit kill. You better have your stealth skills down pat, or else you are in for a good deal of game over screens, and maybe a few broken controllers. As the game manual says, "for the truly possessed". All difficulty levels will become available after completing the main game once. That means BOTH chapters. Also, with the assault rifle, when you lightly press the weapon button the gun will be drawn but the gun will not fire, when you press down fully, then the gun will fire.

While in FPV the shoulder buttons that allow you to strafe and peek will move your character more depending on how hard you push down on the shoulder buttons. This is very useful for gun duels such as the Olga battle. I'm not sure if this counts but the analog sticks can control how fast you move your character.

If you slightly push on the stick your character will slowly walk. When you push it to full tilt the character will break into a quick run. Use the saved frequencies to contact the right person for saving, Tanker-Otacon, Plant-Rose. Select which memory card, and choose the file you want to save in. When you save in a certain area, when you load it you will have the same equipment, ammo, and items as when you entered. If you want to keep what you have gathered in a room then exit the area and then renter and then save.

Then just follow normal saving procedure. Outside of this there is no other way to save in the Snake Tales besides after you complete it. You will be able to play as either Raiden or Snake. Some differences will occur when playing, such as when battling Solidus as Snake you will only be able to use your fists, since Snake can't use a sword.

You will be able to choose three difficulty levels. Easy, Normal, and Hard. The battles are The cutscenes that are available are Snake infiltrates tanker 2. Snake watches the soldiers descend from the Kasatka 3. Snake sticks up Olga 4.

Vamp vs. SEALS 6. Fortune vs. SEALS 7. Solidus vs. They will give you a great idea of what happened there if you haven't played Metal Gear Solid. When you finish one the next one will appear. Is a short overview of the book. Explains quite thoroughly the plot of Metal Gear Solid.

Radar Choose between the three radar modes. Blood Toggle blood appearance on and off. Sound Choose between stereo and monaural sound. Dolby Digital 5. Pretty sweet. Caption Toggle the captions that appear during cut-scenes and during the game on or off. Own-view If you prefer you can reverse the controller directions from the default directions.

Item window Have your Items and Weapons appear in linear form as in MGS or in groupings, which was the preset option in MGS2 Quick change When you do a quick change you can now have it unequip the item or switch with the last equipped one. Screen adjustment Adjust the game screen so that it is centered in your TV screen. Brightness adjustment Adjust the brightness of the game. All of these options can be changed during gameplay by accessing a node during the Plant chapter.

In this walkthrough I will provide for you all of the necessary information related to each and every area. I won't be telling you to pick up ammo and stuff like that, I'm sure you capable of handling that yourself I have been wrong about stuff like that before. The story thus far Following the Shadow Moses incident Ocelot leaked the top-secret specifications for Metal Gear to the public, via the black market.

With he help of his new accomplice, Otacon aka Hal Emmerich, a scientist from Shadow Moses , he infiltrates the transport for the new Metal Gear. The first thing you want to do is take notice of a few basic facts that will come into play in this area. First, it's raining. This means several things. One, Snake can catch a cold if he stays out too long. The length of time for this varies between difficulty levels so stay on your toes. Second, the rain will camouflage some of the noises that would otherwise alert nearby guards.

This means you can be a little more careless with no further consequences. Third, once you go inside you will track water around in your footsteps. We'll deal with that last point later. On either side of this area on the bottom level is a white box. If you want to you can knock out a guard and drag his body into this box. This will trigger a scene where Snake opens the gate and tosses the body into the river. This is useful for disposing of unsightly bodies and effectively covers up all evidence of your passing through the area.

Should the area be put on Alert Mode, the bodies will not be discovered. By now, I imagine you're wondering if I'm going to explain how to get through this area. Indeed, that's what I'm going to do right now. The first thing you want to do is head northeast. There is a guard patrolling up ahead at the top of a flight of stairs. Mosey on over there and halt midway up the stairs. The camera gives you a good view of the guard so wait until he is facing away from you until you make your move.

Once that guard has been taken care of, move up the next flight of stairs and onto the topmost level. This upper balcony has a single guard who patrols the perimeter of it. I suggest that you keep an eye on the radar and watch for where he is coming. You can hold him up as he is about to turn north towards your position. Dispose of him as you see fit.

Now follow this balcony until you come to the opposite set of stairs on the west side. Head down and press up against the wall at the top of the stairs. Keep an eye on this guard. If you want to hold him up, wait until he heads south and then run up behind him. If you want to head directly into the next area just wait until he moves away from the stairs then descend and open up the door using the action button Otacon will call and inform you of how to do this.

There are four doors total in this area: 1 Port level 1 : Opens into Deck A, crew's quarters 2 Port level 2 : Locked 3 Starboard level 1 : Locked 4 Starboard level 2 : Opens into Deck B, crew's quarters If, by chance, you chose to enter on the Starboard level 2 door then pick up the walkthrough at Deck B, crew's quarters. This locker room contains a few doodads that you might enjoy. A close inspection of the corridor will reveal pipes lining the ceiling and extending through the floor.

You can exploit these pipes by shooting them to release their contents steam and thus distracting the area's guard. Doing this is entirely optional. This area's guard patrols on the east side so for now worry about the fact that you just came in from the rain. Snake may or may not have a cold. If he does he'll be sneezing occasionally.

The downside of Snake sneezing and pretty much the only side is that he'll probably wind up sneezing as you're approaching a guard, promptly giving you away. Beware of this possibility. Additionally, you'll want to shake the rain off of Snake by tapping the X button a few times. By doing this, Snake will leave fewer wet footsteps that could give you away. Now you'll want to head south through the corridor since the door closest to you is broken try to open it for a laugh.

If you want to, head into the locker room but otherwise keep your focus on the guard in the eastern part of the corridor. He patrols north-south so stay hidden until he turns north then knock him out and pass through the door in the north. Open the locker and look at the poster in First Person View. Now call Otacon and Snake will be playing with his A main hall with two rooms abutting the east and west ends each containing a staircase, the west staircase has a sleeping guard at the bottom , the main room contains a stairway in the south and a lounge in the north with a bar, tables, a magazine rack, and a small lounge area.

This area contains a few neat odds and ends that I will go over later. For now you can focus on the guard that Snake peeked at in that short cutscene. You have a few options here. First, you can watch the two guards until their patrols allow you an opening to pass through. Your goal is the staircase in the center of the main room. If you don't want to sneak past, your best bet is to simply use jump-out shots when the time is appropriate to knock the guards out.

Either way, make your way to the staircase and enter the door on the right; the door on the left is broken. Another neat thing is the bottles. You can shoot these bottles and watch their contents spill out all over the place. This has little practical use aside from a potential guard distraction if you shoot the bottles while he's near the bar. Talk about detail! In FPV look at the mini-bar itself and you should see an ice bucket on the counter. Shoot it so that its contents spill out, ice. Now, look at the ice for a while and eventually you will see it melt.

Also, the larger pieces of ice will take longer to melt than the smaller ones. Not many games have that kind of detail programmed into them for good reason probably. On the west side of this room there is a small TV screen showing views of the new Metal Gear prototype.

If you shoot it will go blank. A small preview for what you'll be seeing up close later on. When you shoot the magazines on the magazine rack they will explode in a shower of paper shreds. You can continue doing this until the pieces are too small to shoot. Shooting these magazines will also attract the attention of the guards nearby. If you utilize this appropriately you can create an opening to slip through.

Try firing a bullet through the plants in the room and the leaves will shake a little bit. They shake more if you shoot the stems. When you fire bullets into the glass in the middle of the room it will make a small crack in the glass. Fire about 15 shots into the glass and it will completely shatter, showering shards of glass all over the floor. The guard at the bottom of the stairs in the west has flies circling his head, if you stay there for a moment the flies will suddenly become attracted to you, the flies can alert guards to your presence so be careful.

You start on the east side near the middle. You can go either north or south. The small area to the east of your starting point contains a staircase to nowhere and a hatch that leads back onto the aft deck. Going south is the easier of the two options in my opinion presented to you.

Immediately head south and pause as you are about to turn the corner into the east-west portion of the hallway. There is a single guard here. I suggest proceeding one of two ways. When he passes you again run west and then keep going through the hallway until you are able to turn into the next stairwell. Head up the stairs on the west side of the area to the next floor. Please note that if you go north the lights will cast revealing shadows on the ground.

You'll have to take this into account when approaching the guard that patrols the northern part of the area. It's also harder to sneak past this guy, you have to hold him up or knock him out in order to make it through to the other side. Take note of their discussion and think of how you are going to get past the camera.

This area is just a long corridor with two staircases on either side, with the east one blocked by crates. There is a staircase in the middle that goes to Deck D. The only security precautions in this hallway for now is the security camera mounted on the wall, near the ceiling. There are no guards here at the moment but there will be later on in the game, when you come back through here.

All you have to do is watch the camera and sidestep under and past it when you won't be in its field of vision. If you picked up Chaff grenades earlier you can use those here as well. Deck D has a kitchen in the west, a pantry in the east, and a staircase going up to Deck E in the south. There is a hallway that extends from the kitchen to the pantry and wraps around the staircase to the other entryway to the kitchen.

The kitchen is accessed by two sliding doors, one by the staircase that you came up and the other to the south of it, by the staircase to Deck E. The kitchen is monitored by a surveillance camera and two guards, they will patrol in between the tables in a circular pattern you can watch on the radar to see what I mean. In the food pantry there is currently no guard but that will change eventually. The security camera, for now, cannot be destroyed. You can use either a Chaff grenade or stealth to work around it.

The guard that patrols the northern corridor mainly patrols back and forth he will venture into the kitchen eventually he is the second guard. In the southern corridor, where the stairs are, a single guard patrols east west. Your main worry in this area is the hallway that connects the north and south halls. If you look closely you will see two vertical, black strips on the wall. Do you know what they are? Well, if you get close enough to them Otacon will call Snake and explain it to him.

Before you do so, I suggest knocking out the two guards in these hallways to keep your worries to a minimum. Break the beams and you'll be seeing that delightful game over screen. Use either the Cigarettes or the fire extinguisher to reveal the beams. Look near the bottom, no beams. You can crawl past this obstacle.

Once you are past the beams, head up the stairs to the bridge. If you enter the pantry in the eastern part of the area this will trigger a guard coming into the pantry. Stay in the back once you enter the pantry and wait for him to enter. He'll doze off shortly after entering the room so you shouldn't have much trouble getting by him. If you experiment enough you may be able to create a little tune. The pantry may be of some enjoyment. The boxes there contain an amount of fruit and there are some bags of flour.

Shooting these items will break them apart and such. I've always found it fun to have a gunfight in this room once I get the USP. Right now there are only a few things of note, the hatch on the left side and a Ration right next to it. You'll have to use the hatch on the right because the hatch on the right has been disabled for now. Head outside through the hatch. At the start shoot out the tarp strings and the spotlight, this will make things easier later.

Olga will spray bullets at you with her USP and occasionally throw a grenade. The prime time to get a shot off are when she is preparing to throw a grenade, or when she is reloading. An easy way to predict when she will need to reload is to count her shots, when she gets to 15 she will have to reload. After shooting out the light and the tarp go to the right side, behind the boxes. I suggest that you get into first person mode to shoot and use the strafing buttons. When you see her peek out from behind the crate to shoot, try to get a shot off.

You will see her stand up to throw a grenade, when she does this is a good time to get a head shot in to take away a large chunk of her stun bar. You know when she is going to throw a grenade when you hear her say, "Take this! If she does get behind the tarp, on the right, there is a little space between the boxes that you are hiding behind.

Lie down there and aim through there. You should be able to shoot her from there. Also you can wait for the tarp to flap out from in front of here so that you can see where she is and then get a shot off in that location. If you don't feel like going the hard way then I suggest shooting the ropes that are holding it to the rack and it will fly off and be of no more trouble.

Doing this will prevent a lot of frustration. If she shines the light at you, you should be able to knock out the spotlight by hitting it with an M9 bullet. If you can't do that crouch down by the low row of boxes and peek up quickly you should be able to see her briefly before the light distorts your view.

She should be almost directly in front of you so if you miss by a little you should see an exclamation point where she is, aim at that spot. The are also a few ways to distract or confuse Olga. The first is to knock on the boxes. This will attract her attention to that spot and you can move somewhere else to try and get a shot off before she notices you.

You can also let her catch a glimpse of you in one spot then you can crawl to the other side of the area and try to shoot her from there before she realizes where you are. Now you can have at least a fighting chance against any attack teams that you run across. Use it only in very secluded places or in places where there is a lot of background noise.

If you are going for dog tag collection be very careful with who you kill. If you kill a guard who's tag you have not gotten yet then you will be unable to retrieve his tag until you play through the game another time. One good thing that you get with the USP is a flashlight that comes on whenever you draw the gun. This obviously allows you to see in dark places and illuminate places which need lighting.

Of course, when you draw the USP near an enemy it the light will attract his attention. So make sure that you have the guard in a hold up or something like that when you draw it. Otherwise, be prepared to duke it out. It is raining again and once again Snake can catch a cold, so don't stay out too long. There is a place on the opposite side of this area almost identical to the one you fought Olga in. The middle part of this area is raised up about 20 feet above the surrounding area.

On top of this is the crow's nest, there is a ladder here so that Snake can climb up to the top. The area on the left has a few items of interest This item may be useful but it is unlikely considering that it can only be used effectively outside. If you think about it, using a wet box to hide inside while you are indoors will arouse suspicion.

The dry box would be a better bet. Considering that this is the last time you'll be traveling outside Pick it up if you like. Either way, head inside to the bridge through the hatches on either side of the bridge.

Also, if you call Otacon while looking at Olga Snake will mention near the end of their little conversation that he has "had enough of tomboys. You can find it in the Previous Story section of the Special menu. There's little else to do here.

Everything else is the same as last time. The easiest way to approach this is as follows: knock on the wall next to the stairwell on the right and wait inside the stairwell, hidden from sight, for the guard to come around and inspect the noise.

When he does you can deal with him however you want. You can head through the kitchen if you like. The camera can easily be taken out with the USP, just make sure no one is around or conscious when you do so. The guard who enters the pantry after you enter I mentioned him earlier is one of those "tough guy" guards who won't give up his dog tag without further prodding in the form of a bullet to the kneecap.

Now would be the opportune moment to grab his dog tag because you won't be coming through here again. He simply patrols back and forth from one end of the corridor to the other. Not very complicated. The security camera is still the same as last time you were here. Wait against the wall on the side where the locker is and watch the guard coming from the part where the camera is.

