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1) An implement comprising an elongate, tubular body, a mount disposed within said tubular body, an elongate writing element projecting in one direction from said mount and a knife projecting in opposite direction from said mount, the mount being displaceable in said one direction to extend the tip of said elongate writing element from one end of said body, and in said opposite direction to extend said knife from the opposite end of said body. 2) An implement as claimed in claim 1, in which said mount is displaceable to an intermediate position in which both said elongate writing element and said knife are disposed within said tubular body. 3) An implement as claimed in claim 1, further comprising an element extending from said mount through an elongate opening in the side of said tubular body, to enable a user to displace said mount longitudinally within said body. 4) An implement as claimed in claim 3, in which said element includes an elongate clip extending alongside said tubular body. 5) An implement as claimed in claim 1, further comprising means for retaining said mount in said longitudinal position to which it is displaced. 6) An implement as claimed in claim 5, further comprising an element extending from said mount through an elongate opening in the side of said tubular body, to enable a user to displace said mount longitudinally within said body, wherein said elongate opening is formed with serrations along at least one edge thereof, for engagement by at least one tooth provided on said element extending through said opening. 7) An implement as claimed in claim 6, in which said element is displaceable inwardly, against a spring bias, to disengage its said at least one tooth from said serrations. 8) An implement as claimed in claim 1, in which said elongate writing element comprises a pen. 9) An implement as claimed in claim 1, in which said elongate writing element comprises a pencil. |
Fast all-optical switches and attenuators |
A polarizing beam-splitter apparatus, comprising: an input port through which an input beam of lights is provided; a first polarizing beam splitter that receives the input beam and splits the beam into at least a first and second beam, said first beam having substantially a first desired polarization state and said second beam having a second polarization state orthogonal to said first polarization state but possibly admixed with the first polarization state; and an optical system that receives the second beam and provides a third beam having the second polarization state and a smaller admixture of the second polarization state than the second beam. |
1. A polarizing beam-splitter apparatus, comprising: an input port through which an input beam of light is provided; a first polarizing beam splitter that receives the input beam and splits the beam into at least a first and second beam, said first beam having substantially a first desired polarization state and said second beam having a second polarization state orthogonal to said first polarization state but possibly admixed with the first polarization state; and an optical system that receives the second beam and provides a third beam having the second polarization state and a smaller admixture of the second polarization state than the second beam. 2. A polarizing beam-splitter apparatus according to claim 1 wherein the first beam splitter comprises a first planar surface that reflects light having the second polarization state and transmits light having the first polarization state and wherein the input beam is incident on the surface at a first angle. 3. A polarizing beam-splitter apparatus according to claim 2 wherein the first angle is substantially 45°. 4. A polarizing beam-splitter apparatus according to any of the preceding claims, wherein the optical system comprises a polarizing beam splitter that receives the second beam and splits the second beam into the third beam and a fourth beam having substantially the first polarization state. 5. A polarizing beam-splitter apparatus according to any of claims 1-4 wherein the optical system comprises a second beam splitter having a second planar surface that reflects light having the second polarization state and transmits light having the first polarization state and wherein the second beam is incident on the second planar surface at a second angle and light reflected by the second surface from the second beam forms the third beam and light transmitted by the second surface forms a fourth beam. 6. A polarizing beam-splitter apparatus according to claim 5 and comprising an absorber that receives the fourth beam. 7. A polarizing beam-splitter apparatus according to any of claims 5-6 wherein the second angle is substantially 45°. 8. A polarizing beam-splitter apparatus according to any of claims 5-7 wherein the first and second surfaces are substantially parallel as a result of which, the first and third beams are parallel and displaced from each other. 9. A polarizing beam-splitter apparatus according to any of claims 5-7 wherein the first and second surfaces are surfaces formed on a same substrate material substantially transparent to light in the input beam. 10. A polarizing beam-splitter apparatus according to any of claims 1-9 and comprising: at least one controllable polarization rotator positioned to receive one of the first and third beams and operable to change the polarization state of the beam it receives; and a polarizer that receives the beam from the rotator and transmits an amount of optical energy in the received beam responsive to the polarization state of the beam. 11. A polarizing beam-splitter apparatus according to claim 10 wherein the at least one controllable polarization rotator comprises a polarization rotator for each of the first and second beams. 12. A polarizing beam-splitter apparatus according to claim 10 or claim 11 wherein the polarization rotator comprises: at least one volume of PLZT through which light received by the rotator is transmitted; and at least one electrode for applying a voltage to the volume of PLZT, which voltage controls the state to which the rotator changes the polarization of light that the rotator receives. 13. Apparatus according to any of claims 10-12, comprising a pair of polarization rotators arranged around said polarization controller, to rotate polarization of light entering and exiting said controller. 14. Apparatus according to claim 13, wherein an electric field direction of said controller is perpendicular to a plane common to said beams. 15. An optical switch comprising an input port through which the switch receives light and first and second output ports to which the switch selectively directs light that it receives comprising: a first polarization state apparatus that receives light from the input port and provides a light beam having a desired polarization state; a polarizing beam-splitter apparatus according to any of claims 1-9 that receives the light beam from the polarization state apparatus at the beam splitter apparatus input port and generates at least one first beam and/or at least one third beam responsive to the polarization of the light that it receives; and wherein the first output port receives light from the at least one first beam and the second output port receives light from the at least one third beam. 16. An optical switch according to claim 15 and comprising: a polarizing beam splitter that receives light from the input port and generates fifth and sixth spatially separated beams therefrom said fifth beam having substantially a third polarization state and said sixth beam having a fourth polarization state substantially orthogonal to the third state; a second polarization state apparatus that receives the first and second beams of light and changes the polarization state of at least one of the fifth and sixth beams so that the polarization state of both beams is the same; and wherein the fifth and sixth beams are directed to the input port of the beam splitter apparatus, which apparatus generates a first and/or third beam responsive to the fifth beam and a first and/or third beam responsive to the sixth beam. 17. An optical switch according to claim 16 comprising a first polarizer through which light from the first beams from the polarizing beam-splitter apparatus is transmitted and wherein said first polarizer transmits substantially only light having the first polarization state. 18. An optical switch according to claim 16 or claim 17 comprising a second polarizer through which light from the third beams from the polarizing beam-splitter apparatus is transmitted and wherein said second polarizer transmits substantially only light having the second polarization state. 19. An optical switch according to claim any of claims 16-18 comprising: an first optical combiner that combines light in the first beams provided by the beam splitter apparatus responsive to light in the fifth and sixth beams and directs the combined light to the first output port. 20. An optical switch according to claim 19 comprising: a second optical combiner that combines light in the third beams provided by the beam splitter apparatus responsive to light in the fifth and sixth beams and directs the combined light to the second output port. 21. An optical switch according to claim 19 wherein the first optical combiner comprises: a third polarization state apparatus that receives the first beam provided from light in the fifth beam and transmits the light in the third polarization state and receives the light in the first beam provided by light from the sixth beam and transmits the light in the fourth polarization state; an optical joiner that receives light in first beams from the third polarization state apparatus and combines the received light into a single beam that is transmitted to the first output port. 22. An optical switch according to claim 20 wherein the second optical combiner comprises: a fourth polarization state apparatus that receives the third beam provided from light in the fifth beam and transmits the light in the third polarization state and receives the light in the third beam provided by light from the sixth beam and transmits the light in the fourth polarization state; an optical joiner that receives light in the third beams from the fourth polarization state apparatus and combines the received light into a single beam that is transmitted to the second output port. 23. An optical switch according to claim 19 or claim 21 and comprising a first controllable attenuator controllable to attenuate light from the first combiner by a desired attenuation before the light reaches the first output port. 24. An optical switch according to claim 20 or claim 22 and comprising a second controllable attenuator controllable to attenuate light from the second combiner by a desired attenuation before the light reaches the second output port. 25. An optical switch according to claim 22 wherein the first attenuator comprises: at least one controllable polarization rotator positioned to receive the light from the first combiner and operable to change the polarization state of the light it receives; and a polarizer that receives the beam from the rotator and transmits an amount of optical energy in the received responsive to the polarization state of the light. 26. An optical switch according to claim 23 wherein the second attenuator comprises: at least one controllable polarization rotator positioned to receive the light from the second combiner and operable to change the polarization state of the light it receives; and a polarizer that receives the beam from the rotator and transits an amount of optical energy in the received responsive to the polarization state of the light. 27. An optical switch according to claim according to claim 25 or claim 26 wherein the polarization rotator comprises: at least one volume of PLZT through which light received by the rotator is transmitted; and at least one electrode for applying a voltage to the volume of PLZT, which voltage controls the state to which the rotator changes the polarization of light that the rotator receives. 28. A switch array comprising a plurality of switches according to any of claims 15-27, sharing an elongated optical element, said elongation being perpendicular to a plane of each of said switches. 29. A switch according to any of claims 15-27, comprising at least one reflector for folding an optical path of said switch. 30. A compound optical switch comprising at least two optical switches according to any of claims 14-26 wherein the first output port of each optical switch is a same single first shared output port and the second output port of each optical switch is a same single second shared output port. 31. A compound optical switch comprising a cascade of optical switches wherein an n-th tier of the cascade comprises 2n optical switches according to any of claims 15-27 and wherein light from the first and second output ports of an optical switch in the n-th tier is input to the input ports of two optical switches in the (n+1)-st tier. 32. A compound optical switch according to claim 31 wherein each optical switch in the n-th tier receives light from only a single output port of the optical switches in the (n−1)st tier. 33. A compound optical switch according to claim 31 or claim 32 comprising N tiers and comprising an output port that receives light from at least two output ports of the optical switches in the n-th tier. 34. A router-selector optical switching network, comprising: a number of input channels equal to a power of two; a number of output channels equal to the same or a different power of two; a router section for each input channel comprising a binary branching tree of polarizing beam splitters, light paths joining them, and controllable polarization rotators; a selector section for each output channel comprising a binary branching tree of polarizing beam joiners, light paths joining them, and controllable polarization rotators; wherein the controllable polarization rotators operate to control the connection of any output channel to at most one input channel and any input channel to at most one output channel. 35. A router-selector optical switching network according to claim 34, wherein the light paths of each router are co-planar, the light paths of each selector are co-planar, the planes of all the router light paths are parallel to each other, the planes of all the selector light paths are parallel to each other, and the planes of all the router light paths are perpendicular to the planes of all the selector light paths. 36. A router-selector optical switching network according to claim 34 or claim 35, wherein at least one of the polarizing beam splitters or one of the polarizing beam joiners is a polarizing beam splitter apparatus according to any of claims 1-14. 37. A method of aligning a first optical element with a second optical element comprising: mounting the first optical element on a first part of a support comprising first and second parts, wherein the first part is movably coupled to the second part; mounting the second part of the support in a fixed position relative to the second optical element; applying a curable adhesive to the support so that the adhesive contacts both the first and second parts; moving the first part so that the first optical element is aligned with the second optical element, and curing the adhesive to secure the first part in the aligned position. 38. An optical configuration, comprising: a substrate; at least two optical elements that lie in a same path and are coupled to said substrate; and at least one ball and socket joint formed between at least one of said elements and said substrate, such that said one element can be oriented on said joint in a plurality of orientations relative to the other one of said elements. 39. An optical configuration according to claim 38, and including curable adhesive in the bearing. 40. An optical configuration according to claim 39, wherein the curable adhesive is cured by ultraviolet light. 41. An optical configuration according to any of claims 38-40, wherein said adhesive is viscous and prevent slipping of said joint when no external forces are applied to said optical element. 42. An optical configuration according to any of claims 38-41, wherein said one optical element or said substrate is transparent to ultraviolet light. 43. An optical configuration according to any of claims 38-42, wherein said ball is on said substrate. 44. An optical configuration according to any of claims 38-42, wherein said ball is on said element. 45. An optical configuration according to any of claims 38-44, wherein said ball is integral to one of said substrate and said element. 46. An optical configuration according to any of claims 38-44, wherein said ball is mounted on one of said substrate and said element. 47. An optical configuration according to claim 46, wherein said ball is attached using an adhesive to said one of said substrate and said element. |
<SOH> BACKGROUND OF THE INVENTION <EOH>As the size and speed limits of semiconductor technology are approached, optical networks provide an attractive alternative for communications and computation. Optical networks require switches for switching an optical signal between two or more outputs, as well as multicasters for distributing an optical signal to more than one output, and attenuators for reducing the amplitude of an optical signal. It is desirable for these elements to be all-optical, working directly with the optical signal, rather than converting it into an electronic signal and back into an optical signal. U.S. Pat. Nos. 5,363,228, 5,414,541, and 6,041,151, the disclosures of which are incorporated herein by reference, are examples of implementations of an all-optical switch. The device described by U.S. Pat. No. 5,414,541 is a 1×2 switch, in which the light from one input channel is passed to either one of two output channels. The input channel can have any polarization state, and the output channel into which each input channel is directed has the same polarization state as the input channel. The term “polarization state” as used here includes not only any state of pure linear, elliptical or circular polarization, but also unpolarized light, and any degree of partial polarization. The light from the input channel is passed through a polarizing beam splitter, for example a crystal of calcite, and separated into two beams of orthogonal pure polarization states, for example vertical and horizontal linear polarization. One of the beams then passes through an element which changes its polarization to be the same as the other beam. This element, for example, could be a half-wave plate of a birefringent material with its principal axis oriented at an angle of 45 degrees to the polarization of the beam, which will convert horizontally polarized light to vertically polarized light. The two beams, now with the same polarization, are then passed through a controllable polarization rotator, which rotates the polarization by an amount that can be controlled externally. For example, a ferroelectric crystal can be used, which has a degree of birefringence that depends on the electric field that is applied to it. If no electric field is applied, then the controllable polarization rotator is inactive, and light passes through it with no change in its polarization. If an electric field of a particular direction and magnitude is applied to the ferroelectric crystal, then the controllable polarization rotator is active, and the polarization of the light changes its direction by 90 degrees when passing through it. The light is then passed through another polarizing beam splitter, which either allows the light to go straight through, or displaces it to the side, depending on the polarization of the light. If the controllable polarization rotator is inactive, then the light emerging from the second polarizing beam splitter will be directed toward the first output channel. If the controllable polarization rotator is active, then the light emerging from the second polarizing beam splitter will be displaced toward one side, and will be directed toward the second output channel. If an electric field is applied to the controllable polarization rotator, but less than the full field needed to rotate the polarization by 90 degrees, then part of the light from the input channel will go into one output channel, and part of the light will go into the other output channel. The amount of light going into each channel is controlled by controlling the electric field in the controllable polarization rotator. The 1×2 switch is thus used as a multicaster. U.S. Pat. No. 5,363,228 describes an all-optical cross-bar switch, using similar methods, in which each of N input channels can be directed to any of M output channels. This device suffers from various drawbacks. For example, since it uses polarization separation, 2n optical paths are required, where n is the number of input ports. In addition, complicated manufacturing and fine alignment is required. U.S. Pat. No. 6,041,151 describes a bypass-exchange switch which operates in a similar fashion to the 1×2 switch described in U.S. Pat. No. 5,414,541. This bypass-exchange switch has two inputs and two outputs, and the two inputs are always directed to different outputs. |
