text:
|Publication number
|Publication type
|Application number
|Publication date
|Mar 10, 1987
|Filing date
|Mar 18, 1985
|Priority date
|Mar 18, 1985
|Fee status
|Publication number
|John F. Dougherty
|Original Assignee
|Dougherty John F
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (29), Referenced by (30), Classifications (25), Legal Events (5)
|External Links: USPTO, USPTO Assignment, Espacenet
This invention relates generally to fasteners and more specifically to fasteners which removably secure a lace or rope or the like in a position to prevent movement thereof.
Among the many fastening systems that are available at the present time, the primary fastener still remains the use of a line such as a string, twine, rope and the like. The following are illustrations of some of the uses of such a line for fastening systems which are applicable to the present invention.
1. The line may be linked to another line which is already secured in an established position.
2. The line may be linked to another line to form a longer line.
3. Any number of lines may be linked in a certain manner to form netting.
4. The line may be linked to itself so as to form an enclosure for holding articles together.
5. The line may be linked to itself so as to form an enclosure securing another enclosure such as a box or a package.
6. The line may be used in conjunction with lace hooks to fasten various objects such as shoes or boots.
7. The line may be used in conjunction with a dead-man to secure a boat or in conjunction with a tie down to secure a load on a truck.
These and other functions commonly involve the tieing of knots which obviously requires a certain amount of time and, in some cases, requires particular learning skills. The system of the present invention offers advantages of simplification in that it eliminates the need to tie knots while accomplishing the same results in less time and without requiring any particular skill.
U.S. Pat. No. 254,343, issued Feb. 28, 1882 to Lequin and U.S. Pat. No. 730,031, issued June 3, 1903 to Leopold present an alternative to the above discussed methods of lacing. The Lequin patent proposes to use a non-flexible metal hook wherein a spike or the like is secured in the hook at its rear part and extends in a downwardly or outwardly fashion. This hook is used for securing the palm side of a glove. These spikes are designed so as to penetrate the lace and hold it in position.
The Leopold patent has much the same type of proposal wherein a non-flexible metal hook is used with some additional type of material secured therein so as to provide a reduced area at the outer ends thereof with a centrally embedded spike. This device is also intended to penetrate the lace and hold it in position.
Both of the above patents are cumbersome in that they present the problem of manufacture wherein each individual eyelet terminates in a grommet or the like and each grommet is placed within the edge of material on opposite sides of an opening. Since the hooks are metal, there is very little or no flexibility and the unlacing is inhibited due to the fact.
Applicant is unaware of any such proposed type of fastening devices used for any other purpose than that of lacing the boots or gloves as illustrated in the above discussed patents.
Accordingly, it is an object of this invention to provide a device for removably securing a lace or the like having a base with at least one fastener integral therewith.
A further object of the invention is to provide a fastener having a base with a wall flexibly secured thereto terminating in a lip and a spike embedded betwen the wall and the base and extending toward the lip.
A further object of this invention is to provide an integrally molded strip with a plurality of fasteners integral therewith, each of said fasteners having a wall flexibly interconnected to the base and a spike or the like embedded in the base and extending toward the opening created by the wall and the base.
These and other objects of the invention will become apparent from the following description when taken together with the drawings.
FIG. 1 is a perspective view of a basic type of fastener of the present invention;
FIG. 2 is a side elevation view of the fastener of FIG. 1 showing a lace passing into position within the fastener;
FIG. 3 is a partial sectional view of a modified version of the fastener of the present invention;
FIG. 4 is a perspective view of a further modification of the fastener of the present invention;
FIG. 5 is a perspective view of another modification of the fastener of the present invention;
FIG. 6 is a perspective view of a further modification of the fastener of the present invention;
FIG. 7 is a perspective view of the fastener of FIG. 6 as used in a particular environment;
FIG. 8 is a perspective view of a fastener of the present invention together with a base which may be sewn;
FIG. 9 is a front view of a fastener of FIG. 8;
FIG. 10 is a side elevation view of a further modification of the present invention;
FIG. 11 is a perspective view illustrating one of the uses of the fastener of FIG. 10;
FIGS. 12, 13 and 14 illustrate various configurations of the retaining means within the fastener;
FIG. 15 is a side elevation view of a modified retaining means;
FIG. 16 is a side elevation view of a further modification of the fastener having two fastening devices extending from a base;
FIG. 17 is a perspective view illustrating one of the uses of the fastener of FIG. 16;
FIG. 18 is a perspective view of a further modification of the fastener having a base and three fasteners;
FIG. 19 is a perspective view illustrating one of the uses of the fastener of FIG. 18;
FIGS. 20 and 21 are perspective views of further modifications of the fastener of FIG. 18;
FIGS. 22 is a side elevation view of a further modification illustrating two fasteners integrally joined;
FIG. 23 is a perspective view illustrating one of the uses of the fastener of FIG. 22;
FIG. 24 is a perspective view of the use of a strip including a plurality of fasteners as used with a garment;
FIG. 25 is a side elevation view of a further modification illustrating an enlarged fastener which accepts two laces;
FIG. 26 is a perspective view of a further modification of the fastener.
FIG. 27 is a side elevation view of a further modification of the fastener of the present invention showing opposed fasteners extending from a base;
FIG. 28 is a perspective view illustrating the use of the fastener of FIG. 27;
FIG. 29 is a perspective view of a further modification of the fastener of the present invention;
FIG. 30 is a bottom view of the fastener of FIG. 29;
FIG. 31 is a side view of the fastener of FIG. 29;
FIG. 32 is a perspective view illustrating one use of the fastener of FIG. 29;
FIG. 33 is a perspective view further illustrating the use of the fasteners of FIG. 29 and FIG. 32 so as to form a netting;
FIG. 34 is a perspective view of a further modification showing a rod having integral opposed fasteners; and
FIG. 35 is a view illustrating the use of the rod of FIG. 34.
The present invention provides a device for removably securing a line or the like, said device having a base with at least one fastener integral therewith. The fastener has at least one wall extending outwardly therefrom and is, effectively, flexibly secured to the base with the wall terminating in a lip. In one configuration, the base has a ridge opposite the lip with the ridge and lip being substantially adjacent each other and a spike or serrated member which is secured within the fastener and extends in the direction of the lip and the ridge so that a lace or the like may be forced past the lip and ridge and be impaled by the spike, thereby retaining the lace in a fixed position. In a modification of the present invention, one or more walls may extend from the base with each wall terminating in a lip in opposed positions. Thus, the lips of the wall form the opening instead of the lip and ridge of the wall and base. The various modifications of the device are multifunctional in application.
Turning now to the drawings, there is shown in FIG. 1 a perspective view of one of the embodiments of the fastener of the present invention. Base 29 has extending thereabove upper wall 31 with vertical wall 24 providing an interconnection therebetween. Since the entire fastener is integrally molded from a flexible plastic material with memory, wall 24 effectively attaches upper wall 31 flexibly to the base 29 so as to allow wall 31 to be moved in an upwardly and downwardly position. One such plastic is Delrin� which is a Dupont product.
Wall 31 terminates in a downwardly extending lip 33 and base 29 has an upwardly extending ridge 35 opposite lip 33. The lip and ridge are prefereably of a dimension so as to be substantially adjacent each other thereby providing a closed area at the forward part of the fastener.
Spike 43, preferably of metal, is embedded at its base 45 within vertical wall 24. Spike 43 extends in a direction towards lip 33 and ridge 35. As can be seen from FIG. 2, wherein lace 27 is shown in a position passing between lip 33 and ridge 35, wall 31 is raised by the lace so as to allow the lace to pass between the lip and the ridge and be impaled by spike 43. After the lace is impaled on the spike, the memory of the plastic returns the lip and ridge to the position shown in FIG. 1. Removal of the lace reverses the sequence.
With this configuration, lace 27 can be quickly secured with the fastener. Further, it can be quickly removed from the fastener by merely lifting upwardly and outwardly thereby releasing the lace from the spike.
FIG. 3 is a partial cross-sectional view of a modification of the fastener of FIG. 2 wherein upper wall 51 terminates in lip 53 opposite ridge 55 as in FIG. 1. However, in this modification, a groove 57 is formed in the inner part of lip 53 and spike 59 extends outwardly so as to rest within groove 57. This provides a protective sheath for spike 59 since it is not exposed unless and until a lace is forced between lip 53 and ridge 55, raising lip 53 so as to allow the lace to be impaled on the spike. When the lace has entered the interior of the fastener, lip 53 will resume the position shown in FIG. 3, thus preventing any unwanted human contact with spike 59.
Turning now to FIG. 4, there is shown a further modification of the fastener of the present invention wherein base 61 incorporates two walls 62 and 63 extending in opposite directions, terminating in lips 64 and 65 with wall 61 having mating ridges 68 and 69. Additionally, there is shown the use of two spikes rather than one. When these spikes are used, they should be substantially parallel to each other and extend in the direction of the opening created by the lips of the ridge. This type of fastener may be used as as illustrated for securing the terminal end of rope 70.
FIG. 5 discloses a further modification of a fastener of the present invention wherein base 71 has one fastener including one wall extending in a manner parallel to the base and terminating in a lip 75, mating with ridge 76, and a further fastener having upstanding walls extending basically vertically from base 71 and terminating in mating lips 77 and 78. This type of fastener may be used for securing the end of a rope or the like as illustrated in FIG. 4.
FIG. 6 is yet a further modification of the fastener of the present invention wherein base 80 has two upper walls 81 and 82 terminating in lips 83 and 84 and mating with ridges 85 and 86 in much the same fashion as in FIG. 4. However, a double walled enclosure 82 extends upwardly between the two fasteners. This walled structure does not have any piecing means therein. However, it does include parallel ridges for retaining the end of the rope. These ridges are sufficient for holding the end of the rope since no substantial pressure is exerted at the point.
FIG. 7 illustrates one use of the fastener of FIG. 6 wherein it is secured to the side panels of a pickup truck by any well known means so that a load may be secured to the bed of the truck.
Turning now to FIG. 8, there is shown a further modification of the present invention wherein a base 91 includes integral upper wall 93 extending thereabove and terminating in lip 95 which mates with ridge 97 similar to that of FIG. 1. Again, spike 98 extends outwardly from the inner part of a fastener in the direction of the lip and ridge. As illustrated in this figure and in FIG. 9, lip 95 and ridge 97 may be substantially adjacent each other and, preferably, actually touch. This reduces the possiblity of the lip accidentally hooking any foreign object. This is permissible due to the flexbility and memory of the upper wall 93 and the intermediate vertical wall as discussed hereinabove. In the particular configuration of FIG. 8, the base is designed with a thin flange so as to permit it to be sewn on material by stitching 99 or, if so desired, the same broad bases provides an adequate area for attachment to an article using adhesives.
FIG. 10 shows a further modification of the fastener of the present invention wherein base 101 has an upper wall terminating in lip 103 mating with the ridge 105 on the base and spike 107 extending from the inner part of the fastener as discussed hereinabove. The opposite end of the base terminates in a flange 109 having a borehole 111 therethrough.
FIG. 11 illustrates one usage of the fastener of FIG. 10 wherein the flange 109 has a ring 115 passing therethrough so it may be pased over a post 117. In this manner, rope 113 may be passed between the lip and the ridge and impaled by the spikes so as to retain the end of the rope in the position as shown.
FIGS. 12 through 14 illustrate various preferred enlarged configurations of the spike or retaining means. FIG. 12 discloses a base 121 having a spike of substantially constant dimensions extending outwardly thereform. FIG. 13 shows a base 125 having a spike 127 extending outwardly therefrom with an enlarged end 129 which aids in retaining the lace or the like in place. FIG. 14 shows base 131 having a spike 113 extending outwardly therefrom with the geometrical configuration of the spike increasing in cross-sectional area as it approaches the outer end of the spike.
FIG. 15 shows a base 135 being serrated so as to have flattened teeth 137 extending therefrom so as to retain the lace within the fastener. A use for this type of gripping device has been illustrated in FIG. 6.
FIG. 16 shows a further modification of the present invention wherein a base 141 has an upstanding fastener extending from both ends. The fasteners terminate in lip 143 and 149 which mate with ridges 145 and 151 on the vertical part of the base. Additionally, each fastener has spikes 147 and 153 contained therein. The base may also have integral legs 154 and 155.