When he passes you he will be wide open, do whatever you want with him now. You can bypass the camera by using the USP to knock it out, Chaff grenades to disable it or by shuffling underneath it. Head down to Deck B. As with last time, head south until you can press up against the wall and watch the guard in the south do his patrol. You can choose to sneak by him by waiting for him to walk away and then hiding in the niche by the locker or you can wait for him to turn around and then gold him up.

The door that leads to the crew's lounge on Deck A is your goal. Walk a bit down the stairs until you have at least a partially unobstructed view of the magazine rack. Take aim at the rack and fire a bullet into it. The magazine you hit will explode into shreds of paper and this should attract the attention of both guards in the room. Use this opportunity to slip by them and head down the stairwell in the eastern part of the area.

Basically I split it in half, you should be able to figure out where I did it. This is the starboard half of the Engine Room right side but you know that because that's the only way you could have come in. What you should know right off the bat is that it is pretty safe to use the USP in just about any place in the Engine Room since the noise created by the engines of the ship creates a lot of background noise to muffle your shots.

Feel free to use the USP so long as you're not standing directly next to a guard. This room is mainly comprised of catwalks running around the open spaces around the engines so there is a lot of maneuvering room available if you have to fight off an attack team or slip past a guard. You can hang off a lot of the catwalks as well. Feel free to use that fact to your advantage. When you first enter the Engine Room and go north there will be a short little cut-scene where Snake sees Vulcan Raven's Shadow on the wall.

Snake thinks it may be a hint of some sort and believes that he will have to battle him that is if you call up Otacon on the Codec after the scene. If you go north a little bit more then you will see that the shadow is just created by a little Raven doll in front of a flashlight. Nothing to worry about. Search the lockers if you want and then make your way out into the engine room.

The first guard who is standing opposite the door can be approached several ways. You won't be able to get his dog tag though so think before you act. He doesn't really move around too much so you can proceed to the catwalk below and deal with that guard.

This next guard can also be dealt with in a variety of ways. When he does drop down onto his head to knock him out. Be sure to tranquilize him after you knock him out or else when he wakes up he'll send the area into caution mode. Alternatively, you could hold him up as he walks away.

The guard on the bottom level should be easy to deal with. You can tranquilize him when he's not looking or you can hold him up when he turns away. Or you can also ignore him and run by when he looks away from the catwalk to need to proceed on to the next part of the engine room. You want to head for the stairs to the west and somewhat south of where this last guard is.

A warning must however be given. Only a small number of the texts published in this book comes fromnative English speakers. The editors asked authors to contract professional revision, but it is not certain thateveryone did it and it is impossible to warrant the quality of the services contracted by diverse authors.

Notwithstanding thiseffort, it would be impracticable to standardize the entire book in terms of language style and to correct it interms of grammar and syntax. For these reasons, this eBook must be read with some tolerance by those to whomsome mistakes may sound ungentle. We hope that in view of the greater goal such obstacles can be easilyovercome.

Oliveira, M. Wewill start with the notion that radio is going through changes that affect its expressiveness,programming, routines and values. Accordingly, we intend to characterize the uses that Portugueseradio news stations are making of social networks such as Facebook and Twitter. The article will begin by reflecting on these changes in radio and the implications for radiojournalism. One of the most important conclusions is that social networks are used as an excellent platform topromote user participation.

This finding was especially verified in the case of Facebook, wherelisteners frequently post comments, criticize radio policies and suggest news topics. Keywords: radio, journalism, social networks, PortugalIntroductionNews radio is still important, but not as it was before. Today, a consumer looking for news has access to awide range of subjects, formats and channels whether they be television, printed media or the Internet websites,social networks, and blogs.

We access news in multiple formats and from multiple platforms. Radio all over the world is a do pting various forms of multimedia convergence in order to deal with thisnew multiplatform media environment. News radio news is present on the Internet, on cell phones and, in recentyears, has started to make use of the social networks. On the other hand, journalism is seeking new ways and formats to adapt to new technologies and thuscontinue to position itself as a win do w to the world, despite the emergence of other forms of journalism, such ascitizen journalism.

In the digital age, news consumption has changed, and has become a social experience: people share thenews, instead of just receiving them, as they did before. The question is: What is the role of the news radio today? In this paper we intend to analyze the relationship between the Portuguese news radio and the socialnetworks, especially Facebook and Twitter. Starting from the perspective that nowadays radio is a multiplatform medium, our aim is to understandand reflect how the Portuguese news radio is using social networks to disseminate its journalistic content.

The study examines the use of multimedia tools, the presence of comments, the user generated contentand the hypertextuality used in messages on Facebook and Twitter. Nowadays, news radio is one part of the overpopulated media environment. Besides radio, a personlooking for news has many other alternatives, especially on the Internet. Scott, Traditional media which, of course, includes radio, is facing competition from a broad range of otherplatforms that have started to disseminate information.

Traditional news organizations exist in a scenario whereother forms of producing news are operating simultaneously. This competition includes not only news production,but also business models. Indeed, alongside these models comes a huge volume of non-profit and noncommercialenterprises sharing the media landscape with traditional media. In this context, the challenge for radio news stations is to adapt to a new environment, creating policies inorder to maintain their credibility over and above their own legacy.

Since the beginning, radio has always demonstrated an ability to adapt successfully to other changes incommunications technology, but the challenges have never been as huge as they are nowadays. David Hendy makes the point that, traditionally, radio is a time-based medium since its programs emergein a linear flow of time In one way Hendy is right, because this model of radio still exists, but thechallenges imposed by the digital era have created a scenario where it exists simultaneously alongside a newradio.

More interactive, multi-faceted and whose programs do not only exist in the flow of time. Never before has radio been forced to change its expressiveness, becoming a multimedia platform. Soundis no longer the only resource for radio. Also, radio has never been as interactive as it is today, and has never beenso bombarded with requests to make the news available in so many platforms. In the digital era, radio must find the balance between these new characteristics and the need to maintainthe old ones so it continues to be a significant source for listeners in terms of breaking news and public issues.

One of the keys for the future of radio journalism might be in the way that radio will take advantage of itsknowledge regarding immediacy and credibility, which are two very significant attributes constantly sought bythose who search for news on the Internet.

New platforms, and especially the Internet, provide radio with more space not time and this can be anopportunity for more creativeness and alternative ways of covering public issues. Also, radio has become moreinteractive, since the Internet offers more opportunity for debate and user participation about public affairs.

Finally, radio on the Internet surpasses the limits of the traditional broadcast, reaching listeners all over the worldwhich means a challenge for news reporters. The idea is that despite the many and ongoing changes in the ways that people access information, radio isstill an important win do w to the world. In the most competitive media environment ever, radio must a do ptpolicies to enable itself to be an important source of news for people.

In this context, the presence among thenew media platforms is seen as a logical strategy. Accordingly, radio is requested to a do pt strategies in order tofacilitate portable consumption and audience participation in its news content. Thus, social networks are,nowadays, advantageous platforms for the radio news stations. Social networks and journalismShayne Bowman and Chris Willis wrote in their book We Media - How audiences are shaping the future ofnews and information that the profession of journalism finds itself at a rare moment in history.

Indeed, journalism in the digital era has changed, and one of the most important changes stems from theopportunity that is provided by the new platforms which allow consumers the possibility of participating in thenews content. This can be do ne in several ways, by creating weblogs, websites or, most recently, by using thesocial networks. Thus, when we are talking about the relationship between journalism and social networks, participatoryjournalism is frequently highlighted as a benefit.

News consumers can comment on the topics, suggest new approaches or add facts about an eventreported by journalists. The presence of journalism in social networks increases, at least in theory, the possibility ofmore diversity and pluralism in news production once people inside and outside the newsroom are engaged inthe communicating process.

Nowadays, journalism is a blend of producing and consuming information. Social networks contribute to what Dan Gillmor defines as a conversation, which means that peopleand journalists share comments and discuss public affairs. Another characteristic frequently mentioned when we are talking about journalism and social networks isthe speed with which it is possible to distribute information.

Twitter is often mentioned as a useful platform fordisseminating news in a short period of time, and this is seen by journalists as a good way to reach the consumer. It led as the top news story on Twitter for seven weeks in a row — a feat not reached by anyother news story on any of the platforms studied. Consuming news is, at present time, a social experience. Another study revealed that the presence of journalism in social networks leads to a new journalisticdiscourse.

The study looked at the Spanish reality and aimed to understand the relationship between journalismand Facebook. A new form of information to which the media arenot used to" Garcia Alvarez et. Traditional journalism is still themain source of news stories but, as they are on social networks, users stress the importance of a news topic byhighlighting it to other users.

Portuguese radio news and InternetIn Portugal, the last two decades have shown a decrease of investment in news radio. The Portuguese scene is characterized by hundreds of local radio stations, most of them music radiostations, and by other national radio stations which are, in the majority, also music radio stations.

With respect to news radio stations, the Portuguese scene is very small. Antena 1 is the main radio channel of the Portuguese Public Service which also includes six other radiostations. Antena 1, at the time called Emissora Nacional, was the voice of the government during the Portuguesedictatorial regime, which ended in Today, Antena 1 has the biggest radio newsroom in Portugal and itsprogramming includes much journalistic content. TSF is the youngest radio station among the three analyzed in this study.

TSF is an all-news radio and introduced a new style into the Portuguese radiojournalism scene, based on live reporting and breaking news. These three radio stations are, indeed, the most relevant in the context of Portuguese radio news, sincethey dedicate a high percentage of their programming to journalistic content. Also, they represent the biggestnewsrooms among the Portuguese radio news stations.

Despite the center of news production in these three stations still being the traditional radio, the efforts toimprove their presence in various platforms are evident. TSF was the first of the Portuguese media companies, including newspapers and TV, to offer podcasts onits website. The strategy of TSF on the Internet aims to maintain the same imageacquired through its traditional version. Therefore, the website contains news contents, programs and many otherjournalistic formats.

TSF highlights user participation by providing tools that allow for commenting on the newscontent. The website has been inundated in recentyears, especially since , by several videos. The Portuguese Public radio service a do pted a different policy for its presence on the Internet.

Sharing and retweeting sounds — the relationship between radio journalism and social networksthese two mediums. The result is not as positive for the radio section because the most important news contentsavailable on the website are produced by public TV. Despite being a public service, the website do es not promoteuser participation. Radio news and social networksAntony Mayfield in his e-book What is social media?

The Portuguese scholar, Gustavo Car do so 37 , advocates that radio is the medium that competesmore easily with social networks. The author says that radio, like social networks, contributes to the discovery ofnew talent in music.

To do that, suggests Car do so, radio stations should increase their strategies on proximity tothe listeners by creating, for example, communities. Since then,Facebook and Twitter have become a part of the Portuguese radio news stations. In this case, the objective is to suggest the programming of the traditional radio, forexample, by making available the on-air programming schedule. This has a very curious effect, as the users start to comment on the topic, anticipating the debateeven before it goes on the air.

However, the main use that is given to Facebook and Twitter is to link news topics that take the user to thewebsite. Firstly, as a way to promote the debate and criticize the radio station. TSF was accused by the listeners ofhaving promoted the Prime-Minister since those who called in to the program only did compliments. The listenersconsidered that TSF had not do ne its job well and started to criticize the radio station on Facebook. Secondly, as a way to gather news sources by posting messages in which reporters request informationabout a news story they are working on.

In thiscase, the profile of the radio journalists turns out to be part of the radio station. The study addresses the following research questions:Q1: What uses are being made of the social networks by the Portuguese radio news stations? Q2: Do social networks contribute to the increase of multimedia expressiveness on radio?

Q3: How can we define the presence of the Portuguese radio news stations on the social networks? To answer these questions, the study looked at the following aspects analyzing the news topics availableon the Facebook and Twitter accounts: multimediality presence of videos, photos, sounds , interactivity whichtools are used to promote interactivity , the circuit of news contents where do the links take us and usergeneratedcontent do the users participate with comments and the journalists take part in this conversation.

Q1: What uses are being made of the social networks by the Portuguese radio news stations? Facebook andTwitter help to promote the spontaneity of the user and to break do wn barriers to access. Users do not need apassword or a registration to participate. This helps to explain the reason why topics that are available onFacebook receive so many comments, compared with the news available on the websites. Portuguese radio news stations receive many comments from users on news topics posted on Facebookwhich means a renewal of interactivity on the radio.

Indeed, radio has always been interactive, but never has itbeen so easy for listeners to participate in the news content. Users do comment and do suggest, but the only conversation isamong the users. Facebook represents an important platform for spreading user participation in the public sphere. However,it is interesting to note that not all the comments posted by users are contributing to a positive discussion aboutpublic affairs.

Indeed, in some cases, users use Facebook to criticize and offend without any argumentativeposition. Is this true? TSF Radio — Yes, we know. It was reported by Benfica TV and we are checking the facts to report thattoo. Ilive about meters from the works and it is now 01h30 in the morning and right now, as in thepast two days, a jackhammer is working. Facebook, TSF, November, 4th, In this particularly case the user felt he could talk about a situation he was going through.

People seeFacebook as a platform to get in touch with the radio station and to turn personal problems into public affairs. Sharing and retweeting sounds — the relationship between radio journalism and social networksAllowing user participation is, in fact, one of the most important benefits that social networks offerjournalism.

Commenting on the news on Facebook has started to be common practice among users, and because ofthat radio news stations have created rules about participation. For example, TSF explain that comments containing insults or obscene and defamatory language areremoved and Antena 1 wrote on Facebook that users who make offensive comments, appealing to verbal orphysical violence or provoke discussions are removed from the page.

Social networks, especially Facebook, are seen as an important win do w to opening radio to interactivityand participation by promoting the discussion of public affairs. In some cases, social networks appear to be theonly way to do that. For example, the Portuguese public radio service still do es not provide any other accesspoints on digital platforms for user participation. Q2: Do social networks contribute to the increase in multimedia expressiveness on radio? Radio has become a multimedia medium and because of this one of the most important questions aboutthe future of radio is to understand what the role of sound will be in a multiplatform radio.

On social network sites, sound takes second place. On Twitter and Facebook it is possible to post sounds,but this is a rare practice among the Portuguese radio news stations as the norm is to post sounds linked to theaffiliated website instead of posting the sound on the profile. Our observations also revealed that social networks are used by the Portuguese radio news stations to postphotos about the stories reported by the journalists.

Indeed, we found that posting photos is one of the mostintensive practices do ne by radio news stations on Facebook. On Facebook, photo galleries are frequently posted which are not on theaffiliated website. Posting videos is not very common practice. Rarely do news topics posted on Facebook contain videos. Regarding multimedia expressiveness, our observations revealed that the most common practice is theposting of text and photos.