<SOH> SUMMARY OF INVENTION <EOH>An aspect of some embodiments of the invention concerns a polarizing beam splitter in the form of a periscope. A periscope is a structure with a place for light to enter, a place on the other side for light to exit, and two reflectors, generally parallel to each other, which the entering light reflects from, so that the light exits the periscope going in the same direction as it entered, but displaced. In the case of a polarizing beam splitter periscope, all of the light is not reflected, but some of it is transmitted through the reflectors. Light entering the periscope at the bottom strikes a transparent plate with an optical coating, for example a multi-layer optical reflective coating (which is often wavelength dependent), oriented at 45 degrees to the direction of the light, and either is reflected upward, or goes straight through and exits the periscope, depending on its polarization. The plate could be oriented at an angle other than 45 degrees, in which case the reflected portion will be deflected by an angle other than 90 degrees, and prisms and other optical elements could be used, and multiple beams could be created, but a simple coated plate at 45 degrees is best for many applications. The light reflected upward strikes another plate, optically coated as before, and generally parallel to the plate at the bottom. Although the second plate need not be parallel to the first plate, if it is parallel then the beam reflected from the second plate will be parallel to the light entering the periscope. Because this light is already polarized, most of it reflects from the second plate, and emerges from the periscope at the top, traveling parallel to the light that emerges at the bottom. A small amount of light that is polarized in the wrong direction will not reflect from the plate on the top, but will go straight through it, and will be lost. Optionally, an absorber mounted on or above the top of the periscope absorbs this light, to prevent it from being scattered and having some of it eventually getting back into the system. Hence the light emerging from the periscope at the top will have an even higher degree of polarization than the light reflected from the first plate. In an exemplary embodiment of the invention, the amount of contamination is 1%, 5%, 10%, 20% or any smaller, greater or intermediate percentage of the desired light power. The degree of reduction may be, for example, 50%, 80%, 90%, 95%, 99% or any smaller, intermediate or greater percentage. The periscope serves the same function as a calcite crystal, splitting a beam of light into two beams of orthogonal linear polarizations. But the periscope is more compact and has a smaller footprint, since calcite only displaces light (of one polarization) by a small angle, and a long calcite crystal is needed to produce a reasonable lateral displacement of the light. On the other hand, the optically coated plates used in the periscope only work well for a limited range of wavelengths, while a calcite crystal (or a synthetic crystal with similar properties, such at yttrium vanadate) works over a broader range of wavelengths. This limitation depends on the embodiment used, and a periscope works over a broader range of wavelengths if unwanted paths are blocked, as described below. Optionally, instead of using a second optically coated a plate, a mirror is used to direct the reflected beam in the same direction as the transmitted beam. Optionally, after reflecting from the mirror, the beam passes through a polarizer to block the small amount of light that is polarized in the wrong direction. Optionally, the light that is transmitted through the optically coated plate at the entrance to the periscope also passes through a polarizer, to block the small amount of that light which is of the polarization that should be reflected. A periscope, like any polarizing beam splitter, can be used in reverse, to combine two beams of different polarizations to form a single beam. In an exemplary embodiment of the invention, pans of beams with different initial polarizations may be converted to beams with same polarization that can travel close together so they can share optical elements. Another aspect of some embodiments of the invention concerns an all-optical variable attenuator, or an all-optical switch which includes a variable attenuator. If the light in one of the output channels of a 1×2 switch is absorbed or otherwise discarded, then the device is a variable attenuator, modulating the intensity of the light beam at frequencies up to the maximum operating frequency of the controllable polarization rotator. To build a 1×2 switch with controllable attenuation of each output channel, one could attach two such stand-alone variable attenuators to the output channels of a 1×2 switch. However, there is an alternative embodiment in which a variable attenuator is incorporated into a 1×2 switch, such as that described in U.S. Pat. No. 5,414,541, which is simpler than adding two stand-alone variable attenuators to the output. Instead, an extra controllable polarization rotator is added in front of each output channel, before the two polarization states are recombined into a single beam by a polarizing beam splitter. If the extra controllable polarization rotator changes the direction of polarization by 90 degrees, then, when the light goes through the final polarizing beam splitter, it will be displaced to a different position than it would have been if its polarization hadn't been changed, and it will miss the output channel completely. If the extra controllable polarization rotator rotates the direction of polarization of the light by less than 90 degrees (in general it will make the light elliptically polarized in this case), then some of the light will change its polarization direction by 90 degrees, and some of it will remain in the original polarization state. Then, when the light goes through the final polarizing beam splitter, some of the light will recombine into a beam which is directed into the output channel, and some of it will be displaced by a different amount and will miss the output channel. By controlling the electric field on each of the controllable polarization rotators, the light going into each output channel can be attenuated by a controlled amount. Another aspect of some embodiments of the invention concerns an optical switch in which unwanted paths are blocked, in order to reduce cross-talk. Cross-talk can occur between different channels if the polarizing beam splitter does not completely separate the two polarization states, or if the controllable polarization rotator, when it is supposed to be fully active, does not convert an entering light beam into a completely orthogonal polarization state, but leaves a small component of the original polarization state. Cross-talk can also result from scattering of light. Any of these conditions will generally result in a small amount of light that was intended to go into one output channel ending up in the wrong output channel. In order to reduce cross-talk, additional controllable polarization rotators are placed in front of each output channel, before the polarizing beam splitter where the two beams of pure orthogonal polarization states recombine into one beam. Each of these additional controllable polarization rotators is set so that light that is supposed to enter that output channel is allowed through with no change in its polarization, while light that is not supposed to enter that output channel (i.e. cross-talk) has its polarization state converted to the orthogonal state (rotated 90 degrees, in the case of linearly polarized light) so that it cannot enter that output channel, but is displaced off to the side when it passes through the polarizing beam splitter. Alternatively or additionally, polarizing filters, which let through light or one polarization and either absorb or scatter light of the orthogonal polarization, are placed anywhere in the optical path after the light has been directed toward one or the other output channel, and before the two beams going toward each output channel have been recombined into one beam. These polarizing filters are oriented so that they only let through light of the polarization that would be expected at that point, if all the polarizing beam splitters and controllable polarization rotators worked perfectly. Alternatively or additionally, controllable polarization rotators are placed anywhere in the optical path, even after the two beams going toward each output channel have been recombined into one beam, and polarizing filters are placed in the optical path after the controllable polarization rotators. These polarizers play the same role that the final polarizing beam splitters play, keeping light of the wrong polarization from going into the output channel. This configuration is useful if the final polarizing beam splitters are already being used for another purpose, such as variable optical attenuation (described below), so cannot also be used for blocking unwanted paths. In some optical switching networks, unwanted paths are necessarily blocked, in order to make it possible to connect all the desired input channels to each output channel. This is true, for example, in the selector part of the router-selector optical switching network shown in FIG. 7 . There is a controllable polarization rotator at the exit of each periscope (which functions as a beam combiner rather than a beam splitter) in the selector section of the network, where light from all the input channels going into a given output channel is repeatedly combined in a binary tree. The controllable polarization rotator must be active, rotating the polarization of light going through it by 90 degrees, whenever the light from the left side of one periscope enters the right side of the periscope above it, or vice versa, in the schematic view in FIG. 7 , and this automatically blocks the unwanted light from the other side of that lower periscope from entering that upper periscope. In other optical switching networks, blocking of unwanted paths is not a necessary part of the network, but is an optional added feature which improves performance by reducing cross-talk. That is true, for example, in the 2×2 switch shown in FIG. 6 . Blocking unwanted paths increases the range of wavelengths that periscopes operate at, and decreases the tolerances for manufacture of periscopes. Thus two of the significant disadvantages of using periscopes as polarizing beam splitters are at least partially overcome, and it becomes possible to take advantage of the desirable features of periscopes, such as their small size. Another aspect of some embodiments of the invention concerns optical switches in which the parts of the switch are arranged in a three-dimensional configuration which is compact easy to manufacture, or otherwise advantageous. One way to accomplish this is to use half-wave plates, with principle axes oriented at an oblique angle, before and after one or more of the controllable polarization rotators. This makes it possible to change the orientation of the principle axes of the controllable polarization rotator, in order to make the controllable polarization rotator fit better into the layout of the switch. In particular, if the controllable polarization rotator is a ferroelectric crystal, or a ceramic using the Kerr effect, then an electric field needs to be applied to it along one of the principle transverse axes, and a large uniform electric field is most readily applied if the controllable polarization rotator is short in that dimension, and has large, flat electrodes attached to its sides. (Similarly, if the controllable polarization rotator uses the Faraday effect, then a large uniform magnetic field is most readily applied if the controllable polarization rotator is short in the direction of the field.) If the polarizing beam splitter displaces the beam in a direction parallel to the direction of polarization of the displaced beam, then it is often most convenient to use a layout for the switch whose envelope has a rectangular cross-section, with principle axes parallel and perpendicular to the direction of polarization of the displaced beam. But the electric field in the controllable polarization rotator, in the case of a ferroelectric crystal or electro-optic ceramic, is at a 45 degree angle to the direction of polarization of the light that passes through it. By placing half-wave plates, oriented with their principle axis 22.5 degrees to the direction of beam displacement, before and after the controllable polarization rotator, the principle axes of the controllable polarization rotator can be aligned with the principle axes of the rectangular cross-section of the envelope of the switch, and in particular the short dimension of the controllable polarization rotator can be aligned with the short dimension of the envelope. Another aspect of some embodiments of the invention concerns an optical switch or another optical configuration in which parts are mounted on rotatable bearings, such as ball and socket bearings. The parts are aligned, for example by hand or machine, and when alignment is achieved, ultraviolet light is applied to a UV cured adhesive in the bearing, curing the adhesive and fixing the bearing in place. The adhesive can be applied to the bearing before or after the parts are aligned. Optionally, if the adhesive is applied before the parts are aligned, it is viscous enough so that the alignment will not slip spontaneously before the adhesive is cured, but is not so viscous that it is difficult to perform the alignment. Another aspect of some embodiments of the invention concerns all-optical router-selector networks. Such a network is built up of polarized beam splitters (which can be, for example, periscopes, as described above, or calcites), and controllable polarization rotators. The aspects of the invention described previously, especially the techniques used to make a compact three-dimensional layout, are especially useful in a network with a large number of inputs and outputs. As used herein, “calcite” means any material with similar birefringent properties, including synthetic materials such as yttrium vanadate (YVO 4 ). Another aspect of some embodiments of the invention concerns all-optical switches with controllable polarization rotators using lead lanthanum zirconate titanate (PLZT), an electro-optic ceramic (using the Kerr effect) which has a response time of only 10 to 100 nanoseconds, much faster than the response time of ferroelectrics and nematic liquid crystals. However, the change in index of refraction in PLZT is proportional to the square of the electric field, in contrast to ferroelectrics where it is linear, and the effect is rather weak. Since operating at high voltage causes increased scattering of light, PLZT is often used at moderate voltages (20 to 80 volts), in which case a longer interaction length is needed (compared to ferroelectrics) to rotate the polarization by 90 degrees. Another aspect of the invention concerns the ability to scale up a 1×2 or 2×2 optical switch to have many parallel input channels, each connected to its own one or two output channels. This can be done when the layout of the single 1×2 or 2×2 switch is essentially two-dimensional, for example the 1×2 switch design shown in FIG. 2 or the 2×2 switch shown in FIG. 6 . Calcites, periscopes, and half-wave plates can simply be extended in a direction perpendicular to the plane of the drawing. Even controllable polarization rotators can be extended in this way if their electric field is in the plane of the paper, i.e. vertical in the case of a ferroelectric or an electro-optic ceramic, and horizontal (in the direction of propagation of the light) in the case of a twisted nematic liquid crystal. Then an arbitrary number of input and output channels can be lined up side by side. There is thus provided in accordance with an exemplary embodiment of the invention, a polarizing beam-splitter apparatus, comprising: an input port through which an input beam of light is provided; a first polarizing beam splitter that receives the input beam and splits the beam into at least a first and second beam, said first beam having substantially a first desired polarization state and said second beam having a second polarization state orthogonal to said first polarization state but possibly admixed with the first polarization state; and an optical system that receives the second beam and provides a third beam having the second polarization state and a smaller admixture of the second polarization state than the second beam. Optionally, the first beam splitter comprises a first planar surface that reflects light having the second polarization state and transmits light having the first polarization state and wherein the input beam is incident on the surface at a first angle. Optionally, the first angle is substantially 45°. In an exemplary embodiment of the invention, the optical system comprises a polarizing beam splitter that receives the second beam and splits the second beam into the third beam and a fourth beam having substantially the first polarization state. Alternatively or additionally, the optical system comprises a second beam splitter having a second planar surface that reflects light having the second polarization state and transmits light having the first polarization state and wherein the second beam is incident on the second planar surface at a second angle and light reflected by the second surface from the second beam forms the third beam and light transmitted by the second surface forms a fourth beam. Optionally, the apparatus comprises an absorber that receives the fourth beam. Alternatively or additionally, the second angle is substantially 45°. Alternatively or additionally, the first and second surfaces are substantially parallel as a result of which, the first and third beams are parallel and displaced from each other. Alternatively or additionally, the first and second surfaces are surfaces formed on a same substrate material substantially transparent to light in the input beam. In an exemplary embodiment of the invention, the apparatus comprises: at least one controllable polarization rotator positioned to receive one of the first and third beams and operable to change the polarization state of the beam it receives; and a polarizer that receives the beam from the rotator and transmits an amount of optical energy in the received beam responsive to the polarization state of the beam. Optionally, the at least one controllable polarization rotator comprises a polarization rotator for each of the first and second beams. Alternatively or additionally, the polarization rotator comprises: at least one volume of PLZT through which light received by the rotator is transmitted; and at least one electrode for applying a voltage to the volume of PLZT, which voltage controls the state to which the rotator changes the polarization of light that the rotator receives. In an exemplary embodiment of the invention, the apparatus comprises a pair of polarization rotators arranged around said polarization controller, to rotate polarization of light entering and exiting said controller. Optionally, an electric field direction of said controller is perpendicular to a plane common to said beams. There is also provided in accordance with an exemplary embodiment of the invention, a n optical switch comprising an input port through which the switch receives light and first and second output ports to which the switch selectively directs light that it receives comprising: a first polarization state apparatus that receives light from the input port and provides a light beam having a desired polarization state; a polarizing beam-splitter apparatus as described above that receives the light beam from the polarization state apparatus at the beam splitter apparatus input port and generates at least one first beam and/or at least one third beam responsive to the polarization of the light that it receives; and wherein the first output port receives light from the at least one first beam and the second output port receives light from the at least one third beam. Optionally, the switch comprises: a polarizing beam splitter that receives light from the input port and generates fifth and sixth spatially separated beams therefrom said fifth beam having substantially a third polarization state and said sixth beam haying a fourth polarization state substantially orthogonal to the third state; a second polarization state apparatus that receives the first and second beams of light and changes the polarization state of at least one of the fifth and sixth beams so that the polarization state of both beams is the same; and wherein the fifth and sixth beams are directed to the input port of the beam splitter apparatus, which apparatus generates a first and/or third beam responsive to the fifth beam and a first and/or third beam responsive to the sixth beam. Optionally, the switch comprises a first polarizer through which light from the first beams from the polarizing beam-splitter apparatus is transmitted and wherein said first polarizer transmits substantially only light having the first polarization state. Alternatively or additionally, the switch comprises a second polarizer through which light from the third beams from the polarizing beam-splitter apparatus is transmitted and wherein said second polarizer transmits substantially only light having the second polarization state. Alternatively or additionally, the switch comprises: an first optical combiner that combines light in the first beams provided by the beam splitter apparatus responsive to light in the fifth and sixth beams and directs the combined light to the first output port. Optionally, the switch comprises: a second optical combiner that combines light in the third beams provided by the beam splitter apparatus responsive to light in the fifth and sixth beams and directs the combined light to the second output port. In an exemplary embodiment of the invention, the first optical combiner comprises: a third polarization state apparatus that receives the first beam provided from light in the fifth beam and transmits the light in the third polarization state and receives the light in the first beam provided by light from the sixth beam and transmits the light in the fourth polarization state; an optical joiner that receives light in first beams from the third polarization state apparatus and combines the received light into a single beam that is transmitted to the first output port. Alternatively or additionally, the second optical combiner comprises: a fourth polarization state apparatus that receives the third beam provided from light in the fifth beam and transmits the light in the third polarization state and receives the light in the third beam provided by light from the sixth beam and transmits the light in the fourth polarization state; an optical joiner that receives light in the third beams from the fourth polarization state apparatus and combines the received light into a single beam that is transmitted to the second output port. In an exemplary embodiment of the invention, the switch comprises a first controllable attenuator controllable to attenuate light from the first combiner by a desired attenuation before the light reaches the first output port. Alternatively or additionally, the switch comprises a second controllable attenuator controllable to attenuate light from the second combiner by a desired attenuation before the light reaches the second output port. In an exemplary embodiment of the invention, the first attenuator comprises: at least one controllable polarization rotator positioned to receive the light from the first combiner and operable to change the polarization state of the light it receives; and a polarizer that receives the beam from the rotator and transmits an amount of optical energy in the received responsive to the polarization state of the light. In an exemplary embodiment of the invention, the second attenuator comprises: at least one controllable polarization rotator positioned to receive the light from the second combiner and operable to change the polarization state of the light it receives; and a polarizer that receives the beam from the rotator and transmits an amount of optical energy in the received responsive to the polarization state of the light. In an exemplary embodiment of the invention, the polarization rotator comprises: at least one volume of PLZT through which light received by the rotator is transmitted; and at least one electrode for applying a voltage to the volume of PLZT, which voltage controls the state to which the rotator changes the polarization of light that the rotator receives. There is also provided in accordance with an exemplary embodiment of the invention, a switch array comprising a plurality of switches as described herein, sharing an elongated optical element, said elongation being perpendicular to a plane of each of said switches. In an exemplary embodiment of the invention, the switch comprises at least one reflector for folding an optical path of said switch. There is also provided in accordance with an exemplary embodiment of the invention, a compound optical switch comprising at least two optical switches as described herein where the first output port of each optical switch is a same single first shared output port and the second output port of each optical switch is a same single second shared output port. There is also provided in accordance with an exemplary embodiment of the invention, a compound optical switch comprising a cascade of optical switches wherein an n-th tier of the cascade comprises 2 n optical switches as described herein and where light from the first and second output ports of an optical switch in the n-th tier is input to the input ports of two optical switches in the (n+1)-st tier. Optionally, each optical switch in the n-th tier receives light from only a single output port of the optical switches in the (n−1)st tier. Alternatively or additionally, the switch comprises N tiers and comprising an output port that receives light from at least two output ports of the optical switches in the n-th tier. There is also provided in accordance with an exemplary embodiment of the invention, a router-selector optical switching network, comprising: a number of input channels equal to a power of two; a number of output channels equal to the same or a different power of two; a router section for each input channel comprising a binary branching tree of polarizing beam splitters, light paths joining them, and controllable polarization rotators; a selector section for each output channel comprising a binary branching tree of polarizing beam joiners, light paths joining them, and controllable polarization rotators; wherein the controllable polarization rotators operate to control the connection of any output channel to at most one input channel and any input channel to at most one output channel. Optionally, the light paths of each router are co-planar, the light paths of each selector are co-planar, the planes of all the router light paths are parallel to each other, the planes of all the selector light paths are parallel to each other, and the planes of all the router light paths are perpendicular to the planes of all the selector light paths. Alternatively or additionally, at least one of the polarizing beam splitters or one of the polarizing beam joiners is a polarizing beam splitter apparatus as described herein. There is also provided in accordance with an exemplary embodiment of the invention, a method of aligning a first optical element with a second optical element comprising: mounting the first optical element on a first part of a support comprising first and second parts, wherein the first part is movably coupled to the second part; mounting the second part of the support in a fixed position relative to the second optical element; applying a curable adhesive to the support so that the adhesive contacts both the first and second parts; moving the first part so that the first optical element is aligned with the second optical element; and curing the adhesive to secure the first part in the aligned position. There is also provided in accordance with an exemplary embodiment of the invention, comprising: a substrate; at least two optical elements that lie in a same path and are coupled to said substrate; and at least one ball and socket joint formed between at least one of said elements and said substrate, such that said one element can be oriented on said joint in a plurality of orientations relative to the other one of said elements. Optionally, the configuration comprises curable adhesive in the bearing. Optionally, the curable adhesive is cured by ultraviolet light. In an exemplary embodiment of the invention, said adhesive is viscous and prevent slipping of said joint when no external forces are applied to said optical element. Alternatively or additionally, said one optical is transparent to ultraviolet light. In an exemplary embodiment of the invention, said ball is on said substrate. Alternatively, said ball is on said element. In an exemplary embodiment of the invention, said ball is integral to one of said substrate and said element. Alternatively, said ball is mounted on one of said substrate and said element. Optionally, said ball is attached using an adhesive to said one of said substrate and said element. |