FIG. 17 shows one use of the fastener 118 wherein a box having varoous items contained therein uses ropes 159 to maintain the items in the box. Since the fasteners are connected by base 141, made of flexible plastic, the device may be used for boxes having widths up to the dimension of the distance between the two fasteners.
FIG. 18 discloses a further modification of the present invention wherein a base 161 includes three fasteners extending upwardly therefrom with two of the fasteners 163 and 165 being in opposing configuration and the third fastener 167 facing those two fasteners but being removed by 90�.
FIG. 19 discloses how such a device may be used for securing a package by placing the base on one of the faces of the package and securing the line as disclosed. The base may have some adhesive to hole it in place on the box while the rope is being secured.
FIG. 20 discloses a modification of FIG. 18 where two fasteners 171 and 173 extend from the base in opposed fashion.
FIG. 21 discloses a further modification of the fastener of FIG. 18 wherein four fasteners are used all facing each other and spaced about the base at approximately 90� intervals. Either FIGS. 20 or 21 could be used in a manner similar to that illustrated in FIG. 19 as well as for other purposes.
FIG. 22 discloses a further modification of the present invention wherein two fasteners have an integral base including sections 183 and 185 with the fasteners facing in opposite directions. Each of the fasteners has a spike extending from the inner part as described hereinabove.
FIG. 23 illustrates one usage of the fastener of FIG. 22 wherein, as can be seen, the fastener provides for the securing of the ends of two different ropes or the ends of a single rope.
FIG. 24 illustrates a further usage of the fastener of the present invention wherein two strips 191 and 193, having a plurality of fasteners 195 and 197 are secured to the edges of wearing apparel for interconnection by a lace. Such a garment may be a back brace 199 which may be secured with varying tension levels according to individual need by the lacing method using lace 200. Fastener 203, of the type discussed relative to FIG. 25 may be used to secure the ends of lace 200.
FIG. 25 illustrates a further modification of the present invention wherein base 201 has a lengthened upper wall 203 extending thereform with a lengthened spike 205 so as to accommodate a double lace with the spike penetrating both laces as previously indicated, one example of the use of this fastener is shown in FIG. 24.
FIG. 26 illustrates a further embodiment of the present invention which may be used with a shoe. A substantially smooth elongated base 202 having a substantially smooth profile includes angular channels 206 extending through the upper wall so as to provide a plurality of upper walls 204 and 204' similar to those discussed hereinabove. Walls 204' form fasteners which include lace penetrating means (not shown) so that the shoe may be laced as illustrated. These fasteners are flexible so that the shoe may be easily laced and unlaced.
FIG. 27 discloses a further modification of a fastener of the present invention wherein base 207, two upper walls 209 and 211 extending thereabove terminating in lips 213 and 215 which are in opposing configuration. Both lips extending above the associated ridge on the base with each having associated spikes 217 and 219. As shown in FIG. 28, this type of fastener may also be used to connect the ends of two separate lines 208 and 210, or the ends of the same line.
FIGS. 29, 30 and 31 illustrate a further modification of the present invention. FIG. 29 is a perspective view, FIG. 30 is a bottom view and FIG. 31 is a side view of the fastener. The fastener has a base 221 from which four walls extend outwardly therefrom and terminate in lips 223, 225, 227 and 229. Spike 231 extends centrally outwardly in a direction between the lips.
FIG. 32 shows one use for the type of fastener shown in FIG. 29 wherein the spike is of sufficient length so that two lines may be secured therein at substantially right angles.
FIG. 33 discloses how the fastener of FIG. 29 may be used. In this configuration, fasteners 237 are integrally formed with strip 239. Strip 239 is secured at its edges to backing member 241 so as to leave an area through which line 243 may be passed. As can be seen, by using separate lines in a 90� fashion, a netting of any size, shape or configuration may be formed.
FIG. 34 discloses a further modification of the fastener of the present invention which may be used with the fastener as illustrated in FIG. 33, particularly for decorative purposes. Rod 251 includes integrally formed fasteners 253 and 255 on opposite sides thereof. As shown, more fasteners 255 are provided per linear foot than fasteners 253. It should be noted that the number and spatial relationship of the fasteners may be used in accordance with any preselected pattern. Additionally, the rod could be curved if desired.
Use of the rod fastener of FIG. 34 is illustrated in FIG. 35. Rod 251 is shown as hanging from a mesh, as disclosed in FIG. 33, of a particular size using fasteners 253. Because of the increased number of fasteners 255, a smaller mesh 259 may be hung below the rod. This provides a decorative means as well as a means for privacy at lower levels.
The following advantages are now apparent from the above description and drawings.
The function of linking one line into the position established by another fastener is illustrated in FIG. 11 where the opening 111 secures the fastener to the established link. Another line 113 can swiftly be attached and detached at the other end of the fastener. Such fastener could be used in a gymnasium situation where successive groups of players bring their own nets attaching and detaching them to the established fasteners in turn.
The function of linking line to line as illustrated in Fig. 23, could be used if, for example, a common clothes line were cut or broken in its mid section. Ordinarily, it would be necessary to untie theline at either end to free enough line to allow the tying of a knot reuniting the broken halves. If there were no line to spare, it would be necessary to replace the entire line. However, through the use of the fastener of FIG. 23, the line could be repaired without untying or tying anything and the exact tension desired reestablished in seconds, a fraction of the time by any other solution and without the expense of a new line.
The fastener, as disclosed in FIGS. 29-33 secures, at the point of intersection, two lines crossing on a common plane at an angle of 90 degrees. This fastener allows the easy formation of a net of any size and uniquely, of any shape, (not merely squares and rectangles). It eliminates any need for weaving lines together or for tying knots at intersections.
Further, in conjunction with the rod fasteners of FIG. 34, multi-mesh nets may be formed as illustrated in FIG. 35. Nets used today, such as fishing nets, form part of the interior decoration of many restaurants, stores and private homes. Employing the system of the present invention this decorative technique can be greatly expanded. For example, multi-mesh nets could be used as booth dividers in restaurants; a tight mesh at head level, a looser mesh rising toward the ceiling. Again, a net could easily be made to provide a room dividing "wall" with arched doorway or the like. Such a net could be employed as a trellis for climbing plants with heavy meshes where heavier foilage is desired. Such techniques offer designers and decorators unlimited versatility. Further, such structures would be very low in material cost, be easily assembled on site or elsewhere and require no special skill or tools. The system allows an even quicker, simpler disassembly with no loss of material.
The fastener of FIG. 16 may be used with a single line to form an enclosure for holding articles together. FIG. 17 illustrates two such fasteners in use.
The three configurations of the fastener disclosed in FIGS. 18, 20 and 21 may be used with a line to secure a container or the like. FIG. 19 illustrates a box or package thus secured. The method most commonly used today to secure a box with a line ideally requires three hands, the third hand to prevent movement of the line and loss of tension while the other two hands knot the line. Through the use of such fasteners as illustrated, especially if they were provided with "stick-on" bottoms, a line could secure a box quickly, easily and with only one hand.
The most commonly used system for fastening footware comprises laces and eyelets, but this system has some well known drawbacks. For example, a considerable amount of time is generally required for the lacing and unlacing process. After lacing, more time is often required to tighten the different levels to achieve a desirable tension and, finally, knotting is necessary to maintain such tension insofar as possible. In untying, the knot must be undone so that the tension is released enough to allow the withdrawal of the foot. This usually requires that the lace be completely taken out of the upper levels of eyelets. Even more time is required when the lace must be completely removed for washing or to allow the cleaning and polishing of the footware. When lace ends are frayed, the lacing process becomes almost impossible. Further, although it might be possible to establish a comfortable tension at every lacing level during the lacing process, it cannot be maintained during use since the lace moves between eyelets in response to pressure from the foot or leg. This very common problem leads countless boot and shoe users to leave the upper levels of lacing eyelets unused, thus losing the intended support in that area. Countless others suffer varying degrees of pain and discomfort in order to obtain the full support of their footware. Still others, despairing of finding the style of footware they want with the comfort they require, restrict their activities or press unsuitable footware into unintended uses, like wearing western boots for climbing hills.
A less commonly used alternative lacing system makes use of metal hooks. This system allows quick and easy installation and removal of a lace but it has serious drawbacks. Because the hooks protrude well above the surface of the footware and because they are made of hard metal formed like a hook open on three sides, they are a constant hazard to snare any foreign object such as a pant leg, table cloth or another lace, hence they can cause tearing and tripping. Additionally, if brought into contact with furniture or painted surfaces, they can cause pitting or scratching. Further, if any force is brought against the lacing area of such footware, the protruding hard metal hooks will intercept it and transfer it directly to the foot or leg with consequent pain and possible injury. Attempting to minimize this hazard adds bulk, weight and cost to footware. Todays lace hooks are manufactured singly, one at a time and must be handled, positioned and attached one at a time; such labor-intensive work necessarily adds notably to cost. Further, protruding, angular metal hooks cannot be blended stylistically with a smooth leather finish. As with the eyelet system, systems using lace hooks as manufactured today, offer no tension control of the lace since it moves between hooks in response to pressure from foot or let rather than in response to the requirements of comfort.
The system as illustrated in FIG. 26 elminates every single problem encountered in the lacing systems discussed above. Swift and easy lacing is simultaneous with establishing the desired tension at each lacing level. Once set, the tension cannot alter, so that maximum comfort is achieved and maintained until the footware is removed. Unlacing inswift and easy. Since the lace need not pass through narrow eyelets, frayed lace ends are of no connsequence. Force brought against the lacing area is now even less dangerous than with the eyelet system since the broad based flexible lacing strips as shown will absorb and distribute it. These strips may be strong and lightweight, which are easily and inexpensively produced with automatic equipment. Further, the streamlined design, low profile and closed mouths of the fasteners present no hazard to extraneous objects. Finally, being plastic, the lacing strips of FIG. 26 can be produced with any color and finish, thus lending themselves to easy stylistic blending with any footware. Also the fasteners could include a soft plastic layer on the outer surface thereof if a more gentle surface is needed or required.
Referring to FIGS. 4, 5 and 6, the precise combination of fasteners to be chosen depends, of course, on the particular purpose to be served and the composition of the rope being used. Ordinarily, when mooring a boat or stabilizing a load on a small truck, ropes are fixed to some type of cleat by the tying of knots. Care and skill are required lest a poorly knotted rope release and a valuable boat or load be lost. A knot improperly tied might also be very difficult to untie, leading to frustration and lost time.
The fasteners of FIGS. 4, 5 and 6 allow the almost instantaneous secure attachment of a rope with the simultaneous establishment of any desired tension. Referring to FIG. 7, the rope need simply be drawn to the desired tension alongside the mouth of either the fastener on the right or left, then, while maintaining that tension the rope is moved sideways through the mouth of the fastener and into its body where it is impaled on the spikes. The hands now no longer need to grip the rope with any particular force, but can, in an easy manner, rotate the free portion of the rope and attached it to the opposite fastener without leaving any slack. The rope is now doubly secured and the loose end can be looped in any fashion then fixed in the central retainer, thus presenting a neat, finished appearance. Of course, any later tightening that might become necessary can be accomplished with equal ease. Removal of the rope requires the simple reversal of the procedure.
As will now be obvious, the present invention provides a fastening device for various uses relating to lacing and securing of lines and ropes. It is to be understood that the above description and drawings are illustrative only and that modifications could be made in any of the fasteners described without departing from the scope of the invention which is to be limited only by the following claims.
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|U.S. Classification
|International Classification
|Cooperative Classification
|European Classification
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|Fee payment
Year of fee payment: 4
|Sep 9, 1994
|Fee payment
Year of fee payment: 8
|Sep 29, 1998
|Maintenance fee reminder mailed
|Mar 5, 1999
|Fee payment
Year of fee payment: 12
|Mar 5, 1999
|Surcharge for late payment