Sounds and videos are very rare. According to our study, social networks are a complement to the on-air radio. The center of productionremains the traditional radio. Journalists think first about broadcasting news. Only after this task is completed,news are first posted on the affiliated website and finally on Facebook and Twitter. This means that the newsroomis still organized according to the traditional routines and values of radio journalism.

This scenario is the key to understanding how radio news stations are using social networks. Facebook and Twitter are mostly used by Portuguese radio news stations as a way to suggest and promotenews available on their affiliated website.

The circuit of the news posted on the social networks is as shown in thefollowing diagram:ECREA: Radio Evolution: technology, contents, audiences — conference Figure 1 — Circuit of newsOur observations revealed that the news go on the air, then go to the website and finally are posted on thesocial networks. Finally, the link on Facebook or Twitter takes the user back to the website. Twitter is only used to promote the news available on the websites.

In short, the breaking news pointed outby some authors as an advantage for journalism is not a common practice among the Portuguese radio newsstations. Twitter is only used as a platform to promote the radio, following the basic idea that if people are onTwitter, the radio should be there too. Also, through the social networks, listeners can participate in the phone-in programs.

Both give the listener the opportunity to leave acomment about an issue which will be read by the journalist on the air. Good morning. This morning, TSF Forum talks about corruption. Bureaucracy is seen as a fertile ground forcorruption and this is one of ten themes of the challenge that TSF has released to the leaders of major parties. Wewanted to know what actions they propose to reduce bureaucracy and make the fight against corruption moreeffective.

The Forum also wants to hear your opinion. What can be do ne to stop the phenomenon of corruption? Doyou believe that existing laws are sufficient? Facebook, TSF, 3rd June, In this way, despite the comment being available on the Facebook profile, the host of the phone-inprogram can choose the ones he considers more relevant to the discussion and read them on the air.

Facebook, inthis case, acts as an extension of the on-air programming and also contributes to increasing interactivity with thelisteners. ConclusionSocial networks did not invent interactivity on the radio, but they have transformed it. In fact, socialnetworks represent a strategy to open the radio to its listeners. This is the most important conclusion we havereached in this study. Facebook is a very useful platform for user participation. Taking advantage of its spontaneity, users caneasily get in contact with the radio station and with the journalists and participate in the discussion of publicaffairs.

Thus, Facebook contributes to increase the values of democracy in the sense that the presence of morevoices, at least in theory, represents more diversity and pluralism. Traditional radio, with its temporal constraints, isprevented from opening its programming to constant participation. The exceptions are the phone-in programs. With the Internet, and particularly with social networks, radio opens itself to discussion and criticism. Listeners can, as we saw, participate in the news content, but they also can also criticize the radio and suggest newstrategies.

Sharing and retweeting sounds — the relationship between radio journalism and social networksDespite this openness, we have observed that the discussion still remains incomplete, as journalists are stillreluctant to get involved in the conversation. The fact that the Portuguese newsrooms are becoming increasinglysmall can also be an argument in understanding why journalists do not take part in this discussion.

Journalists are constantly occupied with the tasks of news production. Our study also revealed a reduced use of some of the potential of social networks. Social networks are part of the global radio news station policy, but in these early years, despite being anexcellent platform to increase interactivity, Facebook and Twitter are just used to promote the traditional radio. Indeed, we have observed that although the three main Portuguese radio news stations are in the social networks,much of the content available in their Facebook and Twitter pages is simply repurposed material from offlineprogramming.

ReferencesAlves, Rosental Tese de do utoramento. In Car do so, Gustavo et. Quimera: Lisboa. Fenton, Natalie New Media, Old News. How deeply have such changes affected the relationshipbetween radio producers and listeners? The paper will analyse the case of three Italian radioprogrammes which have developed an intense interaction with their listeners through Facebook. We will conduct a quantitative observation of the social media activity of these programmes overan entire week and show the different social media strategies implemented by the selectedprogrammes, as well as their different dramaturgical relations with the listeners.

Inthat famous book, Radio. A flight notfrom images themselves, but from the mechanisms of visual perception. Radio listening still maintains some elements of blindness, but the way in which we now experience this medium isno longer totally disembodied and immaterial. Technological implants on the body of the radio medium, from the telephone to SNS, can also be read as acoming-of-age novel, in which we follow the main character, the public, through the various stages of its growthand development.

With SNS the novel ends: the main character has passed puberty. The presence of the publicwithin radio programmes goes from zero grade — the telephone — which implies only the presence of a voice,invisible and disembodied, to the most advanced stage so far — Facebook — in which the public has a face, a name,a personal space for discussion the Wall , a bio-cultural profile the Info section , a collective intelligence theHome Page , a general sentiment Arvidsson It is the end of the public as a mass that is blind it cannot seethe source of the sound , invisible it cannot be seen by the transmitter , passive it cannot take part in theconversation and insensitive it cannot manifest its emotions towards the speaker.

Tiziano Boninibody of the radio medium renders the immaterial capital made up by the listeners public and tangible. While untilrecently the audience was invisible to radio and was confined to its private sphere except in the case of phonecalls during a programme, today listeners linked to the online profile of a radio programme are no longer invisibleor private, and the same goes for their opinions and emotions. And if emotions and opinions are no longerinvisible or private, they are measurable.

The new communications model that derives from the short-circuit between radio and social media is ahybrid model, partly still broadcast, partly already networked. Radio is still a one-to-many means ofcommunication. However, telephone already made it partly a one-to-one medium phone interview and many-toone open mic, phone talk radio ; to this we have to add SNS, which are at once a one-to-one chat , one-to-many tweets, FB notes or posts , many-to-many FB Home, Twitter hashtags , many-to-one FB comments kind ofmedia.

Both types of relation areapproaching a dynamic typical of peer-to-peer culture. At the same time, the relation between listeners is similarly changing. While before SNS the concept of radio public was a purelyabstract entity, which could be understood sociologically and analysed statistically, today this community is nolonger only an imagined one Anderson People who listen frequently to a radio programme and are its fanson FB have the opportunity, for the first time, to see and recognise each other, to communicate, to create newlinks while bypassing the centre, in other words the radio programme itself.

This is how the value production process in radio works inthe era of SNS: listeners enact their tastes online, the radio author increasingly a producer, as Benjamin predictedin his essay The author as Producer re-interprets and re-elaborates them, providing the audience with adramaturgically constructed listening experience in which it finds its contents mixed together. On the SNS stage everyone, radio makers and listeners alike, is ableto perform, to take part, to alternatively play the role of the actor contributing with contents and of the audience contributing with comments and liking.

How much the listeners take part in this production process is still controlled byradio makers and this has to be taken into account when designing a new radio or cross media format. Results of a comparative study on the use of social media by three Italian successful radio programmes. In the second part of this paper we will present the results of a comparative study on the use of socialnetworks by three Italian radio programmes.

The observation of the Facebook fan page of the three programmestook place in the week between 1 and 6 March Source: Facebook internalstatistics Over 55Europe: activeusers RaiTunes is a programme with a younger audience, while the Facebookpublic of Caterpillar and Io Sono Qui is in line with the average public of the radio stations that air them. Schedule: Monday-Friday, His name is a benchmark in music radio formats and his voice represents thereal brand of the programme.

The music format see below is addressed to urban listeners and a young audience. Music is mixed live by DJ Frankie B, an international music producer and sound designer well-known on the clubscene. The presenter, himself a musician and DJ, frequently performs with Frankie B. The programme was chosenbecause of its great appeal to younger generations this is in fact the most important show addressed to them ,for its cross media approach and for its considerable and innovative use of social media.

Once or twice a week usually Tuesdays and Thursdays the playlist is user-generated, that is it is put together by the community oflisteners together with the presenter. The selection is made during the day before the evening show on theFacebook fan page of the programme.

The fans are young and extremely active. They post an average of 60 to You Tube links to music videos every day even on weekends. The page is constantly updated throughout theday. Fans keep on posting at every hour, day and night. It resembles a collective stream of consciousness. Music video posting is the realglue of the RaiTunes community. The listeners of the show are used to music shows, are used to go to concertsand they behave like a concert audience.

The fans who post on the wall demonstrate a wide musical knowledge,perfectly matching the musical choice of the presenter. Before and after the show the fans keep on posting music and making comments about it, but when theshow begins something special happens: they stop posting music and start listening to the programme, leavingthe FB page open on their computers. During the 80 minutes of the show the FB page is updated almost everyminute with comments and questions about the music played by the DJ.

Doing radio in the age of Facebookaverage of 60 to 80 comments on the wall, like a concert audience chatting about what it is listening to. Every daythe same dynamic takes place: when the programme begins, fans stop posting videos it is like when a concertbegins and the audience falls silent : they agree to enter into another dimension, the spectacular one.

The changeof behaviour on the FB fan page marks a ritual passage. Presenter and listeners recognise that they belong to thesame tribe and taking part in the show means celebrating the music they share. Music is the totem around whichpeople gather. Listeners would like to listen to more music, butwhen the show comes to an end, they can only continue to post other music on the FB wall and listen to the musicchosen by their peers.

A few minutes after the end of the show the RaiTunes team publishes a You Tube video playlist on FB, acollection of videos of all the songs played during the episode that has just ended. TwitterTwitter is frequently updated by the production team, to disseminate contents, news and You Tube playlistsavailable on other platforms.

Broadcast use only. WebsiteThe website of the programme contains the archive of all the episodes aired so far, available for listening toin streaming. During the show it is possible to see what is going on in the studio through a mobile webcamsituated on the roof. The webcam frames the presenter and his guests and is remote-controlled by the socialmedia manager, on the other side of the studio. Case history 2: CaterpillarBroadcaster: Radio 2 Rai www.

Conceived as a drive time talk show, it is the most listened-to programme on Italian radio in itstime slot. It provides an independent, tongue-in-cheek take on national and foreign current affairs. Phone talk plays akey role, occupying more than one third of the programme listeners, correspondents, politicians, artists, critics,etc.

Tiziano BoniniPodcasts are easy to access from the Rai iPhone applicationSocial media activityBlogThe blog is updated every day with a synopsis of the latest show and videos of the music played. It is usedas a multimedia archive of the programme. The morning after the show a podcast is available for do wnload also via Facebook. Thepresenters and authors of the programme usually join the conversation and interact directly with their fans.

Fanscan also find relevant excerpts of the show a live music show, poetry by the correspondent Marco Ardemagni,satirical videos available on demand on the fan page, extra contents not aired during the show and extra contentnot suitable for radio video interviews from the correspondents, text notes, collection of photos of special events.

TwitterThe Caterpillar Twitter profile page, however, is more institutional and formal. It is only used as a one-tomanymedium, in order to disseminate to "followers" issues and links related to the programme. It looks more likea newspaper-style homepage. Audience participationFacebook fans of the programme post comments both in real time during the show and after. Fanspublish an average of 7 to 15 wall posts every day. Listeners use Facebook not only to show whether they likesomething that has been broadcast or not, but also to publish news and links they find useful, either for theprogramme or for the "Caterpillar community".

The radio show and the Facebook page are both examples of networked media, since they rely heavily onuser generated content and comments. Every day the presenter tells the story of his life as if it was an22 ECREA: Radio Evolution: technology, content, audiences — conference Doing radio in the age of Facebookaudio diary and shares his experiences with the listeners. The second part of the programme is based on thephone calls of listeners who want to share their private stories.

Fans and friends start to reply to thepresenter's call with comments and long posts, sharing private experiences of everyday life through Facebook. The author of theprogramme has accustomed his Facebook audience to expect one call every morning. He opens the game withthe first post of the day, letting the listeners be the main characters of the play on the Facebook stage. During theweek of observation, the presenter's calls received between 25 and 80 comments, depending on the popularity ofthe topic.

Almost every day the author chooses one story among the best ones appeared on Facebook in the hoursbefore the show. Usually the first phone call of the day comes from a Facebook fan, then the presenter starts to take other livephone calls too. Sometimes fans spontaneously reply to the call of the day by posting an excerpt of a film or asong that reminds them of the topic of the day on the Wall.

The presenter normally edits and uses these contentsembedding them into the radio flow of the episode of the day. One hour before the show the author posts a YouTube link to the video of the song that will be broadcast during his story.

Facebook is conceived as a mine ofrough contents to be chosen, edited and then embedded into the radio production flow. Comments, life stories,links to video or audio contents, are used by the author and his team as material for the production of theforthcoming show.

During the show: fans and friends that are listening to the show through the web post comments about theprogramme on the wall. After the show: fans keep on commenting the show that has just ended and start to post on the wall aphoto shot by them that can represent where they are in that very moment emotionally or geographically. During the observed week fans published an average of 6 to 20 posts per day on the wall of theprogramme, while the programme's team made between 3 and 5 posts per day.

RaiTunes uses the music links suggested by the audience,Caterpillar uses the news links suggested by the audience, Io Sono Qui uses the life stories told by the audienceand its content suggestions photos, video, songs. Tiziano BoniniSocial media Manifesto for radioEven if social media use has entered the production routine of radio only in the last two to three years,turning out to be a crucial tool, but quite often misunderstood and underestimated too, in the case historiesanalysed so far we can note many similar social media practices, which are both effective and innovative.

Thecomparative study reveals that broadcasters have finally started to understand the importance of social media innurturing their relation with audiences, like an umbilical cord connecting listeners to producers while the radio isoff.

As a conclusion, we will try to put together the best practices discovered during the research and write a kin do f Social Media Manifesto, or more simply, a bare bones guide to the ideal social media strategy for broadcasters. Dramaturgic structureSocial media management is an authorial and creative work.

It is similar to the work of a theatre directorand has to do with storytelling. And storytelling has its rules. Social media spaces are not virtual at all, they arelively spaces where people attempt to show themselves at their best, making great efforts to perform one of thecharacters they would love to look like in real life.

As people's FB and TW profiles are nothing but storytellingperformances, programmes' profiles have to address issues of performance and storytelling too. The most successful Facebook and Twitter pages analysed so far all share a specific and clearlyrecognisable dramaturgic structure: frequent, cyclical and regular updates, every day. Facebook and Twitterprovide a flood of data, and posts and tweets will quickly flow off followers' screens.

Tweeting frequently will builda bigger following. Radio producers have to show listeners that they are always alive, always present, and theyhave to convince them to visit their page more often during the day. They have to build expectations among theirfollowers. Posting 15 tweets a day, but all in the same half hour, will not do , as most of the followers will not evensee them.

