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Harmony Touch™ User Guide...
� � � � � � � � � � � � � � � Connecting your Harmony Touch to your Computer � � � � � � Setting up Devices to Control � � � � � � �...
� � � � � � � 30 � � � � � � 30 To Access Your Delay and Speed Settings From Your Harmony Touch Resetting your Remote � � � � � � � � � � �...
Harmony Touch is not Detected by my Computer � � � � � � � � � � � � 38 Devices do not Respond to Harmony Touch at All � � � � � � � � � � � �...
Harmony Touch is an activity-based Infrared (IR) universal remote control. Using the myharmony.com web- based setup, you can control up to 15 different devices. Using its Smart State technology, the Harmony Touch keeps track of the devices it controls, which allows it to always know which devices are on and off, and what input each device is currently set to.
Harmony Touch. By connecting your Harmony Touch to your Mac or Windows computer and following the instructions at myharmony.com, you can enter your device information, create activities and choose your favorites. With this information, myharmony.com will configure your Harmony Touch to control your entertainment system.
* Windows 7 and 8 support both 32 and 64 bit browsers Your Myharmony�com Account To use your Harmony Touch, you will need an account where you can enter the details for your devices, activities and favorites. Using your web browser, go to http://myharmony.com If this is your first time using myharmony.com, click the Create an Account button on the left.
3. When both plugins are installed, your browser will automatically refresh and continue the set up process. Connecting your Harmony Touch to your Computer Whether you are creating a new account or adding Harmony Touch to an existing account, you will be asked to connect Harmony Touch to your computer using the USB cable provided.
Logitech Harmony Touch Setting up Devices to Control The first step to setting up Harmony Touch is to tell myharmony.com what devices you have in your entertainment system that you want to control. Using the Setup Guide that came with your Harmony Touch, gather the manufacturer, model and connection information for each device.
If you have a previous Harmony remote such as Harmony One, Harmony 1100 or Harmony 880, you can reuse the device and activity settings from that remote to program your Harmony Touch. To copy a previous Harmony remote's settings, click the Copy button shown here. You will be asked to enter the ID and password for your previous account.
5. Prepare to use your original device remote to teach your Harmony Touch a few commands.Point the end of the original remote at the sensor on the back of your Harmony Touch, as shown.: 6. F ollow the instructions to push buttons on the original remote to teach commands on the Harmony Touch. Adding an Unknown Device CAUTION: This process should only be used if you cannot find a match for your device.
Logitech Harmony Touch 1. Click the button to confirm the Manufacturer and Model Number for the device and click OK. 2. Click Select your device type to display a list of device types. 3. Select the device type that best describes your device and click Next. Based on your selection, another page displays to get more information about your device.
Logitech Harmony Touch Setting up Activities After you have added devices, you can create activities. Use activities to manage how you use your devices together. For example, if you have a TV, AV receiver, and Cable or Satellite box, you will want to control all of these devices when you watch TV.
Logitech Harmony Touch 9. Choose the TV input from the list displayed. 10. Choose the AV receiver input from the list displayed. 11. Confirm that the devices and inputs are correct for your Watch TV activity and click Finish. Following is an example of the completed Watch TV activity setup.
Use this page to arrange your favorites in the order you want to see them on your Harmony Touch screen. You can drag and drop each channel icon to move or remove it from the screen as desired.
Logitech Harmony Touch Setting up Favorites When Your Provider is not Listed In some cases your provider will not be listed in the favorites setup pages. You can still set up favorites using the channel list and assigning a channel number for each channel manually.
Logitech Harmony Touch Setting up Favorites Manually If no channel provider or channel list is available for your area, you can still set up favorites manually. 1. Select your country. 2. If channel information is not available for your country, the page will remain as is, and Next will be enabled.
Upgrading your Harmony Touch firmware When a new firmware is available for your Harmony Touch, it will automatically be applied the next time you sync your remote. The remote may reboot multiple times during this process. Once the firmware update is complete, your Harmony Touch will sync with myharmony.com as usual.
After completing the first time setup process, you can return to the myharmony.com website to make setup changes and sync them on your Harmony Touch. You can also change basic settings directly on your remote. Follow these steps to manage your remote setup on myharmony.com 1. Login to your myharmony.com account.
Modifying Device Settings You can also modify the way your Harmony Touch will control each of your devices. These topics are covered in the Settings and Advanced Features of this guide.
6. Click Browse to choose an icon from your system or accept the displayed icon. This icon will also be used to represent your activity on your Harmony Touch. 7. Click Next. The list of devices available for this activity is displayed, with suggested devices checked.
Logitech Harmony Touch To Change Activity Settings 1. Click Activities. Your list of Activities displays. 2. Select the activity to change settings. 3. Click Change your settings. The Rename Activity page displays. This is the first page in the process for changing the activity settings.
On the Manage Favorite Channels screen, you can drag and drop your current favorites into the exact order you want them to appear on your Harmony Touch. Here you can also drag a favorite to the trash can icon to...
Logitech Harmony Touch To add a favorite channel that does not appear in the list on the previous screen, select Add a favorite channel to manually enter the name and number for that channel. Optionally, you can upload an icon for that channel from your computer's file system:...
Logitech Harmony Touch Channel Tuning Settings If your favorite channels aren't working properly, you may need to adjust how your remote sends the channel tuning commands to your TV or cable/satellite set top box. Click the Adjust how you change channels link on the main Favorites screen in your myharmony.com account.
For example, to Watch TV, Harmony Touch will turn on your TV, turn on your cable set top box, turn on your AV receiver, set the input on your TV to HDMI 2, set the input on your AV receiver to HDMI 1.
Logitech Harmony Touch Activity Commands When your activity starts, all the buttons on Harmony Touch are now configured to control that activity. For example, the volume button will control your AV receiver while the Play and Pause buttons and the directional navigation will control your cable or satellite set top box.
Logitech Harmony Touch Gesture Control Each activity will include an option to control your devices using simple swipes and taps on the Harmony Touch screen. 5 different gestures are supported: • Swipe up • Swipe down • Swipe left •...
Tap any favorite to tune to that channel. TIP: If your entertainment system is off or if you are in another activity when you select a favorite, Harmony Touch will automatically start the Watch TV activity and tune to the selected channel. Managing Your Favorites Tap the edit button at the bottom right of the Favorites screen to manage your favorites.
Turning your system off The Off button has a special use on the Harmony Touch. Instead of using the Off button to turn off each device separately, you use the Off button to turn off all of the devices for an activity. For example, in a Watch TV activity that uses a TV, Cable Box, and AV Receiver, pressing the Off button will turn off all three devices at the same time.
When you pick up the Harmony Touch from the dock, it turns on automatically. Place the remote back in the dock to charge the battery again after use.
After completing the first time setup process, many settings can be changed on the remote that let you personalize your Harmony Touch or affect how it controls your entertainment system. On the Harmony Touch home screen, swipe all the way to the right, to access the Settings menu.
Choose a different input and tap the "back" arrow. Reorder Activities If you want to rearrange your activities on your Harmony Touch, you can do that here. Tap and hold on each activity, then drag up or down on the screen to move it to a new position.
Logitech Harmony Touch Device Delay Settings Delays and speed settings can be changed to alter the speed with which the Harmony Touch sends commands to your devices. They can be changed from within your myharmony.com account, or on your remote.
Logitech Harmony Touch To Access Your Delay and Speed Settings From Myharmony�com 1. Click Devices 2. Click the device for which you would like to view or modify delays and speed settings 3. Click Change Device Settings. The Device Settings page opens. 4. S elect Device Delay Settings and click Next. The Device Delay Settings page opens.
Logitech Harmony Touch Advanced Features This section covers more advanced settings of your Harmony Touch that let you perfect how it controls your entertainment system. Activity Sequences Adding steps to an activity allows you to customize, for example, your Watch a Movie activity to automatically send the Play command to start your movie.
Logitech Harmony Touch Button Sequences A button sequence is a set of commands that can be grouped together and assigned to one button on your remote, helping to automate certain tasks. They are very similar to Activity Sequences above except you can perform them any time within the activity by pressing the assigned button whereas Activity Sequences are only performed when starting or stopping the activity.
Device Settings Adding/Fixing Device Commands If your Harmony Touch is missing a specific command for one of your devices, it is possible to "teach" this command to your Harmony Touch, using a remote control on which this command is present.
Device Power Settings The power settings feature allows you to modify the way Harmony Touch powers devices on and off, or to configure your remote to leave certain devices on all the time. Certain devices such as projectors may need additional commands to power up properly—you can add or modify these here as well.
Logitech Harmony Touch Device Input Settings The input settings feature allows you to modify the way Harmony Touch accesses inputs on your devices. Input settings also allow you to modify the number and order of inputs for the devices that Harmony controls.
Logitech Harmony Touch To Change or Fix What a Button Does 1. Click Buttons. The change or fix button operations page displays. 2. Under Change or Fix what a button does, point to Select an activity or device—the remote buttons are highlighted to show that the button options will be modified.
Support Troubleshooting Harmony Touch Harmony Touch is not Detected by my Computer If you've connected your Harmony Touch, but it does not appear to be connected, try the following steps. After each step, try plugging in the device again. •...
Always exclusively use your Harmony Touch to control your devices. Do not use other remotes to control your devices, or manually turn them on or off or change inputs. The Harmony Touch is unable to track changes that it does not make and will continue to assume your devices are in the state that it has left them in.
© 2012 Logitech. All rights reserved. Logitech, the Logitech logo, and other Logitech marks are owned by Logitech and may be registered. Blu-ray Disc is a registered trademark of the Blu-ray Disc Association. HDMI, the HDMI logo, and High-Definition Multimedia Interface are trademarks or registered trademarks of HDMI Licensing, LLC in the United States and other countries.


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10 Things You Didn't Know About Paul Simon
Many a musician would give their left arm and any necessary collateral materials to have a career like the one that Paul Simon has enjoyed. He found huge success as one half of Simon & Garfunkel and went on to have an equally fruitful solo career after the pairing dissolved. Today, at age 70, Simon is still creatively juiced, having released his most recent album 'So Beautiful Or So What' to positive fanfare from both critics and fans alike. Here's a look at 10 things you might be surprised to learn about Paul Simon.