text:
hello everybody,
maybe you can help me , i am new to this operating system and i have a question. for example... when i go to download a random file off the net it goes to archieve manager? and never opens anything i download always says error . does anybody have any clue on this one? cause i am a idiot i guess
thanks for your anticipated help

subset: commoncrawl


text:
Regional Workhorse
Long-haul comfort for regional runs
For regional hauls that require some time on the road, the 61-inch sleeper on the VNM 630 offers long-haul comfort by combining convenient, common-sense work and storage space with a 42-inch wide bed that makes those extra nights a little more pleasant. The mid-roof sleeper configuration is well suited for any trailer height, making the VNM 630 a versatile player for a wide range of applications.
Mid-Length hood
61" Mid-Roof Sleeper
If you don't want to compromise on fuel efficiency, driver comfort, or safety, this versatile regional workhorse is the ideal choice for operations that include urban delivery environments.
1250-1550 lb-ft torque
325-425 horsepower
Fuel-efficient Volvo D11 and D13 engines feature a standard common rail fuel injection system that gets the most power out of every drop of fuel. Also available with a Cummins ISL G natural gas-powered engine.
Brilliantly Simple
12-speed Volvo I-Shift
The Volvo I-Shift makes drivers more productive, using intelligent electronics to continuously monitor grade, speed, weight, and engine load, shifting when necessary or holding a gear—whichever saves more fuel. So regardless of experience or training, I-Shift helps every driver become more fuel-efficient.
Our commitment to progress
Volvo Innovation
Volvo is Safety
Since 1927, safety has been the guiding principle for everything we build.Read more


text:
Facebook Brings Comments Box to Mobile
Photo of the author, Jennifer Beese by Jennifer Beese on January 12, 2012
Facebook has launched the Comments Box plugin for mobile, which aims to help media sites interact with readers across the web and mobile devices.
The plugin, which is part of Facebook's Social Plugins, allows an individual to comment on a news site regardless of where he or she is reading it. According to the social network, over 400,000 sites – including the Statesman Journal and the Detroit Free Press – have already integrated the plugin.
If you have already installed Comments Box you will notice that the plugin automatically appears on the mobile web version of your site. Facebook hopes that the latest update will make commenting more social, improve the quality of conversations online, and ultimately drive more traffic to media sites.
The social network has also added a Subscribe button, which lets other readers subscribe to a commenter's public Facebook profile. This could really help to boost referral traffic as frequent commenters publish articles to their News Feeds.
Additionally Facebook hopes that the Comments Box plugin will help to reduce trolling and increase the quality of conversations by making readers log in with their real identity, versus leaving anonymous comments. You can learn more about the plugin's technical details on Facebook's Developer site.
[Via: Mashable]
Jennifer Beese: Jennifer Beese has worked as a community manager and social media strategist. When she's not writing, you can find her studying anatomy and physiology—she literally has a skeleton in her closet—or under the stars with her telescope.


text:
Bank of Baroda Admit Card 2016 bankofbaroda.co.in
Bank of Baroda will soon declare its Bank of Baroda Admit Card 2015 for Bank of Baroda Recruitment 2016 written examination at its official website bankofbaroda.co.in. Candidates who have applied for the posts in Bank of Baroda Recruitment 2015, those candidates are looking for their Admit Card 2016. Candidates may download their admit cards through its main portal.
Details about Bank of Baroda Admit Card 2016:
Bank of Baroda has released the recruitment notification on 4th August, 2016 to recruit the candidates for filling up vacant post like Part Time Medical Consultant Post through this recruitment. Bank of Baroda invites the online applications from interested and eligible candidates through online mode from its official website. Bank of Baroda is the public sector banks. It provides all the banking facilities to the citizens. Bank of Baroda will conduct the written examination for medical consultant posts. Selection process will be based on the candidates performance in the interview. Selected candidates can get the pay scale as Rs.1500/- per month.
Bank of Baroda will soon release the Bank of Baroda Admit Card 2016 at its main portal on or before the 10 to 15 days prior of the examination date. All the applied candidates can download their admit card on or before the examination date. Admit card is very important for every examination. Admit card contains the more information about exam name, exam date, exam time, hall ticket number and name of the candidate, address of the candidate and uploaded scanned photograph and signature of the candidate and etc. Without admit card no one will give the permission to enter into the examination hall to write the examination. Candidates can download their admit card by entering the candidates registration number and date of birth etc through its official website. Candidates can follow the below simple steps to get their admit card easily.
How to download Bank of Baroda Admit Card 2016:
- Go to the official website www.bankofbaroda.co.in.
- Click on the link of Bank of Baroda Admit Card 2016.
- Enter your application number and date of birth etc.
- Click on the submit button.
- Admit card will be generated on the screen.
- Download it and save it and take hard copy it for examination purpose.