Sirna expression system and process for producing functional gene knockdown cell or the like using the same |
The in vivo siRNA expression system according to this invention is a system that intracellularly expresses small interfering (si) RNAs and comprises antisense and sense code DNAs coding for antisense and sense RNAs targeting any region of a target gene mRNA and one or more promoters that function to express the antisense and sense RNAs from the antisense and sense code DNAs, respectively. |
1. An intracellular siRNA expression system comprising an antisense code DNA coding for antisense RNA directed against a region of a target gene mRNA, a sense code DNA coding for sense RNA directed against the same region of said target gene mRNA, and one or more promoters capable of expressing said antisense and sense RNAs from said antisense and sense code DNAs, respectively. 2. The siRNA expression system according to claim 1, wherein a final transcription product of the siRNA expressed by the system is 15 to 49 bp long. 3. The siRNA expression system according to claim 1, wherein a final transcription product of the siRNA expressed by the system is 15 to 35 bp long. 4. The siRNA expression system according to claim 1, wherein a final transcription product of the siRNA expressed by the system is 15 to 30 bp long. 5. The siRNA expression system according to claim 1, wherein a double-stranded RNA region of the siRNA in which two RNA strands pair up contains a mismatch or a bulge. 6. The siRNA expression system according to claim 5, wherein one of nucleotides in the mismatch is guanine, and the other is uracil. 7. The siRNA expression system according to claim 5, wherein the siRNA contains 1 to 7 mismatches. 8. The siRNA expression system according to claim 5, wherein the siRNA contains 1 to 7 bulges. 9. The siRNA expression system according to claim 5, wherein the siRNA contains both 1 to 7 mismatches and bulges. 10-27. (Canceled) 28. An intracellular siRNA library expression system comprising a double-stranded DNA coding for siRNA comprising an arbitrary sequence having the length of the siRNA to be expressed, and two promoters facing to each other with said DNA coding for siRNA—in between which are capable of expressing the mutually complementary RNAs from respective strands of said double-stranded DNA. 29. An intracellular siRNA library expression system comprising a stem-loop siRNA producing unit in which an antisense code DNA and a sense code DNA complementary to said antisense code DNA are linked in the opposite direction via a linker, and a promoter capable of expressing the stem-loop siRNA at either side of said unit. 30. The siRNA library expression system according to claim 28, wherein a final transcription product of the siRNA expressed by the system is 15 to 49 bp long. 31. The siRNA library expression system according to claim 28, wherein a final transcription product of the siRNA expressed by the system is 15 to 35 bp long. 32. The siRNA library expression system according to claim 28, wherein a final transcription product of the siRNA expressed by the system is 15 to 30 bp long. 33-42. (Canceled) 43. The siRNA expression system according to claim 1, wherein said promoter is a pol II or pol III promoter. 44. The siRNA expression system according to claim 1, wherein said pol III promoter is U6 promoter. 45. The siRNA expression system according to claim 1, wherein said promoter is an inducible promoter. 46. The siRNA expression system according to claim 1, wherein said promoter is separately located upstream of said antisense and sense code DNAs. 47. The siRNA expression system according to claim 1, wherein said system comprises loxP sequences in the form of any one of the following (a) to (c) so that the expression can be controlled: (a) the promoter comprises distal sequence element (DSE) and proximal sequence element (PSE) with a space there between, and in the space two loxP sequences, one in the vicinity of DSE and the other in the vicinity of PSE; (b) the promoter comprises DSE and PSE that are located to maintain the promoter activity, a loxP sequence there between, and another loxP sequence either upstream of DSE or downstream of PSE; and (c) two loxP sequences are located so as to interpose the antisense code DNA or sense code DNA. 48. The siRNA expression system according to claim 1, wherein antisense and sense code DNAs are maintained in the same vector DNA molecule, or separately in different vector DNA molecules. 49. The siRNA expression system according to claim 1, wherein the promoter is located at the one side of a unit in which the antisense and sense code DNAs are connected in the opposite direction via a linker. 50. The siRNA expression system according to claim 49, wherein said system comprises loxP sequences in the form of any one of the following (a) to (d) so that the expression can be controlled: (a) the promoter comprises DSE and PSE with a space there between, and in the space two loxP sequences, one in the vicinity of DSE and the other is the vicinity of PSE; (b) the promoter comprises DSE and PSE that are located to maintain the promoter activity, a loxP sequence there between, and another loxP upstream of DSE or downstream of PSE; (c) two loxPs are located so as to interpose the antisense code DNA or sense code DNA; and (d) two loxPs are arranged so as to interpose a linker comprising a stop sequence. 51. The siRNA expression system according to claim 49, wherein the antisense and sense code DNAs are maintained in a vector molecule. 52. The siRNA expression system according to claim 48, wherein said vector is a plasmid vector. 53. The siRNA expression system according to claim 48, wherein said vector is a viral vector. 54. The siRNA expression system according to claim 48, wherein said vector is a dumbbell-shaped DNA vector. 55. A cell maintaining the siRNA expression system according to claim 1. 56. The cell according to claim 55, wherein said cell is a mammalian cell. 57. An individual organism maintaining the siRNA expression system according to claim 1. 58. A composition comprising the siRNA expression system according to claim 1. 59. The composition according to claim 58, wherein said composition is a pharmaceutical composition. 60. A method for producing a cell in which the target gene expression is silenced, wherein said method comprises: (a) introducing the siRNA expression system according to claim 1 into cells, and (b) selecting cells in which said siRNA expression system is introduced. 61. The siRNA library expression system according to claim 28, wherein a double-stranded RNA region of the siRNAs in which two RNA strands pair up contains a mismatch or a bulge. 62. The siRNA library expression system according to claim 28, wherein said promoter is a pol II or pol III promoter. 63. The siRNA library expression system according to claim 28, wherein said promoter is an inducible promoter. 64. The siRNA library expression system according to claim 28, wherein siRNAs expressed by the system are composed of random RNA strands. 65. The siRNA library expression system according to claim 28, wherein said system is an assembly of multiple siRNA expression vectors that each targets a gene sequence comprising a coding region and/or a non-coding region. 66. The siRNA library expression system according to claim 28, wherein siRNAs expressed by the system are composed of RNA strands encoded by DNA fragments of any cDNA or genomic DNA, said fragment has the length of the siRNA to be expressed. 67. An assembly of the siRNA library expression systems according to claim 28, wherein different siRNAs are expressed by each system in said assembly. 68. A method of searching for a functional gene, the method comprising: (a) introducing the siRNA library expression system according to claim 28; (b) selecting cells into which said siRNA library expression system or said assembly has been introduced; and (c) analyzing the phenotype of the cells thus selected. 69. A method of searching for a functional gene, the method comprising: (a) introducing the assembly of siRNA library expression systems according to claim 67 into cells; and (b) selecting cells into which said siRNA library expression system or said assembly has been introduced; and (c) analyzing the phenotype of the cells thus selected. 70. The method of searching for a functional gene according to claim 68, wherein said method further comprising a step of screening for a functional gene based on the sequence of DNA coding for siRNA in the cell whose phenotype has been found altered as the result of the phenotype analysis. 71. A method for selecting a highly active siRNA, the method comprising: (a) introducing the siRNA library expression system according to claim 28, and (b) measuring the expression level of a specific gene or protein in the cells into which said siRNA library expression system or said assembly is introduced. 72. A method for selecting a highly active siRNA, the method comprising: (a) introducing the assembly of siRNA library expression systems according to claim 67 into cells; and (b) measuring the expression level of a specific gene or protein in the cells into which said siRNA library expression system or said assembly is introduced. 73. The siRNA expression system according to claim 5, wherein said promoter is a pol II or pol III promoter. 74. The siRNA expression system according to claim 5, wherein said pol III promoter is U6 promoter. 75. The siRNA expression system according to claim 5, wherein said promoter is an inducible promoter. 76. The siRNA expression system according to claim 5, wherein said promoter is separately located upstream of said antisense and sense code DNAs. 77. The siRNA expression system according to claim 5, wherein said system comprises loxP sequences in the form of any one of the following (a) to (c) so that the expression can be controlled: (a) the promoter comprises distal sequence element (DSE) and proximal sequence element (PSE) with a space therebetween, and in the space two loxP sequences, one in the vicinity of DSE and the other in the vicinity of PSE; (b) the promoter comprises DSE and PSE that are located to maintain the promoter activity, a loxP sequence therebetween, and another loxP sequence either upstream of DSE or downstream of PSE; and (c) two loxP sequences are located so as to interpose the antisense code DNA or sense code DNA. 78. The siRNA expression system according to claim 5, wherein antisense and sense code DNAs are maintained in the same vector DNA molecule, or separately in different vector DNA molecules. 79. The siRNA expression system according to claim 5, wherein the promoter is located at the one side of a unit in which the antisense and sense code DNAs are connected in the opposite direction via a linker. 80. The siRNA expression system according to 79, wherein said system comprises loxP sequences in the form of any one of the following (a) to (d) so that the expression can be controlled: (a) the promoter comprises DSE and PSE with a space there between, and in the space two loxP sequences, one in the vicinity of DSE and the other is the vicinity of PSE; (b) the promoter comprises DSE and PSE that are located to maintain the promoter activity, a loxP sequence there between, and another loxP upstream of DSE or downstream of PSE; (c) two loxPs are located so as to interpose the antisense code DNA or sense code DNA; and (d) two loxPs are arranged so as to interpose a linker comprising a stop sequence. 81. The siRNA expression system according to claim 50, wherein the antisense and sense code DNAs are maintained in a vector molecule. 82. The siRNA expression system according to claim 79, wherein the antisense and sense code DNAs are maintained in a vector molecule. 83. The siRNA expression system according to claim 80, wherein the antisense and sense code DNAs are maintained in a vector molecule. 84. The siRNA expression system according to claim 78, wherein said vector is a plasmid vector. 85. The siRNA expression system according to claim 51, wherein said vector is a plasmid vector. 86. The siRNA expression system according to claim 81, wherein said vector is a plasmid vector. 87. The siRNA expression system according to claim 78, wherein said vector is a viral vector. 88. The siRNA expression system according to claim 78, wherein said vector is a dumbbell-shaped vector. 89. A cell maintaining the siRNA expression system according to claim 5. 90. A cell maintaining the siRNA expression system according to claim 76. 91. The cell according to claim 89, wherein said cell is a mammalian cell. 92. The cell according to claim 90, wherein said cell is a mammalian cell. 93. An individual organism maintaining the siRNA expression system according to claim 5. 94. An individual organism maintaining the siRNA expression system according to claim 76. 95. A composition comprising the siRNA expression system according to claim 5. 96. A composition comprising the siRNA expression system according to claim 76. 97. The composition according to claim 95, wherein said composition is a pharmaceutical composition. 98. The composition according to claim 96, wherein said composition is a pharmaceutical composition. 99. A method for producing a cell in which the target gene expression is silenced, wherein said method comprises: (a) introducing the siRNA expression system according to claim 5 into cells, and (b) selecting cells in which said siRNA expression system is introduced. 100. A method for producing a cell in which the target gene expression is silenced, wherein said method comprises: (a) introducing the siRNA expression system according to claim 76 into cells, and (b) selecting cells in which said siRNA expression system is introduced. 101. The siRNA library expression system according to claim 29, wherein a final transcription product of the siRNA expressed by the system is 15 to 49 bp long. 102. The siRNA library expression system according to claim 29, wherein a final transcription product of the siRNA expressed by the system is 15 to 35 bp long. 103. The siRNA library expression system according to claim 29, wherein a final transcription product of the siRNA expressed by the system is 15 to 30 bp long. 104. The siRNA library expression system according to claim 29, wherein a double-stranded RNA region of the siRNAs in which two RNA strands pair up contains a mismatch or a bulge. 105. The siRNA library expression system according to claim 30, wherein a double-stranded RNA region of the siRNAs in which two RNA strands pair up contains a mismatch or a bulge. 106. The siRNA library expression system according to claim 31, wherein a double-stranded RNA region of the siRNAs in which two RNA strands pair up contains a mismatch or a bulge. 107. The siRNA library expression system according to claim 32, wherein a double-stranded RNA region of the siRNAs in which two RNA strands pair up contains a mismatch or a bulge. 108. The siRNA library expression system according to claim 29, wherein said promoter is a pol II or pol III promoter. 109. The siRNA library expression system according to claim 29, wherein said promoter is an inducible promoter. 110. The siRNA library expression system according to claim 29, wherein siRNAs expressed by the system are composed of random RNA strands. 111. The siRNA library expression system according claim 29, wherein said system is an assembly of multiple siRNA expression vectors that each targets a gene sequence comprising a coding region and/or a non-coding region. 112. The siRNA library expression system according to claim 29, wherein siRNAs expressed by the system are composed of RNA strands encoded by DNA fragments of any cDNA or genomic DNA, said fragment has the length of the siRNA to be expressed. 113. An assembly of the siRNA library expression systems according to claim 29, wherein different siRNAs are expressed by each system in said assembly. 114. A method for selecting a highly active siRNA, the method comprising: (a) introducing the siRNA library expression system according to claim 29, and (b) measuring the expression level of a specific gene or protein in the cells into which said siRNA library expression system or said assembly is introduced. 115. A method for selecting a highly active siRNA, the method comprising: (a) introducing the siRNA library expression systems according to claim 29 into cells, and (b) measuring the expression level of a specific gene or protein in the cells into which said siRNA library expression system or said assembly is introduced. |
<SOH> BACKGROUND ART <EOH>RNA interference (hereafter abbreviated as “RNAi”) is the phenomenon (process) capable of inducing the degradation of target gene mRNA so as to silence the target gene expression by introducing into cells a double-stranded RNA (hereafter abbreviated as “dsRNA”) that comprises a sense RNA having the sequence homologous to the target gene mRNA and antisense RNA having the sequence complementary to the sense RNA. RNAi, because of its capability to silence the target gene expression, has received considerable attention as a simple gene knock-down method that replaces the conventional gene disruption method relying on the tedious, inefficient homologous recombination, or as a means of gene therapy. The above-mentioned RNAi was originally discovered in nematodes (Fire, A. et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans . Nature 391, 806-811 (1998)). Thereafter, it is also observed in various organisms including plants, round worms, Drosophila , and protozoa (Fire, A. RNA-triggered gene silencing. Trends Genet. 15, 358-363 (1999); Sharp, P. A. RNA interference 2001. Genes Dev. 15, 485-490 (2001); Hammond, S. M., Caudy, A. A. & Hannon, G. J. Post-transcriptional gene silencing by double-stranded RNA. Nature Rev. Genet. 2, 110-119 (2001); Zamore, P. D. RNA interference: listening to the sound of silence. Nat Struct Biol. 8, 746-750 (2001)) Silencing of target gene expression was confirmed by actually introducing exogenous dsRNA in these organisms. This technique has been employed as a method for producing knock-down individuals. Similar to in these organisms, RNAi induction in mammalian cells has been attempted by introduction of exogenous dsRNA. However, in this case, protein synthesis was inhibited by the action of host's protective mechanisms against the virus infection which was triggered by the transfected dsRNA, so that RNAi could not be observed. Recently, Tuschl et al. reported that RNAi can be induced also in mammalian cells by transducing the cells with short dsRNAs of 21 or 22 nucleotide long having a single-stranded 2 or 3 nucleotide 3′ overhang in place of long dsRNAs as those used in other organisms (Elbashir, S. M. et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411, 494-498 (2001); Caplen, N. J. et al. Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems. Proc. Natl. Acad. Sci. USA 98, 9742-9747 (2001)). As described above, RNAi has also been successfully induced in mammalian cells using small interfering double-stranded RNA (hereafter abbreviated as “siRNA”). For the functional analysis and gene therapy based on the gene silencing by RNAi, an efficient introduction of siRNA into cells and its stable intracellular maintenance become essential. Efficiency in introducing exogenous siRNAs into cells varies depending on the cell type, being as low as 1% to 10% in certain cells. Also, exogenous siRNAs introduced into mammalian cells disappear in a few days after introduction, having no sufficient stability required for analysis of gene functions. Furthermore, in gene therapy, administration of siRNA at regular intervals becomes necessary, which increases physical loads of patients. Moreover, it is extremely difficult to induce RNAi exclusively in a specific tissue or at a specific stage of development/differentiation by introduction of exogenous siRNA. In addition, though siRNAs are small in size, synthesis of RNA is markedly so expensive compared to DNA synthesis and the RNAi induction directly by siRNA is not economical. Now that most of the primary DNA sequence of the human genome has been determined, systematic and efficient methods for searching for functional genes has been developed to speedily elucidate gene functions. Gene silencing by RNAi can be utilized for the systematic search for the functional gene based on the phenotypic alteration of cells or individuals to accelerate the finding and analysis of novel functional genes. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG. 1 represents an siRNA expression system using the U6 promoter, and a method for producing siRNA employing the system. (A) shows an siRNA production process. Two U6 promoters produce sense and antisense short RNAs, with adding four uridines (Us) to 3′-ends of RNAs. Sense and antisense RNAs thus expressed are annealed to form the duplex of siRNAs with a 4-nucleotide 3′ overhang. (B) shows a palindrome type siRNA expression system, comprising a double-stranded DNA coding for siRNA comprising sense and antisense code DNAs and promoters at both ends, from which sense and antisense RNAs are expressed. FIG. 2 represents an example of the construction for producing an siRNA expression system using a ribozyme. FIG. 3 represents the EGFP gene silencing effect of an siRNA expression system directed against EGFP when the system was introduced into cells expressing hygromycin/EGFP. Left side panels (A, D, G and J) show the expression of hygromycin/EGFP; middle panels (B, E, H and K) the expression of DsRed; right side panels (C, F, I and L) the results of merged expressions of hygromycin/EGFP and DsRed. FIG. 4 represents the gene silencing effect of an siRNA expression system directed against either sea pansy (Renilla) or firefly luciferase when the system was introduced into HeLa S3 cells having the luciferase activity. In FIG. 4 a , an ordinate value means luciferase activity of the cells, in which the siRNA expression system directed against the firefly luciferase was introduced, normalized based on the sea pansy luciferase activity, or the activity of the cells, in which the siRNA expression system directed against the sea pansy luciferase was introduce, normalized based on the firefly luciferase activity. FIG. 4 b shows the concentration-dependent silencing effect of the siRNA expression systems on firefly or sea pansy luciferase activity when a varied amount of the siRNA expression system directed against each luciferase was introduced into cells. FIG. 5 represents the gene silencing effect using a series of siRNAs or siRNA expression systems directed against different target sites on the same target gene (for firefly luciferase). FIG. 5 a shows the gene silencing effect of siRNA expression vectors directed against various target sites when the vectors were introduced into cells. FIG. 5 b represents the results obtained when the exogenous siRNAs directed against different target sites were directly introduced into cells at different concentrations. FIG. 6 a represents the gene silencing effect of the length of 3′ overhang of siRNA. FIG. 6 b shows the gene silencing effect of siRNA expression systems directed against two target genes or two target sites. In FIG. 6 b , luciferase activity values were normalized based on the activity of β-galactosidase introduced as an internal control. FIG. 7 represents the capability of the siRNA expression system to silence the endogenous β-catenin gene. Panels A, B, and C are for the group transduced with the siRNA expression vector directed against β-catenin (pHygEGFP/iβ-catenin), while panels D, E, and F are for the group transduced with the empty vector (pHygEGFP). These groups were all stained with the anti-p-catenin antibody. Left side panels (A and D) represent the expression of Hygromycin/EGFP; middle panels (B and E) the expression of β-catenin; and right side panels (C and F) the merged image of these two expressions. FIG. 8 represents the comparison of RNAi effects between the tandem and stem-loop siRNAs. siRNA expression vectors pU6tandem19 and pU6stem19 are tandem and stem-loop, respectively. Cont. represents the control (vacant vector). FIG. 9 represents the gene silencing effects of various siRNA expression vectors. FIG. 10 represents the gene silencing effects of siRNA expression vectors containing a cytomegalovirus-derived promoter (CMV promoter), and tRNA promoter. FIG. 11 represents the RNAi induction effects of double-stranded siRNAs containing a mismatch, or a bulge. FIG. 12 is a diagram describing the principle of Tet-ON system. In the absence of tetracycline, the tetracycline repressor protein binds to U6 promoter, resulting in the suppression of transcription, while, in its presence, the tetracycline repressor protein binds to it to be released from U6 promoter so as to initiate transcription. FIG. 13 is a graph representing RNAi inducing effects of siRNA expression vector having the tetracycline-inducible promoter. U6Teti represents an siRNA expression vector containing the tetracycline operator sequence in U6 promoter, and U6i represents an siRNA expression vector not containing the sequence. FIG. 14 is a diagram depicting automatic cleavages in the RNA transcript containing a trimming ribozyme. FIG. 15 is a diagram depicting siRNA production by self-processing of trimming ribozyme. Nucleotide cleavages occur at the positions indicated by black arrowheads to produce siRNAs. FIG. 16 is an electrophoretogram showing siRNA production by RNA self-processing. Bands corresponding to 21 nt siRNA are indicated with an arrow. FIG. 17 is a diagram showing an example of the construction for controlling siRNA expression using the Cre-lox system. FIG. 18 is a diagram representing an example of the preparation of the stem-loop siRNA library expression system. FIG. 19 is a diagram representing an example of the preparation of the siRNA library expression system. {circle over (1)} shows a random DNA fragment having the dephosphorylated blunt ends of 19 to 29 bp long. {circle over (2)} represents the random DNA fragment {circle over (1)} which is ligated with the 5′-phosphorylated hairpin type DNA linker 1 at its both ends. FIG. 20 is a continuation of FIG. 19 . {circle over (3)} shows strand displacement from the Nick site by Bst DNA Polymerase. {circle over (4)} represents the fragment {circle over (3)} which is ligated with DNA linker 2. FIG. 21 is a continuation of FIG. 20 . {circle over (5)} shows strand displacement from the Nick site by Bst DNA Polymerase. {circle over (6)} represents the cleavage of {circle over (5)} by AscI. FIG. 22 is a continuation of FIG. 21 . {circle over (7)} represents an siRNA library expression pre-library. {circle over (8)} shows BspMI cleavage of the siRNA library expression pre-library. In the case of inserting the Loop sequence, TTCG, between the sense and antisense code DNAs, the cleavage proceeds to step®-2 in FIG. 23 . {circle over (9)} represents the completed siRNA library expression system as a result of blunting by Klenow Fragment, removal of DNA linker 1, and self-ligation. FIG. 23 is a continuation of FIG. 22 . {circle over (8)}-2 represents the case of inserting the Loop sequence, TTCG, between the sense and antisense code DNAs in {circle over (8)}. The siRNA library expression pre-library is cleaved by BsgI. {circle over (8)}-3 shows cleavage of the siRNA library expression pre-library by BspMI. The cleaved site by BsgI cannot be attacked by BspMI. {circle over (9)}-2 represents the completed siRNA library expression system as a result of blunting by T4 DNA Polymerase, removal of DNA linker 1, and self-ligation. FIG. 24 is a diagram representing the preparation of EGFP cDNA fragment of approximately 20 to 25 bp long. The final product that is a random EGFP cDNA fragment of approximately 20 to 25 bp long with the dephosphorylated blunt end serves as the random DMA fragment in FIG. 19 {circle over (1)}. FIG. 25 is a diagram representing the preparation of a cloning vector. The promoter is either the human U6 promoter or human tRNA promoter. The cloning vector containing U6 promoter was prepared using BspMI and Klenow, while that containing tRNA promoter was prepared using BseRI and T4 DNA Polymerase. FIG. 26 represents micrographs showing the results of observing the EGFP fluorescence intensity with a confocal microscope. FIG. 27 is a graph representing relative EGFP fluorescence intensities in pUC18, U6 GFP25 siRNA lib-loop-, U6 GFP25 siRNA lib TTCG, tRNA GFP25 siRNA lib loop-, and tRNA GFP25 siRNA lib TTCG, measured 24 and 48 h after the transfection. FIG. 28 is a diagram representing the stem-loop siRNA expression system containing two loxPs that interpose the linker portion containing the stop sequence. FIG. 29 is a graph representing the gene silencing effect of the siRNA expression adenovirus vector. FIG. 30 is a graph representing the gene silencing effect of the siRNA expression HIV vector. FIG. 31 is a graph representing the gene silencing effect of the siRNA expression dumbbell-shaped vector. FIG. 32 is a graph representing the gene silencing effect of the siRNA expression system containing a mismatch or a bulge in the double-stranded RNA region of siRNAs. Numerals in parentheses at the top of each sequence represent SEQ ID NOs. detailed-description description="Detailed Description" end="lead"? |
Partially interconnected networks |
A partially interconnected network has a plurality of topological nodes, each of the topological nodes having at least three direct point-to-point topological links connected to other topological nodes. Each of a proportion of the plurality of topological nodes is connected to one of a group of point-of-presence (PoP) units. The group of PoP units is arranged to provide access to a selected service or services, one or more of each of the at least three direct point-to-point topological links from each topological node not being connected to one of a group of PoP units connecting to one or more than one of the plurality of topological nodes being connected to one of the group of PoP units. There is at least one choice of routing between any two topological nodes, the choice of routing being either via two topological links connected in series at another topological node or a direct point-to-point topological link between the two topological nodes. |
1. A partially interconnected network having a plurality of Topological Nodes, each Topological Node having at least three direct point-to-point Topological Links connected to other Topological Nodes, each of a proportion of the plurality of Topological Nodes having connected thereat one of a group of Point-of Presence (PoP) Units, said group of PoP Units arranged to provide access to a selected service or services, one or more than one of each at least three direct point-to-point Topological Links from each Topological Node not having connected thereat one of a group of PoP Units connecting to one or more than one of the plurality of Topological Nodes having connected thereat one of the group of PoP Units, there being at least one Choice of routing between any two Topological Nodes, a Choice of routing being either via two Topological Links connected in series at another Topological Node or a direct point-to-point Topological Link between the two Topological Nodes. 2. A partially interconnected network as claimed in claim 1, wherein each of a further proportion of the plurality of Topological Nodes has connected thereat one of a further group of PoP Units arranged to provide access to a further selected service or further selected services, one or more than one of each at least three direct point-to-point Topological Links from each Topological Node not having connected thereat one of a further group of PoP Units connecting to one or more than one of the plurality of Topological Nodes having connected thereat one of the further group of PoP Units, there being at least one Choice of routing between any two Topological Nodes, a Choice of routing being either via two Topological Links connected in series at another Topological Node or a direct point-to-point Topological Link between the two Topological Nodes. 3. A partially interconnected network as claimed in claim 1 or 2, where all the Topological Nodes not having connected thereat one of a particular group of PoP Units are each directly connected via direct point-to-point Topological Links to an equal number of Topological Nodes having connected thereat one of that particular group of PoP Units. 4. A partially interconnected network as claimed in claim 1, 2 or 3, wherein the selected service or services or the further selected service or further selected services is/are chosen from an Internet Service Provider (ISP), a video source, a call centre, an International Network Interconnection Point, a further Network Interconnection Point or an Intelligent Network Server, any of which may be accessed with the help of Intelligent Network call control arrangements. 5. A partially interconnected network, comprising: a plurality of topological nodes, each topological node having at least three direct point-to-point topological links connected to other topological nodes, each of a proportion of the plurality of topological nodes having connected thereat one of a group of point of presence (PoP) units, said group of PoP units being arranged to provide access to a selected service or services, one, or more than one, of each of the at least three direct point-to-point topological links from each topological node not having connected thereat one of a group of PoP units connecting to one, or more than one, of the plurality of topological nodes having connected thereat one of the group of PoP units, and at least one choice of routing between any two topological nodes, the choice of routing being either via two topological links connected in series at another topological node or a direct point-to-point topological link between the two topological nodes. 6. The partially interconnected network as claimed in claim 5, wherein each of a further proportion of the plurality of topological nodes has connected thereat one of a further group of PoP units arranged to provide access to a further selected service or further selected services, one, or more than one, of each of the at least three direct point-to-point topological links from each topological node not having connected thereat one of a further group of PoP units connecting to one, or more than one, of the plurality of topological nodes having connected thereat one of the further group of PoP units. 7. The partially interconnected network as claimed in claim 5, wherein all the topological nodes not having connected thereat one of a particular group of PoP units are each directly connected via direct point-to-point topological links to an equal number of topological nodes having connected thereat one of that particular group of PoP units. 8. The partially interconnected network as claimed in claim 6, wherein at least one of the selected service, services, further selected service and further selected services is chosen from an internet service provider (ISP), a video source, a call center, an international network interconnection point, a further network interconnection point, or an intelligent network server, any of which is accessible with the help of intelligent network call control arrangements. |
Arrangement and a method relating to access of applications/services |
The present invention relates to a portal structure for providing end user stations (5) with access to services/applications. It comprises a portal core (1) a connectivity layer via which end user station access is provided and a number of services/applications (25). The services/applications (25) are represented in a generic markup language and the portal core (1) uses said generic markup language for storing at least application/service data information and for communication with said services/applications. It further comprises a presentation arrangement (11) for communication with said applications/services (25) and said end user stations (5). Each service/aaplication (25) represented by generic data in the generic markup language may optionally be provided with a number of metalink tags, such that each service/application (25) is able to generate generic link data in the generic markup language irrespectively of the location of the portal structure and of other applications. The portal core presentation arrangement (11) comprises first translating means (16) for replacing such metalinks with real (public) addresses of the services/applications (25), such that continuous navigation is enabled for end users irrespectively of the location of accessed services/applications (25). |
1-32. (canceled) 33. A portal for providing end user stations with access to services/applications, comprising: a portal core having a number of service enabling means; a connectivity layer via which end user station access is provided; a number of services/applications, wherein the services/applications are represented in a generic markup language, the portal core using said generic markup language for storing at least application/service data information and for communication with said services/applications; and a presentation arrangement for communication with said applications/services and said end user stations, each service/application represented by generic data in the generic markup language optionally being provided with a number of metalink tags, such that each service/application is able to generate generic link data in the generic markup language irrespectively of the location of the portal and of other applications, wherein the presentation arrangement comprises first translating means for replacing such metalinks with real (public) addresses of the services/applications, such that continuous navigation is enabled for end users irrespectively of the location of accessed services/applications. 34. A portal according to claim 33, wherein the portal core comprises rendering means comprising said first translating means. 35. A portal according to claim 34, wherein the presentation arrangement comprises said rendering means. 36. A portal according to claim 34, wherein the rendering means comprises second translating means for translating/rendering service/application data in the generic markup language into a markup language used by the end user station. 37. A portal according to claim 33, wherein the portal core further comprises session handling means for user session management. 38. A portal according to claim 37, wherein said session handling means are separate from, but in communication with, said rendering means. 39. A portal according to claim 33, wherein a number of types of metalinks are defined depending on the location of the linked service/application in relation to the service/application accessed by the portal core. 40. A portal according to claim 39, wherein a first type of metalink (“self”) is defined which refers to the service/application that itself has generated the data, as represented in the generic markup language. 41. A portal according to claim 39, wherein a second type of metalink (“local”) is defined which refers to resources which are local to the current service/application and in that it includes information about the path to the resource in relation to the current service/application. 42. A portal structure according to claim 39, wherein a third type of metalink (“absolute”) is defined which comprises a link to any public or private portal, web-page, resource, application or content and in that it contains a complete URL-address to such portal, resource etc. 43. A portal according to claim 39, wherein a fourth type of metalink (“application”) is defined which refers to an application which is defined in the portal under a given name and in that the metalink contains information about said name. 44. A portal according to claim 33, wherein data representing services/applications expressed in the generic markup language and metalinks are defined in a Document Type Definition Language with an URL-attribute. 45. A portal according to claim 33, wherein said portal supports access by mobile end user stations. 46. A portal according to claim 45, wherein said portal supports access by fixed end user stations, e.g. PC:s, wherein fixed end user stations use a markup language different from the markup language(s) used by mobile end user stations. 47. A portal according to claim 33, wherein a service/application optionally is provided with a number of metainformation tags and in that the rendering means adds the parameters of said metainformation tag(s) to all metalink parameters at least for some type(s) of metalinks. 48. A portal according to claim 47, wherein an application/service comprises a number of links to other applications/services, that metainformation tags optionally can be added to “self” or “application” type metalinks and in that all parameters common to all the links of an application are stored in the portal core per end user and per application instance. 49. A portal according to claim 45, wherein said portal supports access to services/applications by mobile end user stations, e.g. WAP-devices over a mobile communications network with access nodes in the connecting layer and fixed stations WEB-devices, e.g. broadband devices such as PC:s. 50. A portal according to claim 33, wherein the generic markup language is XML. 51. A portal according to claim 50, wherein the XML data and the metalinks are defined in a Document Type Definition language (DTD) and in that a metalink tag in XML comprises information about metalink type, addressing attributes containing service/application location information depending on metalink type and a number of parameters. 52. A portal according to claim 33, wherein the second translating means translates XML data by performing a transformation (XSL) to an output format, i.e. a markup language appropriate for an accessing end user station, e.g. HTML for a fixed end user station or WML for a mobile end user station. 53. A portal for providing end user stations with access to services/applications comprising: a portal core; a number of service enabling means; a connecting and data bearer layer via which end user access is provided; and a number of services/applications, wherein the portal core is XML-based using XML as a markup language for storing data as XML-data and for communication with services/applications, and wherein the portal core comprises means handling presentation on end user stations, that the services/applications generate XML-data, a service/application generating XML-data optionally being provided with metalink tag(s) for referring to other application(s)/service(s)/content, such that each service/application is able to generate XML link data independently of which is the location of the portal core and of other services/applications, and wherein the portal core comprises first translating means for replacing metalinks with real addresses of services/applications, such that continuous navigation is provided for end users independently of the location of accessed services/applications. 54. A portal according to claim 53, wherein it supports end user access by mobile as well as fixed end user stations, e.g. WAP-devices and broadband devices such as PC:s, interactive TV etc. 55. A portal for providing end user access to services/applications comprising: a functional services/applications layer; a user access layer; and an intermediate communication layer for communication with services/applications and end user access means (stations), wherein the intermediate communication layer comprises a presentation arrangement with rendering means and session handling means receiving requests for services/applications by end user stations, forwarding such requests to the requested service/application, and for receiving XML-data information representing requested services/applications, that the services/applications may be provided with metalink tag(s) for referring to other applications/services/contents, said rendering means comprising translating means for replacing metalinks with corresponding real address information relating to services/applications and means for translating such XML-data information to a format usable by the requesting end user station. 56. A method for providing an end user station with access to services/applications via a portal comprising a portal core, a plurality of services/applications and end user connectivity means, comprising the steps of: providing an application/service in a generic markup language with a number of metalink tags for referring to other application(s)/service(s)/content and/or with a number of metainformation tags; receiving in the portal core a request for a service/application from an end user station in the end user language format; forwarding the request to the requested service/application; returning data relating to the requested service/application as represented by the generic markup language to the portal core with possible metalink tags and/or metainformation tags; translating the metalinks defined by the tags to real addresses of the links in the portal core and/or storing the metainformation of the metainformation tags into a portal session for the end user relating to the requested service/application; and providing the service/application to the end user station in a format appropriate for the end user station. 57. A method according to claim 56, wherein the portal core comprises rendering means, said rendering means performing the steps of: detecting if data of a service/application delivered in the generic markup language contains any metalinks; and if yes, processing said metalink(s) and replacing it with the real address of the service/application referred to. 58. A method according to claim 56, further comprising the step of providing services/applications with metalinks of given types depending on where the content of a service/application, to which there should be a link, is located. 59. A method according to claim 58, further comprising the steps of: providing an application/service with a metalink tag referring to the application itself for content provided by the application/service itself; providing a service/application with a metalink tag referring to local content for content provided local to the service/application, which metalink contains a reference to the path to the content relative to the service/application; providing a service/application with a metalink tag referring to any portal, content etc which comprises an attribute with the complete URL address of said portal, content etc.; and providing a service/application with a metalink tag referring to another service/application if said other application/service is known and given a name by the portal, including a reference attribute containing the given name. 60. A method according to claim 56, further comprising the steps of: providing a service/an application with a metainformation tag comprising a number of parameters; adding the metainformation parameters to the metalink parameters; storing the metalink parameters and the metainformation parameters common to all the links of the service/application parameters in common in storing means in or associated with the portal core per user and per service/application instance; and sending the parameters that are different for different links to the requesting user terminal. 61. A method according to claims 56, further comprising the step of translating the service/application data as expressed in the generic markup language into the/a markup language used by the requesting terminal station. 62. A method according to claim 56, wherein the portal is mobile and supports access by mobile end user stations, e.g. WAP-devices as well as fixed end user stations, e.g. PC:s. 63. A method according to claim 56, wherein the generic markup language is XML and in that the rendering means supports translation into e.g. HTML as well as WML. 64. A method of accessing a service/application from an end user station comprising the steps of: accessing a portal by selecting a link with parameters to a desired service/application; performing a look-up to find the real address of the service/application in the portal; adding the link parameters to the real address; examining if any metainformation parameters are stored relating to the requested service/application instance for the requesting end user, if yes, adding the stored metainformation parameters to the real address with added link parameters; sending the request containing at least link parameters to the service/application; delivering service/application data expressed in a generic markup language (XML) including metalink tags and/or metainformation tags to the portal core; replacing metalink address information by real address information and/or; storing received metainformation in storing means associated with the portal core; and processing the service/application data in a generic markup language and converting it to the format used by the end user station. |