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Efforts to develop functional tissue-engineered blood vessels have focused on improving the strength and mechanical properties of the vessel wall, while the functional status of the endothelium within these vessels has received less attention. Endothelial cell (EC) function is influenced by interactions between its basal surface and the underlying extracellular matrix. In this study, we utilized a coculture model of a tissue-engineered blood vessel to evaluate EC attachment, spreading, and adhesion formation to the extracellular matrix on the surface of quiescent smooth muscle cells (SMCs). ECs attached to and spread on SMCs primarily through the α5β1-integrin complex, whereas ECs used either α5β1- or αvβ3-integrin to spread on fibronectin (FN) adsorbed to plastic. ECs in coculture lacked focal adhesions, but EC α5β1-integrin bound to fibrillar FN on the SMC surface, promoting rapid fibrillar adhesion formation. As assessed by both Western blot analysis and quantitative real-time RT-PCR, coculture suppressed the expression of focal adhesion proteins and mRNA, whereas tensin protein and mRNA expression were elevated. When attached to polyacrylamide gels with similar elastic moduli as SMCs, focal adhesion formation and the rate of cell spreading increased relative to ECs in coculture. Thus, the elastic properties are only one factor contributing to EC spreading and focal adhesion formation in coculture. The results suggest that the softness of the SMCs and the fibrillar organization of FN inhibit focal adhesions and reduce cell spreading while promoting fibrillar adhesion formation. These changes in the type of adhesions may alter EC signaling pathways in tissue-engineered blood vessels.
- focal adhesions
- fibrillar adhesions
- tissue engineering
- integrin
- elastic modulus
during the past decade, there have been great strides toward creating a clinically viable tissue-engineered blood vessel (TEBV) (12). While most studies have focused on improving the strength and mechanical properties of the vessel wall, the functional status of the endothelium within these TEBVs has received less attention, despite having a critical role in vasodilation, platelet coagulation and clot formation, immune response, and water and solute permeability (3).
Endothelial cell (EC) function is influenced by interactions between integrins on the basal surface and the underlying extracellular matrix (ECM) (9, 21). The integrins involved with attachment and the proteins that interact with the cytoplasmic domains of these integrins determine the type of adhesion that is formed. These adhesions include nascent focal complexes that form during cell spreading, classical focal adhesions, and elongated and elliptical fibrillar adhesions that arise when fibrillar fibronectin (FN) forms (for a review, see Ref. 32). Late focal complexes and focal adhesions consist of many of the same proteins, such as αvβ3-integrin, vinculin, paxillin, and focal adhesion kinase (FAK); in contrast, fibrillar adhesions lack the aforementioned proteins and are rich in α5β1-integrin and tensin. Tensin can also be found in focal adhesions. Focal adhesions are primarily located at the cell periphery with very low colocalization with FN fibrils, whereas fibrillar adhesions form along FN fibrils and are located in the central portion of the cell (14).
Zamir et al. (33) and others (for a review, see Ref. 6) showed that when cells initially attach to the substrate, integrins cluster and the properties of the substrate determine the type of adhesions that are formed. If the underlying FN matrix is deformable, then fibrillar adhesions will form (14). If the substrate is rigid and contains a nondeformable matrix, then the immobilized adhesion complexes produce high tension that induces the formation of focal adhesions (14). The types of adhesions the ECs create and the integrins involved can affect EC function (9, 21).
In vivo and with TEBVs, ECs adhere to the ECM and underlying SMCs that are much softer than the substrates used for two-dimensional culture in vitro. We (28) developed and characterized a direct coculture system as a mimic of TEBVs. In coculture, SMCs organize FN into fibrils prior to the attachment of ECs. Thus, these substrate conditions in coculture and TEBVs may affect the adhesion of ECs. In this study, we tested the hypothesis that the substrate conditions in coculture reduce EC focal adhesion formation and promote the formation of fibrillar adhesions. To quantitatively assess this hypothesis, we compared EC adhesion on SMCs with EC adhesion on tissue culture plastic or polyacrylamide surfaces of different elastic moduli.
Media composition and coculture formation.
All media were supplemented with 1× antibiotic-antimycotic (GIBCO-BRL, Carlsbad, CA). Human aortic SMCs (Cambrex, Walkersville, MD) were expanded with SMC growth media containing smooth muscle basal media (SmBM; Cambrex) supplemented with SmGM-2 singleQuots (Cambrex). To switch SMCs from a proliferative state to a quiescent state, SMCs were maintained in serum-free quiescent media composed of DMEM-F-12 (GIBCO-BRL) supplemented with 1× insulin-transferrin-selenium (GIBCO-BRL). Human aortic ECs (Cambrex) were expanded in endothelial basal media-2 (EBM-2; Cambrex) supplemented with EGM-2 SingleQuots (Cambrex). Cocultures were maintained in a coculture medium composed of medium 199 (GIBCO-BRL) supplemented with 5.5% human serum (Sigma, St. Louis, MO) and 1× insulin-transferrin-selenium (GIBCO-BRL). All cells were trypsinized with 0.025% trypsin-EDTA (Clonetics) for 5 min at 37°C except when ECs were separated from SMCs in coculture.
To prepare cocultures, SMCs were plated at 75,000 cells/cm2 on tissue culture plastic dishes (Corning, Corning, NY) or polystyrene slideflasks (NUNC, Rochester, NY) that were incubated with 3.3 μg/ml human plasma FN (Sigma) for over 1 h. SMCs (passages 7–11) were grown to confluence in SMC growth media, forming one to three layers. Two days after SMCs had been seeded, media were changed from SMC growth media to SMC quiescent media. Two days after the addition of the quiescent media, human aortic ECs (passages 7–10) were seeded at a confluent density (75,000–100,000 cells/cm2) directly on top of the quiescent SMCs in EC growth media except where noted. After ∼24 h, media were changed to coculture media. All cell cultures were maintained in a tissue culture incubator at 37°C and 5% CO2, and the medium was exchanged every other day.
Imaging ECs and SMCs in coculture.
To visualize cells, ECs were stained with DiI-Ac-LDL (5 μg/ml, 4 h, 37°C; Biomedical Technologies, Stoughton, MA) while in coculture or with CellTracker orange (2 μM, 10 min, 37°C; Invitrogen, Carlsbad, CA) before being plated.
Fluorescent images were obtained with a confocal laser scanning microscope (LSM 510, Carl Zeiss, Thornwood, NY) or an inverted fluorescent microscope (Zeiss Axiovert S-100, Carl Zeiss) connected to a digital camera (Carl Zeiss). The camera was connected via a frame grabber card (Pr-LG3-01 PCI, Scion, Frederick, MD) to a Macintosh G3 computer (Apple Computer, Cupertino, CA).
Blocking EC attachment.
The EC integrins involved with the initial attachment of ECs to SMCs were determined utilizing monoclonal integrin blocking antibodies. Approximately 200,000 ECs, labeled with CellTracker orange, were suspended in 250 μl of Dulbecco's PBS solution (DPBS; GIBCO-BRL) and rotated in a 1.5-ml tube in the tissue culture incubator for 30 min with no antibodies, 5 μl of mouse anti-human α5β1-integrin (Chemicon, Temecula,CA), mouse anti-human αvβ3-integrin (Chemicon), or a combination of the two antibodies. Next, the cell suspension was combined with 1.75 ml of DPBS and added to either quiescent SMCs or 12-well tissue culture plates that were first incubated with 3.3 μg/ml FN for 1 h, rinsed, and incubated with 1% BSA for 1 h to block nonspecific binding. After a 10-min incubation at 37°C, nonadherent ECs were removed with three DPBS rinses, and adherent cells were fixed with 3.7% paraformaldehyde for 15 min at 37°C. Ten random fluorescent images were captured per sample, and numbers of attached ECs were measured with a custom cell counting Matlab (version 7.0, The MathWorks, Natick, MA) program.
EC spreading.
To determine the spreading rate of ECs plated on SMCs or polyacrylamide gels, ECs were stained with CellTracker orange and plated at 20,000 cells/cm2 in the fully supplemented EBM-2 media. After 30, 75, 120, and 165 min of attachment, approximately six fluorescent images were captured. Image J (version 1.36, National Institutes of Health) was used to trace and measure the cell area of at least 30 ECs per time point. A linear regression between EC area and attachment time was performed to determine the rate of EC spreading.
To block EC spreading, the following antibodies, at a dilution of 1:100, were added to the media after ECs attached and spread for 5 min: no antibody (control), mouse anti-human α5β1-integrin (Chemicon), mouse anti-human αvβ3-integrin (Chemicon), or a combination of the two antibodies. After 1 h of spreading, cultures were fixed with 3.7% paraformaldehyde and imaged. The cell areas of at least 50 ECs were measured per experiment using ImageJ.
Cells were fixed with 3.7% paraformaldehyde for 15 min at 37°C and permeabilized with 0.2% Triton X-100 (Sigma) at room temperature for 5 min. Cells were rinsed with DPBS and then incubated with 10% goat serum (Sigma) for 30 min at 37°C to block nonspecific binding. Primary antibodies [vinculin, 1:100 (Sigma); paxillin, 1:400 (Invitrogen); PY20, 1:100; FN, 1:200; and tensin, 1:25 (BD Biosciences-Pharmingen, San Diego, CA)] were incubated with the cells for 1 h at 37°C in 10% goat serum. Cells were rinsed multiple times and then incubated at 37°C for 45 min in 10% goat serum with Alexa fluor 488 or 546 goat anti-mouse secondary antibody (1:500, Invitrogen). The focal adhesion area was measured by capturing at least 6 images/sample of the vinculin immunofluorescence and using ImageJ to trace and measure the area of at least 30 random focal adhesions/sample.
Green fluorescent protein-vinculin and -tensin expression.
Green fluorescent protein (GFP)-vinculin and GFP-tensin plasmids (provided by S. Lin, D. Lin, and K. Yamada) were selected by ampicillin resistance and amplified in Escherichia coli (DH5α, Invitrogen). ECs were transfected with the GFP-plasmid using a liposomal method (Lipofectin Transfection Reagent, Invitrogen) according to the manufacturer's protocols. Briefly, ECs were grown to 50–70% confluence in six-well plates before transfection; 1 μg/ml of plasmid and 7 μl/ml of Lipofectin were mixed and incubated with Opti-MEM I (Invitrogen) for 20 min to form plasmid-Lipofectin complexes. ECs were then incubated for 2.5 h with Opti-MEM I containing the plasmid-Lipofectin complexes. The medium was exchanged with EC growth media, and cells were allowed to recover overnight.
Separation of ECs from SMCs for total RNA and protein isolation.
Cocultures were washed with DPBS without Ca2+ and Mg2+ to remove media proteins. A 5-min wash was then performed with ice-cold DPBS without Ca2+ and Mg2+. This caused the ECs to contract slightly due to the drastic temperature change, thus allowing more of the trypsin to access EC integrins. Trypsin-EDTA (0.25%) at 37°C was then added to the cells for 5 min. Trypsin was neutralized with Trypsin Neutralizing Solution (Clonetics), and the cell suspension was pipetted multiple times to break any remaining bonds between cells. Cells were pelleted and resuspended with 1 ml of 0.1% BSA in DPBS. ECs and SMCs were then separated using magnetic beads.
A volume of 25 μl of CD31 EC Dynabeads (Invitrogen) was mixed with the cell sample for 20 min at room temperature. The bead-cell solution was placed in the Dynal MPC Magnet (Invitrogen), where the EC-bead complex was pulled to the side of the magnet and the supernatant (SMCs in suspension) was removed. EC-bead complexes were resuspended in 0.1% BSA in DPBS. The solution was placed in the magnet, and the supernatant was discarded. ECs were then washed three more times to increase EC purity. The magnetic bead separation technique yielded an EC purity of 98.7 ± 0.4, which was determined by identification of positive Ac-LDL-DiI staining of separated ECs (n = 3). Purified ECs were then lysed with lysis buffer for either RNA or protein isolation.
Total EC RNA was isolated from the EC-bead complex using a commercially available RNA isolation kit (High-Pure Total RNA Isolation Kit, Roche Applied Science, Indianapolis, IN). Total EC protein was isolated with Cell Lytic-M (Sigma) supplemented with Protease Inhibitor Cocktail (1:10, Sigma). The protein lysis buffer was rotated with the EC-bead complexes for 15 min at 4°C, and the cell debris and beads were pelleted with centrifugation so that the lysate could be isolated.
Quantitative real-time RT-PCR.
RNA purity and quantity were measured using a NanoDrop Spectrophotometer (NanoDrop Technologies, Wilmington, DE). Total RNA (50 ng) was reverse transcribed using the cDNA Synthesis Kit (Bio-Rad, Hercules, CA) and a MyCycler (Bio-Rad) thermal cycler. One cycle of 5 min at 25°C, 30 min at 43°C, and 5 min at 85°C was performed. Primers (Integrated DNA Technologies, Coralville, IA), RNAse free water, and IQ SYBR Green Supermix (Bio-Rad) were combined with the cDNA samples and placed in a MyIQ Single Color Real-Time PCR Detection System (Bio-Rad). A two-step cycle configuration was performed with an initial denaturation for 3 min at 95°C and 50 cycles at 95°C for 15 s and 61°C for 1 min. All samples were performed in triplicate for all genes. The 2 method was used to determine relative gene expression (17), where CT is threshold cycle. Primers were selected based on the gene sequence (Pub-Med) of interest and using Primer3 shared software (23). The primers used were as follows: 1) β2-microglobulin, 5′-GGCTATCCAGCGTACTCCAAAG-3′ and 5′-CAACTTCAATGTCGGATGGATG-3′; 2) α-actinin, 5′-GGCAAGATGAGAGTGCACAA-3′ and 5′-AGATGTCCTGGATGGCAAAG-3′; 3) paxillin, 5′-TGATAGGATTTGGGGCAGAG-3′ and 5′-CCAGTGGAGTGGTTTGGACT-3′; 4) tensin, 5′-GGCTTAGAGCGAGAGAAGCA-3′ and 5′-CCCGTCCAGAGAAGAGAGTG-3′; 5) vinculin: 5′-CTTTGCTGCTACAGGGGAAG-3′ and 5′-GGATATGGGACGGGAAGTTT-3′; 6) FAK, 5′-TTATTGGCCACTGTGGATGA-3′ and 5′-TACTCTTGCTGGAGGCTGGT-3′; and 7) RhoA, 5′-AAGGACCAGTTCCCAGAGGT-3′ and 5′-TTCTGGGGTCCACTTTTCTG-3′.
Western blot analysis.
The BCA protein assay (Sigma) was used to determine the protein concentration in the lysate. Protein lysate (20 μg) was boiled with Lamaelli sample buffer (1:1, Bio-Rad) for 5 min. The resulting solution was then added to a 7.5% Tris·HCl Ready-Made gel (Bio-Rad), and electrophoresis was completed at 150 V for ∼35 min. A wet transfer (Towbin buffer) was completed at 100 V and 90 mA for 1 h to transfer vinculin and paxillin from the gel to a polyvinylidene difluoride (PVDF) membrane. Tensin was transferred in Towbin buffer with 20% methanol and 0.05% SDS for 3 h at 500 mA. The membrane was blocked for 1 h with 5% milk. The primary antibody reactive to the protein of choice was incubated with the PVDF membrane overnight at 4°C (vinculin, 1:1,000; paxillin, 1:1,000; and tensin, 1:150) in 1% milk. After multiple washes with Tris-buffered saline (TBS)-Tween solution, the membrane was incubated with a secondary antibody (anti-mouse IgG-horseradish peroxidase, Santa Cruz Biotechnology) in 5% milk at 1:5,000 dilution for 1 h. Enhanced chemiluminescence (Pierce) solution was added to the membrane for 5 min, and the membrane was exposed to CL-XPosure Film (Pierce). Bands were then analyzed using the gel analyzer macro in ImageJ.
Atomic force microscopy.
Samples were probed with an Asylum 1-D Atomic Force Microscope (AFM; Asylum Research, Santa Barbara, CA) in contact mode. A long narrow hydrophilic borosilicate cantilever with a spherical tip (radius = 2.5 μm; Bioforce Nanoscience, Ames, IA) and a nominal spring constant of 60 pN/nm was used to contact the sample. The actual spring constant was determined by a thermal fluctuations method. Samples were probed approximately seven times at random locations. The indentation (δ) was obtained by subtracting the deflection from the movement of the piezoelectric ceramic (z) as follows: δ = z − z0, where z0 is the initial deflection. Force-deflection curves for the polyacrylamide gels and SMCs were obtained using IGOR Pro 5.04B (Wavemetrics, Portland, OR). Since cells are viscoelastic and the mechanical properties vary spatially, the apparent modulus E* was calculated by fitting the force-indentation curves to the following equation: (1) where F is the force, E* is Young's modulus, ν is Poisson's ratio [a value of 0.5 is commonly used for cells (25) and polyacrylamide gels (7)], and R is the radius of the sphere on the tip. To minimize viscous contributions, we indented at 2 μm/s. All plots of force versus δ3/2 had correlation coefficients above 0.98.
Polyacrylamide gel preparation and FN cross-linking.
Polyacrylamide gels were prepared on glass slides as described by Pelham and Wang (22). Briefly, glass slides were treated with aminoproplyltrimethoxysilane and 0.5% glutaraldehyde to create a surface that would firmly adhere to the gel. Mixtures consisting of 5% acrylamide and 0.025–0.1% bis-acrylamide were prepared by mixing 40% acrylamide, 1 M HEPES, 2% bis-acrylamide, and distilled water. Acrylamide solutions were degassed for 20 min to remove oxygen that could inhibit the polymerization of acrylamide. Next, 10% ammonium persulfate and N,N,N′,N′-tetramethylethylenediamine were added to the acrylamide solution to catalyze the cross-linking of the polymer. A volume of 15 μl of the acrylamide solution was pipetted onto the aminosilanized glass slides. A 22-mm-diameter circular glass coverslip was immediately placed over the solution, and gels were allowed to polymerize for 30 min before being rinsed with 50 mM HEPES and removal of the coverslip. Gels were stored in 50 mM HEPES at 4°C and used within 2 wk.
To immobilize FN, a cross-linker, N-sulfosuccinimidyl-6-(4′-azido-2′-nitrophenylamino)-hexanoate (sulfo-SANPAH; Pierce), was added to the surface of the gels at a concentration of 0.5 mg/ml. Sulfo-SANPAH was activated by placing the gels ∼3 in. from four 7.5-W tubes of 302-nm UV light for 8 min. Gels were rinsed, and the UV cross-linking was repeated. Next, gels were incubated with 10 μg/ml FN overnight at 4°C and rinsed with DPBS. To increase the amount of FN matrix, some gels underwent three additional incubations with 10 μg/ml FN for 1 h each at 37°C. FN-coated gels were stored for <1 wk at 4°C before being used.