text:
Some random notes about me:
- I love whimsy. I am sometimes whimsical and mischievous once I get to know someone.
- I'm independent to a fault. I'm still learning to allow myself to rely on others.
- I'm very good when things are very bad. Not that this happens often anymore.
- I'm a cuddler, it's true..
- I like making lists to keep my mind occupied. List of words that are fun to say (wigwam, iambic pentameter,..). List of cool names that have been ruined by product marketing (Ajax, Porsche,..), List of over the top insults (... not suitable ...), things like that.
- I can't spell. No seriously.. I've a math mind.
- With the exception of karaoke, I've found little that I cannot do if I set my mind to it. I'm quite capable, but sometimes struggle with motivation. Often I feel like a 'how' in search of a 'why'.
- As a kid I had a pet raccoon and squirrels. No, I wasn't raised in a Disney cartoon..
- I am INT*, that is, I fluctuate between INTJ and INTP by mood.
- I perceive others through their actions; not what they say, what they think, or how they feel. In my view, one's actions are the clearest indicators of who they are.
- I keep a theoretical construct of how the world works in my head. I whimsically refer to it as WAJ (World According to Jason). Every time the model does not hold, I refine it. It's my own personal mental playground. I go there when my mind wants to run around and play on the monkey bars.
- Ok, so I'm not exactly sane, but I'm not clinically insane either! :)
As a more direct answer:
Software engineering.
Staying in shape is an ongoing battle (desk jobs will kill you!).
I've a plan to live forever that I'm still working out all the kinks in, but with luck I'll have lots of time :)
Oh, and I'm trying to do that dating thing too.
- I'm fairly good at reading people.
- I'm a good listener, though when people bring me their troubles sometimes I have a difficult time identifying when I should be empathizing vs. problem solving. Being in problem solving mode 8+ hours a day makes it difficult to not slip into that mode unintentionally.
Book - Red Mars
Movie - Run Lola Run
Show - Pirates of Penzance (I concede, the ending sucked)
Music - Bejork, Regina Spektor
Food - Sushi
My favorit color is green.
I snack on bananas, oranges, and tomatoes... a lot!
I love storms.
I love to swim, especially in the rain.
I love live performances. I really want to see"The Book of Mormon"
I'm a fan of anything done by Joss Whedon.
Political Theory,
Science (chemistry and physics mostly),
I go through phases, at this moment my obsession is chemistry, and khan academy is feeding my addiction...
I can't sing, and it really bugs me.
My methodically planned strategy for living a drama free life is obviously a FAIL.
This statement is a lie. I don't lie, I just paradox.


text:
|Jamie Slade - Dianne is a consummate professional and communicator. Any...
|Mr. Ben Robert Monro - Dianne is a joy to work with. She is professiona...
Dianne Carpenter
Phoenix, Arizona
If you notice offensive or incorrect information, please contact us.
Mr. Kyle Trauberman
Software Developer@
Apollo Group
Mr. Ben Robert Monro
Manager - Software E@
Theresa Jauregui
SharePoint Analyst@
Apollo Group Inc.