Radio producers have to educate the public, making them feel that their page is constantly updatedwith valuable contents. Second Act - During the show: InterActWe have noticed that successful work and presence on social media generates a continual flow ofcomments and updates from listeners during the show.

Third Act — After the show: the show must go onSuccessful programmes are conceived like multimedia projects. When the radio show comes to an end,the programme continues on the web. Cross media interactionConnect all the platforms and enforce communication flows between them. Doing radio in the age of Facebookin talk shows is to give the same importance to listener feedback, no matter which platform they came from email, phone call, sms, Facebook, Twitter. The debate around the issue of the day starts on social media, thencontinues on air: the presenters keep quoting comments made in real time on social networks.

If people get used to knowing that what happens in the social mediasphere is valuable for the programmetoo, they will participate more. Presenters and authors of the programme have to play at the listeners' level, and to build a fair and straightinteraction with them. Every time you post something on social media you should provide it with a context for itto be properly understood, and personalise information, adding your personal view or feeling.

Every podcast alertyou make has to be accompanied by a quick and personal synopsis of the programme contents, using a catchylanguage, not the cold and standardised language of marketing but the warmer one of true personal engagementwith it. Every post is a little story. Take advantage of General Intellect and realize Walter Benjamin's dreamSocial media are wonderful tools for nurturing and empowering the General Intellect 2. Thanks to theirnetworked structure, social media seem to be making the dream of Brecht and Benjamin 3 come true: listenersbecoming authors UGC.

Among your listeners lie hundreds of experts in different fields willing to take part incontent production. Caterpillar RAI perfectly outsources some reporting to the listeners and takes advantage ofcitizen journalism: its listeners publish suggestions about topics to be discussed and offer themselves as reportersfrom the place they live in. Ask listeners to tweet their reports in real time while travelling. The minds of thelisteners, once connected through social media, can be very powerful and fast.

Share the loveShare, quote, forward, retweet valuable contents. You need to give in order to get. ReferencesAnderson, Benedict La radio. Au microphone: Dr. Walter Benjamin. Tiziano BoniniVol. On the Higher Education context many changes are occurring due to the introduction of newlearning paradigms, many of them take advantage of web 2. Social networks are currently being a do pted in many Higher Education communities as platformsto support the interaction among community members, taking advantage of the potential of thosenetworks to foster strong and meaningful relationships and support the awareness andconsolidation of group identity.

This potential is being explored to promote new possibilities forteaching and learning that include new approaches such as the personal learning environments. This article addresses the potential that radio services have for Higher Education communities in aweb 2. The article explores theperceptions that Aveiro academic members have about webradio potentialities in terms of sense ofbelonging creation and community cohesion.

Keywords: webradio, university, community, social networks Radio as a service of a university communityThe incorporation of the radio in the university field, as well as their potential use by the academiccommunity, is not a recent phenomenon. The first initiative of this kind took place in at the University ofWisconsin Faus, College radio refers to a type of station that operates within an academic community and presentscharacteristics of community radio and educational ones.

These stations can be a global institutional projectinvolving the entire university community or an initiative from a more restricted entity faculty, student union,student-teacher of a specific subject… Sauls, 1. In fact, the phenomenon of college radio has evolved from the first experimental stations and, nowadays,has multiple configurations depending on the technological support broadcast FM, AM, web , audience of aclosed circuit to a wider community of listeners , aims education, outreach, entertainment or managementmodels Sauls, 2.

Characteristics that imply a programming for the college radio, different from commercial ones. This type of stations, to which also belong community radio stations, is characterized by uncommercialobjectives and social vocation. College radio has also a cohesive feature that, combined with the fact that itoperates within an academic community, gives it characteristics of community radio stations.

Indeed the main goal of any college radio is to provide a service to the community, regardless of whetherit is a strictly academic community or a wider community Sauls, 2. The purpose of this paper is to deepen into the perceptions that different audiences in the academiccommunity of Aveiro have about the potential of a college radio for the community cohesion and the promotionof a sense of belonging,. The underlying conception of the university webradio is here a platform with links tosocial networks, a space to share materials among professors-students or students-students, and other kind ofinteraction tools.

Radio transposition to the Internet offers lots of potentialities for the college stations. In fact, since the early college radio web initiatives that took place in the late 90's, thisphenomenon has been expanding. The radiomorphosis. A new paradigm based on the interactionThe mediamorphosis Fidler, in radio renewed the audio product with the addition of componentsinherent to digital system.

Thus, webradio set up a platform where converge multiple features of the conventional media with thosederived from its new multimedia essence like flexibility, ubiquity, synchronous and asynchronous communication,language and interactive multimedia. The phenomenon of radiomorphosis Prata, was reflected primarily on the genres and on theinteraction. Two connected areas that establish the essence of the Internet medium and alter broadcastingconcept nature Cordeiro, From the perspective of interaction, the transfer from terrestrial radio to the web has strengthenedrelations with the user through new forms of relationship.

Interaction that has evolved from participation viaemail, an e-review of wiretapping tradicional model, to other nearish and instantaneous modes like socialnetworks. This is due to the return of listeners, interacting in relation tobroadcast content and also due to the release profile on portals, directories and virtual communities" Kischinhevsky, Radio 2.

An approximation of Aveiro University Members perceptionsThe interaction of these listeners in multiple social networks establishes a relation between them and thepractitioner, a relationship which allows real-time feedback regarding the contents conveyed. This enablesconsolidation of collaborative media based on a single network that combines social networking and various webtools 2. In this context,prosumer figure rises up as a listener consumer and content producer at once Toffler The multimedia nature of the web allows to push the limits established between the radio and its listeners.

A relation marked by the fact that, as Moare stressed inBuffarah Junior, 6 there is no place on the net for passive recipients. The radio experience at that time,new concepts and gain time previously inaccessible devices " Ahmed, This new potential of Internet radio enables its use in a community college with multiple objectives. The characteristics of this digital natives group can be considered convergent with the web broadcastingpotentiality: "nomadism, individualism, customization and personalization, exhibition and voyeurism, public andprivate space, memory of the generation on demand and a young profile in transformation" Rodrigues da Cunha, This convergence should not be dismissed on college radio.

Metho do logyTo accomplish the aim of assessing the perceptions that Aveiro University members from now on AU have about webradio potential for academic community cohesion, an approach to its main audiences has beencarried out: students and professors. An approach that a do pted different samples, analysis tool and metho do logy.

The selection of both convenience samples was do ne according to different criteria. StudentsAccording to Rose and Lenski and Baker students are configured as the primary recipient of auniversity webradio. This is a circumstance of special interest from the point of view of webradios potential foruniversity community cohesion. A test sample of 78 individuals belonging to three different groups of students in the UA was chosen:communication graduate students masters and do ctorates , students coursing other subjects undergraduate andgraduate and foreign students-researchers in various scientific areas.

The selection of the second group of 18 undergraduate and graduate in other scientific areas was due tothe need for a sample of students from the AU whose media consumption, and ideas regarding the potential ofan university webradio, would not be influencied by their proximity to the field.


There, the bridge is now safe. Also, watch out for the guard over looking the bridge, and continue on to the next area, Strut F. Theres nothing for you now, all you have is a level 1 key-card. Anyway, move around and take out all the guards with a M9, and hide them in a locker. Go downstairs and you'll be in large area with lots of boxes and crates and wonderful stuff like that.

Run down the hall and you'll see two doors. Go into the right one and grab whats in there. Don't bother trying to get to the other side. Instead, leave and go to the left door. Open up all the lockers and grab what you find. Now access the node like usual. Go down, and you'll see a duct. Crawl into it and guess what? Anyway, if you wanna know how to use it, check the Weapons section at the top of the Walkthrough.

If not by now you should have heard the "Why are you late with your status report? Hopefully you did what I said and put them in a locker. After they leave, the strut is yours, because everyone's locked out. Now for the actual bomb disposal. Go back to the upper level and to the middle left. Press Triangle and hang over, then drop down with X. Go down to the ground, and you'll see the C4, freeze it. I know what your thinking now. Your saying right now "This is boring and tedious, I don't wanna go around freezing bombs.

Let me battle! Get up and to the other side, now go down, and to the left. You'll see and door. Enter it. Go to the other side, go up the stairs and enter the door, back to Strut A we are. Drag him to the lockers on the left, and open the locker on the right.

Grab the socom bullets, then throw the guard in it. Now enter the Southern room, where the actual pump room is. Make sure the guard is facing away, and tranquilize him, then approach the camera on the Northwest side and shoot it with your SOCOM. Go over to the stairs on the left, up them, then down through the pipes. You'll see a bunch of Pumps. Keep crawling until you see some big yellow pipes, then crawl down them, and go to the right.

Get up and look at the pump. Whip out that Coolant again and defuse the bomb Now you call Stillman. He explains about the Church incident and lets you know that he isn't.. He lied about it because he couldn't bare that he was responsable for not being able to save all the lifes of the people who died in the explosion. If you want, you can to the right set of stairs and get a cardboard box. Anyway, now get out of the pump room, head up and left, to the AB connecting bridge.

You'll see a guard walking the other way, tranquilize him. Enter the door to the right and tranquilze the guard there. Go up towards the door, and you'll see an electrical box with a door opened. Close it and you'll see the C4, freeze it and finally, the last bomb! Raiden then calls Stillman, and you find out some more info about whats going on, and about Fatman's plan. Its vacant again. Go into the kitchen, then into the pantry and grab Sensor B.

Then you'll get a call from Stillman, and guess what? You've gotta get to the Bottom of Strut A and defuse the big one that was triggered. You have seconds. Get going. If not, tranquilize them again. Run down and then make a right turn, onto the bridge. If not, just take them out from a distance. Run to strut A. As your going down, Stillman calls. He fell for it, Fatman has Stillmans number. He used microwaves with a 7 foot detection range that would make the C4 blow up regardless.

Stillman is stuck, he can't make it out in enough time so just gives up. Stillman advises Pliskin to run from Strut H. Raiden dosen't know if he can defuse this bomb now, put Stillman assures him he can do it, and warns him to spray it from far away as possible, just before the the bomb blows up in Shell 2. Grab all the ammo and rations you see.

And continue into the room which you came from. You to the farthest South side, facing the middle of the sub, duck and equip the coolant. You should see the C4, now spray it and end the bomb craze. Raiden calls the Colonel and tells him that he's finally done defusing the bombs. Raiden asks what he's to do now, and is reminded he must rescue the President.

Head back up the elevator. As you enter the stock room, a cutscene occurs. I smell a boss battle. Fortune comes down the elevator with her absolutly HUGE railgun, and smiles thinking that Raiden is an enemy. She fires a shot and Raiden and sees his face. Noticing it's not the person she's looking for, she frowns, but quickly smiles, hoping Raiden should bring her fun. And I'm not saying that because shes hard to beat, I'm saying it because it's true.

You cannot hit he with anything. Just dodge her shots, and try not to get killed. Her railgun is uber-powerful and will end up destroying the whole room. Don't run around yelling "OMG She's destroying the room! Why me! I'm gonna die! After the room looks like a warzone.

Raiden now gets a call from Colonel, and must get to Strut E. To me it's hilarious how he's like "Look, this isn't the best time right now! Anyway, Vamp comes down the elevator and tells Fortune to handle Fatman. Right then, Raiden pops out and blasts a shot with his Socom right at Fortune, which is deflected and goes right through Vamp's skull, killing him. Raiden, pops a few more shots, but they miss Lady Luck, because she cannot be killed.

Raiden, scared, runs up the elevator. Fortune continues sobbing over Vamps death. Then, the most unusual happens, Vamp comes back to life. Vamp says that he has already died once, and can't die twice, is he Don't worry, its enough time. When you reach the gates that lead into Strut A's pump room, crawl, because there are now claymores there. Tranquilize the one standing near the door the the FA connecting bridge.

Take out anyone out there. Tranquilize the guys who are in your way, and head north towards the EF connecting bridge. Cross the bridge? And you'll notice a C4, right in clear view. Freeze it and get ready. Anyway, after freezing the bomb, a cutscene occurs, where you meet the man who killed Stillman just minutes ago, Fatman.

Fatman blades by on rollerblades with some kind of beverage in his hand, and tells Raiden all about himself, eventually leading to Raiden and Fatman getting pretty pissed at each other. Fatman slams his glass on the ground after Raiden calls him a common criminal. Fatman decides to play of game of Plant a bomb with Raiden. And so the battle ensues. Equip the M9, and Sensor A. This really isn't hard.

The first bomb is on the crates by the area it's around, and the second bomb is on the big green box within the Sensor area. Afterwards, his bombs are completly random. Plant a few C4's out, Fatman will hit them alot. Whenever he stops by you, punch him down using Circle.

When he's dizzy and on the ground, shoot him in the head. Eventually, Fatman will fall. Just make sure the always have Sensor A out, and like always, freeze the bombs with your coolant. As I stated before, this battle is only hard in Extreme and European Extreme modes because you have little time to defuse the bombs. After you defeat him, a cutscene ensues. Fatman is now crouched against a crate, and bleeding. He pulls out a trigger and activates it, and guess what, it's the last and most powerful bomb of them all.

Don't panic it's in the general area. Fatman's last words are "Live and Grow Fat. You really only need about twenty. At first I thought you had to shoot the Bejesus out of Fatman and spray him with the coolant. Then I picked him up moving him, and ding, that's where the C4 for. And, In addition, you get Peter Stillman's Dogtag. Take it and defuse the bomb. After a talk with the Colonel, save it and get out of Strut E.

Wait, I forgot, you can't leave just yet. As you go down the stairs, the Ninja appears. Could it be? Is Gray Fox still ticking? You'll find out. Raiden and the Ninja talk for awhile, and Raiden is ordered to get to Shell 1 and find the hostages by the Colonel, but most importantly, he must find Ames. As for the AKsu and D. Mic, Raiden is told to find it on his own. Raiden yells something at the Ninja but he's too late, the Ninja has already dissappeared.

Alright, finally you can leave. Just head to the EF connecting bridge. You'll see a door to the north, go in and search around, eventually you'll find the AKsu. Grab it, and take all the ammo. If you want you can grab all the other stuff in Level 1 and Level 2 doors, seeing as you have the card. Take out all the Cyphers that are out here, if you don't, you'll probably die. Are they gone.. Now head to the left and you'll see a long bridge.

Run across it and don't stop for anything. The panels will fall out. Do not un-equip your AKsu infront of a guard, because they will call their friends and send about 8 more guys into the core, and that is hell to escape from. First, go south and to the left. Enter the little room and open all the lockers for some items. Then go up and grab the Stun Grenade ontop of the crate. Finally, acess the node and go back to where you came into Shell 1 at. Go north and then to the east, where you should see an elevator.