Statistical analysis.
ANOVA was performed when making multiple comparisons and a Tukey-Kramer analysis was used as a post hoc test. Student's t-test was performed when only two groups of data were compared. A value of P < 0.05 was considered statistically significant. Data are expressed as means ± SE.
α5β1-Integrin controls EC attachment and spreading on SMCs.
The numbers of ECs that attached to SMCs or FN-coated plastic were significantly reduced by blocking the α5β1-integrin complex (n = 3–5; Fig. 1A ). In contrast, the addition of a blocking antibody to αvβ3-integrin alone did not affect EC attachment. EC attachment to SMCs or FN-coated plastic was fully inhibited by simultaneously blocking α5β1- and αvβ3-integrin on both substrates. Thus, ECs attached to SMCs with the same integrins involved in the attachment to FN-coated plastic, suggesting that FN on the surface of SMCs (28) is the major ECM protein influencing EC attachment in coculture.
EC spreading on FN-coated plastic was significantly reduced only by simultaneously blocking α5β1- and αvβ3-integrin, whereas blocking only α5β1-integrin significantly reduced EC spreading on SMCs (n = 5; Fig. 1B). Simultaneously blocking α5β1- and αvβ3-integrin did not further reduce EC spreading on SMCs compared with blocking α5β1-integrin alone. These results suggest that ECs used either α5β1- or αvβ3-integrin complexes to spread on FN-coated plastic, but ECs spread on SMCs primarily through the α5β1-integrin complex. Also, after 1 h of spreading, control ECs on SMCs were approximately half the size of control ECs on FN-coated plastic.
ECs form focal complexes on SMCs and on FN-coated plastic.
Since EC spreading on SMCs was significantly reduced compared with EC spreading on FN-coated plastic (28), we hypothesized that there were differences in focal complex formation. The formation of focal complexes was determined by the positive staining of phosphotyrosine, paxillin, and vinculin along the cell periphery (Fig. 2). After 1 h of spreading, ECs that adhered to SMCs had similar amounts of phosphotyrosine and paxillin at the cell periphery as ECs in monoculture, but there was a noticeable decrease in vinculin recruitment at the cell periphery of ECs in coculture. Also, after 1 h of spreading, ECs in monoculture formed some focal adhesions, but focal adhesion formation was not found in coculture.
ECs on SMCs form fibrillar adhesions and lack focal adhesions.
As cell spreading progresses, focal complexes mature into focal adhesions due to the tension generated along the actin filaments within the cell (for a review, see Ref. 24). Consistent with the literature, subconfluent ECs expressing GFP-vinculin formed focal adhesions 4 h after attachment when adhered to FN-coated plastic (Fig. 3A). In contrast, subconfluent ECs adhered to SMCs did not form focal adhesions. To assess whether focal adhesions formed at later times in coculture, we used immunofluorescence to identify focal adhesions 2 days after attachment (not shown). Again, ECs adhered to FN-coated plastic formed focal adhesions. ECs adhered to SMCs did not form focal adhesions, although SMCs in coculture did form focal adhesions to the underlying plastic substrate. ECs in coculture lacked vinculin and paxillin plaques, but staining revealed that both of these proteins were diffusely present within cells.
Due to the presence of FN fibrils on the surface of SMCs (28) and the role of α5β1-integrin in EC spreading, we hypothesized that ECs in coculture formed fibrillar adhesions. Subconfluent ECs expressing GFP-tensin formed fibrillar adhesions after 4 h of spreading when adhered to SMCs, as indicated by the centrally located elongated tensin plaques (Fig. 3A). In contrast, subconfluent ECs adhered to FN-coated plastic did not form fibrillar adhesions after 4 h of attachment. The small tensin plaques present in the monoculture at the 4-h time point resembled focal adhesions and not fibrillar adhesions due to their small size and location at the cell periphery. After 2 days of attachment, fibrillar adhesion formation was found in coculture, as indicated by the elongated pattern of tensin immunofluorescence, whereas EC fibrillar adhesions were not found in monoculture (not shown). ECs in monoculture did seem to have more tensin plaques after 2 days compared with 4 h of attachment. Consistent with this observation, FN between ECs and SMCs in coculture remained in a fibrillar organization (28), whereas ECs in monoculture had only started to reorganize the FN into fibrils (Fig. 3B). Thus, ECs rapidly formed fibrillar adhesions in coculture due to the presence of fibrillar FN on the surface of SMCs.
To assess whether the lack of focal adhesion formation in coculture led to a reduction in focal adhesion proteins, the amounts of vinculin and paxillin within confluent ECs were quantified by Western blot analysis 2 days after the onset of coculture (n = 3; Fig. 4A). Vinculin and paxillin protein expression within ECs in coculture were significantly lower than ECs in monoculture, whereas ECs in coculture expressed a significantly higher amount of tensin. The gene expression of many molecules involved in focal adhesion and fibrillar adhesion formation was determined 3 days after coculture formation (n = 5; Fig. 4B). Consistent with focal adhesion protein expression, ECs in coculture had significantly reduced mRNA levels of molecules associated with focal adhesion formation compared with ECs in monoculture. Interestingly, FAK gene expression was similar within ECs on SMCs and FN-coated plastic. Tensin protein and gene expression within ECs were significantly increased in coculture compared with monoculture.
Substrate rigidity alone does not dictate EC spreading or focal adhesion formation in coculture.
We hypothesized that ECs in coculture had reduced cell areas and lacked focal adhesions due to the stiffness of underlying SMCs. We measured the apparent elastic modulus of SMCs with an AFM at various time points after serum starvation (n = 3–4; Fig. 5A). The apparent elastic modulus did not significantly change from 2 to 9 days of serum starvation. Thus, the data were pooled, and the apparent elastic modulus of SMCs was found to be 1.98 ± 0.26 kPa (n = 10). Polyacrylamide gels were made with 5% acrylamide and various bis-acrylamide concentrations yielding substrates of similar elastic moduli to SMCs (n = 3–7; Fig. 5B). The relationship between elastic modulus and bis-acrylamide concentration was similar to previously published results (7). FN was cross-linked to the surface of gels to promote attachment to polyacrylamide. Since immobilized FN can inhibit fibrillar adhesion formation, some of the gels with cross-linked FN were incubated three additional times with FN to build a denser FN matrix and promote fibrillar adhesion formation.
EC spreading rates on SMCs (n = 5) and gels (n = 5) of similar elastic moduli as SMCs are shown in Fig. 6. The addition of three additional coatings of FN did not alter the spreading rate of ECs; thus, we pooled the data. As expected, the EC spreading rate increased as the stiffness of the gel increased, but the EC spreading rate on SMCs was significantly lower than the spreading rate on gels of similar elastic moduli, as indicated by the separate populations shown in Fig. 6. The linear regression curves formed from the gel data were used to predict the EC spreading rate when adhered to a substrate with an elastic modulus of 1.98 kPa (SMC elastic modulus). The predicted spreading rates were significantly greater than the actual EC spreading rates found in coculture. The EC spreading rate on SMCs was equivalent to gels with an elastic modulus about one-half the value obtained with SMCs. Thus, the SMC rigidity is not the only contributor to the reduced EC spreading on SMCs.
We also hypothesized that the lack of focal adhesions within ECs in coculture was due to the "softness" of the underlying SMCs. To test this hypothesis, we performed vinculin and paxillin immunostaining on subconfluent ECs that had spread for 4 h (Fig. 7) or fully confluent ECs after 2 days of attachment on polyacrylamide gels. The vinculin plaque size was measured to determine the focal adhesion area (n = 3; Fig. 8). Consistent with the literature (22), we found that focal adhesion size increased as the stiffness of the gel increased. Subconfluent spreading ECs formed larger focal adhesions than fully confluent ECs. Although small in size, focal adhesions formed within ECs adhered to the softest gels (0.41 and 0.86 kPa), whereas ECs that adhered to the more rigid SMCs (1.98 kPa) lacked focal adhesions (Fig. 3). Therefore, the softness of SMCs alone does not explain the lack of EC focal adhesions found in coculture.
In this study, we found that ECs in coculture lacked focal adhesions and had a reduced spreading rate compared with ECs adhered to polyacrylamide gels of similar stiffness as underlying SMCs in coculture. ECs in coculture attached and spread along SMCs primarily through the α5β1-integrin complex. Interactions between α5β1-integrin and fibrillar FN on the surface of SMCs induced the rapid formation of fibrillar adhesions and led to a reduction in focal adhesion protein expression. The lack of focal adhesions and abundance of fibrillar adhesions in coculture was not a transient observation, as indicated by immunostaining after 2 days of culture. Despite this shift in the adhesion mechanism, we (28) have previously found that ECs attached to SMCs can resist shear stresses as high as 300 dyn/cm2.
The results from this study illustrate that the FN receptor, α5β1-integrin, is the primary integrin complex responsible for EC attachment and spreading in coculture. While αvβ3-integrin aided EC attachment in coculture, it did not appear to affect cell spreading in coculture. The role of α5β1- and αvβ3-integrin in EC attachment and spreading on FN-coated plastic was very similar to that of ECs in coculture, indicating that FN on the surface of SMCs is the major ECM protein that controls EC attachment and spreading in coculture. TEBVs, like native arteries, produce significant amounts of FN mRNA and protein (11). Thus, EC interactions with FN in coculture may be relevant to TEBVs and native arteries. These results, along with previous findings (28), indicate that α5β1-integrin binds to fibrillar FN on the surface of underlying SMCs and directs ECs to spread along the surface of SMCs orienting in the direction of fibrillar FN. Katz et al. (14) found that classical focal adhesions containing αvβ3-integrin did not coincide with fibrillar FN. The reduced role of EC αvβ3-integrin in coculture could be due to the fibrillar organization of FN as well as the threefold lower surface protein expression of αvβ3-integrin compared with α5β1-integrin after trypsinization (data not shown).
The extracellular integrin domains bind to the ECM on the substrate, whereas the cytoplasmic domains interact with various adhesion proteins. As the cell spreads, the cytoplasmic integrin domains interact with adhesion proteins to form focal complexes at the leading edge. Focal complexes mature and form focal adhesions during cell spreading. The maturation process involves a temporal recruitment of different adhesion proteins. The proteins recruited to these sites first include αvβ3-integrin and phosphotyrosine, with paxillin following shortly thereafter (31). Later in the maturation process, vinculin is recruited (31), and if the cell generates enough tension along the actin filaments, then focal adhesions form. We found that ECs adhered to FN-coated plastic or SMCs recruited similar amounts of phosphotyrosine and paxillin to the leading edge, but there seemed to be less vinculin recruited to the leading edge of ECs in coculture than in monoculture. In addition, ECs that adhered to FN-coated plastic rapidly formed focal adhesions, whereas ECs in coculture did not form focal adhesions. Thus, the lower levels of vinculin recruitment in coculture could lead to an inhibition of the maturation process to form focal adhesions. The limited maturation of focal complexes in coculture could be due to the lack of αvβ3-integrin interactions with the ECM and the lower substrate stiffness in coculture.
We found that ECs in coculture have lower cell spreading rates compared with ECs on polyacrylamide gels with a similar stiffness as SMCs. Although ECs plated on SMCs (1.98 kPa) did not form any focal adhesions, focal adhesions formed in ECs plated on polyacrylamide gels with an elastic modulus between 0.41 to 2.66 kPa. After 2 days in culture, ECs in coculture had reduced levels of paxillin and vinculin gene and protein expression compared with ECs in monoculture. Although vinculin-deficient cells spread to half the size of wild-type cells (8), ECs that attached to FN-coated plastic or SMCs in the present study likely had similar levels of vinculin during the initial stage of spreading.
Results from several studies, all of which were performed on synthetic substrates, have suggested that substrate rigidity plays a major role in cell spreading and focal adhesion formation (7, 13, 22, 30). Our spreading and focal adhesion results on polyacrylamide gels agree with the literature, but our results of EC spreading and adhesion formation on SMCs indicate that substrate rigidity plays a limited role in spreading and focal adhesion formation in coculture. Thus, there are additional interactions between the cells in coculture that reduce cell spreading and inhibit focal adhesion formation. The fibrillar organization of FN on the surface of SMCs could limit the interactions of αvβ3-integrin with the ECM, thus reducing focal adhesion formation. In addition, the uneven topography of the underlying fibrillar FN and SMCs could further decrease cell spreading and limit focal adhesion formation. It has been shown that the uneven topography (islands with 2-μm diameter and 95-nm height) created on synthetic surfaces reduced cell spreading and focal adhesions compared with a flat surface (4, 5). The topography on the surface of SMCs could have local variations of several micrometers, thus decreasing EC spreading and focal adhesion formation. Altering glucose concentrations may modulate the effect of FN topography and alter EC spreading rates (10, 15). A functional TEBV must have mechanical properties similar to native vessels, and to increase strength of the vessel wall through collagen production, researchers have exposed the vessels to cyclic strains (for a review, see Ref. 19). Thus, stretch preconditioning of SMCs could also alter the organization of the ECM, thus altering EC spreading rates and adhesion mechanisms in coculture.
Focal adhesions may be affected by metabolic products from SMCs. For example, 12-hydroxyeicosatetraenoic acid [12(S)-HETE], a lipoxygenase metabolite of arachidonic acid, decreased αvβ3-integrin- and vinculin-containing focal adhesions within ECs up to 4 h after 12(S)-HETE treatment (26, 27), but 24 h after the removal of 12(S)-HETE, ECs recovered their αvβ3-integrin-containing focal adhesions (26). SMCs maintained in medium containing lower glucose concentrations (5 mM) produced significantly less 12(S)-HETE than those maintained in media with higher glucose concentrations (23 mM) (1). Therefore, the high glucose concentration (17.5 mM) in the quiescent media could stimulate SMCs to produce a significant amount of 12(S)-HETE (1), which could decrease initial EC focal adhesion formation in coculture. The effect of glucose concentration on 12(S)-HETE production and reduced focal adhesion formation does not explain the lack of focal adhesions after 2 days in coculture since the glucose concentration in coculture media is 5.6 mM. The effect of 12(S)-HETE on fibrillar adhesion formation is not known.
Focal adhesion formation depends on the cell's ability to generate the necessary contractile forces within the cell. Contractile forces are generated by interactions between actin and myosin due to myosin ATPase activity. Myosin ATPase activity is regulated by myosin light chain phosphorylation, which has been found to be enhanced by Rho (for a review, see Ref. 29). Clustering of β3-integrins can activate RhoA (2), whereas clustering of β1-integrins suppresses RhoA activation (20). Our results suggest that the elastic modulus of cellular substrates is only one of many factors, such as eicosaniod mediators, surface topography, and ECM organization, that modulate cell behavior. Therefore, we hypothesize that SMC rigidity, topography, eicosaniod production, and surface ECM organization limit EC spreading by inhibiting β3-integrin clustering and promoting β1-integrin clustering, which, in turn, suppresses RhoA production and activity to a point where the actin cytoskeleton cannot generate enough tension to form focal adhesions and continue spreading. In turn, there is a reduced recruitment of paxillin and vinculin to these adhesions, leading to their downregulation.
The lack of focal adhesions and abundance of fibrillar adhesions found within ECs adhered to SMCs may have important functional implications. Tensin, the major protein in fibrillar adhesions, may be involved with signal transduction since it contains a Src homology 2 (SH2) domain and can be phosphorylated on tyrosine, serine, and threonine residues (for a review, see Ref. 18). FAK also contains a SH2 domain, which can act as a docking site of many different signaling molecules. For example, blockade of FAK-dependent signaling in coronary arterioles reduced endothelial nitric oxide synthase (eNOS) activity, and it was proposed that FAK interacts with the well-established shear stress-induced kinase cascade of Src, phosphoinositide 3-kinase, Akt, and eNOS (16). Therefore, the composition of the proteins at EC adhesion sites has the potential to regulate many important signaling mechanisms controlling EC function.
National Institutes of Health Grants R21-HL-72189 and F31-EB-006298 supported this project.
The authors thank Steven Reich for the assistance with optimizing the EC-SMC separation protocol as well as Melissa Brown for determining the number of integrin complexes on the surface of ECs. Also, we give special thanks to Dr. Shin Lin, Dr. Diane Lin (University of California, Irvine, CA), and Dr. Kenneth Yamada (National Institutes of Health) for supplying the GFP-vinculin and GFP-tensin plasmids.
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
- Copyright © 2007 by the American Physiological Society