text:
An optimized protocol for microarray validation by quantitative PCR using amplified amino allyl labeled RNA
Validation of microarrays data by quantitative real-time PCR (qPCR) is often limited by the low amount of available RNA. This raised the possibility to perform validation experiments on the amplified amino allyl labeled RNA (AA-aRNA) leftover from microarrays. To test this possibility, we used an ongoing study of our laboratory aiming at identifying new biomarkers of graft rejection by the transcriptomic analysis of blood cells from brain-dead organ donors.
qPCR for ACTB performed on AA-aRNA from 15 donors provided Cq values 8 cycles higher than when original RNA was used (P < 0.001), suggesting a strong inhibition of qPCR performed on AA-aRNA. When expression levels of 5 other genes were measured in AA-aRNA generated from a universal reference RNA, qPCR sensitivity and efficiency were decreased. This prevented the quantification of one low-abundant gene, which was readily quantified in un-amplified and un-labeled RNA. To overcome this limitation, we modified the reverse transcription (RT) protocol that generates cDNA from AA-aRNA as follows: addition of a denaturation step and 2-min incubation at room temperature to improve random primers annealing, a transcription initiation step to improve RT, and a final treatment with RNase H to degrade remaining RNA. Tested on universal reference AA-aRNA, these modifications provided a gain of 3.4 Cq (average from 5 genes, P < 0.001) and an increase of qPCR efficiency (from -1.96 to -2.88; P = 0.02). They also allowed for the detection of a low-abundant gene that was previously undetectable. Tested on AA-aRNA from 15 brain-dead organ donors, RT optimization provided a gain of 2.7 cycles (average from 7 genes, P = 0.004). Finally, qPCR results significantly correlated with microarrays.
We present here an optimized RT protocol for validation of microarrays by qPCR from AA-aRNA. This is particularly valuable in experiments where limited amount of RNA is available.
Gene expression profiling using microarrays has rapidly become an analytical tool of choice for translational research laboratories. Genome-wide or more dedicated microarrays are generally used as a fishing expedition to identify candidate genes or pathways that can be used either for their prognostic performance and/or for their therapeutic potential in many diseases. The technique relies on the relative quantification of mRNA expression in cells or tissues. Circulating blood cells can be used as an alternative to tissue biopsies when these are not available. This alternative nevertheless assumes that a systemic biosignature of the pathological state exists and can be assessed through gene expression profiling of blood cells. Consistently, while biosignatures of blood cells were originally reported to be a useful prognostic tool for acute myeloid leukemia
In addition to the type of blood collection, every steps of the microarray technique can influence the quality of the results. When minute starting amounts of RNA are available, additional steps of amplification have to be performed
Results and Discussion
An alternative to total RNA to perform microarrays validation by qPCR
We took profit of an ongoing protocol of our lab which aimed at identifying new prognostic biomarkers of renal graft rejection. Hypothesizing that inflammation in the organ donnor conditions the success of transplantation
Amplification and amino allyl labeling of RNA inhibits qPCR
To compare the effectiveness of qPCR from total RNA and AA-aRNA, we used samples from the 15 donors for which we had both AA-aRNA and remaining total RNA. 1 μg RNA and 100 ng AA-aRNA were reverse transcribed and resulting cDNAs were diluted 10-fold before amplification by qPCR using primers specific for the ACTB gene. The rationale for using different amounts of RNA and AA-aRNA is that AA-aRNA is generated from only mRNA (a T7-Oligo(dT) primer being used during first strand DNA synthesis in the amplification protocol) whereas RNA includes all RNA species. The choice of 1 μg RNA and 100 ng AA-aRNA, as already used by others
Figure 1
Amplification and amino allyl labeling inhibit qPCR
Amplification and amino allyl labeling inhibit qPCR. Total RNA extracted from whole blood cells of 15 healthy brain-dead organ donors was subjected to amplification by T7 polymerase and amino allyl labeling to generate AA-aRNA. 1 μg of un-amplified and un-labeled RNA and 100 ng AA-aRNA (amplified and amino allyl labeled RNA but not coupled with fluorescent dye) were reverse transcribed using SuperScript II, resulting cDNAs were diluted 10-fold and subjected to qPCR using primers specific for ACTB. (A) Cq obtained from the 15 samples are represented in a box plot. A statistically significant difference was detected between RNA and AA-aRNA (t = -25.23; P < 0.001; paired t-test). (B) Cq obtained with RNA and AA-aRNA for each of the 15 patients were strongly correlated (R2 = 0.85; P = 0.00006; Pearson product moment correlation). Similar results were obtained in two independent experiments.
Amplification and amino allyl labeling decrease qPCR sensitivity and efficiency
Following experiments were performed with a universal reference RNA - the same as used for microarrays - because of its high quality and availability. This RNA is a commercially available mix of total RNA from 10 human cell lines. We first aimed to reproduce the inhibition of qPCR on AA-aRNA reported in Figure
Figure 2
Amplification and amino allyl labeling decrease qPCR efficiency and sensitivity
Amplification and amino allyl labeling decrease qPCR efficiency and sensitivity. One μg RNA and 100 ng AA-aRNA from human universal reference were reverse transcribed with SuperScript II. (A) Ten-fold dilutions of resulting cDNAs were subjected to qPCR using primer pairs specific for VEGFB. The amplification chart shows higher Cq values for AA-aRNA compared with RNA. (B) cDNAs obtained from reverse transcription of RNA and AA-aRNA were serially diluted and qPCR was performed to evaluate qPCR efficiency using VEGFB-specific primers. qPCR efficiency calculated from standard curves (E = [10-1/slope]-1) is down-regulated by amplification and amino allyl labeling.
Table 1
Cq values obtained by qPCR using RNA and AA-aRNA
Mean ± SD
20.7 ± 4.9
25.6 ± 7.0 *
qPCR was performed on universal reference RNA and AA-aRNA using primer pairs specific for the indicated genes. ND: not detectable. * P = 0.02 vs RNA (one way repeated measures ANOVA).
Optimized protocol for reverse transcription (RT)
In an attempt to circumvent the loss of sensitivity of qPCR performed with AA-aRNA, we modified the RT protocol (Figure
Figure 3
Scheme depicting RT protocol optimizations
Scheme depicting RT protocol optimizations.
Protocol optimization improves RT yield
To test RT protocol optimization, we used the universal reference RNA either under its native form (RNA) or after amplification and amino allyl incorporation (AA-aRNA). These 2 RNAs were reverse transcribed either with the original RT protocol, the optimized RT protocol without RNase H treatment, or the optimized RT protocol with RNase H treatment. This dichotomy allowed us to evaluate the effect of RNase H
Figure 4
Protocol optimization improves RT yield
Protocol optimization improves RT yield. One μg RNA (A) and 100 ng AA-aRNA (B) obtained from universal reference RNA were used as inputs to either the original RT protocol (black bars), the optimized RT protocol without RNase H treatment (white bars), or the modified RT protocol with RNase H treatment (hatched bars). Resulting cDNAs were subjected to qPCR using primer pairs specific for VEGFB, MMP9, TFRC, HAMP and GAPDH. Threshold Cq values for each gene and the mean ± SD of the 5 genes are indicated. RT protocol optimization decreased Cq values and RNase H did not induce a further decrease. * P < 0.001 vs original RT protocol (one way repeated measures ANOVA). ND: not detectable.
RNase treatment improves qPCR efficiency
We next investigated whether the optimized protocol and RNaseH treatment alter RT efficiency and linearity. It was indeed reported that RNase H may allow for amplifying certain genes which may not be accessible to PCR
Figure 5
RNaseH improves qPCR efficiency
RNaseH improves qPCR efficiency. Different quantities of AA-aRNA from universal reference RNA (100-1000 ng) were used as inputs into either the original RT protocol or the optimized RT protocol, with or without RNase H treatment. Resulting cDNAs were diluted 10-fold and subjected to qPCR using primer pairs specific for VEGFB, MMP9, TFRC, HAMP and GAPDH. Cq values were plotted against the Log of the concentration of the AA-aRNA used for RT and linear regression was applied. qPCR efficiency (E) was calculated by the slope of the regression line. A slope of -3.2 indicates optimal efficiency. qPCR linearity (R2) corresponds to the correlation coefficient of the regression line. A coefficient R2 of 1 indicates optimal linearity. (A) Representative experiment using VEGFB primers. (B) Plots representing qPCR efficiency as a function of linearity for the 5 genes tested.
Table 2
Effect of protocol optimization and RNase H on qPCR efficiency and linearity
Mean ± SD
Efficiency (slope)
- RNase H
-1.96 ± 1.10
+ RNase H
-3.43 ± 0.30
- RNase H
-1.36 ± 0.70
+ RNase H
-2.88 ± 0.29
Linearity (R 2 )
- RNase H
0.80 ± 0.18
+ RNase H
0.98 ± 0.01
- RNase H
0.90 ± 0.11
+ RNase H
0.98 ± 0.01
This table gathers the results obtained from experiments shown in Figure 5 and shows statistical analyses.
These data show that RNase H treatment improves qPCR efficiency. This result is consistent with previous data
Importance of the distance from 3' end in the design of qPCR primers
Random hexamers used in traditional RT protocol bind anywhere in the RNA, allowing RT of all RNA independently of their size. In contrast, the RT of the amplification procedure that generates a AA-aRNA for microarrays is performed with T7 oligo(dT) primers that anneal only to the poly(A) tails of mRNA. This is an important limitation of this protocol since only mRNA below a certain length (typically around 1000 bp) may be correctly reverse transcribed, and therefore detectable by qPCR. To address this issue, we designed 5 pairs of primers located at different distances from the 3' end of the hif1a gene (Table
Table 3
List of primers used in this study
cDNA length (bp)
Genbank accession number
Forward primer
Reverse primer
Distance from 3' end (bp)
Primers were designed with the Beacon Designer software. All PCR products were sequenced to confirm specificity.
Figure 6
Choice of primer location for qPCR from AA-aRNA
Choice of primer location for qPCR from AA-aRNA. One μg of universal reference RNA and 100 ng AA-aRNA were reverse transcribed using our optimized RT protocol. Resulting cDNAs were diluted 10-fold and subjected to qPCR using 5 pairs of primers located at different distances of the 3' end of the hif1a gene. Cq values were plotted against these distances. Linear regression lines illustrate the effect of increasing the distance from 3' end. Moving away primers from the 3' end increased Cq values when performing qPCR with cDNA generated from AA-aRNA.
Validation of microarray results by qPCR using AA-aRNA
As stated before, the ultimate goal of performing qPCR on AA-aRNA is to validate microarray results. We therefore tested whether the implementations described in this paper improved this validation. For this purpose, we used AA-aRNA obtained from blood cells of the same 15 brain-dead organ donors described earlier. On one hand, these 15 AA-aRNA were analyzed by genome-wide microarrays. Among the 25,000 genes represented on the microarrays, 5 genes were selected based on their significant association with renal graft rejection (analyses not shown here): the cytokine Tumor Necrosis Factor-α (TNF-α), the accessory protein LY96, the neutrophil elastase ELANE, the adhesion molecule annexin A1 (ANXA1), and the transcription factor Signal Transducer and Activator of Transcription 3 (STAT3). Expression levels obtained by microarrays were calculated as the log ratio sample vs universal reference RNA. On the other hand, expression levels of these genes were assessed by qPCR starting with the same 15 AA-aRNA that were used for microarrays. 100 ng of each AA-aRNA was reverse transcribed either with the original RT protocol or with the optimized RT protocol. The resulting two types of cDNA were subjected to qPCR using primers specific for ACTB, GAPDH, LY96, ELANE, ANXA1, STAT3 and TNF-α. Two pairs of primers were used for TNF-α, the first one (TNF-α-1) located 1243 bp away from the 3' end of the tnf-α gene and the second one (TNF-α-2) located 119 bp away from the 3' end (Table
Figure 7
RT protocol optimization decreases qPCR Cq values
RT protocol optimization decreases qPCR Cq values. Total RNA extracted from blood cells of the same 15 brain-dead organ donors used in Figure 1 was amplified and amino allyl labeled to generate AA-aRNA. 100 ng of each of these AA-aRNA were then reverse transcribed to cDNA using either the original or the optimized RT protocol, and qPCR was applied to assess expression levels of ACTB, GAPDH, LY96, ELANE, ANXA1, STAT3 and TNF-α. Results are represented as Cq values for each gene (mean ± SD of the 15 donors) and as the mean ± SD of the 7 genes. The optimized RT protocol decreased Cq values. ND: not detectable. * P < 0.001 and # P = 0.004 vs original RT protocol (paired t-test).
We next determined whether the optimized RT protocol affected the correlation between microarray results and qPCR data. For this purpose, expression levels obtained by qPCR were normalized to GAPDH and plotted against expression values obtained by microarrays (Figure
Figure 8
Correlation between microarrays and qPCR
Correlation between microarrays and qPCR. Gene expression profiles of blood cells from 15 brain-dead organ donors were analyzed by microarrays covering 25,000-genes. Among these, the following were considered in these experiments: TNF-α, LY96, ELANE, ANXA1 and STAT3. Expression levels of these 5 genes obtained by microarrays were calculated as the log ratio sample vs universal reference RNA. 100 ng of AA-aRNA remaining from microarray experiments were reverse transcribed using either the original or the optimized RT protocol and assessed for expression levels of these genes by qPCR. GAPDH was used as reference gene for normalization. For each gene, expression data from microarrays (log ratio gene/reference) were plotted against expression data from qPCR calculated with the formula (2Cq gene - Cq GAPDH). Correlation between microarrays and qPCR, either using the original (●) or the optimized (○) RT protocol, were calculated by Pearson product moment correlation and are summarized in Table 4. Linear regression lines are displayed for correlations between microarrays and qPCR data obtained with the original (full lines) or the optimized (dotted lines) RT protocol. RT protocol optimization did not modify the correlation between microarrays and qPCR.
Table 4
Correlation between microarrays and qPCR
Microarrays vs original RT protocol
Microarrays vs optimized RT protocol
This table gathers correlation coefficients (R2) and
We have implemented an optimized protocol for the validation of microarrays data by qPCR. This protocol allows using AA-aRNA leftover from microarray experiments when limited amount of RNA is available. It can aid in the quantification of low-abundant genes and provides a significant level of correlation between microarrays and qPCR. In addition, this protocol provides high-quality material that can be used to validate expression of relevant genes that may be highlighted by other approaches than microarrays. Such approaches, some of them being increasingly used in the field of biomarker or therapeutic targets discovery, include bioinformatic analysis of functional networks or signaling pathways
RNA samples
Two types of RNA were used in this study. First, RNA was extracted from whole blood cells of 22 brain-dead organ donors. Median age was 50 (36-64), 14 donors were males, and the initial events that led to brain death were cerebrovascular accident (n = 10), brain trauma (n = 6), acute onset of brain hypoxia (n = 2), suicide (n = 2) and trauma (n = 2). According to the French legislation, studies on brain-dead patients do not require informed consent. The French "Agence de la Biomédecine" approved the protocol and blood was withdrawn after signature of next of kin for all scientific studies of the brain-dead patient. Diagnosis of brain death was established according to the criteria of the French "Agence de la Biomédecine"
RNA amplification and amino allyl labeling
Messenger RNAs were amplified using the Amino Allyl MessageAmp® kit (Ambion, Cambridgeshire, United Kingdom) according to the manufacturer's protocol, starting with one μg of total RNA. This protocol is based on the Eberwine RNA amplification procedure
Reverse transcription of RNA and AA-aRNA
The same RT protocol was applied to RNA (un-amplified and un-labeled RNA) and AA-aRNA (amplified and amino allyl labeled RNA but not coupled with fluorescent dye). 1 μg of RNA and 100 ng of AA-aRNA were reverse transcribed into cDNA using the SuperScript II reverse transcriptase (Invitrogen, Merelbeke, Belgium) with the following protocol: RNA or AA-aRNA was mixed with the 5× RT buffer, random hexamers, dNTPs and DTT in a total volume of 19 μl. Samples were then heated to 42°C for 2 min, and 1 μL of SuperScript II was added to a total volume of 20 μl. Final concentrations were: 50 mM Tris-HCl, 75 mM KCl, 3 mM MgCl2, 0.5 mM dNTPs, 10 mM DTT, 200 U of SuperScript II, 180 ng of random hexamers (Invitrogen). RT was allowed for 50 min at 42°C and was followed by enzyme inactivation at 70°C for 15 min. The absence of contaminating DNA was checked using no RT assays.
Optimized RT protocol. RNA or AA-aRNA were mixed with 180 ng random hexamers and dNTPs to a total volume of 13 μL, heated to 65°C for 5 min and rapidly chilled on ice for 5 min. 4 μl of 5× RT buffer and 2 μl DTT were then added and samples were incubated for 2 min at 25°C. Then 1 μL of Superscript II was added and samples were pre-incubated for 10 min at 25°C before RT for 50 min at 42°C. RT was stopped by heating to 70°C for 15 min. Finally, 1 μL (2 U) of RNase H (Ambion) was added, and incubation was continued for 20 min at 37°C. Reagent concentrations were the same as above.
Quantitative real-time PCR (qPCR)
cDNAs obtained from RT of RNA or AA-aRNA were diluted 10-fold and 4 μL were mixed with 16 μL of SYBR®Green Master Mix (Biorad, Nazareth, Belgium) containing 300 nM of each primer (final volume 20 μL). Amplification was carried out in the IQ5 thermal cycler (BioRad) under the following conditions: heating for 3 minutes at 95°C, 40 cycles of denaturation for 30 seconds at 95°C, followed by an annealing/extension for 1 min. A negative control without cDNA template was run in every assay and measures were performed in duplicates. Primers were designed with the Beacon Designer Pro 7.0 software (Premier Biosoft) and their characteristics are indicated in Table
Additional file 1
MIQE_checklist. Minimum information for publication of quantitative real-time PCR experiments guidelines.
Click here for file
Transcriptomic profiles of whole blood cells from 22 brain-dead organ donors were obtained using oligonucleotide microarrays representing 25,000 genes
Statistical analysis
Results are presented as mean ± SD or as median (interquartile range) for description of demographic characteristics. Comparisons between two groups were performed with two-tailed t-test for Gaussian data and Mann-Whitney test for non Gaussian data. Comparisons between multiple groups were performed with one way ANOVA for Gaussian data and Kruskal-Wallis one way ANOVA on ranks for non Gaussian data. Paired data among multiple groups were compared with one way repeated measures ANOVA and all pairwise multiple comparison procedures (Holm-Sidak method). Correlation tests were performed using the Pearson product moment correlation method. Statistical significance tests were generated with the SigmaPlot v11.0 software and the SigmaStat software (for Windows version 3; SPPS Inc. Chicago, Illinois, USA). A
Authors' contributions
CJ and YD jointly designed the study and developed the optimized RT protocol. CJ carried out the experiments. YD drafted the manuscript. DL and PMM participated in the design of the study and clinical sample collection. DRW provided a critical review of the manuscript. All authors participated in the drafting of the manuscript. All authors read and approved the final manuscript.
This study was supported by grants from the "Agence de Biomédecine" (France), the Centre de Recherche Public-Santé (Luxembourg), and the Ministry of Culture, Higher Education and Research (Luxembourg).
Prognostically useful gene-expression profiles in acute myeloid leukemia
Use of gene-expression profiling to identify prognostic subclasses in adult acute myeloid leukemia
Using peripheral blood mononuclear cells to determine a gene expression profile of acute ischemic stroke: a pilot investigation
Correlation of Peripheral-Blood Gene Expression With the Extent of Coronary Artery Stenosis
Whole blood and leukocyte RNA isolation for gene expression analyses
Stabilization of mRNA expression in whole blood samples
Failure of the PAXgene Blood RNA System to maintain mRNA stability in whole blood
Strategies for microarray analysis of limiting amounts of RNA
Amplified RNA synthesized from limited quantities of heterogeneous cDNA
Comparability of microarray data between amplified and non amplified RNA in colorectal carcinoma
Efficacy of RNA amplification is dependent on sequence characteristics: implications for gene expression profiling using a cDNA microarray
Amplification biases: possible differences among deviating gene expressions
The T7-primer is a source of experimental bias and introduces variability between microarray platforms
Microarray analysis of defined Mycobacterium tuberculosis populations using RNA amplification strategies
Validation of array-based gene expression profiles by real-time (kinetic) RT-PCR
The role of postischemic reperfusion injury and other nonantigen-dependent inflammatory pathways in transplantation
Comparison of microarray-based mRNA profiling technologies for identification of psychiatric disease and drug signatures
Angiogenesis and endothelial cell repair in renal disease and allograft rejection
RNase H and its effects on PCR
Improvement of reverse transcription PCR by RNase H
The search for new cardiovascular biomarkers
Gene expression levels assessed by oligonucleotide microarray analysis and quantitative real-time RT-PCR -- how well do they correlate?
Microarray validation: factors influencing correlation between oligonucleotide microarrays and real-time PCR
Identification of potential targets in biological signalling systems through network perturbation analysis
Computational biology for cardiovascular biomarker discovery
Aspects historiques, réglementaires et épidémiologiques de la mort encéphalique, Prise en charge des sujets en état de mort encéphalique dans l'optique du prélèvement d'organes et de tissus
An open-access long oligonucleotide microarray resource for analysis of the human and mouse transcriptomes