<SOH> TECHNICAL FIELD <EOH>The present invention relates to a portal structure for providing end user stations with access to services/applications. Particularly it relates to an Internet portal, and specifically it relates to a portal structure providing an end user with improved navigation support. Most particularly it relates to a portal structure facilitating access to applications/services irrespectively of the location of such services and irrespectively of which is the type of the end user station. |
<SOH> SUMMARY OF THE INVENTION <EOH>By using a generic markup language in a portal, content of applications and services can be stored independently of end user station or user device, and before showing the content of an application or a service, the content can be transformed into the format, i.e. the markup language, that can be understood by the end user device. One example of such a generic markup language is the XML (Extended Markup Language). As a standard for navigation in an XML-based portal is the XLink specification used, which allows elements to be inserted into XML documents in order to create and describe links between resources. XLink provides a framework for creating both basic unidirectional links and more complex linking structures. It allows XML documents to assert linking relationships among more than two resources, associate metadata with a link and to express links residing in a location separate from the linked resources. However, continuous navigation between portals and particularly external content providers can not be achieved through simply using XML and XLink. What is needed is therefore generally a portal structure providing an end user with fast and easy access to services and applications irrespectively of whether the services and applications (content) are located within the portal structure itself or whether the applications/services and the content thereof reside outside the portal (i.e. are provided by external providers). Particularly a portal structure is needed which is capable of providing an end user with quick and simple access to content of applications and services, also if the wanted services/applications require special access rights, or more generally a portal structure supporting handling of dynamic issues, like access control etc. A portal structure is also needed through which service/application providers are able to provide the same navigation features in the content they supply as the portal structure itself does, i.e. as it does for applications/services residing within the portal. A portal structure is also needed which is able to provide a common “look and feel” irrespectively of where applications/services reside or by whom they are provided. Particularly a portal structure is also needed which is capable of storing content independently of user access device or user station, and supporting transformation of content of applications and services to the format of the user device or a format that the user device (i.e. the end user station) is able to understand. Even more particularly a portal structure is needed through which the number of parameters that have to be sent to applications/services (content providers) from the browser of an end user is reduced as compared to hitherto known structures. Particularly a portal structure is needed which allows for connection of a large number of internal as well as external service and application providers, or content providers, while still providing the same navigation features to external providers as to internal providers without requiring an extension or additional programming of the portal implementation. Even more particularly a portal structure is needed which is mobile, i.e. which allows access by mobile end user stations, and specifically a portal structure is needed which allows mobile station as well as fixed station access. Still further a portal structure is needed which is end user friendly and easy to use and which allows user personalization or customization such as to suit the specific needs and preferences of different end users. Particularly a portal structure is needed which in an attractive way integrates Internet and WAP (Wireless Application Protocol) based services so that access is enabled from any Internet connected PC, WAP-device or any other mobile device using various mobile access networks such as for example GSM (Global System for Mobile communications), GPRS (GSM General Packet Radio Service), WCDMA (Wideband Code Division Multiple Access), UMTS (Universal Mobile Telecommunications), SMS (Short Message Service), broadband also allowing access by PDAs (Personal Digital Assistant), i.e. technology independent access. In the following some further concepts used herein will be described or defined. A portal is generally a non-physical entity in the Internet domain, which can be described as an “electronic publishing space” which is owned by an individual or an organization, and which provides either direct access to information and services, or links to other entities in the Internet or private intranet domains, providing information and services to authorized end users. A portal is in its simplest form a regular home page or list of links, whereas in more advanced forms it may offer interactive services not only to those who consume what is published, but also to those who are granted the right by the editor to publish on the portal, as well as to the editor himself, regarding different aspects on how the portal is used. Wireless end users are given access through a “service” portal. Such a “service” portal is different from a traditional fixed Internet portal for PCs and end users demand personalized services delivered to and presented on their mobile terminal at least as an option. In this document the concept portal (structure) is used. It can be both a conventional Internet portal or a “service” portal, a mobile portal. An application is one or several cooperating software entities, the functional focus being user interaction and usefulness for the end user. An application platform is a defined combination of software and hardware entities used to implement applications of a certain kind, which are characterized by the functionality and quality of its constituent parts. By portal infrastructure is, in general terms, meant the software and hardware entities needed to either host or produce or generate a specific portal. Specifically it contains a portal core, an IP infrastructure and service enabling means. A service enabling means, here also denoted a service enabler, is a support functionality accessed via APIs (Application Programming Interface) raising the abstraction level and simplifying the application developers task. The portal core, as referred to above, is the core of a portal infrastructure. By a service network is generally meant an IP-based network which consists of nodes hosting application servers, service capability servers, application support servers, IP infrastructure servers etc. Application support servers interface with service network resources or other external resources than core networks, whereas service capability servers interface with resources and functionality in core networks. In the present application a portal structure is intended to mean a portal core, a plurality of services and applications with their content and service enabling means (service enablers). Generally may also the connectivity and data bearer functionality be seen as included. Therefore, in order to solve one or more of the problems referred to earlier in the document, the invention discloses a portal structure, for providing end user stations with access to services/applications, i.e. the content of services and/or applications. It comprises a portal core, a number of service enabling means, connectivity means acting as data bearer and via which end user station access is provided, and services/applications (providers). The services/applications use or generate a generic markup language to represent a content, the portal core uses said generic markup language for storing application/service data information and for communication with said services/applications. The portal core also comprises a presentation arrangement for communication with the applications/services and with said end user stations. Each service/application as represented by generic data in the generic markup language may be provided with one or more metalink tags referring to other services, applications or content (internal or external) such that each service/application is able to generate generic link data by means of the generic markup language irrespectively of the location of the portal core and of other applications/services. The portal core presentation arrangement comprises first translating means for replacing such metalinks with real (public addresses) of the services/applications content referred to, such that continuous navigation is enabled for end users irrespectively of the location of services/applications (content). The portal core presentation arrangement particularly comprises rendering means which includes said first translating means. Alternatively the rendering means are provided separately in the portal core and only comprises second translating means for translating (rendering) service/application data presented in the generic markup language into a format, or a markup language, as understood or used by the end user station having requested a service or an application. A portal core further comprises session handling means for user session management. Particularly service logic is kept separate from presentation related logic. Said session handling means are also separate from the rendering means. According to the invention a number of different types of metalinks can be defined such that an application or a service can be provided with the appropriate kind(s) of metalink(s) depending on where the content to be referred to resides. In one embodiment four different kinds of metalinks are defined, although the inventive concept of course not is limited to the definition of four different kinds of metalinks or to the specifically exemplified metalink types. However, according to one embodiment the first metalink, which may be denoted a metalink of type “self” is defined which refers to the service/application that itself has generated the data as represented by the generic markup language. A second metalink type “local” may also be defined which refers to resources which are local to the service/application. It includes information about the path to the resource in relation to the service/application. A third metalink type “absolute” is defined which comprises a link to any public or private portal, WEB-page, resource, application or content and it contains a reference with the complete URL (Uniform Resource Locator) to such resource, content etc. Finally a fourth metalink type denoted “application” is defined, according to this embodiment, which refers to an application defined in the portal structure under a given name and it contains information about said name. Particularly data representing services/applications as expressed in the generic markup language and metalinks are defined in a Document Type Definition (DTD) language with an URL-attribute. DTD is e.g. an XML document describing all the elements and their attributes which are allowed in all the documents belonging to the particular DTD. In a most advantageous embodiment the portal structure is mobile, i.e. it supports access by mobile end user stations over a mobile communication network such as for example GSM (Global System for Mobile communications), GPRS (GSM General Packet Radio Service), UMTS (Universal Mobile Telecommunications System), Bluetooth (which is a short range radio technology), WAP (Wireless Application Protocol) etc. Advantageously the portal structure supports access by broadband devices such as PCs or more generally fixed devices as well as mobile devices such as WAP-devices. In other terms the portal structure supports access by fixed as well as mobile end users stations using different access formats or using different markup languages for communication with the portal structure. Detection of the type of a requesting end user station may be done in any appropriate manner. However, it may with advantage be performed as disclosed in the copending Swedish patent application “An Arrangement and a Method Relating to End User Station Access of a Portal” which was filed on the same date as the present application, by the same applicant, and the content of which herewith is incorporated herein by reference. In that application it is disclosed how an end user station can get access to the portal structure itself, also if the type of the end user station is not known to the portal structure, as long as the class to which the type belongs is known to the portal, which adaptively will get to know new types, i.e. it is generally adaptive to new types. According to one particular embodiment a service or an application may optionally be provided with one or more metainformation tags. The rendering means adds the parameters of such a metainformation tag to all metalink parameters at least for some types of metalinks. Particularly the metainformation tags can optionally be added to “self” or “application” type referring metalinks and all parameters common to all the links of an application are stored in the portal core per end user and per application instance. All parameters common to all the links in the application will then be defined in one place and can be stored by the portal. The stored parameters can then be sent to the application when the end user accesses the application next time (in the same session). The parameters will, as referred to above, be stored per end user and per application instance which means that different application instances can have different global parameters. The advantage thereof is that the addresses (URLs), of applications that need to be sent to the end user, get much shorter. This is particularly attractive in case the end user accesses the portal using a mobile end user station, i.e. a WAP-device. Only parameters that differ in different links will have to be sent to the end user station, i.e. when different links within one and the same application or service have different parameters. It is also possible to add such metainformation tags to other links than metalinks, e.g. XLinks. The functioning will be the same as that described above referring to metalinks. Thus, addition of metainformation tags to some kind of links is generally advantageous also in case the concept of metalinks is not implemented, e.g. since shorter URLs can be used to e.g. a mobile end user station. Although the invention is not limited thereto, in a particularly advantageous implementation, the generic markup language used by or generated by the services/applications and the portal core, particularly the presentation arrangement of the portal core, is XML. The XML-data and the metalinks are defined in a Document Type Definition language (DTD) and a metalink tag in XML-data comprises information about metalink type, a reference and addressing attribute (URL) containing service/application location information. The second translating means (of the rendering means) translates XML-data by performing a transformation (XSL), i.e. the XML-data is processed with a transformation stylesheet (XSL transformation) to produce an output format, i.e. a markup language, appropriate for the accessing end user station, e.g. HTML for a fixed end user station and WML for a mobile end user station. XML is e.g. described in Extensible Markup Language (XML) 1.0 (Second edition), W3C Recommendation 6 Oct. 2000. XSL/XSLT is described in XSL Transformations (XSLT) Version 1.0 W3C Recommendation of 16 Nov. 1999 and XSL Transformations (XSLT) Vers. 1.1., W3C Working draft, 12 Dec. 2000. These documents are herewith incorporated herein by reference. A portal structure is particularly disclosed which provides end user stations with access to applications/services. The portal structure comprises a portal core, a number of service enabling means, a connectivity and data bearer layer via which end user access is provided and a number of services/applications (providers). The portal core is typically XML-based and uses XML as a markup language for storing data as XML-data and for communication with services/applications. The portal core further comprises means for presentation on end user stations. The services/applications are represented by XML-data in the portal core and each service/application in XML-data may be provided with/generate one or more metalink tag(s) such that each service/application is able to generate XML link data independently of which is the location of the portal core and of the services/applications. The portal core further comprises first translating means for replacing metalinks with real addresses of the services/applications (content) referred to. The portal structure is, according to an advantageous implementation, mobile and supports end user access by mobile as well as fixed end user stations, e.g. WAP-devices and broadband devices such as PC:s, (or rather the used browser) interactive TV etc. A portal structure for providing end user access to services/applications is provided which comprises a functional services/applications layer, a user access layer and an intermediate communication layer for communication with services/applications and with the end user via the access layer. The intermediate communication layer comprises a presentation arrangement with rendering means and session handling means receiving requests for services/applications by end user stations, forwarding said requests for services/applications by end user stations to the service/application layer, receiving XML-data information representative of the requested services/applications, and comprising means for converting such XML-data information representative of requested services/applications to a format usable by the requesting end user station. The services/applications may be provided with metalink tag(s) and said presentation arrangement comprises translating means for replacing metalinks with corresponding real address information of the service(s)/application(s) referred to by the metalink(s). Instead of XML any other generic markup language with similar properties may be used. The invention also discloses a method for providing end user stations with access to services/applications via a portal structure comprising a portal core, services/applications and end user connectivity means, which method particularly includes the steps of; receiving in the portal core a request for a service/application from an end user station in an end user station markup language; forwarding the request to the requested service/application; receiving data relating to the requested service/application as represented by a generic markup language which may include one or more metalink tags referring to (other) applications/services/content in the portal core from the service/application; translating the metalink tag(s) to real addresses of the application(s)/service(s) referred to in the portal core; providing the data of a requested service/application to the end user station in a format (markup language) appropriate for the end user station. The portal core particularly comprises rendering means which perform the steps of; detecting if data of a service/application in the generic markup language contains one or more metalinks; if yes, processing said metalink(s) and replacing it/them with (a) real address(es) of the service(s)/application(s) referred to. Particularly the method includes the steps of; providing a service/application with a metalink of a given type depending on where the requested content of a service/application referring to is located. The method may comprise the steps of; providing an application/service with a metalink tag referring to the application itself if the content of the application/service referring to is provided by the application/service itself; providing a service/application with a metalink tag referring to local content if the content referring to is provided local to the service/application, which metalink contains a reference to the path to the content in relation to the service/application; providing a service/application with a metalink tag referring to a link to any portal, content etc. which comprises an attribute with a complete URL address of said portal, content etc. and/or providing a service/application with a metalink referring to another service/application if the content to be referred to is associated with an application/service known and given a name in the portal structure, including a reference attribute containing the given name. The method with advantage includes the steps of; providing a service/application with a number of metainformation tags comprising a number of parameters; adding the metainformation parameters to the metalink parameters; storing the metalink parameters and the metainformation parameters which are common to all the links of the service/application in common in storing means of the portal core per user and per service/application instance; sending the parameters that are different for different links to the requesting end user station. The method preferably includes the steps of; translating the service/application data expressed in the generic markup language into the markup language used by the requesting terminal station. Still further, particularly the portal structure is mobile and supports access by mobile end user stations, e.g. WAP-devices, as well as fixed end user stations, e.g. PC:s. In a particular implementation the generic markup language is XML and the rendering means supports translation into e.g. HTML (HyperText Markup Language) as well as WML (Wireless Markup Language). The invention also discloses a method of accessing a service/application from an end user station comprising the steps of; accessing a portal structure by selecting a link with parameters to a desired service/application; performing a look-up to find the real address of the service/application in the portal structure; adding the link parameters to the real address; examining if any metainformation parameters are stored relating to the requested service/application instance for the requesting end user; if yes, adding the stored metainformation parameters to the real address with added link parameters; sending the request containing at least link parameters to the service/application; delivering application/service data expressed in a generic markup language (XML) and including possible metalinks and metainformation to the portal core; replacing metalink address information by real address information; storing any received metainformation in storing means associated with the portal core; processing the service/application data in a generic markup language by converting it into the/a format (markup language) used by the end user station. It is here supposed that applications/services, irrespectively of where they can be found, are expressed as e.g. XML-data, optionally including metalink tags and/or metainformation tags. The present invention discloses a dynamic conversation selection (based on arbitrary/complex criteria) and it shows a system/method that can handle a plurality of heterogeneous client devices. The inventive concept also allows easy extension for new devices or device types. However, according to the invention the source of specific content (application/service/provider) is determined dynamically, and thus not known to neither users nor content providers themselves. Further, content providers can address own/other content without need to know: Source, means service/applications-specifics. Location and configuration of portal infrastructure especially if content is located inside or outside portal provider infrastructure. In addition, client accesses content without knowledge of physical storage locations. Still further it provides for device-specific content request adaptation, content request and reply adaptation according to device capabilities, implied security feature, since sensitive parameters are not disclosed to user, while still sent in actual content request towards application/service and particularly user-specific/personalized content presentation. According to the invention locations are not exposed, and hence they can be changed during normal operation. |