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Dragoon Hoodies
Main Tag
This here is my FFXIV Dragoon icon design. Big fan of Final Fantasy XIV and always loved all the different class logos/icons. Wanted to capture the color scheme of each class' soul crystal in a unique way. All of my designs can also be found in poster form on my Etsy store: https://www.etsy.com/shop/StromDesigns
Tags: fantasy, xiv, finalfantasy, drg, eorzea


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The Totally NEW Team Corwin Thread
View Single Post
May 25th, 2012, 22:15
Right now I don't quite understand the new DDO Market.
My understanding so far is as following :
It is ONLY for online buying of
- Turbine Points
- The Expansion Pack
for now. I almost expext that other packs might follow there one day.
Now, since this Market is newly opened, there appears to be a discount of 10 % there (using the coupon code of DDOMK10).
The questions I have now are :
- Do these 10 % apply to the current Bonus POints thing (that goes until May, 31st), too ?
Edit : No, these Bonus Points seem only to apply for the IN-game DDO Store.
As far as I have understood it, NOT to the new Market.
- Do these 10 % apply to the Expansion Packs, too ? Because then they would get a bit cheaper.
However, I think that they are still a bit too far away from me, level-wise. It is very tempting to buy at least the base pack, sine with these 10% off it reaches almost the same price as reguilar Turbine Point Cards I can buy here in Gamestop shops.
But on the other hand I'm not sure what kind of payment is accepted there. And you know how conservative I am
Edit : Could it be that the Expansion will one day perhaps cost the same amount of Turbine Points (roughly) than Turbine Point Cards cost ? I just thought this, because the prices of the Base Edition and the 20 TP Cards are so much similar …
Any intelligent fool can make things bigger, more complex, and more violent. It takes a touch of genius and a lot of courage to move in the opposite direction. (E.F.Schumacher, Economist,
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Jun 30, 2009
Ok so it might look it but believe it or not the engine wasn't running and the car was being pushed!
Cal,that is very good
Can't take credit for it when I was sitting in it while Annie pushed the car and fired the camera remotely!...lol
Looking good dude!
Rig shot?, how long was the exposure?.
Looks sweet
200 MPH Pah......Brilliant pics. Good work both
Hi Calvin, that's very clever!
Yep......30 seconds....longer exposure will cancel out camera shake
Very nice, I've been serously considering getting a rig....more money!.
Take up Crack Coccaine mate...cheaper and less addictive than photography!
Ha, yeah I know what you mean. My 'camera bags' got well over a grands worth of stuff in there. I get lens envy.....expensive!.
Wow, looks really good that!
She did well to push the car to 200 mph lol!
Feel your pain dude......one of our lenses is £1200....lol
I'm certainly no photography guy or crack head lol, so I'd like to know how you accomplish this effect please mate, like to learn something & this is intriguing asseen many a time this kind of photo effect but never knew how they got it like this.
That looks wicked!
Me too want to know how ?
its called a "rig" shot......here are the basics.......a long pole.....roughly 5 metres is attached to the car via suction cups and or clamps. The camera is attached to the end of this pole.
As you can imagine, a 5 mtre pole with a huge camera on the end will bounce and flex like a bloody bungee cable!.....so it needs to be attached in a way that it eliminates some of this. An invisible tether is used (fishing wire.....shhh trade secret there)
then....start to push the car....dont have it running as even engine vibrations will cause camera shake.
when car is moving, remotely trigger the camera or set it on self timer with a long exposure setting....for this one we used 30 secs!
Canon L glass is calling me....I can hear it saying 'empty your wallet and buy me'
Must resit though!
My bird thought I was mad when I got my Sigma 10-20mm......"couldnt you buy a really good camera for that much though?"
Is this the same car you lowered so you can climb into it?
You slave driver!
Like this....camera attaches to the end of the pole..
You do a long exposure shot which leaves the shutter open for a longer time then push the car slowly. Anything thats in frame and static will stay in focus, anything thats moving such as the back ground, lights etc will be blurred. Photoshop out the rig after and job done!.
Long exposures are great fun...
Same principle as this shot I took ages ago when I first tried long exposures..
Its just that in the lovely pic of the Audi the cars the still bit in the frame and the back grounds blurring.....opposite to my shot but same principle.
thanks for explaining it Tom,I always wondered how yet didn't feel the need to ask how.
Looks good i like that style of picture
great pic mate both shots looks awsome
both shots are now gonna get used as my desktop background, fantastic work lads
Very nice shot mate and doesn't look like much movement in the rig. i've recently bought some cups and clamps to make a rig but am struggling with flex in the pole, My friend is in the process if making me one from ally box section which will hopfully overcome this somewhat. I will try the fishing line trick though, good tip.
very cool shoot!
the exposures is charming!
I just don't get with these rig shots how such a long exposure doesn't result in any blur!
Good job!
Awesome shot's but i'd be terrified to put my camera on "A rig "but it definetly looks worth it.
Ive saw alot smaller rig's for wheel shot''s and stuff, What price do these usually sell for?
Its a good way to upgrade your camera if it falls off!!! Claim on the insurance LOL!!!
Tom if you want to try some good glass..... try .....
I used to work in London and a couple of times hired some L series glass from them.....
if youre looking for some advice they're a good bunch!
I hired a f2.8 70-200mm twas REALLY nice to use - I'd get shot if I'd bought it lol got some cracking pics with it cars rolling....
It's only a matter of time bud... I just bought this one and this one. New body, another L series and some new filters on the way too. ...And here was me thinking the A3 was a pricey toy :s
I've told myself I'm not putting the new lenses on anything until the new body arrives. I'm struggling to keep to it.
The best of luck bud!
Lads...lol...Im sure the wife will be pleased with that comment....She's the photographer, i'm just the forum whore...lol
stunning pictures
I love how you made that work. Did you use photoshop? Anyway it likes very well done. thanks for posting
~ Sam
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By Wendy
Everyone is welcome at the Nasher Museum.
Even if you are 1 foot tall, blue and two-dimensional. And mute.
The Flat Blue Devil made an appearance recently in our permanent collection gallery.
He was dwarfed by the nostril hairs of Ron Mueck's sculpture, "Mask (Self-Portrait)."
Flat Blue Devil has also been seen shopping and lurking around campus.
Read more about his misadventures on Duke Today.
IMAGE: Flat Blue Devil with Ron Mueck's 1997 work, "Mask (Self-Portrait)."
Polyester resin and mixed media. Collection of Mr. and Mrs. J. Tomilson Hill.