text:
This is the story of Harvey Free D4 male tribute in the zombie apocolipse.
January 17th 2084
My name is Harvey Free and i may be the last man alive.
I´ve been on the move for so long, running from those things, Those monsters that used to be human. They were bakers, programmers, teachers, neighbors. Now they sniff the air like wolves and their dead eyes watching endlessly for their prey. Me.
I´m down to a few supplies and my trusty wrench. i´m so tired but i have to get to the city before nightfall.
January 21st 2084
I´m not alone! I finally made it to district 4, and was so excited that i almost got myself killed on my way to their fort. They´ve walled off 4 blocks of the city, but they dont know what to do next. i´ve decided to join them, lead them, to put my foot down here and stop running.
Taking back this city is going to be challenging, but we´re prepared to do what it takes to reclaim this place in the name of humanity.
Today we begin to rebuild.
Day 1
They have maneged to wall off 4 blocks of D4. A suburb with 2 small houses ( where i am currantly living), A Two floor apartment, A police station with with basic weapons and three police cars with no gas and a Farm.
There are 6 survivers including me. Three soldiers, Mohammed Henderson, Sam Young and Valentino Martin.One scavenger called Raphael Campbell and one Builder Calham black (To who i have given my luckey Wrench).
We have 50 food stockpiled with the farm producing and extra 3 every day. We are losing 3 food every day so if we want more people to join us we will need to either send out some scavengers to get food or rebuild another farm house.
Currantly there are 16 zombies surrounding our Fort. To the south there is a mall that we could reclaim to help with the defense. There is a farm to the North east of us so we will try and reclaim it.
Missons: i am sending Raphael and the three soldiers to The near by farm to scavenge for food and weapons, but there are two zombies there but i dont think it will be a problem.
Me and Calem Black are going to the suburb to try and recruit a survivor.
Day 2
Raphael Met a scruffy, hungry looking pooch while scavenging. Despite her scrawniness the old girl had so much energy it cheered us right up. We named her Marmaduke. Since Raphael Found her he can keep her considering they seem to like each other. they also found 17 food.
Our recruitment was a success! This scientist Ryder Butler was so desparate for some solid walls between him and the zombies, he just about jumped into our arms Welcome aboard!
Missions: We are sending Raphael And Marmaduke to get the rest of Ryders Supplies from the suburbs.
Sam and Valentino are going to The farm house to kill the zombies there.
Me and Mahammed are going to the church to recruit two survivors.
Calham Black is going to scout the mall. Ryder Is staying in the base today as it would be unfair to send him out after he just got here.
Day 3
Scavenged for supplies: Thank god for beef jerky. It´s the first real meat i´ve eaten in three months. Cocktail wieners don´t count.
Recruitment Failed: The woman we were trying to recruit wouldnt lower her gun the entire time we talked to her. said she isn´t convinced its safe at the fort and that she would be better of by herself. maybe we will try again later.
Mall Scouted: The mall looks like a shitty place to live, but there is at least one guy trying to do that. we should invite him to join us and give him deceent digs. there is some food there as well.
Sam and valentino still havent returned from they´re zombie killing mission.
Missions: Me and Raphael are going to the mall we scouted to recruit the man there.
Mohammed, Ryder and Calham get the day off.
Day 4
21 Zombies attacked our fort. Another easy victory thanks to some well placed traps. Its a good thing there were people at the fort.
Zombies Killed: Zombies love fireworks. I saw this in a film once and it really works! The buggers just stand there gaping at the sky mezmorized, making them easy picking.
Recruitment success: Always good to have another scientist to help out. Claudio Carmel seems pretty handy and eager to get to work.
Calham Black is going to the Farm to reclaim it, but it will take four days.
I have been thinking. We should start looking for a Lab for our scientists so that we can resurch Zombie vitals or radio or electricity.
Missions: Black Reclaiming the Farm.
Raphael, Sam and marmaduke are going back to the mall to get Claudios supplies.
Two survivors have been spoted at the church but they have no food and the zombies are starting to surround them so I am going in to recruit them.
Valentino and Mahummed are going to clean out the suburbs to the north.
Claudio Carmel is going to scout the church. And Ryder is going scouting at the All-Mart, leaving nobody at the Fort.
Day 5
Survivors Spotted! There is a group of people camped out in the church. They´ve got enough food for now but without our help i dont know how long they will last.
Recruitment success: I met a scientist in the church. He agreed to come with us but on the condition that his husband come with him two. The Gay couple are Billiam and Diego Fisher. Billiam is a Scientist and Diego is A builder. We now have no more space for survivors.
Scavenged for supplies: Found a little bonus in the mall: Two boxes of old 1980s porno mags. This should keep the boys entertained though it seems that mostly all these girls had going for them was their hair. Morale increased after this along with 8 food.
Ryder reported back from his scouting mission and said " There is a shit ton of food in the AllMart. Makes my mouth water just thinking about it."
Our food stockpile is at 67 but we are losing 7 food every day but after our scavengers get that food from the AllMart food wont be a problem for quite a while.
Missions: Raphael, Sam and Marmaduke are heading to the AllMart to get the supplies and will be back tommorow.
Billiam, Claudio, Ryder and diego are scouting on the outside of the Fort, while i stay on guard duty.
23 Zombies attacked but the fort held. I had to get my hands dirty this time, but nobody was Killed. We killed 3 Zed but the rest fled.
Day 6
Zombies killed: Valentino reports that he saw a hoarde of zed coming for the fort so him and the boys took a few down. Good thing too because if there had been more i dont think i could have stopped them.
AllMart Scavenged: We dug up some sweet loot at the AllMart. 17 food and an awesome cowboy hat. That Just earned Sam a place as a leader and a scavenger.
Survivors spotted: Claudio-"I scouted the big farm and was suprised to find people there! They were aggro so i left them alone, but they are probubly just as scared as everyone else.
I have Decided that seen as we have no more space for survivors that the least we can do is clear out the zombies around them so they can at least sleep in peace.
Missions: Raphael, Same and marmaduke are going to the big farm but i have given them weapons just in case.
Muhammed and Valentino are Killing zombies at the hospital. We are also hoping to reclaim it and reaserch a Anti-Zombie Medicine.
The three scientists are going to scout the lab to the east of out fort. they hope to find something there.
Me and Diego get the day off and will spend it in the fort.
21 Zombies attacked the fort but it held. Zed made a mess out there but we both kept our cool. We killed 5 Zed in total.
Day 7
Scavenged The Big Farm: 15 food and we found at treat. Mash potatoes with gravey,Roast Turkey and Pudding. Of course it will be cold because we have no electricity but you cant be picky.
Survivors spotted: Claudio " I met some stray survivors living at the farm. They were scared as hell and wanted to stay put but i bet Harvey Or one of the other Leaders could recruit them."