Press Triangle and get in. When you get it, go to the left and down, you should now see the floors which are: 1, B1, B2. Go down to B2 and press Circle. And go down into the actual computer room. Don't rush it, you might bump into a guard. Inside the actual computer room is the D. Mic and a box. When you get those two, access the node and go back to the elevator. With all of that done, now you have to go to B1.

Biometrics, crap. Before anything, go to the right and into a small room, and access the node. Now to get into the Conference room where the hostages are being held. This shouldn't be hard though. Walk up behind a guard casually and enequip your AK, then choke him and drag him. Bring him to the retinal scanner, and a small scene will occur with Raiden using the sentry to get in.

Listen to him, because if don't this will be alot harder. Once the Colonel finishes up, equip the AK again. Now you have to find Ames. But how are you going to find a man without a description? Well, go up to a man and put the D. Mic near him, if his heart beat seems irregular if you hear a beeping sound , press triangle to ask him if he's Ames.

If not, equip your AK and go to the next guy. If the guard sees you he will come to you ask examine you. Just equip your AK and stand there, you will get a Game Over if you are caught. Have you found Ames yet?

Richard Ames is his full name, and he was sent in to keep tabs on the President. Ames asks about how he knew about him, then explains about the Big Shell. He tells you what the Sons of Liberty plan to do, and reveals something about the Big Shell. In the middle of the conversation, this so called "Solid Snake" appears in a different room.

Ames tells you to listen in, but you can skip this if you want. Point your D. Mic to the left and you can here him, looks like he still hasn't gotten over that sickness, even after 4 years. After its all over, Ocelot walks towards you. You have 10 seconds to equip your Ak. When you do, don't move, don't breath, don't do anything, or Raiden will be pumped full of bullets in a second.

Ocelot speaks with Ames, who ends up dead, apperantly his pacemaker went out on him. Afterwards, Raiden is reveal, and just as Ocelot is about to send him to the grave, the Ninja comes down and secures a path for Raiden. As Raiden get out, Caution mode is entered. Ok, it's time to get back to the game. Go towards the little room where the node it, putting the soldier guarding that room to sleep with the M9.

Put him in one of the lockers, and hide yourself in one until Caution is gone. Then run for the elevator, head up the first floor and plan on heading to Shell Connecting Bridge Most importantly, if you didn't get the PSG-1 from the Strut F warehouse when you went to get the AKSu, get it along with everything else you can find, then make start heading to Shell Connecting Bridge. If you see an item, then grab it. Anyway, head up and to the left, or Northwest.

Either way you should end up on the DE Connecting Bridge. This room is still heavily guarded. Make your way to the North door, and if you want get the ration in the center of the room, it's right behind the tank. When your through in here, head to the north door that I spoke of before, which will lead to Shell Connecting Bridge. He was out for awhile, but now he's back. After a conversation, he tells Raiden that he has to take out the Control units by gunfire, because if he gets too far in, the IR sensors will detect him and blow the Big shell out of the water.

Alright, IF you are on Normal mode, there are 10 different control units you have destroy. Look up towards the door leading back to Shell 1, you'll see it, its on the wall. On a tank right in the first center tank. Left side of the bridge, near a tank. Look to the right of the center tank, it will be there. Go to the right, and equip your PSG Pop in a pentazemin look towards a flock of birds. Shoot at them and they will fly off. It you hit the unit straight on, it'll be gone.

If you just shot a bird, then destroy it. Eventually the Colonel and Rose will call and scold you. On the far left, behind a tank. On the right side of Shell 2's door. On the left side of Shell 2's door. Look at the Sons of Liberty flag. Shoot at the center of it with your PSG This one is often hard to get, and you wont be sure if you got it, but just fire crazily at the flag because no Semtex is around.

You should hit it. Use a pentazemin or 2. Look up and you'll see a Cypher. Shoot the control unit and only the control unit, if not, you will blow the big shell out of the water. The control unit is on the top of the cypher. Once again, be careful. When you get halfway, Pliskin meets up with you in the Kasatka with his partner "Otacon". Raiden looks towards Shell 2 and sees the man who claims to be Solid Snake.

But, it is really Solidus Snake, the third Snake. Pliskin fires at Solidus angrily yelling that Solidus is not Solid Snake. Solidus smiles, and addresses Pliskin as his "Brother" after a few scenes. Solidus jumps in the harrier, Raiden is given the Stinger Missle Launcher, and now, you, Raiden, must take down the Harrier single handedly..

Although the difficulty is normal, this is still hard. It took me plenty of time to beat this on Extreme, and I have yet to win on European Extreme. For the Actual fight, grab the stinger, the bullets, and anything else Pliskin throws down at you. Equip the newly aqquired Stinger missle launcher and attack the harrier. Watch out for it's many attacks. When the multiple bullets come, hide on the bottom floor, crouch, and hide behind the green metal box. This should keep you safe.

Also hide, when he sends the motherload of missles at you. Here's a few tips.. You need as much as you can get. Take a shot whenever you can, you don't want to die. When your bleeding, hide out for awhile. Some people find this battle incredibly hard, some people find it pretty easy.

This battle varies for me, but from popular opinion, I've deemed it hard. With the battle over, the harrier crashes down to the ground, taking a part of the Shell Connecting Bridge with it. Some more cutscene's happen and you find out that Pliskin is actually After everything has happened, you gain control of Raiden on the ruined bridge. First, save if you haven't already, then run over to the fire infront of you and stop it with your coolant.

Go down the stairs quickly and grab the ration, because those stairs will eventually fall off. Now you have two paths to go. You can jump across the bridge and get the Ak- surpressor, or go to the right, climb down and go across safely.

If you plan to get the AK, like I suggest you do, start far away and right when you hit the end, jump across the bridge. This might take a few tries, but you saved already, so just reset it if you die. When you get there, take out the fire with your Coolant and grab the AK- Surpressor. If you took the other way, just watch out for the birds, they've been crapping all over, and if you slip, you'll fall into the ocean. Now, go to the right and then up across the bridge. Don't stop because some parts of the bridge fall off Jeez, this whole place is falling apart, eh?

Now, you'll be at a dead end, enter hanging mode over the little path, and drop down. When you reach the ground, you'll see two guards come out to investigate. If you have a silenced weapon, use it to take them out. If not, just use the M9.

With the guards taken care of, advance onward. You'll notice two places where there is no ground. The first one you can run and jump over. The second one is very hard to do so, so just enter the hanging mode and move across it. When you get through with that, climb up the ladder and you'll be off the the next Strut. Be careful, because alot of guards look through here, AND the floor will break on you. If you are caught, just dispatch of all the cyphers.

Crawl quickly across this, and if you're too slow, you'll fall right with them. Keep on going until you get to the end of the path then turn up. Once you reach this, you will see a gap where it seems crossing is not possible. Flatten against the wall and press X to crouch, and move across the thin strip.

When you get to the end, get back up. Halfway down the path, a guard comes up and starts to uh.. Stand there and let Raiden get pee'd on, call the Colonel on the Codec. Hilarity ensues. Laugh for awhile, then continue on, and you'll come to a dead end. Enter hanging mode, and drop down.

Take out all the Cyphers here, and the guard too. Continue on to the left, and that should leave you in Shell 2's flooded core. You can listen in, or just skip it, I advise you listen if you want to here the story. The President is in this room, but it won't be so easy to get to him because Olga turned on an Electric Current on the floor leading to the room, thus, making it impossible to get in and save him. Now, you must find a Nikita Missle Launcher to disable the floor and save the president.

Sounds easy, right? You've got to find it, while underwater. It's on B1, and thats flooded! Alright, you've got control of Raiden again, head for the elevator which is to the north. Also, go to the little lounge by the elevator and activate the node, and pick up and little items you see.

If you don't have the RGP 6 yet, go to the left corridor, past the elevator, and you will find it. Now, head into the elevator. Your destination: B1. Gosh, this is very hard to explain where to get, so I will try my best. You have to go underwater and get to the most southern part of the room, and be in the center.

That is where the Nikita missle launcher is. If I'm not clear enough, just search around. This is pretty hard to explain. When you found it, head back up to the first floor. Take a step in and shoot both of the Machine guns that are there so their inactive. Now go in and crawl ontop of the box. Face north, and shoot a missle into the duct right infront of you. Shortly, it will be in the President's room. Be careful not to hit him, regardless of being the President, he has the almost non-existant intelligence right now.

The control unit is the big metal panel right at the right part of the room, there's 3 of em, but you only have to hit one, and it has a railing surrounding it. You can't miss em. With that done, head to the President's room. A whole lot of cutscene's occur, and take a very long time to get through. I won't say anything but it involves two things.

First, the President grabs Raiden's manhood, and second, the President is killed by Ocelot. Also, you learn about Arsenal Gear, and about how the Big Shell is just a cover-up for it. With all those cutscenes done, proceed back down to B1, the President wants you to stop Arsenal Gear, but to do that, you must find Emma Emmerich. Yes, that is Otacon's younger sister. If you've played the original Metal Gear Solid 2, you should remember it, and you won't need my help on this part, but if your a rookie, listen in.

Head into the water and submerge. At the first pathway, turn right and head right and head right until you can't head right anymore. Now, go down, and take the next path left, and then next path down. You will get to a watertight door. Open it, and you will find Peter Stillman's dead body, along with the next room. Also, don't forget about getting oxygen, Raiden's not a fish you know!

Onto the next room. Look for an open path along all of the fallen debris. It shouldn't be too hard. When you hit the end, you'll find another watertight door, open it. Now you'll be in the last room. See the set of stairs that are right there, take em up. Finally, your out of the water. Now onto the next room, Heal yourself, its time for a boss battle! And thats not all, Arsenal has just finished it's final check. Vamp tells you a bit about Aresenal Gear because he figures you won't live to tell anyone.

After a bit of talking, it's time to battle! You can finish him pretty fast. Right when you enter the battle, Vamp will be standing on the water for a minute, right then, shoot him in his bullet wound with the M9, it should do some damage. Now, Vamp should submerge underwater. Pull out your Stinger and shoot right into the water, it dosen't matter where Vamp is, it'll lower his Stun Gauge.

Right after, he'll get up and stand there for a seconds or two. Take this oppertunity and shoot him in the head, he'll run around and try to slash you, or dive back into the water. Afterwards he'll go up to the seconds floor and throw knifes at you, slowly at first. Shoot him in the head and duck right when he throws it. The only hard part is when he gets low health, even then, he shouldn't be that hard. Set a Claymore on each side, and he'll walk into them.

Done and done. After defeating Vamp he slowly falls into the water, bleeding like crazy. Raiden wonders if he's finished for good, and continues on. When you have control of Raiden again, grab all the items in the aream, and go up. Grab the Ration right there and jump into the water. Swim to the left on the first path, and take the second last door. You'll now be in a locker room. Activate the node and search all the lockers. You'll find Emma.

When you do, a cutscene occurs. She's just like her brother She gets so scared she pee's herself. At first she dosen't believe Raiden will help her. But later, she is convinced, and they leave. I didn't mention it in this walkthough, but if you've been paying attention to the game you'll know Emma hates water because of an incident that happened when they were just kids.

Ok, so, you've gotta get Emma to Shell 1's Core. This is gonna be hard because Emma can barely walk, Ocelot injected something into Emma that made her legs completly numb. Just swim back the way you did, emerge from water at every point you see. You don't want Emma dying do you? When you get back to the elevator, spray all the bugs with coolant, because Emma hates bugs, just like she hates water. Guards have now come to the room? Why on earth would they jack a place full of guards when it's gonna collapse and kill them all pretty soon anyway.

This is fun though. Grab your AKsu and kill every guard around here. Just make sure to keep out of sight. Don't grab Emma just yet. Go to the exit, and 1 more bastard guard will come in. Pop him full of lead, and go back to get Emma, and lead her to the KL connecting bridge. This is the weirdest part, you have to throw Emma across the bridge Just kidding! I bet you believed me for a minute there didn't you?

Get into the doorway and take out the cypher, and the guard on the other side of the bridge. Go back and grab Emma, and take her up to the main part of the bridge, and down to where the fire is. Whip out the coolant and stop the fire. Continue down with Emma, and you'll reach a door. At first, Raiden will be pissed because he doesn't have a level 5 Keycard. However, Emma does. Emma unlocks the door for Raiden, and they continue on. Go down into the little room and take care of the guards however you'd like.

Kill them, tranquilize them, whatever floats your boat. After their taken care of, take Emma over to the watertight door, open it and open the hatch over to the side of the room. Now you have to climb down a long ladder. What the hell does this girl like? Forget it, just continue down. Now you have to cross the bridge. But it can't support two people, so Emma has to walk across it first. Take out all the claymore on all the paths first, scan the bridge throughly for them.

Take out the cyphers, and the enemy sentrys. Sometimes into it, Snake will call and say he can provide assitance. Call him if you need help, but I suggest you do, he helps out alot. When Emma gets near the end, Vamp jumps out and holds Emma. Snake can't take the shot, so you have to. Great, the fate of this depends in your hands. This does not even qualify as a boss battle, that's how easy this is.

Pop in a pentazemin, take off you thermal goggles, and shoot Vamp with your PSG five times. If you fail this, turn off you game and go cry, thats how easy this is. Vamp finally dies, but he got Emma. Emma has some internal bleeding, and will probably die. Snake takes out two cyphers and takes Emma off to help her. Raiden then runs across the bridge. Forget about the guard chilling and listening to music, he seems like a cool guy.

Run to the elevator and get it. When you get in, you'll end up on B2's computer room, and Emma will be near death. The disk must have been altered when it left the President's hands. Emma assures it will still work somewhat, and her and Otacon talk, and reconcile. Right before you dies, she asks Otacon to call her Emma.

Otacon never does, he just asks what wrong with E. E right before she dies. Otacon begins crying uncontrollably. Snake tells Otacon it'll be alright, and the group continues on towards Arsenal. Otacon is told to rescue the hostages, because he is the only on who can do it. Snake and Otacon do a real cool handshake, and Otacon leaves. The Ninja comes out. Snake and the Ninja knock out Raiden and capture him.

And all along, the Ninja was Olga. Later in a cutscene, Otacon continues crying, but still saves the hostages. The big shell is sunk, because Arsenal uses it to go active. Right above it, Otacon is flying the rest of the hostages out of there and safely to Manhattan. Raiden wakes up restrained to something. When he comes to, he sees Solidus and Ocelot. Solidus reveals that Raiden is his foster child. After a breif talk, they leave, but not without taking all of Raiden's stuff.