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Welcome to TLVstyle – the expert who can take you behind the scenes of Israel's most creative fashion talents
Israel is globally recognized as a dynamic and thriving fashion center with the city of Tel Aviv leading the way.
Brimming with passion and creative energy, Israeli designers are creating bold new statements
in shared spaces and 'apartments-turned- studios' off the beaten-path of Tel Aviv's bustling streets.
Led by fashion maven and curator Galit Reismann, TLVstyle offers you diverse and exciting
ways to connect to the pulse of the emerging Israeli fashion design scene.
With tailor-made tours, special projects and events, each service we provide is uniquely designed
to crack the code of the Israeli fashion and design industries.
It takes a true insider and an expert to navigate this exciting fashion hub.
The stories behind the scenes, the faces behind the brands, TLVstyle provides with insider's insights about Israel's most riveting designers.
Whether you're a design enthusiast, or you simply want to experience Tel Aviv off the beaten track and be inspired by its most creative people, TLVstyle provides exceptional personalized tours that keep travelers coming back for more.
TLVstyle curates content events and exhibitions, serving as a catalyst for creativity, innovation and partnerships in the Israeli design arena.
Each event and exhibition shares stories and narratives carried by garments, possible only in
Tel Aviv's one-of-a kind atmosphere, mix of people
and cultural influences.
From lectures and workshops to fashion shows, hosting international missions and creating education programs, TLVstyle has initiated and collaborated across a broad spectrum of design-centric events and projects. All projects are held with a common ambition: to leverage, celebrate and educate about Israeli fashion and design.
"One of the most memorable tours ever had in Israel"
"I was part of a Jewish National Fund Mission for Art and Fashion in Israel. We felt extremely lucky to have Galit as our guide. Her knowledge and feel for fashion and fashion history was heartfelt and excellent. Her tour helped us to appreciate the incredible creativity and dedication of fashion designers in Tel Aviv" - Cannon S
"A Magical Look Into Israeli Fashion"
"It was fascinating to meet the charming young designers and to hear from them about their design philosophy and creative process. It felt like a once-in-a-lifetime experience and was absolutely inspirational (plus the shopping was fabulous)" - Anastasia T
"An Immersion Into The fashion Industry"
"TLVstyle Tour was nothing less than amazing. Galit takes into consideration exactly what each client is looking for and develops the tour around that. She was able to help me with my own personal style and how to make the most of it. The tour is so much more than going on a shopping trip. It is an immersion into the fashion industry and the beating heart behind it.
It was so fun meeting the designers and seeing where and how they work. Everyone was so welcoming and nice. Galit is extremely knowledgeable in the fashion industry and in the cultural background in Tel Aviv. Besides being fun, it was very informational.
I recommend this tour for anyone" - Skruege
Interested in a special tour? Whether you're an individual traveler, a group or a company interested in an unmatched experience, we can offer you a close encounter with Israel's design forces.
For more information:
Contact Us
If you'd like to collaborate on different levels,
let us know. We're always open to innovative projects.