Transition metal complexes with proton sponges as ligands |
Transition metal complexes with quino[7,8-h]quinolines and cyclopentadieno[1,2-h:4,3-h′]diquinolines as proton sponge ligands and a process for their preparation. These complexes are suitable as catalysts, e.g. the palladium complexes for a Heck reaction, as well as for amination and C—H activation reactions. The platinum and palladium complexes are potentially good cytostatic agents. |
1. A transition metal complex of formula III or IV: wherein X=halogen, hydrogen, alkoxy, OH, nitro or amino group, wherein the two X substituents may be the same or different; Y=hydrogen, carboxy, carboxylate, alkyl or functionalized alkyl group, 011- or amino group, wherein the two Y substituents may by the same or different; L=any ligand, wherein the L, substituents may be the same or different; M=a metal selected from Periodic Table groups 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; Z=hydrogen, alkyl or aryl group, wherein the two Z substituents may be the same or different, or the two substituents together are ═O. R=any substituent, wherein the substituents R may be the same or different, and two R substituents may together form part of a ring system; n=from 0 to 6. 2. The transition metal complex according to claim 1, wherein M is a metal selected from. Periodic Table groups 7, 8, 9, 10, 11 or 12. 3. The transition metal complex according to claim 1, wherein L=halogen, alkyl, carbonyl or carboxy late group. 4. The transition metal complex according to claim 1, having one of formulas V, VI, VII or VIII: 5. A process for preparing a transition metal complex of claim 1 by reacting ligand I or II with a precursor complex in the presence of a solvent, wherein a precursor complex is employed which contains the transition metal M and the substituents L bonded to M and in which at least two coordination sites of M are occupied by weekly coordinating ligands; wherein X=halogen, hydrogen, alkoxy or amino group, nitro or OH group, wherein the two X substituents may be the same or different; Y=hydrogen, carboxy, carboxylate, alkyl or functionalized alkyl group, OH group or amino group, wherein the two Y substituents may be the same or different; L=any ligand, or halogen, alkyl, carbonyl or carboxylate group, wherein the L substituents may be the same or different; M=a metal selected from Periodic Table groups 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12, Z=hydrogen, alkyl or aryl group, wherein the two Z substituents may be the same or different or may be ═O; R=any substituent, wherein the substituents R may be the same or different, and two R substituents may together form part of a ring system; n=from 0 to 6: 6. The process according to claim 5, wherein the solvent, a CO group or a π system are used as said weakly coordinating ligands. 7. The process according to claim 5, wherein halogenated hydrocarbons or THF are employed as said solvent. 8. The process according to claim 7, wherein dichloromethane or chloroform arc employed as said solvent. 9. The process according to claim 5, wherein a transition metal carbonyl halide is employed as said precursor complex. 10. The process according to claim 9, wherein di-(μ-chloro)dichlorobis(ethylene-platinum(II)), cis-dichloro(1,5-cyclooctadiene)palladium(II), dimeric tetracarbonylrhenium(I) bromide or pentacarbonylmanganese(I) bromide is employed as said precursor complex. 11. A process in the form of a “Heck reaction” for the catalyzed preparation of olefinated aromatics or heteroaromatics, characterized in that transition metal complexes of claim 1 with Pd as said transition metal M are employed as catalysts. 12. A process for catalytic amination, characterized in that a transition metal complex of claim 1 with Pd or Pt as said transition metal M are employed as a catalyst. 13. A process for catalytic C—H activation characterized in that a transition metal complex of claim 1 is employed as a catalyst. 14. The process according to claim 13, wherein methanol derivatives are prepared by the oxidation of methane. 15. Method of exerting a cytostatic effect comprising administering to a patient in need thereof an effective amount therefor of the transition metal complexes of claim 1. 16. Method according to claim 15, wherein M in complex III or IV is platinum. 17. Method according to claim 16, wherein the effectiveness or selectivity of the transition metal complex as a cytostatic agent is adjusted by selecting the X substituents in complex III or IV. 18. The transition metal complex according to claim 1, wherein each R is a substituent independently selected from the group consisting of hydrogen, halogen, alkyl and derivatives, aryl and derivatives, sulfonic acid group, carboxylate group and amino group. 19. The process according to claim 5, wherein each R is a substituent independently selected from the group consisting of hydrogen, halogen, alkyl and derivatives, aryl and derivatives, sulfonic acid group, carboxylate group and amino group. 20. The transition metal complex according to claim 1, wherein each L is a ligand independently selected from the group consisting of halogen, alkyl, carbonyl and carboxylate group; and R is a substituent independently selected from the group consisting of hydrogen, halogen, alkyl and derivatives, aryl and derivatives, sulfonic acid group, carboxylate group and amino group. 21. The process according to claim 5, wherein each L is a ligand independently selected from the group consisting of halogen, alkyl, carbonyl and carboxylate group; and R is a substituent independently selected from the group consisting of hydrogen, halogen, alkyl and derivatives, aryl and derivatives, sulfonic acid group, carboxylate group and amino group. |
Veterinary dermatologic composition |
This invention relates to the use of an active compound selected from the group comprising a sphingoid base, a sphingoid base derivative or a mixture of two or more these compounds for manufacturing a cosmetic composition suitable for topical application with animals having an at least partly fur covered skin for maintaining and/or repairing the keratoseborrheaic condition of the skin and/or fur. |
1. Use of an active compound selected from the group comprising a sphingoid base, a sphingoid base derivative or a mixture of two or more of these compounds for manufacturing a cosmetic composition suitable for topical application with animals having an at least partly fur covered skin for maintaining and/or repairing the keratoseborrheaic condition of the skin and/or fur. 2-15. (Canceled) 16. A method of maintaining and/or repairing a keratoseborrheaic condition of the skin and/or fur of an animal having an at least partly fur covered skin comprising administering by topical application to the animal a composition comprised of an active compound selected from a sphingoid base, a sphingoid base derivative or a mixture of two or more of said compounds. 17. The method of claim 16, wherein the active compound is selected from the group of sphingosin, sphinganin, phytosphingosin or a mixture of two or more of these compounds. 18. The method of claim 16, wherein the sphingoid base derivative is a salt of phytosphingosin selected from the group of a α-hydroxy alkanoic acid salt, a β-hydroxy alkanoic acid salt, an α,β-dihydroxy alkanoid acid salt, an alkanedioic acid salt, a mineral acid salt or a lipophilic organic acid salt. 19. The method of clam 16, wherein the sphingoid base derivative is a lactic acid, glycolic acid, malic acid, pyruvic acid, succinic acid, fumaric acid, ascorbic acid, gluconic acid, pyroglutamic acid, a hydrochloric acid, nitric acid and/or phosphoric acid salt of phytosphingosin. 20. The method of claim 16, wherein the sphingoid base derivative is a N-substituted derivative of phytosphingosin. 21. The method of claim 20, wherein the sphingoid base derivative is selected from the group of N-lactyloyl-phytosphingosine, N-salicyloyl-phytosphingosine or N-retinoyl-phytosphingosine. 22. The method of claim 16, wherein the composition contains a cationic surfactant. 23. The method of claim 16, wherein the composition contains 0.001-20 wt. % of the active compound. 24. The method of claim 16, wherein the composition contains 0.005-10 wt. % of the active compound. 25. The method of claim 16, wherein the composition contains 0.01-5 wt. % of the active compound. 26. The method of claim 16, wherein the composition comprises an additive capable of exhibiting a complementary activity, the additive being selected from the group of alphahydroxy acids, lactic acid, citric acid, glycolic acid, beta-hydroxy acids, salicylic acid and derivatives of the aforementioned hydroxy-acids, cytokines, anti-inflammatory steroids and non-steroids, vitamins A, C, D, E, PP, biotine and B-type vitamins, hormones, benzoyl peroxide, various emollients, ureum, reductants, anti-moth agents, antibiotics, anti-fungal agents or disinfectants. 27. The method of claim 16, wherein the composition is in the form of a shampoo, foaming base, spray, spot on, lotion, gel or emulsion of the active compound. 28. The method of claim 16, wherein a single dose of the composition contains 1-500 mg of the active compound. 29. The method of claim 16, wherein the composition takes the form of an emulsion with a mean particle size of between 50-200 nm. 30. A pharmaceutical composition suitable for topical application on animals having skin at least partly covered with fur, the composition comprising an active compound selected from a sphingoid base, a sphingoid base derivative or a mixture of two or more of said compounds. 31. The composition of claim 30, wherein the active compound is selected from the group of sphingosin, sphinganin, phytosphingosin or a mixture of two or more of these compounds. 32. The composition of claim 30, wherein the sphingoid base derivative is a salt of phytosphingosin selected from the group of a α-hydroxy alkanoic acid salt, a β-hydroxy alkanoic acid salt, an α,β-dihydroxy alkanoid acid salt, an alkanedioic acid salt, a mineral acid salt or a lipophilic organic acid salt. 33. The composition of claim 30, wherein the sphingoid base derivative is a lactic acid, glycolic acid, malic acid, pyruvic acid, succinic acid, fumaric acid, ascorbic acid, gluconic acid, pyroglutamic acid, a hydrochloric acid, nitric acid and/or phosphoric acid salt of phytosphingosin. 34. The composition of claim 30, wherein the sphingoid base derivative is a N-substituted derivative of phytosphingosin. 35. The composition of claim 34, wherein the sphingoid base derivative is selected from the group of N-lactyloyl-phytosphingosine, N-salicyloyl-phytosphingosine, or N-retinoyl-phytosphingosine. 36. The composition of claim 30, wherein the composition contains a cationic surfactant. 37. The composition of claim 30, wherein the composition contains 0.001-20 wt. % of the active compound. 38. The composition of claim 37, wherein the composition contains 0.005-10 wt. % of the active compound. 39. The composition of claim 37, wherein the composition contains 0.01-5 wt. % of the active compound. 40. The composition of claim 30, wherein the composition comprises an additive capable of exhibiting a complementary activity, the additive being selected from the group of alphahydroxy acids, lactic acid, citric acid, glycolic acid, beta-hydroxy acids, salicylic acid and derivatives of the aforementioned hydroxy-acids, cytokines, anti-inflammatory steroids and non-steroids, vitamins A, C, D, E, PP, biotine and B-type vitamins, hormones, benzoyl peroxide, various emollients, ureum, reductants, anti-moth agents, antibiotics, anti-fungal agents, or disinfectants. 41. The composition of claim 30, wherein the composition is in the form of a shampoo, foaming base, spray, spot on, lotion, gel or emulsion of the active compound. 42. The composition of claim 30, wherein a single dose of the composition contains 1-500 mg of the active compound. 43. The composition of claim 30, wherein the composition takes the form of an emulsion with a mean particle size of between 50-200 nm. 44. The method of claim 16, wherein the condition comprises anti-microbial, anti-fungal, anti-inflammation activity, inflammatory pathologies related to stress, parasitoses, dermatoses, atopic dermatitis, superinfection folliculites, modifications of the flora and/or contact eczema, for correcting and/or treating cutaneous affections related to troubles with the microflora of the animal skin and/or fur. 45. The method of claim 16, wherein the condition comprises infection associated with the presence of Staphylococcus Intermedius and/or Malassezia Pachadermatis. 46. The method of claim 16, wherein the topical application prevents or corrects cutaneous affections related to problems with the microflora of the animal skin. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The skin and/or fur of an animal form a barrier to the environment, this barrier being capable of adapting itself to varying environmental conditions. Besides this, the skin and/or fur play an important physiological role in providing physical protection, assisting in the thermal regulation of the organism and providing a metabolic, sensorial and storage function. The physical barrier function of the skin is mainly exerted by the outermost lipid layer or stratum corneum. The ceramides present in the stratum corneum ensure protective properties towards the environment and form a lipid filter limiting evaporation of moisture from and controlling permeation of external substances into the skin. Besides this physical barrier function, the skin also exerts a chemical barrier function as its surface is colonised by a wide variety of microorganisms that assist in maintaining a natural equilibrium of the skin. However environmental action and contact of the skin with cleansing or other products may involve distortion of this natural equilibrium, an uncontrollable outgrowth of certain micro-organisms within the skin microflora and the consequential impaired lipid barrier function and dermatologic infections. It has been found in the recent years that also with animals, in particular furry animals, domestic animals as well as farm animals, the number of cutaneous infections where over-populations of microorganisms intervene is severely increasing. In particular, the occurrence of primitive infections, dermatoses over-infections and parasitoses has been more frequently observed. It is believed that the changing living conditions of the animals, the ensuing occurrence of bacterial and fungal overpopulation and a consequential weakening of the fur, may be possible causes thereof. In this respect it has for example been observed with animals like cats and dogs that dermatoses and parasitoses induce severe scratching of the animal, which in turn involves alteration of the microbial microflora residing on the fur and skin, inflammatory reactions and keratinisation troubles. The latter are mostly treated through application of antibiotics and anti-inflammatory agents associated in an emollient. These products however have been found to be inconvenient because of their aggressiveness tot he skin and fur and because they do not respect the ecoflora naturally occurring with the animal. It becomes apparent that malfunctioning of the cutaneous barrier may involve widely varying immunologic reactions with the animal, in particular an augmentation of the microbial and fungal sensitivity, a bacterial proliferation and inflammatory reactions. In particular, with animals like cats and dogs, localised or general keratoseborrheaic phenomena resulting from problems associated with sebum secretion or keratinisation anomalies, constitute a characteristic reaction for furry animals. The observed increase of the occurrence of primitive infections, dermatoses over-infections and parasitoses thus obviates the need to a composition, suitable for topical application on animal skin and/or fur, the composition being capable of treating these phenomena. State of the Art From WO 98/49999 it is known that sphingoid base containing formulations show growth-inhibitory activity against gram-negative as well as against gram-positive bacteria, when present in a concentration of at least 0.005 wt. %. Antimicrobial activity of sphingoid bases has been found against bacteria, yeasts and fungi. Typical applications include treatment of acne, dandruff, mycoses, i.e. cosmetic and/or dermatological applications on skin and hair. Bibel et al in Clinical and Experimental Dermatology 1995, 20, 395-400 express their doubt about the suitability of sphingosin and sphinganin in clinical uses for their anti-microbial and anti-fungal activity. Formulations containing sphinganin were effective against Candida albicans and Staphylococcus aureus , in test experiments where inflammated, depilated guinea pig skin are used as simulators for human skin. Sphingosin had little effect. However, anti microbial activity against staphylococcus present on human tissue does not necessarily entail activity against specific staphylococci appearing with animals. DE-A-196 02 108 and DE-A-196 02 111 relate to the use of sphingosin or phytosphingosin based sphingolipids as a deodorant. The object of using sphingosin or phytosphingosin is to obtain deodorant which is capable of selectively fighting those bacteria that are responsible for the unwanted sweat odour with human beings, leaving the normal microbial population and the human temperature regulating system unaffected. WO95/03028 relates to a skin-renewal-stimulating cosmetic composition for frequent and repeated topical application to normal skin. The composition contains skin-renewal-stimulating acids for improving the appearance and condition of the skin. The long-term irritation induced by topical application these skin-renewal-stimulating acids is reduced through incorporation of a sphingosin material into the formulation. The above-described applications are mainly limited to the field of cosmetic products, the use of sphingoid bases in veterinary applications not being touched. |
<SOH> SUMMARY OF THE INVENTION <EOH>It is the aim of the present invention to provide a cosmetic and pharmaceutical composition, for topical application on animal skin and/or fur, the composition being capable of regulating anomalies occurring with sebum secretion by the skin, regulating/maintaining the processes of desquamation and physiological keratinisation of the skin, regulating and treating primitive infections, dermatoses over-infections and parasitoses, micro-organism population of skin and/or fur and of enhancing the recovery capacity of the skin and/or fur, preventing, correcting and/or treating cutaneous affections related to troubles with the microflora of the animal skin. detailed-description description="Detailed Description" end="lead"? |
Information processing method and apparatus program storage medium, program and information recording medium |
The characteristic playback domain of an AV stream is to be reproduced simply and reliably without complicating the management. As marks representing characteristic points of an AV stream, a mark of the type indicating a point and a mark of the type indicating a section (duration) are provided. The mark of the type indicating a point designates a point with a time stamp corresponding to a preset playback timing of the AV stream. The mark of the type indicating a section specifies the domain based on the time stamp of the playback start point of the preset playback domain of the AV stream and on the duration as from the playback start point until the playback end point. |
1. An information processing apparatus comprising: detection means for detecting a moving picture of a characteristic playback domain from an input AV stream; generating means for generating a time stamp of a playback start point of said playback domain and a clip mark having the duration of said playback domain; and recording means for recording said clip mark on an information recording medium. 2. The information processing apparatus according to claim 1 further comprising: acquisition means for acquiring extension data; wherein said generating means appends said extension data acquired by said acquisition means to said clip mark. 3. An information processing method comprising: a detection step of detecting a moving picture of a characteristic playback domain from an input AV stream; a generating step of generating a time stamp of a playback start point of said playback domain and a clip mark having the duration of said playback domain; and a recording step of recording said clip mark on an information recording medium. 4. A program storage medium having stored thereon a computer-readable program, said computer-readable program comprising: a detection step of detecting a moving picture of a characteristic playback domain from an input AV stream; a generating step of generating a time stamp of a playback start point of said playback domain and a clip mark having the duration of said playback domain; and a recording step of recording said clip mark on an information recording medium. 5. A program for having a computer execute: a detection step of detecting a moving picture of a characteristic playback domain from an input AV stream; a generating step of generating a time stamp of a playback start point of said playback domain and a clip mark having the duration of said playback domain; and a recording step of recording said clip mark on an information recording medium. 6. An information recording medium having an AV stream recorded thereon wherein said information recording medium having recorded thereon a time stamp for a playback start point of a characteristic playback domain detected from said AV stream and a clip mark having the duration of said playback domain. 7. An information processing apparatus for reproducing an information recording medium having recorded thereon an AV stream, a time stamp for a playback start point of a characteristic playback domain detected from said AV stream and a clip mark having a duration of said playback domain recorded thereon, said apparatus comprising: acquisition means for acquiring said clip mark from said information recording medium; and control means for controlling the processing of the information recorded on said information recording medium, based on said clip mark acquired from said acquisition means. 8. An information processing method for an information processing apparatus configured for reproducing an information recording medium having recorded thereon an AV stream, a time stamp for a playback start point of a characteristic playback domain detected from said AV stream and a clip mark having a duration of said playback domain, said method comprising: an acquisition step of acquiring said clip mark from said information recording medium; and a control step of controlling the processing of the information recorded on said information recording medium, based on said clip mark acquired from said acquisition step. 9. A program storage medium having stored thereon a computer-readable program for an information processing apparatus for reproducing an information recording medium having recorded thereon an AV stream, a time stamp for a playback start point of a characteristic playback domain detected from said AV stream and a clip mark having a duration of said playback domain recorded thereon, said computer-readable program comprising: an acquisition step of acquiring said clip mark from said information recording medium; and a control step of controlling the processing of the information recorded on said information recording medium, based on said clip mark acquired from said acquisition step. 10. A program for having a computer execute a program, said computer being configured for controlling an information processing apparatus reproducing an information recording medium having recorded thereon an AV stream, a time stamp for a playback start point of a characteristic playback domain detected from said AV stream and a clip mark having a duration of said playback domain, said program comprising: an acquisition step of acquiring said clip mark from said information recording medium; and a control step of controlling the processing of the information recorded on said information recording medium, based on said clip mark acquired from said acquisition step. 11. An information processing apparatus comprising: first acquisition means for acquiring optional playback domain, as instructed by a user, from a playlist which gives a definition of a combination of preset domains in an AV stream; generating means for generating a playlist mark comprised of a set of a time stamp of a start point of said playback domain and a duration of said playback domain; and recording means for recording said playlist mark on said information recording medium. 