Olga comes in and tells you about some stuff, more importantly, that she was actually here to help you, and that Raiden had to be captured for Snake and him to gain access to Arsenal. After awhile, Raiden is punched in the nuts by Olga, who leaves and then releases his restraints. Great, Raiden's naked. Naked as the day God made him. So now, Raiden can't enter the hanging mode, choke out guards, and he can only punch with one hands.

I don't think you want Raiden's 'how do ya do' flopping all over the place. Oh brother, this is going to be crazy. Open up the locker and grab some medicine. Arsenal Gear is very cold, and with Raiden being Butt-Naked, its very easy to catch a cold. Active the Node, and go on into the next room. They're cooler, faster, more agile, and stronger. Also, if you want to be proper about it, their called "Tengus".

Now you'll get a codec call from Colonel, and he seems sort of odd. Raiden shrugs this off, but continues. Get seen by a guard, run back into Arsenal Gear, stomach. Press up against the tourture machine and wait for a guard to come in. Watch as he says some things to you. Go to the stairs that are to the left, and when you get to the top of them, the Colonel calls you. But its not just a regular call. He tells you to turn the game console off. Don't actually listen to him. Continue on up for awhile, it dosen't matter if you get caught, because you only need to get into one room, and alert mode is done for.

Keep answering the Colonel's strange calls, and when you reach the end of the path and see a way to go on the right, turn to the right, and enter the next room. And get the ration at the far south part. Eventually Rose will call you, and she will be acting sort of weird too. With that, Snake comes and laughs at your nakedness, and gives you your gear back, along with a High Frequency blade.

Snake teaches you how to use it. Pratice with the blade for a bit, and then go over to the end of the room. Also, knock out Snake and grab his Dog Tag. Damn cyphers. Whip out your HF blade and play Cannon fodder. Stand infront of Snake and hold L1, thus, you will deflect most of the bullets while Snake kills everyone. When you get near the end you'll have to go around killing people because there will be too many for Snake to handle.

More BS. Just continue on, where some more Tengus ambush you. Its time for another fight, bring out the H. You'll get a weird "Fission Mailed" screen in the battle once or twice. Don't stop fighting, continue fighting. This is just a Mindgames that the game gives you and is supposed to represent what Raiden feels. This is alot easier than the other battle.

Eventually, the guards will stop coming. Every last soldier has died. After the carnage, Snake and Raiden look at all the bodies. And out comes Fortune. She claims Solid Snake killed her father, Scott Dolph. However, Snake knows it was Ocelot, and they engage in a fight. Ok, thats enough. Watch the cutscene. Oh crap No pun intended.

Only if you don't pay attention. If your paying attention to the Metal Gears, its pretty easy. All you really have to do is Shoot one in the knee, and then the mouth. Repeat until dead. Then repeat on every metal gear until you win. Move around every so often, and watch out for it's attacks. Also, don't let them get too close. After you battle, Raiden falls down exhausted and gives up. A bunch of cutscenes occurs, and you have to endure choking from Solidus again.

Press Triangle really fast for a long time, and you'll survive. If you do, watch the millions of cutscenes, and 1 by 1, everyone leaves by death, or gets off of Arsenal. Eventually, its just Raiden and Solidus. Needless to say if you listen to eventhing it takes about 15 minutes. Afterwards, Solidus tells Raiden that he was the one who killed his parents, upon other reasons.

They must fight.. Or Raiden.. Solidus' creation? Just watch out for his flame attacks, and equip a ration. You don't really need help on this, seeing as its such an easy battle. When you beat Solidus, sit back and watch the ending. Answer: This one really wasn't explained well. Otacon told Raiden that he had a small boat waiting for Snake when he was told about the Tanker sinking. Otacon must have done some fast work, or already have known about what was going on.

If you pay attention to the whole sceen, you see about a second where Snake emerges from the water, and is in the water, not in the tanker. Then when you see the Tanker sinking, you can see a shadowy boat coming near it. Question: Why do I have to play with that loser Raiden!?!?!?! Snake r0x0rz j00r b0x0rz!!!! Answer: Yes, Snake does rock my boxers. You have to play as Raiden because that's how the story went. Don't ask me, ask Hideo Kojima. Answer: To be quite blunt, I don't jam pack it full of useless stuff that the reader dosen't care about.

I just give the person the basic info, and let them figure it out. The herbal medicine IMOD has been shown to strongly modulate immune responses in several animal studies as well as in clinical trials. However, little is known about the mechanisms of IMOD to modulate immunity. Insight into the immunomodulatory effect of herbal medicine IMOD may provide innovative strategies to affect the immune system and to help combat various diseases.

Soluble antigens from group B streptococci induce cytokine production in human blood cultures. Group B streptococcal antigens stimulated tumor necrosis factor alpha TNF-alpha , interleukin-1 IL-1 , and IL-6 production in human blood cultures in a concentration- and time-dependent fashion. The minimal concentrations of type-specific polysaccharides, lipoteichoic acid, and group-specific polysaccharide required to produce these effects were, respectively, 0.

Cell separation experiments indicated that monocytes were the cell type mainly responsible for cytokine production. Time course studies indicated that TNF-alpha was released before the other cytokines. IL-6 levels were moderately but significantly decreased by anti-TNF-alpha. These data indicate that several products from group B streptococci are able to directly stimulate human monocytes to release TNF-alpha, IL-1beta, and IL These findings may be clinically relevant, since proinflammatory cytokines can mediate pathophysiologic changes during sepsis.

Sirtuin inhibition attenuates the production of inflammatory cytokines in lipopolysaccharide-stimulated macrophages. Highlights: Black-Right-Pointing-Pointer Lipopolysaccharide-stimulated macrophages were treated with cambinol and sirtinol.

Black-Right-Pointing-Pointer Cambinol and sirtinol decreased lipopolysaccharide-induced cytokines. Black-Right-Pointing-Pointer Sirtuins are an interesting target for the treatment of inflammatory diseases. Sirtuins are class III histone deacetylases that modulate the activity of several transcription factors that are implicated in immune responses. This work shows that sirtuin pharmacologic inhibitors are a promising tool for the treatment of inflammatory conditions.

Photodynamic therapy induced production of cytokines by latent Epstein Barr virus infected epithelial tumor cells. Photodynamic therapy PDT is a method to treat cancer or non-cancer diseases by activation of the light-sensitive photosensitizers. Epstein Barr virus EBV has been implicated in the development of certain cancers such as nasopharyngeal carcinoma and B cell lymphoma.

CD1d-mediated activation of iNKT cells by this molecule results in the rapid secretion of a range of pro-inflammatory Th1 and regulatory Th2 cytokines. Polarization of the cytokine response can be achieved by modifying the structure of the glycolipid, which opens up the possibility of using CD1d agonists as therapeutic agents for a range of diseases. Lactic acid bacteria inhibit TH2 cytokine production by mononuclear cells from allergic patients. Among factors potentially involved in the increased prevalence of allergic diseases, modification of the intestinal bacteria flora or lack of bacterial stimulation during childhood has been proposed.

Lactic acid bacteria LAB present in fermented foods or belonging to the natural intestinal microflora were shown to exert beneficial effects on human health. Recent reports have indicated their capacity to reduce allergic symptoms. The purpose of this investigation was to determine the effect of LAB on the production of type 2 cytokines , which characterize allergic diseases. PBMCs from patients allergic to house dust mite versus those from healthy donors were stimulated for 48 hours with the related Dermatophagoides pteronyssinus allergen or with a staphylococcal superantigen.

This effect was dose dependent and was observed irrespective of the LAB strain used. No significant inhibition was induced by the control, gram-negative Escherichia coli TG1. The inhibition mechanism was shown to be dependent on antigen-presenting cells ie, monocytes and on the involvement of IL and IFN-gamma.

The tested LAB strains were demonstrated to exhibit an anti-T H 2 activity, and thus different strains of this family might be useful in the prevention of allergic diseases. Docosahexaenoic diet supplementation, exercise and temperature affect cytokine production by lipopolysaccharide-stimulated mononuclear cells. Acute exercise induces changes in peripheral mononuclear cells' PBMCs capabilities to produce cytokines.

The aim was to investigate the effect of docosahexaenoic acid DHA diet supplementation on cytokine production , by lipopolysaccharide LPS -stimulated PBMCs after exercise, and the in vitro influence of temperature. Fifteen male soccer players were randomly assigned to a placebo or an experimental group.

The experimental group consumed an almond-based beverage enriched with DHA 1. The profile of affected cytokines differs between temperature and exercise, suggesting a different PBMC activation pathway. DHA diet supplementation only attenuated cytokine production after exercise and not that induced by temperature. HSP70 stimulates cytokine production through a CDdependant pathway, demonstrating its dual role as a chaperone and cytokine. Here, we demonstrate a previously unknown function for the kDa heat-shock protein HSP70 as a cytokine.

HSP70 bound with high affinity to the plasma membrane, elicited a rapid intracellular calcium flux, activated nuclear factor NF -kappaB and upregulated the expression of pro-inflammatory cytokines tumor necrosis factor TNF -alpha, interleukin IL -1beta and IL-6 in human monocytes. These findings indicate that CD14 is a co-receptor for HSPmediated signaling in human monocytes and are indicative of an previously unrecognized function for HSP70 as an extracellular protein with regulatory effects on human monocytes, having a dual role as chaperone and cytokine.

Effect of calcitriol on in vitro whole blood cytokine production in critically ill dogs. Hypovitaminosis D has been identified as a predictor of mortality in human beings, dogs, cats and foals. However, the immunomodulatory effects of vitamin D in critically ill dogs has not been evaluated. The aim of this study was to evaluate the effect of calcitriol on cytokine production from whole blood collected from critically ill dogs in vitro.

Twelve critically ill dogs admitted to a veterinary intensive care unit ICU were enrolled in a prospective cohort study. There was no significant difference in whole blood cytokine production capacity between survivors and non-survivors at the time of discharge from the ICU or 30days after discharge.

These data suggests that calcitriol induces an anti-inflammatory phenotype in vitro in whole blood from critically ill dogs. Environmental factors are supposed to play a decisive role in the pathogenesis of inflammatory bowel diseases [IBDs]. Increased dietary salt intake has been linked with the development of autoimmune diseases, but the impact of a salt-enriched diet on the course of IBD remains unknown.

In this study, we examined whether high salt intake alters mucosal cytokine production and exacerbates colitis. For in vivo experiments, a high dose of NaCl was administered to mice 15 days before induction of trinitrobenzene-sulfonic acid [TNBS]-colitis or dextran sulfate sodium [DSS]-colitis. Transcription factors and effector cytokines were evaluated by flow-cytometry and real-time PCR. Mice receiving the high-salt diet developed a more severe colitis than control mice, and this effect was preventable by SB Our data indicated that exposure of intestinal mononuclear cells to a high-NaCl diet enhanced effector cytokine production and contributed to the exacerbation of experimental colitis in mice.

Published by Oxford University Press. For permissions, please email: journals. Mislocalization of SLP leads to aberrant inflammatory cytokine and autoantibody production. Central and peripheral tolerance is required to prevent immune responses to self-antigens. MTS mice also spontaneously develop autoantibodies at an early age.

The peripheral MTS-expressing T cells demonstrate skewed cytokine responses when transferred into lymphopenic hosts. Thus, the abnormal effector T-cell phenotype still occurs in the presence of preserved central and peripheral tolerance, suggesting that diminished T-cell receptor signaling can promote skewed T-cell responses.

We compared the production of 19 humoral factors in mixed cultures of mesenchymal stromal cells from Wharton's jelly and allogenic peripheral blood lymphocytes. For evaluation of the specificity of immunosuppressive activity of mesenchymal stromal cells, comparative analysis of the production of these humoral factors in mixed cultures of lymphocytes and epithelial BxPC-3 cells was conducted.

The major difference of mesenchymal stromal cells from epithelial BxPC-3 cells was 7-fold higher production of IL, which can explain the immunosuppressive effect of mesenchymal stromal cells. Using flow cytometry a positive identification of cytokine producing cells in a mixed culture may be achieved. Recently, the ability to assess cytokine production following a wholeblood activation culture has been described.

We compared whole blood culture to standard PBMC culture and determined the individual cytokine secretion patterns for both T cells and monocytes via flow cytometry. In addition, a four-color cytometric analysis was used to allow accurate phenotyping and quantitation of cytokine producing lymphocyte populations. A three-color flow cytometric was used to assess two cytokines in conjunction with CD IL and. Altered cytokine production by specific human peripheral blood cell subsets immediately following space flight.

In this study, flow cytometry was used to positively identify the specific lymphocyte subsets exhibiting space flight-induced alterations in cytokine production. Whole blood samples were collected from 27 astronauts at three points one preflight, two postflight surrounding four space shuttle missions. Absolute levels of peripheral granulocytes were significantly elevated following space flight, but the levels of circulating lymphocytes and monocytes were unchanged.

Lymphocyte subset analysis demonstrated a decreased percentage of T cells, whereas percentages of B cells and natural killer NK cells remained unchanged after flight. Serum and urine stress hormone analysis indicated significant physiologic stresses in astronauts following space flight.

Altered peripheral leukocyte subsets, altered serum and urine stress hormone levels, and altered T cell cytokine secretion profiles were all observed postflight. In addition, there appeared to be differential susceptibility to space flight regarding cytokine secretion by T cell subsets. These alterations may be the. Inhibition of early T cell cytokine production by arsenic trioxide occurs independently of Nrf2.

Nuclear factor erythroid 2-related factor 2 Nrf2 is a stress-activated transcription factor that induces a variety of cytoprotective genes. Nrf2 also mediates immunosuppressive effects in multiple inflammatory models. Upon activation, Nrf2 dissociates from its repressor protein, Keap1, and translocates to the nucleus where it induces Nrf2 target genes.

The Nrf2-Keap1 interaction is disrupted by the environmental toxicant and chemotherapeutic agent arsenic trioxide ATO. The purpose of the present study was to determine the effects of ATO on early events of T cell activation and the role of Nrf2 in those effects. However, ATO also decreased production of these cytokines in activated splenocytes from Nrf2-null mice, suggesting the inhibition is independent of Nrf2.