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Millennium Pipeline Company, L. L. C.
Original Volume No. 1
Contents / Previous / Next / Main Tariff Index
Effective Date: 12/22/2008, Docket: CP98-150-011, Status: Effective
Original Sheet No. 193 Original Sheet No. 193
14.9 Refunds.
(a) For all refunds other than those described at paragraph (b)
immediately below, the original Releasor of any capacity shall receive from
Transporter any reservation charge-related refunds associated with the
assigned capacity, including any refunds related to the reservation charge
portion of payments under a one-part volumetric rate. The Replacement
Shipper holding the assigned right to service at the time of the overpayment
shall receive from Transporter its share of any commodity charge-related
refunds, including any refunds related to the commodity portion of payments
under a one-part volumetric rate, associated with the assigned capacity.
Refunds owed by Transporter will be made by Transporter directly to
Replacement Shipper, or indirectly through the Releasor if Replacement
Shipper has appointed Releasor as its agent for billings pursuant to Section
14.8(a) above.
(b) The refund obligation of Transporter set forth in paragraph (a)
shall be modified where Releasor has released capacity at a rate in excess of
that owed by Releasor to Transporter for that capacity ("Releasor's Margin").
To the extent that Releasor's margin equals or exceeds the amount of any
refund obligation, Transporter shall not be obligated to make refunds to
Releasor. (Any refunds ultimately paid to a Replacement Shipper in that
event shall be paid by Releasor.)
14.10 Fees. Transporter shall not charge a fee for posting of a Release
Notice or a Request to Purchase on its EBB. Transporter shall be entitled to
charge a reasonable fee if Releasor and Transporter agree that Transporter
shall receive a fee for actively marketing the capacity Releasor seeks to
14.11 Term. The term a Releasing Shipper imposes may not conflict with any
provision of the Service Agreement, Rate Schedule or General Terms and
Conditions of Millennium tariff. In the event of such conflict, Millennium
may withdraw the Shipper's notice from Posting.
14.12 Termination. If the Releasing Shipper fails to pay any monthly bill in
accordance with the provision of its service agreement and of Section 10 of
the General Terms and Conditions, Transporter may commence suspension and
termination of service procedures as set forth in Sections 3 and 10 of the
General Terms and Conditions. Any service agreement(s) of a Replacement
Shipper(s) for such capacity shall be terminated as well, unless (1)
Replacement Shipper agrees to pay Transporter the currently effective maximum
rates for service under the Tariff, or (2) Transporter and Replacement
Shipper mutually agree upon a discounted rate or negotiated rate for service
under this Tariff, or (3) Replacement Shipper elects to continue service at
the contract rate between the Releasor and Transporter.


text:
Jeremy Whelehan
Kevin Spacey
Maureen Anderman
Stephen Lee Anderson
Jeremy Bobb
Jack Ellis
Hadyn Dwynne
Chukwudi Iwuji
Isaiah Johnson
NOW: In the Wings on a World Stage (NR)
Documentary 93 min. May 2, 2014
Early on, we hear Spacey's Richard bark, "Now is the winter of our discontent," but Jeremy Whelehan's film, distractible as a puppy in a room full of tennis balls, then jumps to something else, apparently suffering from the misapprehension that watching stage crews mount the sets or audiences file into theaters must be more edifying for audiences than hearing one of the world's great actors speak more than the very first line of one of the world's great plays.
Only one speech from the whole of Richard III sneaks into the hurly burly of smiling castmembers bonding in Beijing, Qatar, and other spectacular places. Guess what? Touring the world for a year with a difficult play and a top-flight cast is both a pleasure and a hardship.
The movie is more the latter, although it occasionally honors the production itself with discussion of the specifics of performance or design. More often it's devoted to the un-urgent business of letting us know that the cast did enjoy seeing the Great Wall of China. In the end, all NOW reveals is that talented people did a difficult thing in far-off places -- and that now they have a video scrapbook.
Jeremy Whelehan Kevin Spacey, Maureen Anderman, Stephen Lee Anderson, Jeremy Bobb, Jack Ellis, Hadyn Dwynne, Chukwudi Iwuji, Isaiah Johnson Abramorama Entertainment


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eGov Services • 1300 Perdido St, New Orleans, LA 70112 • (504) 658-4000
Home » Parks and Parkways » Event Rules & Regulations » Book An Event
A permit is required to use public parks for events in the City of New Orleans. The Department of Parks and Parkways is responsible for the following parks: Jackson Square, Brechtel, Washington Square, Mississippi River Heritage Park, Lafayette Square, Coliseum Square, Armstrong Park and other smaller parks. Click here to see a map to help you determine who has the jurisdiction over other parks in the City of New Orleans.
Full information on hosting events in public parks, including a schedule of fees (by park), rules and regulations, and potential additional permits needed can be found on the Parks & Shelters Permits page.
Citizens wishing to book a park space should call the department (504) 658-3201. or email firstname.lastname@example.com to ask if the date is available.
Once confirmation of date availability is confirmed, requestors can file a booking request form by fax (504) 658-3227 or by email.
Last updated: 3/28/2017 2:40:39 PM


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The House Foreign Affairs Committee will look at religious freedom, democracy and human rights in Asia, and the House Permanent Select Committee on Intelligence will hold a closed briefing on Israel/Palestinian Authority issues.
More details on these and other full committee hearings follow:
House Budget
June 2, "Fannie Mae, Freddie Mac & FHA: Taxpayer Exposure in the Housing Markets," Deborah Lucas, Congressional Budget Office Assistant Director testifies
10:30 a.m., 210 Cannon House Office Building
House Financial Services
June 1, to hear testimony from Geithner on the state of the international financial system
10 a.m., 2128 Rayburn House Office Building
June 3, "Oversight of HUD's HOME Program"
9:30 a.m., 2128 Rayburn House Office Building
House Foreign Affairs
June 2, "Religious Freedom, Democracy, Human Rights in Asia: Status of the Implementation of the Tibetan Policy Act, Block Burmese JADE Act, and North Korean Human Rights Act," Deputy Assistant Secretary of State for Democracy, Human Rights and Labor Dan Baer testifies
10 a.m., 2172 Rayburn House Office Building
House Natural Resources
June 1, "American Energy Initiative: Identifying Roadblocks to Wind and Solar Energy on Public Lands and Waters, Part II"
10 a.m., 1324 Longworth House Office Building
House Small Business
June 1, "Access to Capital: Can Small Businesses Access the Credit Necessary to Grow and Create Jobs?"
1 p.m., 2360 Rayburn House Office Building
House Veterans' Affairs
June 1, "Putting America's Veterans Back to Work
10 a.m., 334 Cannon House Office Building
House Ways & Means
June 2, on how business tax reform can encourage job creation
10 a.m., 1100 Longworth House Office Building
House Permanent Select Committee on Intelligence
June 2, Israel/Palestinian Authority Update (closed hearing)10 a.m., HVC-304


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WASHINGTON — In another setback for President Barack Obama's health care initiative, the administration has delayed until 2015 a significant consumer protection in the law that limits how much people may have to spend on their own health care.
The limit on out-of-pocket costs, including deductibles and co-payments, was not supposed to exceed $6,350 for an individual and $12,700 for a family. But under a little-noticed ruling, federal officials have granted a one-year grace period to some insurers, allowing them to set higher limits, or no limit at all, in 2014.
The grace period has been outlined on the Labor Department's website since February but was obscured in a maze of legal and bureaucratic language that went largely unnoticed. When asked in recent days about the language — which appeared as an answer to one of 137 "frequently asked questions about Affordable Care Act implementation" — department officials confirmed the policy.
The discovery is likely to fuel continuing Republican efforts this fall to discredit the president's health care law.
Under the policy, many group health plans will be able to maintain separate out-of-pocket limits for benefits in 2014. As a result, a consumer may be required to pay $6,350 for doctors' services and hospital care, and an additional $6,350 for prescription drugs under a plan administered by a pharmacy benefit manager.
Some consumers may have to pay even more, as some group health plans will not be required to impose any limit on a patient's out-of-pocket costs for drugs next year. If a drug plan does not currently have a limit on out-of-pocket costs, it will not have to impose one for 2014, federal officials said Monday.
The health law, signed more than three years ago by Obama, clearly established a single overall limit on out-of-pocket costs for each individual or family. But federal officials said that many insurers and employers needed more time to comply because they used separate companies to help administer major medical coverage and drug benefits, with separate limits on out-of-pocket costs.
In many cases, the companies have separate computer systems that cannot communicate with one another.
A senior administration official, speaking on condition of anonymity to discuss internal deliberations, said: "We knew this was an important issue. We had to balance the interests of consumers with the concerns of health plan sponsors and carriers, which told us that their computer systems were not set up to aggregate all of a person's out-of-pocket costs. They asked for more time to comply."
Health plans are free to set out-of-pocket limits lower than the levels allowed by the administration. But many employers and health plans sought the grace period, saying they needed time to upgrade their computer systems.