12. The information processing apparatus according to claim 11 further comprising: second acquisition means for acquiring extension data; wherein said generating means appends said extension data, acquired by said second acquisition means, to said playlist mark. 13. An information processing method comprising: an acquisition step of acquiring an optional playback domain, as instructed by a user, from a playlist which gives a definition of a combination of preset domains in an AV stream; a generating step of generating playlist mark comprised of a set of a time stamp of a start point of said playback domain and a duration of said repay domain; and a recording step of recording said playlist mark on said information recording medium. 14. A program storage medium having stored thereon a computer-readable program comprising: an acquisition step of acquiring an optional playback domain, as instructed by a user, from a playlist which gives a definition of a combination of preset domains in an AV stream; a generating step of generating a playlist mark comprised of a set of a time stamp of a start point of said playback domain and a duration of said playback domain; and a recording step of recording said playlist mark on said information recording medium. 15. A program for having a computer execute: an acquisition step of acquiring an optional playback domain, as instructed by a user, from a playlist which gives a definition of a combination of preset domains in an AV stream; a generating step of generating a playlist mark comprised of a time stamp of a start point of said playback domain and a duration of said playback domain; and a recording step of recording said playlist mark on said information recording medium. 16. An information recording medium having an AV stream recorded thereon, wherein the information recording medium has further recorded thereon a playlist mark comprised of a set of a time stamp of a start point of an optional playback domain as instructed by the user from a playlist which gives a definition of combination of preset domains in said AV stream, and a duration of said playback domain. 17. An information processing apparatus for reproducing an information recording medium having recorded thereon an AV stream and a playlist mark comprised of a set of a time stamp of a start point of an optional playback domain as instructed by a user from a playlist which gives a definition of combinations of preset domains in said AV stream, and a duration of said playback domain, said apparatus comprising: acquisition means for acquiring said playlist mark from said information recording medium; and control means for controlling the processing of the information recorded on said information recording medium based on the playlist mark acquired by said acquisition means. 18. An information processing method for an information processing apparatus reproducing an information recording medium having recorded thereon an AV stream and a playlist mark comprised of a set of a time stamp of a start point of an optional playback domain as instructed by a user from a playlist which gives a definition of combinations of preset domains in said AV stream, and a duration of said playback domain, said method comprising: an acquisition step of acquiring said playlist mark from said information recording medium; and a control step of controlling the processing of the information recorded on said information recording medium based on the playlist mark acquired by said acquisition step. 19. A program storage medium having stored thereon a computer-readable program for an information processing apparatus reproducing an information recording medium having recorded thereon an AV stream and a playlist mark comprised of a set of a time stamp of a start point of an optional playback domain as instructed by a user from a playlist which gives a definition of combinations of preset domains in said AV stream, and a duration of said playback domain, said program comprising: an acquisition step of acquiring said playlist mark from said information recording medium; and a control step of controlling the processing of the information recorded on said information recording medium based on the playlist mark acquired by said acquisition step. 20. A program for having a computer execute a program, said computer being configured for controlling an information processing apparatus reproducing a recording medium having recorded thereon an AV stream and a playlist mark comprised of a set of a time stamp of a start point of an optional playback domain as instructed by a user from a playlist which gives a definition of combinations of preset domains in said AV stream, and a duration of said playback domain, said program comprising: an acquisition step of acquiring said playlist mark from said information recording medium; and a control step of controlling the processing of the information recorded on said information recording medium based on the playlist mark acquired by said acquisition step. |
<SOH> BACKGROUND ART <EOH>Recently, a variety of optical discs have been proposed as disc-shaped recording mediums that are recordable and that can be dismounted from a recording and/or reproducing apparatus. These recordable optical discs have been proposed as large-capacity mediums of several GBs and are felt to be promising as mediums for recording AV (audio visual) signals, such as video signals. Among the encoding systems for digitally compressing digital video signals, there is an MPEG (Moving Picture Experts Group) 2 system. The MPEG2 is also finding application in recording digital video signals on recording mediums. For example, in recording digital video signals on a recording medium, the video signals are encoded in accordance with the MPEG2 system to record an encoded bitstream on a recording medium. In the digital television (TV) broadcast, which recently made its debut, a video program, encoded in accordance with the MPEG2 system, is transmitted in accordance with the format termed a transport stream. In recording the digital broadcast on an information recording medium, such a system is used in which the transport stream is recorded in the form of digital signals, without decoding or re-encoding. If AV signals are recorded on a recording medium, random access reproduction, such as locating or fast-feed reproduction of AV signals, can be realized readily. Using this feature, it is possible for a user to select an optional reproduction domain, specified by an IN point (in-point) and OUT point (out-point) from the recorded AV signals to formulate the playback route of the AV signals freely. This function is routinely termed a play-list reproduction. For example, in the DVD recording standard (DVD specifications for Rewritable/Re-Recordable Discs, Part 3 Video Recording, Version 1.0, September 1999), the playlist reproduction is prescribed as PGC or Original PGC. The user is also able to select a start point of a favorite scene to record the reproduction information on the locating point information. This function is termed a characteristic point or a mark and, in, for example, the DVD video recording standard, the function is prescribed as being the Movie Cell Entry Point Information (M_C_EPI). If, in case the user has selected an optional reproduction domain (start and end points) from a given playlist, a function which affords a particular meaning to the reproduction domain is to be implemented, the following two methods may be contemplated. The first method is such a method in which, in case a user has selected the specified playback domain from the playlist, a separate playlist, having the playback domain as a playback route, is prepared. This method suffers from a drawback that the number of playlists is increased even although the user is not desirous to prepare separate playlists. The second method is such a method in which, in case a user has selected the specified playback domain from the playlist, the domain so selected is indicated using two marks, namely a start mark and an end mark. This method, however, suffers from a drawback that not only the number of the mark information is increased but also the reproducing or editing operation employing the mark is complicated because the two marks representing the domain must be managed as a pair. If a moving picture of an optional characteristic moving picture is selected from the input AV stream, and a function which accords a particular meaning to the domain is to be implemented, such a method may be envisaged in which the domain is indicated by, for example, two marks, namely a start mark and an end mark. This method, however, suffers from a drawback that the number of the mark information is increased, and that, since the two marks indicating the domain must be managed as a pair, the management method for mark recording and/or reproduction tends to be complicated. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG. 1 illustrates a simplified structure of an application format on a recording medium employed in a recording and/or reproducing system. FIG. 2 illustrates a mark on the PlayList. FIG. 3 illustrates a mark on the Clip. FIG. 4 illustrates the syntax of the PlayListMark( ). FIG. 5 illustrates the syntax of the ClipMark( ). FIG. 6 is a block diagram showing the structure of a moving picture recording and/or reproducing apparatus. FIG. 7 is a flowchart for illustrating the method for formulating the ClipMark. FIG. 8 is a flowchart for illustrating the method for formulating the PlayListMark( ). FIG. 9 is a flowchart for illustrating the method for reproducing the PlayList using the PlayListMark/ClipMark of the section indicating type. detailed-description description="Detailed Description" end="lead"? |
Process for preparing a laminated, thermoformed film |
A process for preparing a laminated, thermoformed film which comprises: a. blowing a film-forming composition to form a bubble; b. collapsing the bubble to form a film stack comprising a plurality of layers; and c. heating said film stack in a mould to thermoform said film stack and to adhere the layers together. |
1. A process for preparing a laminated, thermoformed water-soluble film which comprises: blowing a film-forming composition of poly (vinyl alcohol) to form a bubble; collapsing the bubble to form a film stack comprising a plurality of layers; and heating said film stack to a temperature of from 90 to 130° C. in a mould to thermoform said film stack and to adhere the layers together. 2. A process according to claim 1 wherein the thickness of each film layer in the film stack is 20 to 125 μm. 3. A process according to claim 2 wherein a forming pressure of 60 to 130 KPa is applied. 4. A process according to claim 1 wherein the film stack consists of two layers. 5. A process for producing a water-soluble container containing a composition which comprises: blowing a film-forming composition of poly (vinyl alcohol) to form a bubble; collapsing the bubble to form a film stack comprising a plurality of layers; thermoforming at a temperature of from 90 to 130° C. and simultaneously laminating the film stack to produce a pocket; filling the pocket with the compositions placing a water-soluble film on top of the filled pocket; and sealing the films together. 6. A process according to claim 5 wherein the thickness of each film layer in the film stack is 20 to 125 μm. 7. A process according to claim 6 wherein a forming pressure of 60 to 138 KPa is applied. 8. A process according to claim 5 wherein the film stack consists of two layers. 9. A process according to claim 5 wherein the composition is a liquid composition. 10. A process according to claim 5 wherein the composition is a dishwashing, water-softening laundry or detergent composition for a trigger-type spray. 11. A process according to claim 5 wherein the composition is an agrochemical composition. 12. A process according to claim 2 wherein a forming vacuum of 0 to 4 KPa is applied. 13. A process according to claim 6 wherein a forming vacuum of 0 to 4 KPa in applied. 14. A process according claim 6 wherein the film stack consists of two layers. 15. A process according to claim 6 wherein the composition is a liquid composition. |
Scheduling and decision system |
The inventive subject matter herein is directed toward improved scheduling and planning system (200) in which computer implemented software uses a hierarchical selection list to select at least one of a plurality of unconnected users and contact the selected user(s) as a function of an event (110). Another aspect of the inventive subject matter includes an improved decision and/or scheduling system (100) that has soft fields for describing resources. Further aspects utilize a hierarchical soft field configuration. |
1. An improved scheduling system, wherein the improvement comprises: a hierarchical selection list; computer implemented software that uses the list to select at least one of a plurality of unconnected users, and contacts the selected user(s) as a function of an event. 2. The system of claim 1, wherein the selection list correlates the plurality of users with a severity of a problem. 3. The system of claim 1, wherein the selection list correlates the plurality of users with a plurality of contacting methods. 4. The system of claim 3, wherein the selection list correlates the plurality of contacting methods with a severity of a problem. 5. The system of claim 1, wherein the unconnected user is outside the system. 6. The system of claim 1, wherein the unconnected user is a supplier. 7. The system of claim 1, wherein the unconnected user is a support person. 8. An improved decision system, wherein the improvement comprises a data model that includes soft fields for describing resources. 9. The decision system of claim 1 wherein the soft fields are associated in hierarchical manner. 10. The decision system of claim 1 wherein the soft fields are sortable and searchable. 11. The decision system of claim 1 further comprising additional soft fields for locations. 12. The decision system of claim 1 further comprising additional soft fields for processes. 13. The decision system of claim 8 wherein at least some of the resources are materials. 14. The decision system of claim 8 further comprising a computer implementation of a planning algorithm that assumes limited requirements. 15. The decision system of claim 8 further comprising a computer implementation of a scheduling algorithm that assumes limited requirements. 16. A method of scheduling, comprising: creating a hierarchical selection list comprising an event, at least one unconnected user identification associated with the event, and at least one contact method for the unconnected user; receiving notice of the occurrence of the event; selecting the unconnected user and the contact method using the hierarchical selection list; and notifying the unconnected user using the contact method. 17. The method of claim 16, further comprising correlating a severity of a problem with the hierarchical selection list. 18. The method of claim 16, wherein the step of notifying occurs within 10 seconds of the step of receiving notice. 19. The method of claim 16, further comprising the step of automatically sensing the event. |
<SOH> BACKGROUND OF THE INVENTION <EOH>In manufacturing and other applications, scheduling and decision making systems use computers to mange the increasing quantity of data. Traditionally, the cost of data storage was relatively expensive, but today the cost of storing data is so low that in most instances the quantity of data is not cost prohibitive. What can be cost prohibitive, however, is the time and effort required to filer the data and to get relevant information to the appropriate user in a timely manner. In batch systems, filtering of data can be accomplished either automatically or manually. Automatic filtering may take the form of an instruction hard coded in a program, an input parameter to a program (e.g. a selection criteria), or a routing instruction that instructs the system to route particular output to a particular user. Manual filtering of data is generally performed by a person and can be as simple as discerning the appropriate user to receive a computer generated report and giving the report to that user. Systems which utilize manual filtering may also utilize bins or mail boxes that store hard copy reports for an associated user. A persisting problem, however, with batch systems, is that massive reports are often created and much of the information in the report is either superfluous, out-dated, or intended for more than one user. Online reporting and display of data alleviated some of the problems associated with massive hard copy reports and timeliness of data by allowing users to view real time or near real time data using a display screen. Additionally, online systems empowered users with the ability to make decisions that more accurately reflected current conditions. Still, there were problems getting the right information to the right person at the right time. At times, the right person was not available or access to the right information was limited. Even still, there were instances in which too much information was available and this added to the time needed to make a timely decision. There remains a need for improved scheduling and decision making systems and methods which facilitate getting the right information to the right people at the right time. |
<SOH> SUMMARY OF THE INVENTION <EOH>The inventive subject matter herein is directed toward improved scheduling and planning system in which computer implemented software uses a hierarchical selection list to select at least one of a plurality of unconnected users and contact the selected user(s) as a function of an event. Unconnected means that the user is not physically situated such that online access to the scheduling system is readily available. For example, an unconnected user may be a user of the scheduling system who is physically located outside the local network, a supplier of a product used by the scheduling system, or a customer of the entity controlling the scheduling system. With regard to hierarchy, a selection list advantageously correlates a plurality of users with a severity of a problem. It is contemplated that an event may cause a scheduling or decision making problem. Such an event may include the failure of a machine, the absence of an employee, the shortage of a raw material, and so on. Correlation of a user with a severity of a problem may include selecting a user that is most appropriate to solve a particular problem. For instance if the problem is a shortage of a raw material that will cause a delay in production of a product for an important customer, it may be appropriate to select a purchasing manager who can leverage his position with vendors to get expedited delivery of a product. A selection list may alternatively and/or additionally correlate users with contacting methods. By adding a further layer of correlation, it may be easier to contact the appropriate person to solve a particular problem or address a particular event. Thus, hierarchical selection may include multiple layers (tiers) of correlation in which users are correlated with type of problem, severity of problem, and contacting method. Another aspect of the inventive subject matter includes an improved decision and/or scheduling system that has soft fields for describing resources. A soft field is a field that is created dynamically by a user. Thus, not only do users input values for fields, but they actually create fields. In some embodiments, soft fields will be sortable and/or searchable enabling users to further define the type and extent of the data they wish to receive. It should be noted that soft fields may be used for locations, processes, materials, or most any resource. Hierarchical soft field configuration can be useful in almost any planning and/or scheduling system, especially those that assume limited requirements such as ERP (enterprise resource planning), MRPII (manufacturing resource planning), and so on. |
Expander device |
Apparatus (9) for radially expanding an expandable member, the apparatus including a plurality of radially moveable fingers (22). The radially moveable fingers (22) can move between two positions; in a first position, the moveable fingers (22) are radially extended to a form a cone to facilitate radial expansion of the expandable member; in the second position, one or more of the radially movable fingers (22) can move radially inward so that restrictions in the path of the apparatus can be by-passed. |