Combined immunomodulator and antimicrobial therapy eliminates polymicrobial sepsis and modulates cytokine production in combined injured mice. Elliott, Thomas B. David; Kiang, Juliann G. Purpose: A combination therapy for combined injury CI using a non-specific immunomodulator, synthetic trehalose dicorynomycolate and monophosphoryl lipid A STDCM-MPL , was evaluated to augment oral antimicrobial agents, levofloxacin LVX and amoxicillin AMX , to eliminate endogenous sepsis and modulate cytokine production.

Bacteria were isolated and identified in three tissues. Incidence of bacteria and cytokines were compared between treatment groups. Antimicrobial therapy increased survival in radiation-injured RI mice. Combination therapy increased survival after RI and extended survival time but did not increase survival after CI.

Sepsis began five days earlier in CI mice than RI mice with Gram-negative species predominating early and Gram-positive species increasing later. Treatments significantly modulated 12 cytokines tested, which pertain to wound healing or elimination of infection.

Conclusions: Combination therapy eliminates infection and prolongs survival time but does not assure CI mouse survival, suggesting that additional treatment for proliferative-cell recovery is required. Problem Apoptosis is a normal constituent of trophoblast turnover in the placenta; however in some cases, this process is related to pregnancy complications such as preeclampsia.

Recognition and engulfment of these apoptotic trophoblast cells is important for clearance of dying cells. The aim of this study was to show the cross talk between human endometrial endothelial cells HEECs and apoptotic trophoblast cells in an in vitro coculture model and its effect on cytokine production by HEECs.

Method of study Fluorescent-labeled HEECs were cocultured with fluorescent-labeled apoptotic human trophoblast cells. Confocal microscopy and flowcytometry were used to show the interaction between these two types of cells. Cytokine profiles were determined using multiplex analysis.

Results HEECs are capable to phagocytose apoptotic trophoblasts. This activity is inhibited by the phagocytosis inhibitor cytochalasin B. Phagocytosis of apoptotic trophoblast cells induced the secretion of the proinflammatory cytokines interleukin-6 and monocyte chemoattractant protein-1 by HEECs. Conclusion This study provides the first evidence that HEECs have an ability to phagocytose apoptotic trophoblasts. Furthermore, we demonstrated an inflammatory response of HEECs after phagocytosing the apoptotic trophoblast cells.

This event may contribute to the inflammatory response in both normal pregnancy and pathologic pregnancy such as preeclampsia. A combination therapy for combined injury CI using a non-specific immunomodulator, synthetic trehalose dicorynomycolate and monophosphoryl lipid A STDCM-MPL , was evaluated to augment oral antimicrobial agents, levofloxacin LVX and amoxicillin AMX , to eliminate endogenous sepsis and modulate cytokine production.

Combination therapy eliminates infection and prolongs survival time but does not assure CI mouse survival, suggesting that additional treatment for proliferative-cell recovery is required. However, the signaling pathways targeted by the bacilli to inhibit production of these cytokines are not fully understood. In this study, we demonstrate that the PPE18 protein of M.

Also, this study highlights the possible mechanisms by which the M. In addition, the use of a non-steroidal anti-inflammatory drug, diclofenac sodium DCF , is associated with a significant increase in the mortality rate of influenza-associated encephalopathy. Activated astrocytes are a source of nitric oxide NO , which is largely produced by inducible NO synthase iNOS in response to proinflammatory cytokines.

Therefore, we investigated whether DCF enhances nitric oxide production in astrocytes stimulated with proinflammatory cytokines. In conclusion, this study demonstrates that iNOS and NO are induced in astrocyte cultures by proinflammatory cytokines. The enhancement of DCF on NO production may explain the significant increase in the mortality rate of influenza-associated encephalopathy in patients treated with DCF.

Immunomodulatory action of Copaifera spp oleoresins on cytokine production by human monocytes. Copaifera spp oleoresins have been used in folk medicine for centuries; nevertheless, its immunomodulatory action has not been investigated. Cell viability was assessed by MTT method after incubation of cells with Copaifera spp. Noncytotoxic concentrations of oleoresins were incubated with human monocytes from healthy donors, and cytokine production was determined by ELISA.

HPLC-MS analysis for terpenes allowed the identification of six diterpene acids and one sesquiterpene acid. Oleoresins exerted no cytotoxic effects on human monocytes. Copaifera oleoresins seemed to exert an activator profile on human monocytes without affecting cell viability.

Such effect may be due to the presence of either diterpene or sesquiterpene acids; however, further studies are necessary to determine the involvement of such compounds in Copaifera immunomodulatory effects. Allergen-induced cytokine production , atopic disease, IgE, and wheeze in children. The early childhood allergen-induced immune responses associated with atopic disease and IgE production in early life are not well understood.

We investigated whether distinct cytokine patterns associated with atopic disease can be detected in immune responses of children. Atopic disease was defined as physician- or nurse-diagnosed eczema or hay fever. In young children at risk for asthma or allergy, decreased allergen-induced IFN-gamma secretion is associated with atopic disease and, in some cases, with increased IgE levels.

Increased allergen-induced IL secretion is most strongly associated with early life increase of IgE. Ozone effect on respiratory syncytial virus infectivity and cytokine production by human alveolar macrophages. AM were first exposed to O3 or to filtered air and then infected with RSV at multiplicities of infection m.

The percentage RSV-infected AM and the amount of infectious virus released by the cells were determined at Days 2 and 4 after infection. No difference in the percentage infected AM or in the amount of infectious RSV produced was found between control and O3-exposed cultures. No difference in IL-1 was seen with m. IL-6 levels were also decreased, but only after infection with m. TNF production was unaffected by O3 at all multiplicities of infection. Statistical analysis of the O3 effect on AM cytokine production induced by the different multiplicities, however, revealed no significant effect of O3.

Based on these observations it appears unlikely that O3 alters susceptibility of AM to infection with RSV, nor does O3 dramatically alter cytokine production in response to RSV since effects on IL-1 and IL-6 secretion were only found with the lowest levels of infection which induced cytokine release. TNF promotes a regulated form of necrosis, called necroptosis, upon inhibition of caspase activity in cells expressing RIPK3.

Because necrosis is generally more pro-inflammatory than apoptosis, it is widely presumed that TNF-induced necroptosis may be detrimental in vivo due to excessive inflammation. However, because TNF is intrinsically highly pro-inflammatory, due to its ability to trigger the production of multiple cytokines and chemokines, rapid cell death via necroptosis may blunt rather than enhance TNF-induced inflammation.

Similarly, necroptosis also suppressed LPS-induced pro-inflammatory cytokine production. Consistent with these observations, supernatants from TNF-stimulated cells were more pro-inflammatory than those from TNF-induced necroptotic cells in vivo.

Thus necroptosis attenuates TNF- and LPS-driven inflammation, which may benefit intracellular pathogens that evoke this mode of cell death by suppressing host immune responses. Laquinimod dampens hyperactive cytokine production in Huntington's disease patient myeloid cells. Huntington's disease HD is a neurodegenerative condition characterized by pathology in the brain and peripheral tissues. Evidence suggests targeting immune disruption may slow disease progression.

Laquinimod had beneficial effects on inflammation, brain atrophy and disease progression in multiple sclerosis MS in two phase III clinical trials. Monocytes from manifest manHD and pre-manifest preHD HD gene carriers and healthy volunteers HV were treated with laquinimod and stimulated with lipopolysaccharide. No differences were observed between laquinimod-treated and untreated conditions.

These results provide evidence that laquinimod dampens hyper-reactive cytokine release from manHD and preHD monocytes, with a much reduced effect on HV monocytes. Evidence suggests targeting CNS and peripheral immune disruption may slow Huntington's disease HD neurodegenerative processes.

The effects of laquinimod, an orally active immunomodulator, on. Analgesic activity of piracetam: effect on cytokine production and oxidative stress. Piracetam is a prototype of nootropic drugs used to improve cognitive impairment. However, recent studies suggest that piracetam can have analgesic and anti-inflammatory effects.

Inflammatory pain is the result of a process that depends on neutrophil migration, cytokines and prostanoids release and oxidative stress. We analyze whether piracetam has anti-nociceptive effects and its mechanisms. Per oral pretreatment with piracetam reduced in a dose-dependent manner the overt pain-like behavior induced by acetic acid, phenyl-p-benzoquinone, formalin and complete Freund's adjuvant.

Piracetam presented analgesic effects as post-treatment and local paw treatment. These results demonstrate that piracetam presents analgesic activity upon a variety of inflammatory stimuli by a mechanism dependent on inhibition of cytokine production and oxidative stress.

Considering its safety and clinical use for cognitive function, it is possible that piracetam represents a novel perspective of analgesic. Multiple effects of TRAIL in human carcinoma cells: Induction of apoptosis, senescence, proliferation, and cytokine production.

TRAIL is a death ligand that induces apoptosis in malignant but not normal cells. Recently the ability of TRAIL to induce proliferation in apoptosis-resistant normal and malignant cells was reported. TRAIL at low concentrations preferentially induced cell proliferation. In conclusion, in cultured human carcinoma cells, TRAIL therapy results in three functional outcomes, apoptosis, proliferation and senescence.

TRAIL-induced proapoptotic and prosurvival responses correlate with the strength of signaling. TRAIL-induced cytokine production is responsible for its proliferative and prosurvival effects. Social role conflict predicts stimulated cytokine production among men, not women. To assess whether perceived role conflict is associated with stimulated pro-inflammatory cytokine production and glucocorticoid sensitivity, and whether these associations are moderated by sex.

Subsequently, participants underwent blood draws and leukocyte response to microbial challenge and glucocorticoid sensitivity were assessed by incubating whole blood with lipopolysaccharide LPS in the presence or absence of hydrocortisone. Dimethyl sulfoxide DMSO is currently used as an alternative treatment for various inflammatory conditions as well as for cancer. Despite its widespread use, there is a paucity of data regarding its safety and efficacy as well as its mechanism of action in human cells.

Specifically, we found that between 0. Thus, while we cannot confirm the efficacy of DMSO as an anti-cancer agent, the use of DMSO in arthritis warrants further investigation to ascertain its therapeutic potential. The CC chemokine ligand 5 CCL5; regulated on activation, normal T expressed and secreted is known to recruit and activate leukocytes; however, its role in altering the responses of host cells to a subsequent encounter with a microbial pathogen has rarely been studied.

Recombinant guinea pig rgp CCL5 was prepared, and its influence on peritoneal and alveolar macrophage activation was examined by measuring cytokine and chemokine mRNA expression in cells stimulated with rgpCCL5 alone or exposed to rgpCCL5 prior to lipopolysaccharide LPS stimulation. Bioactive TNF-alpha protein concentration was measured using the L bioassay.

Both macrophage populations displayed significant enhancement of all the genes and TNF-alpha protein levels when stimulated with rgpCCL5, except for CCL2 in alveolar macrophages. These results further support a role for CCL5 in macrophage activation in addition to chemotactic properties and suggest a role in regulating the inflammatory response to LPS in the guinea pig by modulating the production of proinflammatory cytokines by macrophages.

High dilutions of antimony modulate cytokines production and macrophage - Leishmania L. In previous results mice treated with high dilutions of antimony presented reduction of monocyte migration to the site of infection with increase in B lymphocytes population in the local lymph node.

To know the mechanisms involved, a series of in vitro studies was done, using co-cultures of macrophages RAW The assays were performed in quadruplicate. Cells treated with AC 30cH 10 M and AC cH 10 M presented a temporary reduction of the spreading after 02h of incubation, followed by increase after 48h, being the most significant increase observed after the AC cH treatment.

However, the percentage of internalized parasites at 48, 96 and h of incubation was also higher in cells treated with AC cH. It is suggested that the AC cH improves the ability of phagocytes to internalize the parasites, but not to digest them. The cytokines -chemokines panel corroborated these results. In addition, the production of MIP-1 beta CCL4 , a chemokine involved in chronic inflammation, was also reduced after h.

Since this cytokine is an important chemokine for monocytes, it explains the results obtained formerly in vivo. The morphology of macrophages after acridine orange staining revealed that the treatment with AC 30cH reduced substantially the acid vacuoles in the cytoplasm, indicating a certain inability of these cells to digest the parasites.

On the other hand, a large. Concurrent detection of secreted products from human lymphocytes by microengraving: cytokines and antigen-reactive antibodies. Bradshaw, Elizabeth M. Cell surface determinants, cytokines and antibodies secreted by hematopoietic cells are used to classify their lineage and function. Currently available techniques are unable to elucidate multiple secreted proteins while also assigning phenotypic surface-displayed markers to the individual living cells.

These data demonstrate the utility of microengraving for interrogating multiple phenotypes of single human cells concurrently and for detecting rare populations of cells by their secreted products. Necroptotic cells release find-me signal and are engulfed without proinflammatory cytokine production.

Although phagocytosis of apoptotic cells has been extensively investigated, how necroptotic cells are engulfed has remained elusive. Here, we investigated how necroptotic cells attracted and were engulfed by macrophages. We found that necroptotic cells induced the migration of THP-1 cells in a transwell migration assay. Further analysis showed that ATP released from necroptotic cells acted as a find-me signal that induced the migration of THP-1 cells.

We also found that Annexin V blocked phagocytosis of necroptotic cells by macrophages. Furthermore, necroptotic cells were shown to be silently cleared by macrophages without any proinflammatory cytokine production. These data uncover an evolutionarily conserved mechanism of the find-me signal in different types of cell death and immunological consequences between apoptotic and necroptotic cells during phagocytosis. Peerzada, Habibullah; Ghandi, Jay A. Additionally, METH adversely impacts immunological responses, which might contribute to the higher rate and more rapid progression of certain infections in drug abusers.

However no studies have shown the impact of METH on inflammation within specific organs, cellular participation and cytokine production. Using a murine model of METH administration, we demonstrated that METH modifies, with variable degrees, leukocyte recruitment and alters cellular mediators in the lungs, liver, spleen and kidneys of mice. Our findings demonstrate the pleotropic effects of METH on the immune response within diverse tissues.

These alterations have profound implications on tissue homeostasis and the capacity of the host to respond to diverse insults, including invading pathogens. Visner, Gary A. In this study, we tested the hypothesis that pirfenidone has immune modulating activities and evaluated its effects on the function of T cell subsets, which play important roles in allograft rejection.

Additionally, pirfenidone inhibited TCR-induced production of multiple pro-inflammatory cytokines and chemokines. The extract and essential oil of clove Syzygium aromaticum are widely used because of their medicinal properties. Eugenol is the most important component of clove, showing several biological properties.

Concentrations that inhibited the production of cytokines were used before or after incubation with lipopolysaccharide LPS , to verify a preventive or therapeutic effect. Culture supernatants were harvested for measurement of cytokines by enzyme-linked immunosorbent assay. Moore, Julie M. Pregnant women are at an increased risk for malarial infection.