text:
Rahul Sundaram wrote:
Chris Weyl wrote:It's hardly a "small additional cost", especially when looked at in aggregate; this would be an additional non-automateable step required for every update release that is of very questionable value/utility tothe vast majority of our users.I wouldn't say that. Currently it is of questionable value currently but it would be much more valuable without just version bumping and more careful thought put into justifying the need for updates. Every update adds to a large cost in terms of build time, bandwidth, potential regressions etc.
There are distros for people that feel that way. I don't think any of them are named "Fedora".
Then there are distros that believe in following upstream and shipping their bug-fixes and feature enhancements. I'm using my favorite one of those :-). Such distros, incidentally, are great for developers (/me waves) who need to follow upstream (due to dependencies, or just desire to watch upstream) but who need something more stable than rawhide.
Should we not release any updates without a Fedora bug being filed asking to upgrade to the latest upstream?
-- Matthew Please do not quote my e-mail address unobfuscated in message bodies. -- find / -user your -name base -print0 | xargs -0 chown us:cats -- Unknown


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Rapita Systems announces the availability of the RTBx real-time data logger family. RTBx is designed to make it easy to collect execution time data from virtually any embedded, real-time target with minimal impact on the execution time of the application. The high data capacity of RTBx means that timing data can be collected from even the longest running test cases.
Execution time data can be used in a variety of ways in analyzing and optimizing applications for embedded, real-time targets. For example, it can be used with Rapita's execution time analysis tool, RapiTime, to measure minimum and maximum execution times, to verify code coverage, to obtain a predicted worst-case execution time, or to find performance hotspots within the code as optimization candidates.
"The RTBx family provides an effective, target-independent approach for logging execution time data for real-time applications" commented Dr Andrew Coombes of Rapita Systems. "The capacity of the RTBx is astonishing – our experiments have shown that running a fully-instrumented version of the Dhrystone benchmark on a 96MHz ARM9, an RTBx can collect trace data for up to 151 days."
The RTBx is available in two variants: the RTBx1210 captures data up to 16 bits wide and the RTBx1220 captures data at higher rates, in larger volumes and up to 32 bits wide.
For further information on RTBx, visitwww.RapitaSystems.com/RTBx
About Rapita Systems Ltd
Rapita Systems is a specialist in on-target timing analysis of real-time embedded systems. Its RapiTime tool suite performs real-time timing analysis of embedded systems including avionics, telecommunications, space and automotive electronics. RapiTime provides a unique solution for analyzing worst-case execution time and performance profiling. RapiTime provides real-time timing analysis for complex software running on most microcontrollers and microprocessors. A privately owned company, Rapita Systems was founded in 2004 to commercialise worst-case execution time analysis and simulation technology developed in the Real-time Systems Research Group at the University of York.


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Women's Gray 'Cruise' Woven Slip-On Sneaker
Elevate your casual line-up with Munro's gray Cruise sneakers. The woven upper, XL Ultralite sole, signature cushioning and rubber sole combine to make these minimalist sneakers a fail-safe addition to your weekend wardrobe. Simply partner this pair with your favorite jeans for no-fuss style.


text:
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November 24, 2016
We are currently seeking a QA Tester that has 5+ years of Web API Testing experience.
All candidates must sit onsite daily in Winston-Salem and not require sponsorship.
If you are qualified and interested, please apply to this posting immediately.
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Join TEKsystems®, a leading IT staffing, IT talent management and IT services firm, and get your career on the fast track. We have more than 100 offices worldwide, and we partner with over 6,000 clients and place over 80,000 consultants per year. At TEKsystems, we seek to understand our consultants' skills, goals and interests, allowing us to present targeted job opportunities on a contract, contract-to-hire or direct placement basis. TEKsystems' leadership in the market stems from our sincere and personal commitment to driving the success of our customers, consultants and each other.
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To reflect current and future market requirements, the payments industry recognised the need to create a new 3-D Secure specification that would support app-based authentication and integration with digital wallets, as well as traditional browser-based e-commerce transactions. This led to the development of - EMV 3-D Secure – Protocol and Core Functions Specification. The specification takes into account these new payment channels and supports the delivery of industry leading security, performance and user experience.
To learn more about the EMV 3-D Secure Specification, watch the EMVCo demo, listen to the webcast and read the FAQ.
The EMV 3DS Specification received input from EMVCo Technical Associates throughout its creation. EMVCo welcomes new participants who are interested in contributing to the evolution of the specification to engage with EMVCo and get involved in the discussion to ensure their long-term requirements are considered. Learn more.