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Grassroots support critical for state avy programs
By Summit Voice
FRISCO — The Friends of the CAIC sure knows how to time their events, with this year's CAIC Benefit Bash scheduled to be held Saturday evening, just as the season's first significant winter storm is forecast to drop the first good batch of powder on the Colorado high country.
This year's Benefit Bash will once again be held at the Breckenridge Riverwalk Center and features live music, tasty food, beer from New Belgium Brewery, and great people all enjoying the massive silent auction and a seemingly endless stream of door prizes.
The goodies include a Monarch season pass, snowcat skiing trips, all sorts of backcountry gear and even a spot on an Alaska heli guide school trip.
The bash has grown to be one of the most important grassroots fundraisers for the CAIC, which relies in part on donations to maintain a robust backcountry forecasting and avalanche awareness program. Last year the event raised about $72,000 and organizer Joe Vandal said the goal this year is $90,000.
Even at the sizable Riverwalk Center venue, the bash has filled up completely the last few years, so the best option is to get there early. The fun starts at 5 p.m. and ends around 10 p.m. Tickets are priced the same as last year; $30 in advance and $35 at the door.
Colorado has one of the most dangerous snowpacks in North America and typically accounts for one third of all avalanche fatalities in the United States. The purpose of the CAIC is to minimize the economic and human impact of snow avalanches on the recreation, tourism, and commerce industries.
Last year's snowpack was especially treacherous, resulting in an above-average season in terms of avalanche accidents, according to CAIC director Ethan Greene. There were seven deaths and a total of 2,200 reported slides, with 50 people reported caught, 17 partial burials and 11 full burials.
There were also two inbounds avalanche deaths at Colorado ski areas (Vail and Winter Park) last season, both on the same day. Greene said last season saw a continuation in the trend of people getting killed by slides in the sidecountry, after leaving controlled ski area terrain, and a number of people died in "fairly small" avalanches, he added.


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Mets unable to back Harvey in loss to Cards
Mets unable to back Harvey in loss to Cards
The New York right-hander drew rave reviews from his manager after battling back from a rough start to keep his club close in a 5-1 loss to the Cardinals on Tuesday night.
But Harvey left Busch Stadium with his head down.
Harvey (3-4) gave up three runs on four hits in the tell-tale second inning, then shut the door the rest of the way.
"I was impressed with how he went about his job," Mets manager Terry Collins said. "[Pitching coach] Dan [Warthen] and I were sitting on the bench after the three-run inning, and we kind of looked at each other and said, 'Now we'll see what we've got. Let's see if he shuts this sucker down.' He did. He had to work hard, but he did it. You looked up in the fifth inning, and it's still 3-0."
That battle-back ability brought a smile to the face of Collins.
But not the University of North Carolina product, who was extremely displeased with his five-inning performance.
"What I did tonight, in my opinion was [bad]," Harvey said. "I'm not happy about it. I want to go out there and go deeper in the game. I've got to get quicker outs and I've got to keep the pitch count down, and I didn't do that tonight."
Harvey -- who has thrown 157 1/3 innings and will likely have one, maybe two more starts this season due to an innings limit -- gave up three earned runs on six hits. He struck out four and walked three. Harvey had given up two runs or fewer in his last four starts with a 2-1 mark and a 2.08 ERA in that span.
Despite the loss, Collins was pleased with Harvey's courage and determination. He held the Cardinals scoreless on two hits in four of his five innings. He did not allow a hit in either of his last two frames.
"He's an impressive guy to be around, " Collins said. "He's upset that he didn't pitch good tonight. But he pitched OK."
Cardinals manager Mike Matheny was happy to bunch together three runs in the second inning against Harvey.
"You can see why they're excited about him," Matheny said. "He has very good stuff."
New York (64-72) lost for the second time in as many nights after winning three successive games in Miami.
The Mets had won three consecutive series for the first time this season before losing the first two games of this three-game set.
Harvey wasn't the only Mets pitcher to turn in a gutsy effort on Tuesday.
New York right-hander Jeurys Familia, called up on Tuesday, made his Major League debut, entering the game at the start of the eighth inning. He gave up no runs and one hit. Familia struck out pinch-hitter Lance Berkman with a 97-mph fastball to lead off the inning. He gave up a single to Jon Jay, but then got Matt Carpenter to ground into a inning-ending double play. Familia threw nine pitches.
"I felt good warming up in the bullpen," Familia said. "I prepared my mind and tried to focus."
New York managed nine hits off St. Louis left-hander Jaime Garcia (4-6), who went 7 1/3 innings.
Trailing, 5-0, the Mets pushed across a run in the eighth. Pinch-hitter Justin Turner and Ruben Tejada singled before St. Louis shortstop Daniel Descalso booted a potential inning-ending double-play ball to load the bases with one out. Pinch-hitter Mike Baxter brought in Turner with a sacrifice fly. Cardinals left fielder Adron Chambers ended the threat by making a running catch on a deep drive from Daniel Murphy.
The Cardinals used four successive hits to plate three runs in the second. Yadier Molina, David Freese, Skip Schumaker and Descalso singled to give the Cards a 2-0 lead. Carpenter drove in Schumaker with a sacrifice fly to left.
Mets reliever Justin Hampson, recalled on Tuesday, came on at the start of the sixth and allowed one run in one inning. The 32-year-old appeared in three games earlier in the season without allowing a run. Hampson, who grew up in nearby Edwardsville, Ill., was making his first appearance in St. Louis in his 83rd Major League game.
New York, which is 0-for-14 with runners in scoring position in the series, fell to 5-3 on the current nine-game trip.
Steve Overbey is a contributor to MLB.com. This story was not subject to the approval of Major League Baseball or its clubs.


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Russian Research Reactor Fuel Return Program
When the war was cold, the fuel was hot. Hot as in radioactive. During the 1950s, both the U.S. and Russia provided highly-enriched uranium (HEU) fuel to countries around the world. The fuel powered research reactors for experiments in medicine, physics, and energy.
Today, the use of HEU fuel for research reactors is being replaced by proliferation-resistant low-enriched uranium. And since 2002, an international team of nonproliferation experts has been working to locate and repatriate stockpiles of HEU fuel. Three members of the team work for Idaho National Laboratory.
During the Cold War, both the U.S and the former Soviet Union ran programs to encourage peaceful nuclear research through the exchange of nuclear fuels. But when the Soviet Union collapsed in 1991, the countries that had received Russian fuel were left without a process for returning it. Most were unable to provide secure long-term storage for the fuel, and it became a national security risk for the U.S.
In response, the National Nuclear Security Administration (NNSA) established the Russian Research Reactor Fuel Return Program to reduce this global threat. Members of the team eliminate stockpiles of highly enriched uranium by managing the packaging, transportation, and return of HEU fuel from the original receiving country to Russia. Once there, the fuel is permanently converted into safe low-enriched uranium.
The multiyear initiative is both complex and sensitive. Experts from NNSA, the International Atomic Energy Agency (IAEA), the Department of State, Russia, and the host nation are involved during the process.
Quick Facts
- The Russian Research Reactor Fuel Return Program transports Russian-origin highly enriched uranium from research reactors in 14 countries to the Russian Federation.
- A series of well coordinated steps by many international parties ensures the fuel is safely and securely shipped to Russia for down blending and reprocessing.
- As of June 2010, more than 1,300 kilograms (more than 50 nuclear weapons) of HEU fresh and spent fuel has been returned to Russia.
- To date, fuel has been removed from Bulgaria, the Czech Republic, Germany, Hungary Kazakhstan, Latvia, Libya, Poland, Romania, Serbia, Uzbekistan, and Vietnam.
- As a result of the RRRFR Program, Latvia, Bulgaria, and Romania now have no remaining HEU.


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The Home Ranch is available for weddings, family reunions, clubs and groups during April, May, October and November. We require a two night stay with a minimum of 12 people. If you are interested in learning more about these programs please contact Selina or Johnny, 970-879-1780.
Spring & Fall Ranch Weddings & Retreats
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SAN DIEGO ---- Jared Gaither was in uniform, on the practice field Wednesday afternoon.
No, really. I swear.
The Chargers left tackle had a helmet on and everything, working out with his teammates during individual drills, the only period open to the media.
The extent of his participation won't be known until later this afternoon, when the team must detail his activity level as part of their injury report. Regardless, it is his first practice of any kind since July 27, when he was diagnosed with back spasms. That in itself is a big deal.
The man who signed a 4-year $24.5 million contract this offseason has been practically invisible since, showing up occassionally to watch practice or work on a side field. When he last spoke to the North County Times a week ago, Gaither said he "had to be able to function," even if he wasn't at 100 percent.
After what coach Norv Turner described Monday as good workouts last week, Gaither has taken the next step towards returning to game action.
That's a good sign considering the offensive line's struggles last week, and replacement left tackle Mike Harris' difficulties protecting for the pass and especially the run in back-to-back weeks.
If Turner's formula holds true, Gaither will be eased back into practice. Due to the nature of his back injury, physical conditioning has been difficult to maintain. Returning players are typically limited for a few days at least, before they are truely tested as full participants. Only when they pass those test will Turner consider activating a returning player.
That suggests a return the following week at New Orleans on Oct. 7. But, given the Chargers struggles up front, that process may be accelerated. Only time will tell.