Plasmodium falciparum accumulates in the placenta and is associated with dysregulated immune function and poor birth outcomes. Malarial pigment hemozoin also accumulates in the placenta and may modulate local immune function. In this study, the impact of hemozoin on cytokine production by intervillous blood mononuclear cells from malaria-infected placentas was investigated. The findings suggest that the influences of hemozoin accumulation and high-density parasitemia on placental cytokine production are not equivalent and may involve different mechanisms, all of which may operate differently in the context of HIV infection.

Cytokine production dysregulated by accumulation of hemozoin or high-density parasitemia may induce pathology and impair protective immunity in HIV-infected and -uninfected women. Invasion of human aortic endothelial cells by oral viridans group streptococci and induction of inflammatory cytokine production.

Oral viridans group streptococci are the major commensal bacteria of the supragingival oral biofilm and have been detected in human atheromatous plaque. Atherosclerosis involves an ongoing inflammatory response, reportedly involving chronic infection caused by multiple pathogens.

The aim of this study was to examine the invasion of human aortic endothelial cells HAECs by oral viridans group streptococci and the subsequent cytokine production by viable invaded HAECs. The invasion of HAECs by bacteria was examined using antibiotic protection assays and was visualized by confocal scanning laser microscopy.

The production of cytokines by invaded or infected HAECs was determined using enzyme-linked immunosorbent assays, and a real-time polymerase chain reaction method was used to evaluate the expression of cytokine messenger RNA. The oral streptococci tested were capable of invading HAECs. The number of invasive bacteria increased with the length of the co-culture period.

After a certain co-culture period, some organisms were cytotoxic to the HAECs. These results suggest that oral streptococci may participate in the pathogenesis of atherosclerosis. Glucocorticoid effects on sheep peripheral blood mononuclear cell proliferation and cytokine production under in vitro hyperthermia. The present experiment aimed at understanding the effects of cortisol levels on sheep peripheral blood mononuclear cell PBMC proliferation and cytokine production during hyperthermia.

Bromodeoxyuridine assay was performed on cells to determine cell proliferation. The interaction of hyperthermia and cortisol level affected both cell proliferation and IL production. These data confirmed that cortisol concentration simulating stress condition could have a role in the immune system of sheep via mediating cellular homeostasis in the condition of hyperthermia. The negative effects of. Enrofloxacin in therapeutic doses alters cytokine production by porcine PBMCs induced by lipopolysaccharide.

The effect of enrofloxacin on cytokine secretion by porcine peripheral blood mononuclear cells PBMCs was studied. Pigs from group E received enrofloxacin at therapeutic dose for 5 consecutive days. Production of cytokines and stimulation of resistance to viral infection in human leukocytes by Scutellaria baicalensis flavones.

Extracts of Scutellaria baicalensis display a wide spectrum of antiviral activity. It was of great interest to check the effect of baicalein and wogonin preparations on two important mechanisms of innate immunity: the secretion of cytokines and the natural resistance of human leukocytes to viral infection.

To study the effect of S. Four pulverized preparations obtained from roots of Scutellaria and a Sigma-Aldrich preparation of purified baicalein were used in the study. Extract from S. Recent studies have shown that some flavonoids are modulators of proinflammatory cytokine expression. Gefitinib and pyrrolidine dithiocarbamate decrease viral replication and cytokine production in dengue virus infected human monocyte cultures.

The epidermal growth factor receptor EGFR and nucleotide-binding and oligomerization-domain containing 2 NOD2 are important in cancer and in microbial recognition, respectively. These molecules trigger intracellular signaling pathways inducing the expression of inflammatory genes by NF-kB translocation. The role of JAK-3 in TLR-mediated innate immune responses is poorly understood, although the suppressive function of JAK3 inhibition in adaptive immune response has been well studied.

In this study, we found that JAK3 inhibition enhanced TLR-mediated immune responses by differentially regulating pro- and anti- inflammatory cytokine production in innate immune cells. At the transcriptional level, JAK3 knockout lead to the increased phosphorylation of STATs that could be attenuated by neutralization of de novo inflammatory cytokines.

Additionally, higher neutrophil infiltration, IL expression, and intestinal epithelium erosion were observed in JAK3 knockout mice. These findings demonstrate the negative regulatory function of JAK3 and elucidate the signaling pathway by which JAK3 differentially regulates TLR-mediated inflammatory cytokine production in innate immune cells. In this study, we have attempted to combine standard immunological assays with the cellular resolving power of the flow cytometer to positively identify the specific cell types involved in spaceflight-induced immune alterations.

The duration of these missions ranged from 10 to 18 days. Absolute levels of peripheral granulocytes were significantly elevated following spaceflight, but the levels of circulating lymphocytes and monocytes were unchanged. Lymphocyte subset analysis demonstrated trends towards a decreased percentage of T cells and an increased percentage of B cells. Nearly all of the astronauts exhibited an increased CD4:CD8 ratio, which was dramatic in some individuals.

Although no significant differences were seen in the expression of the cellular activation markers CD69 and CD25 following exposure to microgravity, significant alterations were seen in cytokine production in response to mitogenic activation for specific subsets. Cytokine production of the neutrophils and macrophages in time of phagocytosis under influence of infrared low-level laser irradiation. As the object of analysis we used peritoneal macrophages from white mice and neutrophils from peripheral blood of healthy donors.

We used the laser diod with spectrum maximum of nm with doses , and mJ exposition , and s respectively; capacity - 5 mW. We found dynamics in production of the cytokines , which was different for the neutrophils and macrophages. We showed that the infrared LLLI has significant stimulating activity on the proinflammatory cytokines production by neutrophils and macrophages. Moreover we revealed dynamics changing in the Il-8 and Il-4 production. Anesthesiologists at work: an increase in pro-inflammatory and Th2 cytokine production , and alterations in proliferative immune responses.

Anesthesiologists are a population at high risk of alcohol and drug abuse, depression, suicide, and psychiatric hospitalization. The impact of their working milieu on specific immune indices has scarcely been studied, and it is assumed that immune perturbations may contribute to some of the above risks. This study took advantage of an unplanned, 3-month long strike of anesthesiologists, and explored its relations to specific immune measures.

We assessed induced cytokine production and lymphocytes proliferative responses in blood samples taken from 10 anesthesiologists just before the strike and at its end, after a long period of markedly reduced workload. The results indicated that the proliferative responses to phytohemagglutinin PHA and concanavalin A Con A were significantly lower at the end of the strike.

At this time point, we observed a significant decrease in the production of interleukin-6 IL-6 , IL and IL1ra levels, and a significant increase in IL-2 production. A strong trend towards a decline in tumor necrosis factor-alpha TNF-alpha levels was evident, while levels of IL-1beta were unchanged. These findings suggest that the working conditions of anesthesiologists are associated with specific immune alterations, including a shift towards a Th2 cytokines ' dominance, and an elevated pro-inflammatory cytokine response.

A reduced Th1 profile has been related to increased susceptibility to infections, and high pro-inflammatory cytokine levels were recently proposed as etiological factors in cardiovascular diseases and in depression. Cytokine production in patients with papillary thyroid cancer and associated autoimmune Hashimoto thyroiditis.

Hashimoto thyroiditis HT is the most frequent thyroid autoimmune disease, while papillary thyroid cancer PTC is one of the most common endocrine malignancies. The aim of this study was to analyze cytokine profiles in patients with PTC accompanied with autoimmune HT in comparison with those in patients with PTC alone or HT alone and healthy subjects.

Cytokine levels were determined in supernatants obtained from phytohemagglutinin PHA -stimulated whole blood cultures in vitro. Suppressed cytokine production in whole blood cultures is related to iron status and is partially corrected following weight reduction in morbidly obese pre-menopausal women.

Assess ex vivo whole-blood cytokine production and its association with iron status in obese versus non-obese women. Determine the change in ex vivo whole-blood cytokine production six months after restrictive bariatric surgery in the obese group. Subjects were 17 obese BMI: Costa, Pedro A.

The function and regulation of the immune response triggered during malaria is complex and poorly understood, and there is a particular paucity of studies conducted in humans infected with Plasmodium vivax. While it has been proposed that T-cell-effector responses are crucial for protection against blood-stage malaria in mice, the mechanisms behind this in humans remain poorly understood.

Experimental models of malaria have shown that the regulatory molecules, cytotoxic T-lymphocyte attenuator-4 CTLA-4 , lymphocyte activation gene-3 LAG-3 , and programmed death-1 PD-1 are involved in the functional impairment of T cells during infection. Our goal was to define the role of these molecules during P. We demonstrate that infection triggers the expression of regulatory molecules on T cells. Importantly, simultaneously blockade of the CLTA-4, PD-1, and T-cell immunoglobulin and mucin—3 signaling restores the cytokine production by antigen-specific cells.

These data support the hypothesis that upregulation of inhibitory receptors on T cells during P. In particular, IgA has an O-linked sugar chain similar to that in the group-specific component Gc protein, a precursor of the Gc protein-derived macrophage-activating factor GcMAF.

John G. Delinassios , All rights reserved. Both variants are contemporarily expressed by human T cells. This might represent an internal regulatory pathway of TH cell responses and might contribute to the modulation of chronic inflammatory reactions, including allergy. Markel, Troy A. End-organ ischemia is a common source of patient morbidity and mortality.

Stem cell therapy represents a novel treatment modality for ischemic diseases and may aid injured tissues through the release of beneficial paracrine mediators. Female bone marrow mesenchymal stem cells MSCs have demonstrated a relative resistance to detrimental TNF receptor 1 TNFR1 signaling and are thought to be superior to male stem cells in limiting inflammation.

After 24 h, supernatants were collected and measured for cytokines. As a result, male, but not female stem cell cytokine production may be mediated through TNFR2 signaling cascades. Major depressive disorder MDD is frequently diagnosed in military personnel returning from deployment. Literature suggests that MDD is associated with a pro-inflammatory state. To the best of our knowledge, no prospective, longitudinal studies on the association between development of depressive symptomatology and cytokine production by peripheral blood leukocytes have been published.

The aim of this study was to investigate whether the presence of depressive symptomatology six months after military deployment is associated with the capacity to produce cytokines , as assessed before and after deployment. Depressive symptoms and LPS- and T-cell mitogen-induced production of 16 cytokines and chemokines in whole blood cultures were measured before T0 , 1 T1 , and 6 T2 months after return from deployment.

Exploratory structural equation modeling ESEM was used for data reduction into cytokine patterns. Individuals with depressive symptoms after deployment showed higher T-cell cytokine production before deployment. Moreover, pre-deployment T-cell cytokine production significantly predicted the presence of depressive symptomatology 6 months after return.

There was an increase in T-cell cytokine production over time, but this increase was significantly smaller in individuals developing depressive symptoms. T-cell chemokine and LPS-induced innate cytokine production decreased over time and were not associated with depressive symptoms. These results indicate that increased T-cell mitogen-induced cytokine production before deployment may be a vulnerability factor for development of depressive symptomatology in response to deployment to a combat-zone.

In addition, deployment to a combat-zone affects the. Sampey, Gavin C. HIV-1 infection results in a chronic illness because long-term highly active antiretroviral therapy can lower viral titers to an undetectable level. However, discontinuation of therapy rapidly increases virus burden. Moreover, patients under highly active antiretroviral therapy frequently develop various metabolic disorders, neurocognitive abnormalities , and cardiovascular diseases. Collectively, these results imply that exosomes containing TAR RNA could directly affect the proinflammatory cytokine gene expression and may explain a possible mechanism of inflammation observed in HIVinfected patients under cART.

CD38 exacerbates focal cytokine production , postischemic inflammation and brain injury after focal cerebral ischemia. Converging evidence suggests that inflammatory processes significantly influence brain injury and clinical impairment in ischemic stroke.

Although early studies suggested a key role of lymphocytes, recent data has emphasized the orchestrating function of innate immunity, i. The bifunctional receptor and ectoenzyme CD38 synthesizes calcium-mobilizing second messengers e. Therefore, we investigated the dynamics of CD38 in stroke and the impact of CDdeficiency on cytokine production , inflammation and cerebral damage in a mouse model of cerebral ischemia-reperfusion.

We show that the local expression of the chemokine MCP-1 was attenuated in CDdeficient mice compared with wildtype mice after focal cerebral ischemia and reperfusion. In contrast, no significant induction of MCP-1 expression was observed in peripheral blood after 6 hours.

Flow cytometry analysis revealed less infiltrating macrophages and lymphocytes in the ischemic hemisphere of CDdeficient mice, whereas the amount of resident microglia was unaltered. Finally, we demonstrate that CDdeficiency decreases the cerebral ischemic injury and the persistent neurological deficit after three days of reperfusion in this murine temporary middle cerebral artery occlusion tMCAO model.

CD38 is differentially regulated following stroke and its deficiency attenuates the postischemic chemokine production , the immune cell infiltration and the cerebral injury after temporary ischemia and reperfusion. Therefore CD38 might prove a therapeutic target in ischemic stroke. Copyright c by Academic Press, Inc.

Organic UV filters exposure induces the production of inflammatory cytokines in human macrophages. Organic ultraviolet UV filters, found in many personal care products , are considered emerging contaminants due to growing concerns about potential long-term deleterious effects. We investigated the immunomodulatory effects of four commonly used organic UV filters 2-hydroxymethoxybenzophenone, BP-3; 4-methylbenzylidene camphor, 4-MBC; 2-ethylhexyl 4-methoxycinnamate, EHMC; and butyl-methoxydibenzoylmethane, BDM on human macrophages.

After exposure to the UV filters, a significant 1. Our results therefore demonstrate that exposure to the four organic UV filters investigated may alter human immune system function. It provides new clue for the development of asthma or allergic diseases in terms of the environmental pollutants.

Effects of trans-stilbene and terphenyl compounds on different strains of Leishmania and on cytokines production from infected macrophages. Most of the antileishmanial modern therapies are not satisfactory due to high toxicity or emergence of resistance and high cost of treatment. Previously, we observed that two compounds of a small library of trans-stilbene and terphenyl derivatives, ST18 and TR4, presented the best activity and safety profiles against Leishmania infantum promastigotes and amastigotes.

In the present study we evaluated the effects of ST18 and the TR4 in 6 different species of Leishmania and the modifications induced by these two compounds in the production of 8 different cytokines from infected macrophages. We observed that TR4 was potently active in all Leishmania species tested in the study showing a leishmanicidal activity higher than that of ST18 and meglumine antimoniate in the most of the species.

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