text:
On 9/15/06, Rex Dieter <rdieter math unl edu> wrote:
Stephen John Smoogen wrote: > I would like to suggest the following plan for Enterprise Extras: > > 1) There are 4 channels per {Scientific/Centos} Enterprise Linux > extras > extras-devel > extras-testing > extras-updates Frankly, *way* too much complexity here. Heck, we don't even have a -testing repo for FE yet. When/if we figure out -testing for *all* of Extras, then EE will naturally benefit from it as well.
+1 This project will take off or it won't, we should probably let items like this mature on their own as time goes on and the need for them becomes more apparent. No need to add overhead to something that doesn't even exist yet ;-) -Mike


text:
"Sing A Christmas Carol" is the opening song from The Stingiest Man in Town and its animated adaptation.
Introduction version
Chorus: Holly-Ho Holly Ho-Ho-Ho
Holly-Ho Holly-Ho,
Sing a Christmas Carol written long ago.
B.A.H. Humbug Esq.: Written long ago.
The Merriest of Merry Christmases to you!
What a lovely place to meet.
Welcome to the merry household of the
kindest and most generous man in town,
Ebenezer Scrooge. You're surprised that
I called Scrooge kind and generous?
Well, you could take my word for it.
Who am I? The London Humbug, of course.
B.A.H. Humbug, to be more precise. True,
Mr. Scrooge wasn't always so jolly.
There was a time back when he was as mean and
miserable as the counting house in which
he conducted his business
- The Offices of Scrooge and Marley.
Now, Marley was dead to begin with,
but old Scrooge will never pay to have
the name painted out on the office sign.
There was a mean and
stingy man named Ebenezer Scrooge,
His heart was hard and cold because
he was the deviled stooge,
While Scrooge sat down to count
the gold that his mind
could never leave,
Young people in the
square outside celebrated Christmas Eve.
"Humbug" used to be his favorite word.
My mom heard it so often, that's how
I got my name. Oh, but
Scrooge was a tight-fisted,
squeezing, grasping, clutching,
covetous, old sinner! Until that
Christmas Eve, when the ghosts came.
Ladies and gents, boys and girls,
a ghost story for Christmas -
The Stingiest Man In Town!
That night when Ebenezer Scrooge
lay dreaming in his room,
He heard the sound of rattling
chains come clanking
through the gloom,
And why he lay there
shivering in a icy wind of fear,
The Ghost of Scrooge's partner,
Jacob Marley, did appear.


text:
Ocean City
Spring Golf Event set
The Ocean City Board of Realtors has announced its Spring Golf Event on May 8 at the Greate Bay Golf and Country Club in Somers Point.
Funds raised will benefit local high schools for scholarships and scholastic programs. The day includes lunch and dinner with an awards ceremony, silent auction, raffle and more. Prizes will be awarded for tournament contests: tournament winner, runner-up, closest to the pin, hole-in-one, putting challenge, and longest drive - man and woman.
Unique to this tournament is the chance to sponsor a Commemorative Pin Flag to honor graduates of Ocean City High School. The flag can be personalized with graduate's name and class year and after the event, sponsors can take the hole flag home. Individual hole sponsorships are available, as well as other sponsorship opportunities. The Early Bird Rate for golfers ends April 25. Tickets also are available for dinner only. Proceeds benefit local high school scholarships and scholastic programs. Last year's tournament funded three college scholarships and made it possible to sponsor school programs such as After Prom and Booster Club.
To purchase tickets for golf, sponsorships or dinner, see ocbor.com or call 609-399-0128.


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Mary Lynn Watkins, who for more than two decades trained Mardi Gras royalty and their courts in Carnival protocol, died Dec. 4 at Tulane Medical Center after a long battle with lupus. She would have turned 57 on Dec. 5.
Watkins, who was known as Lynn, graduated from St. Martin's Episcopal School and attended LSU before earning her undergraduate degree from Mount Vernon College for Women in Washington, D.C.
She lived in Connecticut, New York, South Carolina, Massachusetts, Maryland and Texas before returning to New Orleans in 1992.
Since her return to her hometown, Watkins spent much of her time training Carnival kings and queens in royal protocol and preserving the traditions of the celebration.
"Mardi Gras was her life," said businessman John P. "Jack" Laborde, who reigned as Rex this year.
Laborde said Watkins' illness prevented her from working with him this year, though he wanted her to. He said he went to her house to let her know he would be Rex, and Watkins started crying.
"We kind of didn't get anything done that day because she was emotionally spent," Laborde said. He later found out that she was hospitalized the day after his visit and so would be unable to work with him for his reign as king of Carnival.
But Laborde also recalled an earlier time when Watkins was of immense help. When he was named captain of the Krewe of Osiris several years ago, Watkins was "the first lady I went to," he said.
Laborde said he told Watkins he wanted to create a new Osiris ball from scratch.
"I said, 'I want a whole new deal,' " he said. Watkins and her father, a former krewe captain, worked with him to create the new ball he wanted, Laborde said.
"She was delightful," he said. "She helped me a lot personally."
Peggy Scott Laborde, who co-anchors WYES-TV's annual Mardi Gras broadcast of the meeting of the Rex and Comus courts, said Watkins "wanted to make sure everybody had fun."
Peggy Laborde, who is not related to Jack Laborde, said she remembers watching Watkins at work during a rehearsal for the Rex ball. Ball participants included people of all ages, from adults to young children.
"Boy, she just kept everybody in line," she said. "But she was never overly stern."
"It's a great honor for their families, and you want to look like you're doing this the correct way," she said. "They made it look very seamless."
Jack Laborde recalled watching her work during the Osiris ball.
"She would sit in the back with flashlights, and the girls (members of the court) would react off her flashlight flashes," he said. He said Watkins wouldn't reveal to him what her signals meant, but the ball participants knew what they were supposed to do when they saw the lights.
Watkins worked with more than a dozen Carnival organizations.
She was a member of the Orleans Club, the New Orleans Country Club, Chi Omega Fraternity and the Junior League. She was founding member of Ladies in Red, a group of friends who celebrate each year at the Rex luncheon.
Survivors include her parents, Marylin and Eben T. "Buddy" Watkins III, and a sister, Lois Smitherman.
A service was held last week. Lake Lawn Metairie Funeral Home was in charge of arrangements.


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Spotter presents his new solution: "SpotterDashboard"
The increasing quantity of data, the need to step back in the face of the acceleration of daily life and the need for analysis to grasp the essential issues fuel the demand for a new generation of tools. Spotter has taken up this challenge.
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Integrated in Spotter's SaaS platform the new module allows the viewing, administration and personalisation of interactive dashboards based on decision modelling.
Accessible on the iPad and other tablets, the dashboards are made up of three kinds of widgets : graphics, syntheses and scores. They all allow access to the underlying project data at a mouse-click.
SPOTTERDASHBOARD allows a multi-level analysis based on integrated dashboards. Parametrised individually for each project, the interface can be personalised according to the user's needs.
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SPOTTERDASHBOARD is one of the three modules that make up the Spotter media monitoring and analysis platform together with SPOTTERANALYTICS and SPOTTERSTUDIO. It contains all the power and high quality of Spotter technology.
With the next generation of its platform, Spotter is offering its clients a real workshop of analysis tools which go beyond monitoring to support decision-making.
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Spotter integrates not just Internet (online media, websites) and social media (forums, blogs, social networks, opinion forums, comments on online articles, etc) but also traditional media (print, television, radio) according to project needs for a 360° vision.
Monitoring and analysis are reliable even at very high volumes of non-structured data from heterogeneous sources through the combination of detection and classification through text mining followed by quality control and adjustment by an expert.
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To find out more about SPOTTERDASHBOARD – the dynamic decision support dashboard – or about our modules SPOTTERANALYTICS and SPOTTERSTUDIO contact us :
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Telling stories of resistance is itself an act of resistance.
When theology fails
After Ruth Everhart was raped, she had to rebuild her beliefs about God's will.
Survivors are in our pews—and pulpits.
This man picked a very good moment to remind me of his power.
Attaining justice for victims of sexual assault cannot be a matter of belief or disbelief. They are individuals, not symbols of a cause.
Arthur Remillard sees the best of football's warrior culture as a man training his body into subjection for the protection of the weak and the advancement of all righteous causes. And maybe it's because I know so little about football, but I don't see it. How does throwing a ball around a field protect the weak? How does sucking all the money from educational institutions advance righteous causes? How does making a touchdown make a man more righteous?
Astonished, by Beverly Donofrio
Beverly Donofrio had just been "looking for a monastery to join, for Christ's sake." She had closed her laptop, having bookmarked religious communities she might write to, then had fallen into a deep sleep. During the night she was raped at knife point in her home in Mexico.
If you haven't realized the urgent need for an expanded Violence Against Women Act, read today's New York Times, where novelist Louise Erdrich restates the theme that runs through her powerful novel The Round House (reviewed in a previous post): Native American women are being battered and raped by non-native men, and they have no legal support for pursuing justice—because non-natives are immune from prosecution by tribal courts.
It's hard to imagine a more efficient way to rack up diverse denunciations than Rep. Todd Akin's approach in an interview on Sunday, when in one breath he both promoted a foul bit of junk science alleging that rape victims don't generally get pregnant (and thus don't need abortion services) and coined the term "legitimate rape." Pretty much everyone everywhere has condemned his comments, and rightly so. A number of rape victims have written responses, including Shauna Prewitt, whose post at xoJane went viral and taught a lot of us something appalling that we didn't know.


text:
Small business owners: Did you know that 97 percent of Internet users look online for local products and services?
Are you succeeding in finding customers and boosting sales through online marketing?
Dayton Get Your Business Online is a morning summit designed to help your small business establish and/or improve sales through websites, e-commerce and social media.
It is scheduled for Tuesday, April 29, from 8:00 a.m. to noon, at the University of Dayton River Campus, 1700 S. Patterson Blvd.
Dayton Mayor Nan Whalen is encouraging small businesses to particiapte.
Ohio Business Portal
Starting a Business in Ohio
Career Cruising
Dayton Minority Business Assit. Center
The US Small Business Administration
Dayton Area Chamber of Commerce
Dayton Chamber Events"
Ohio Small Business Development Centers
- At The Entrepreneurs Center
- At Wright State University
Try our Business Databases