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|March 7th, 2010
I'll keep it simple, you need to divide any given sum by the amount of forces you have, the kind of equipment they posses, the infrastructure they run.
US total military budget is 680~ billion USD total for just over 1.3 milion troops, Chinese military budget is only 70~ billion USD.
The difference? More then 50% of Chinese hardware is over 30 years old.
Now Russia has over a million troops, a much larger fleet, much more ground equipment to keep then both China and US combined and its budget is 50 billion USD.
This translates to an average Russian unit receiving less then 1/20th funding of an average Western European unit and less then 1/10th of an average central European unit.
To give you a picture, just recently russian navy got boiled eggs for breakfreast which was hailed as a major feeding reform.
Of course there are some units that are better funded but even the elites are well below what the Western world considers as average.
|March 9th, 2010
Regarding training - lot of that could be trained without large expenses; I do not see any obstacles to train tactics and hand-to-hand combat while lacking financial resources.
What comes to small arms ammunition - I guess they still have supplies produced during USSR
|March 11th, 2010
For example the invasion in Georgia showed that Russian NCOs cant read maps, that marksmanship is horrendously low and tactics non existent so even the cheap stuff is neglected but this is more due to low discipline and military culture then money.
|March 12th, 2010
In general, I can agree in certain elements Russian soldiers are trained worse than their Western colleagues, however the difference is not so high as you are trying to state.
Besides - you forgot about combat bears
|March 12th, 2010
50000 troops?
and russia has 1 million troops, so when you break it down there isn't much of the pie to go around...
even when taking into consideration, the fact that costs would be lower , it isn't hard to tell that the Russian military is under funded, and this isn't exactly a secret. even the Russians admit this...
|March 12th, 2010
Of course they are underfunded! And becasue of it they decrease number of troops (for example, former divisions have being reorganized in smaller units such as brigades or even regiments), are not able to provide pilots flight hours necessery to maintain their qualification, and so on and so on.
However, it doesn't mean their army do not train at all or jog instead of tactical exercises.
|March 12th, 2010
i would have to say that the quality of tactical training does have a lot to do with what you can afford. its like comparing professional sprts clubs and non professional clubs they all train but the professional one will have better routines and methods and will be more up to date techniques.
so in this respect the Russian military must be suffering. it is impossible to say that it wouldnt effect quality of troops if you cant afford proper facilities up to date training modern equipment etc. etc.
it will all effect their quality.
|March 12th, 2010
Several points of interest here, note that instead of relatively modern T-80s or T-90s they use old T-64s even without ERA, note the AK-47 carried by a soldier on the left of the PKM gunner.
And since you seem to be a fan of images and i too believe they speak louder then words.
This picture is fun, a mechanized unit in Georgia, these guys wear "Ushanka" hats that are about 40 years old and have no protective uniforms, in case there's a fire they're going to be living torches also note the cool diving goggles (no they're not standard issue) with a custom rubber strap on the driver!
|March 12th, 2010
ok guys, I might get involved in to "half row" myself
When I have served for my Country I have had quite few feelings, that are present today, regarding anything Russian anything. Yeas appearance etc is of some contest, but by no means ever I would discount SpecNaz (awfully generalized name) as for any of they abilities. Yeas, there are better troops for most likely any potential scenario around the world, but Russian SF are not bad or really badly trained. Differently- yes, but still capable (please hold me not to start another row regarding some opps) and very importantly also- financially maybe neglected, but not to the extent of ridiculing here by some here- they have decent force!... regrettably...


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Reader Approved
wikiHow to Say "Good Morning" in Japanese
Two Methods:InformalFormalCommunity Q&A
The phrase "good morning" is a common greeting used in Japan, and is considered the respectful way to greet both strangers and friends before 10 a.m. There are two ways to say "good morning" in Japanese: the casual, informal way and the polite, formal way.
1Say "ohayo." Ohayo literally means "good morning," and is pronounced "oh-high-yoh," similar to the pronunciation of the state "Ohio."
2Perform a small head nod when saying "good morning" informally to friends and family members. This movement is considered the casual way to bow if you are from the United States or another country, and aren't familiar with traditional Japanese bowing etiquette.[1]
1Say "ohayo gozaimasu." This phrase is pronounced "oh-high-yoh go-zah-ee-moss," with the letter "u" silent.[2]
2Accompany the phrase "good morning" with a deep bow between 30 and 90 degrees at the waist if greeting someone formally and politely, or when greeting a superior. This is the appropriate way to deliver the greeting "good morning" in formal situations when spending time in Japan.[3]
Community Q&A
How do you say the numbers 1–10 in Japanese?wikiHow ContributorIchi, ni, san, yon, go, roku, nana, hachi, kyuu, juu. In characters: 一、二、三、四、五、六、七、八、九、十。いち、に、さん、よん、ご、ろく、なな、はち、きゅう、じゅう。
How do I say "How are you" in Japanese?wikiHow ContributorThere are many ways to say it, but one of the most common is "O Genki desu ka?" お元気ですか?
In Japanese,what does "kornichi wah" mean?wikiHow ContributorIt's not "kornichi wah," it's "konichiwa." It means hello.
Ask a Question
200 characters leftSubmit
- Make a habit of saying "good morning" to all people you interact with when spending time in Japan or with people of Japanese origin. Japanese culture considers it rude not to greet someone when the moment presents itself, or to greet someone in an offhand, lazy manner.
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Categories: Japanese
In other languages:
Italiano: Dire "Buongiorno" in Giapponese, Русский: пожелать доброго утра на японском языке, Português: Dizer Bom Dia em Japonês
Thanks to all authors for creating a page that has been read 38,808 times.
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WHISTLEBLOWING is not for the faint-hearted. Careers can be ruined, marriages strained, and pariah status is almost guaranteed. Nevertheless, a whistleblower from software company Autonomy decided to step forward and tell their new bosses at Hewlett Packard about alleged financial irregularities in the business.
HP has since written off $8.8bn from the value of Autonomy which it bought for $10bn less than a year earlier, and informed the FBI and Securities and Exchange Commission.
As for the whistleblower, rather than facing the sack, he or she could be entitled to a multimillion-dollar payout thanks to new US laws. Rules introduced by the Wall Street regulator mean a whistleblower can get up to a 30 per cent cut of any fine handed out for wrongdoing.
But former whistleblowers are less convinced such incentives are necessary and say they acted on their conscience rather than the thought of any financial reward.
Paul Moore, the former head of group risk for HBOS who was sacked after warning the board it was taking excessive risks, turned whistleblower in 2009 having taken a payoff and signing a confidentiality agreement.
He said: "Money was never a consideration and I don't think it would have been. I was earning hundreds of thousands a year as one of the biggest compliance officers in the country. By comparison, last year I earned £15,000.
"I took the decision to speak out on the night Lehman Brothers went down. I was breaching a confidentiality agreement but I thought the greatest good for the greatest number of people was to speak up because it would feed into the policy debate." But the fallout left him unable to work in banking again, turning to drink and putting a strain on his family life.
The US tax authorities have similar reward schemes. Earlier this year a former UBS banker, Bradley Birkenfeld, was given $104m for assisting the US tax authorities with its investigation into Swiss bank accounts. He was paid the money out of a $780m fine for the bank.
However, former Olympus chief executive Michael Woodford, who exposed a £1bn fraud at the company before he was sacked, warned against such large payouts. "I agree with the principles of having some financial protection for people going forward," he said, "but there should be a limit of around $2m.
"Look at the UBS whistleblower who got $100m. That's like winning the Euromillions and it's not morally justified to pay such a huge amount. That money should have gone to the state imposing the fines."
There are no state-sanctioned financial incentives for whistleblowers in the UK, and the City regulator is unlikely to follow America's lead.
Despite this, a survey by risk advisers Kroll found UK employees were responsible for nearly one in four tip-offs the SEC gets from foreign employees, suggesting financial incentives could increase the number of whistleblowers coming forward to the UK's Financial Services Authority.
But while the UBS whistleblower, Birkenfeld, is counting his millions, sometimes going public can have a different outcome.
Former Enron vice-president Sherron Watkins was sacked from the company for raising issues around fraud. She turned whistleblower, but never worked in the sector again and now gives talks on corporate governance, having written books on her experience with Enron.
Another famous whistleblower, Karen Silkwood, was immortalised by Meryl Streep in the 1983 film Silkwood. She died in mysterious circumstances after she turned whistleblower on the Kerr-McGee plutonium plant where she worked. She suffered radiation poisoning, leading to her investigating wrongdoing at the company, but on her way to meet a journalist, she died in an unexplained car crash.
Dr Rita Pal, a British whistleblower in the tax-funded National Health Service who exposed patient neglect in a North Staffordshire hospital, said: "The main consequence of whistleblowing is that the pariah effect makes you essentially unemployable due to the perceptions people have. I do have regrets on occasion when I look at the personal consequences. I am a human being after all and the devastation to my life has been incredible."
But Woodford, formerly of Olympus, believes the situation is changing. "Since I became a whistleblower, I've been offered the presidency of a Japanese firm and chairmanship of a British company, so I hope that encourages others to speak out. As the world sees scandal after scandal, people shouldn't fear standing up." — The Guardian, London


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Detectives from the Scioto County Sheriff's Office arrested two Wheelersburg men and charged them with burglary.
The Sheriff's office received a call on July 17, 2013, that someone had broken into a home and taken several items including cash and jewelry.
Detectives were able to locate one of the suspects at his house in Wheelersburg. They recovered one ring; later the second suspect was located at another house.
Jeffery Scott Oppy, 28, and Josh M. Greathouse, 20, both of Wheelersburg, are each charged with one count of burglary.
Both men are being held on a $75,000 bond and will appear in Portsmouth Municipal Court on July 22, 2013.
The investigation is ongoing, according to Sheriff Marty V. Donini.
350 Quarrier Street
Charleston, WV 25301
Main (304) 343-1313
Fax (304) 343-6138


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I've been up to my ears in sorting and cataloging the abundant entries we received for this year's DiabetesMine Design Challenge, which closed for submissions last Friday. So much so that I haven't had time to think about — or write about — much else. A quick numerical overview of the outpouring of creativity we've seen:
152 = total number of contest submissions received this year, once all the 'outliers' were properly located (some weren't originally added to the proper group on Scribd)
47 = number of video entries
105 = number of distinct paper entries (some people loaded multiple pages as separate docs)
19 = number of concepts integrating diabetes care with the iPhone in some way
16 = number of concepts for new non-invasive glucose monitoring technologies
8 = number of concepts integrating gaming or toys into D-education
4 = number of entries focusing on programs for patient motivation/support
3.5 = number of hours I've spent not in my office since the contest closed
2 = number of videos that made me chuckle out loud (in a good way — it's all about the presentation).
Have a look at those two:
(NOTE - being featured here has nothing to do with these applicants' chances of winning)
And now enough about that until May 18, when the full judges' panel will have made its determinations, and the winners will be announced. Applicants, please note: I'm just as nervous as you are, believe me.