1. Apparatus for expanding an expandable member, the apparatus comprising a first member, at least one radially movable portion, a second member, and a force isolating mechanism acting between the first and second members. 2. Apparatus according to claim 1, wherein the first member comprises a housing with a blind bore. 3. Apparatus according to claim 1, wherein the second member comprises a shaft having a cone that bears against the at least one radially movable portion. 4. Apparatus according to claim 3, wherein the shaft and cone can move axially with respect to the first member in and out of engagement with the at least one radially movable portion. 5. Apparatus according to claim 1, wherein the at least one radially movable portion is coupled to the first member so that it can move in a radial and/or axial direction. 6. Apparatus according to claim 3, wherein the force isolating mechanism comprises a spring. 7. Apparatus according to claim 6, wherein the at least one radially movable portion is held in a radially expanded position by the cone on the second member moving axially with respect to the first member to a first position in which the spring is contracted. 8. Apparatus according to claim 7, wherein the second member can move axially under an axial pulling force, and the cone can move to a second position that allows the at least one radially movable portion to move radially inward to bypass a restriction. 9. Apparatus according to claim 8, wherein as the restriction is passed, the axial pulling force drops below a biasing force of the spring so that the spring contracts, and the cone moves into engagement with the at least one radially movable portion causing it to move radially outward to the radially expanded position. 10. Apparatus according to claim 8, wherein the engagement of the at least one radially movable portion with the restriction can cause it to move inwards against the cone thereby moving it to the second position in which the spring is extended. 11. Apparatus according to claim 1, wherein the at least one radially movable portion is pivotally coupled to the first member. 12. Apparatus according to claim 1, wherein an outer face of the at least one radially movable portion defines a cone. 13. Apparatus for expanding an expandable member, the apparatus comprising a body, at least one radially movable portion, and a force isolating mechanism acting between the body and the at least one radially moveable portion. 14. Apparatus according to claim 13, wherein the force isolating mechanism provides a biasing force to the at least one radially moveable portion. 15. Apparatus according to claim 14, wherein a force required to move the at least one radially moveable portion inwards is greater than the biasing force of the force isolating mechanism. 16. Apparatus according to claim 14, wherein a radial position of the at least one radially movable portion is at least partially controlled by the biasing force of the force isolating mechanism. 17. Apparatus according to claim_14, wherein force applied to the body can be isolated from the at least one radially moveable portion by the force isolating mechanism. 18. Apparatus according to claim 13, wherein the force isolating mechanism comprises a resilient member that allows relative movement between the body and the at least one radially moveable portion. 19. Apparatus according to claim 18, wherein the relative movement between the body and the at least one radially moveable portion is in an axial direction. 20. Apparatus according to claim 18, wherein the resilient member has a biasing force that is greater than a maximum load that will be applied to the apparatus. 21. Apparatus according to claim 13, wherein the force isolating mechanism includes a fluid chamber that is in communication with the at least one radially moveable portion, the fluid chamber being in fluid communication with a spring mechanism. 22. Apparatus according to claim 21, wherein the spring mechanism comprises a first chamber, a floating piston in communication with the first chamber, and a second chamber in communication with the piston. 23. Apparatus according to claim 22, wherein the first chamber contains fluid and is in fluid communication with the fluid chamber that is in communication with the at least one radially moveable portion, and the second chamber includes a spring. 24. Apparatus according to claim 23, wherein as the at least one radially moveable portion is forced inward due to a restriction, it acts on the fluid in the fluid chamber, forcing the fluid into the first chamber, wherein the displacement of fluid causes the floating piston to compress the spring in the second chamber and this allows the at least one radially moveable portion to move inwards, thus passing the restriction. 25. Apparatus according to claim 24, wherein once the restriction has been passed, the spring extends forcing fluid in the first chamber to be transferred to the fluid chambers, thus forcing the at least one radially moveable portion outwards. 26. Apparatus according to claim 13, wherein the force isolating mechanism comprises a hydraulic spring. 27. Apparatus according to claim 26, wherein the hydraulic spring includes an inflatable element that is in fluid communication with a fluid chamber. 28. Apparatus according to claim 27, wherein the fluid chamber is filled with a fluid that is incompressible. 29. Apparatus according to claim 27, wherein the fluid in the fluid chamber acts on a floating piston that is located in a second chamber. 30. Apparatus according to claim 29, wherein the second chamber is filled with a gas. 31. Apparatus according to claim 29, wherein as the at least one radially moveable portion is forced inwards due to a restriction, it acts on the fluid in the inflatable element, forcing fluid into the fluid chamber, and the displacement of fluid into the fluid chamber acts on the piston, causing it to compress the fluid in the second chamber. 32. Apparatus according to claim 31, wherein once the restriction has been passed, the fluid in the second chamber expands, forcing the piston to act on the fluid in the fluid chamber, the fluid being transferred to the inflatable element, thus forcing the at least one radially moveable portion outwards. 33. Apparatus according to claim 13, wherein the at least one radially moveable portion is pivotably mounted to the body. 34. Apparatus according to claim 13, wherein the at least one radially moveable portion comprises at least one finger. 35. Apparatus according to claim 34, wherein an outer face of the at least one finger defines a cone. 36. An apparatus for expanding an expandable member, comprising: a housing having at least two apertures in a circumference thereof; at least two radially movable segments disposed at least partially in the apertures; a shaft axially moveable between a first position and a second position relative to the housing, wherein in the first position an enlarged diameter portion of the shaft contacts an inside surface of the segments to retain the segments in an extended position, and in the second position the enlarged diameter portion displaces to permit inward movement of the segments; and a resilient member for biasing the shaft to the first position for axial forces applied to the shaft up to a predetermined axial force. 37. The apparatus of claim 36, wherein the at least two radially movable segments are pivotally coupled to the housing. 38. The apparatus of claim 36, wherein the enlarged diameter portion includes a sloping portion for contacting a mating sloping portion of the inside surface of the segments, an angle of the sloping portion and mating sloping portion is selected relative to the predetermined axial force. 39. The apparatus of claim 36, wherein the resilient member is a spring. 40. An apparatus for expanding an expandable member, comprising: a housing having at least two apertures in a circumference thereof; at least two radially movable segments disposed at least partially in the apertures; and a piston assembly for selectively biasing the segments in an extended position against inward forces applied to the segments up to a predetermined inward force, the piston assembly comprising: at least two fixed pistons associated with each segment to move the segments between the extended position and a retracted position; a reservoir of pressurized fluid acting on a surface of the pistons; and a damping piston having a damping spring on one side of the piston and communication with the reservoir on another side of the piston. 41. The apparatus of claim 40, wherein the at least two radially movable segments are pivotally coupled to the housing. 42. An apparatus for expanding an expandable member, comprising: a body; an inflatable element, wherein the inflatable element is pressurized to outwardly extend an expandable member of the body to an extended position; and a hydraulic absorber for selectively pressurizing the inflatable element against reactive inward forces applied to the inflatable element up to a predetermined inward force. 43. The apparatus of claim 42, wherein the hydraulic absorber comprises: a reservoir in fluid communication with the inflatable element; and a floating piston movable within the hydraulic absorber, the floating piston having a gas accumulator on one side of the piston and communication with the reservoir on another side of the piston. 44. The apparatus of claim 43, wherein the gas accumulator is pressurized downhole. 45. The apparatus of claim 42, wherein the expandable member comprises at least two blades pivotally coupled to the body. 46. A method of expanding an expandable member, comprising: locating an expander device proximate the expandable member; supplying a first axial force to the expander device to translate the expander device across the expandable member and expand the expandable member; and supplying an increased second axial force to the expander device to cause an extendable member of the expander device to at least partially move radially inward to overcome a restriction. |
Self cleaning shredding device having movable cleaning rings |
A self cleaning cutting assembly is provided, having an array of counter rotating cutters in which a plurality of cutters rotating in a first direction are mounted on a first shaft and a plurality of second cutters are mounted on a second shaft for rotating in an opposite direction. The array of cutters is provided with a plurality of clearing or cleaning rings located alternately with the cutters on each shaft. The cutting array is kept clear of debris or partially cut material by movement of the clearing or cutting rings against adjacent cutters. Movement of the clearing or cleaning rings is a combination of rotary motion and linear motion, in which the linear motion is imparted by the cutter located opposite the cleaning ring striking the cleaning rings so as to push the cleaning ring beyond the position of the widest part of the cutter to completely dislodge material from the cutter. |
1. A self cleaning cutting assembly, including a first driven cutter provided on a first shaft for driving movement in a first rotary direction, and a second driven cutter mounted on a second shaft for driven movement in a counter rotating direction to the first cutter, wherein said first and second cutters are located substantially adjacently opposite each other so that material introduced into the cutting assembly between the first and second cutters is cut by the cooperative interaction of the two cutters undergoing counter rotation with respect to each other, so as to produce relatively smaller pieces of material, the cutting assembly further including a floating non-driven cleaning means freely mounted on the first shaft and capable of undergoing a third movement, wherein said cleaning means is located adjacent the first cutter so that movement of the first cutter imparts a first component of the third movement to the cleaning means and the cleaning means is located substantially opposite the second cutter, so that movement of the second cutter imparts a second component of the third movement to the cleaning means so that the third movement of the cleaning means is a combination of the first and second components of the third movement imparted by the first and second cutters respectively, whereby movement of the cleaning means with respect to the first cutter caused by contact from the second cutter maintains the first cutter in a condition substantially free of severed, relatively smaller sized pieces of material cut by the two cutters, thereby reducing or substantially eliminating the relatively smaller sized particles of material from interfering with or disrupting operation of the first cutter by wrapping round or remaining in contact with the first cutter during use of the cutting assembly, thereby providing the self cleaning cutting assembly. 2. A self cleaning cutting assembly according to claim 1, wherein the cutting assembly comprises a shredding device. 3. A self cleaning cutting assembly according to claim 2, wherein the shredding machine shreds material into elongate strips of material, preferably elongate strips of material that have a regularly repeating profile or shape. 4. A self cleaning cutting assembly according to claim 1, wherein the material is a rubber or other resilient material. 5. A self cleaning cutting assembly according to claim 1, wherein said first and second cutters comprise rotary cutting blades and the outer cutting surfaces of the cutters are provided with surface irregularities or discontinuities. 6. A self cleaning cutting assembly according to claim 5, wherein the surface irregularities or discontinuities are selected from teeth, grooves, cleats, recesses, rebates, spaces, and gaps. 7. A self cleaning cutting assembly according to claim 5, wherein the cutting surfaces are provided with a retaining means. 8. A self cleaning cutting assembly according to claim 7, wherein the retaining means is selected from a hook, enlarged teeth, projections, spearhead, tip, nose, finger or other protuberance. 9. A self cleaning cutting assembly according to claim 1, including a plurality of projections located at regularly spaced apart intervals around the circumference of the cutting surface of said first and second cutters. 10. A self cleaning cutting assembly according to claim 1, in which the cutting surface of said first and second cutters includes two cutting edges located at the sides of the blades and defining a central section therebetween. 11. A self cleaning cutting assembly according to claim 10, in which the projections are located on the central section of the cutting surface between the side sections. 12. A self cleaning cutting assembly according to claim 1, in which cleaning means comprises a spacer. 13. A self cleaning cutting assembly according to claim 12, in which the spacer comprises a ring. 14. A self cleaning cutting assembly according to claim 13, in which the ring is freely mounted on the first shaft and has first roller for floating movement in accordance with movement of the first and second cutters and the ring is not substantially driven by the shaft. 15. A self cleaning cutting assembly according to claim 13, wherein the spacer comprises a ring having a reinforcing, internally located, radially directed flange, and an outer circumferential surface. 16. A self cleaning cutting assembly according to claims 13, wherein the ring includes one or more of an inner ring, an intermediate ring or an outer ring. 17. A self cleaning cutting assembly according to claim 1, wherein the third movement is an eccentric movement which is a combination of a rotary movement imparted by the first cutter and a linear movement imparted by the second cutter. 18. A self cleaning cutting assembly according to claim 17, wherein the linear movement is a longitudinal movement in a radial direction. 19. A self cleaning cutting assembly according to claim 13, wherein the movement of the spacer ring against one or more cutters, cleans the one or more cutters and prevents cut pieces or strips of material from remaining against, or being adhered to the one or more cutters. 20. A self cleaning cutting assembly according to claim 17, wherein the linear movement is superposed on th e rotary movement of the spacer ring a plurality of times each revolution of the spacer ring. 21. A self cleaning cutting assembly according to claim 1, characterised in that the number of linear movements imparted to the cleaning means is in accordance the number of projections on adjacent cutters. 22. A self cleaning cutting assembly according to claim 13, wherein said first and second cutters includes a plurality of projections and the outer surface of the spacer ring is struck by the tip of the projection of the cutter located opposite the spacer ring. 23. A self cleaning cutting assembly according to claim 22, in which the cutter having the projection for striking the ring, is located in opposed relationship to the spacer ring so that the cutter is located on the second shaft and the ring is located on the first shaft, said first and second shafts being arranged in opposed relationship to each other. 24. A self cleaning cutting assembly according to claim 22, in which movement of the spacer is along the sides of the cutter so that movement with respect to the cutter cleans material from the cutter, and prevents any severed material from wrapping around the cutter as it rotates. 25. A self cleaning cutting assembly according to claim 24, in which the cutting blade or spacer is provided with a wear portion in which the wear portion. 26. A self cleaning cutting assembly according to claim 25, characterised in that the wear portion is a sacrificial wear portion made from a material which is at least as hard as the material of the cutter or spacer. 27. A self cleaning cutting assembly according to claim 26, in which the sacrificial wear material, quietens operation of the device by reducing the noise of impact of the cutters with the spacer ring or against the sides of the cutters. 28. A self cleaning cutting assembly according to claim 1, wherein the cutters located on the first shaft have different widths. 29. A self cleaning cutting assembly according to claim 1, wherein the cutters in opposed relationship to each other are substantially of the same size. 30. A self cleaning cutting assembly according to claim 1, including two or more different cutting zones located axially along the length of the first and second cutters. 31. A self cleaning cutting assembly according to claim 1, further including a trommel for recycling partially cut items back to the cutting assembly for further cutting. 32. A self cleaning cutting assembly according to claim 31, wherein the trommel comprises a wheel-like framework having a plurality of baskets for receiving partially cut material for recycling the material by reintroducing the material for subsequent severing in the cutting assembly. 33. A method of reducing in size material using a self cleaning cutting assembly comprising introducing the material to be cut into the cutting assembly, cutting the material between two counter rotating cutters located on opposed shafts, and preventing the cut material from wrapping around the cutters by operation of a cleaning means, wherein movement of the cleaning means is effected by rotation of the cutters. 34. (Cancel) A self clearing cutting assembly, substantially as hereinbefore described with reference to the accompanying drawings. 35. (Cancel) A method of cutting a material using a self clearing cutting assembly, substantially as hereinbefore described with reference to the accompanying drawings. |
<SOH> SUMMARY OF THE INVENTION <EOH>According to the present invention there is provided a self cleaning cutting assembly, particularly a cutting assembly adapted for use with a shredding device for shredding material into relatively smaller sized pieces, including: a first cutting means capable of undergoing a first movement; a second cutting means capable of undergoing a second movement; and a cleaning means capable of undergoing a third movement. wherein said first and second cutting means are located substantially adjacently opposite each other so that material introduced into the cutting assembly between the first and second cutting means is cut by the cooperative interaction of the two cutting means with each other to produce the relatively smaller sized pieces of material, said cleaning means being located adjacent the first cutting means so that movement of the first cutting means imparts a first component of the movement to the cleaning means and substantially opposite said second cutting means so that when said second cutting means moves the second cutter means can contact the cleaning means to impart a second component of the movement of the cleaning means wherein the third movement of the cleaning means is a combination of at least the first and second components of movement imparted by the first and second cutter means, whereby movement of the cleaning means with respect to the first cutter means caused by contact from the second cutter means maintains the first cutter means in a condition substantially free of the severed relatively smaller size pieces of material thereby reducing or substantially eliminating the relatively smaller sized pieces of material from interfering with or disrupting operation of the first cutter means by wrapping around or remaining in contact with the first cutter means during use of the cutting assembly so that movement of the cleaning means provides self cleaning of the cutting assembly. Typically the cutting assembly of the present invention is a shredding machine or shredding device. More typically the shredding device shreds material into elongate strips of material. Even more typically the elongate strips of material have a regularly repeating profile or shape, and are typically curved out of the plane containing the lengthwise extending axis of the material. Even more typically, the shredded pieces have a tendency to curl or wrap around the cutters, particularly due to the shape of the shredded pieces being curved or similar, particularly curved about their lengthwise axis. Although one preferred material being treated by the shredder of the present invention is rubber or other resilient material, such as for example, from used motor vehicle tires, including heavy duty truck and bus tires, it is to be noted that the present invention can be used to cut or comminute material of any type. Typically the cutting means is a rotary cutting blade arrangement, typically having one or more cutting surfaces located around the periphery of the blade. More typically, the outer cutting surface of the blade is provided with surface irregularities or discontinuities, such as for example, projections and troughs in the form of teeth, grooves, cleats or the like. More typically, the cutting surface is provided with a retaining means or holding means. Typically the retaining means is a hook, enlarged teeth, projection, spear head or tip or the like. More typically there are two, three, four or more such projections located at regularly spaced apart intervals around the circumference of the cutting surface. More typically, the cutting surface comprises three parts, two cutting edges or sections located at or towards the sides of the blades and a central section located between the side sections. More typically, the holding means is located in or about the central section. Typically there is a plurality of rotating blades arranged to extend in a line or row of similar blades. Typically, the blades rotate in unison. Typically the first movement of the first cutter is a rotation. Typically the second movement of the second cutter is a rotation. Even more typically the direction of rotation of the first cutter is opposite to the direction of rotation of the second cutter. More typically the first and second cutters rotate in opposite directions at the same speed and in unison with each other. Alternatively, the two rows of cutters can rotate at different speeds. The speeds of rotation can be variable. Typically, all the cutters are mounted on shafts. More typically, there are two shafts, each shaft having a plurality of cutters. Even more typically, the cutters on each shaft are all the same size. Alternatively, the cutters on the one shaft can be of different sizes. Even more typically, the cutters differ in widths. Typically, there are cutters of two or three sizes mounted on the one shaft. More typically, there are different cutting zones defined by the different sized cutters. More typically, the cutters located substantially opposite each other are of the same size as each other so as to define different zones of the cutting array. More typically, there is a first zone of cutters in which the cutters are about 100 mm in width, typically there is a second zone of cutters in which the cutters are about 70 mm in width, and typically there is a third zone of cutters in which the cutters are about 40 mm in width. The cutters are readily replaceable on the shafts. Typically, the cutters are replaceable in blocks in accordance with their wear. Typically the clearing or cleaning means is a spacer. Typically the spacer is a ring arrangement. More typically the spacer is a simple ring having a reinforcing internally located radially directed flange and an outer circumferential surface. Even more typically, the ring is of a complex structure having one or more inner ring portions, preferably provided with a flange arrangement, an intermediate ring portion, preferably of a material having different mechanical properties or characteristics or an outer ring portion surrounding the intermediate portion. Typically, the third movement is an eccentric movement being a combination of rotation and linear movement. More typically, the linear movement is longitudinal or transverse movement in a radial direction or tangential direction. More typically, movement of the spacer ring against one or more cutters cleans and/or clears the cutter or cutters and prevents cut pieces or strips of material from remaining against or being associated with the cutter or cutters or being adhered to the cutters. Even more typically, the linear movement, typically radial movement, is superposed on the rotary movement of the ring two, three, four or more times each revolution of the spacer ring. Typically, the projection of the second cutter means strikes the outer surface of the ring to move the ring with respect to the first cutter to clean or clear the first cutter. More typically, the outer surface of the spacer ring is struck by the tip of the projection of the cutter or the like. Even more typically, the linear movement of the ring is such that the ring extends beyond the tip of the projection so as to push the material away from an adjacent cutter so as to completely clean the cutter. Typically the spacer ring is not driven by the driving shaft but rather is driven by contact with the first cutter located opposite the ring, or by the cutters on either side of the spacer ring in combination with contact by the oppositely located cutter. Typically, movement of the spacer is against or along the sides of the cutter so that the movement with respect to the cutter cleans material from the cutter, particularly the cutting surfaces of the cutter, and prevents any severed piece of material from wrapping around the cutter as it rotates by knocking the severed material away from the cutter. More typically, the extent of movement of the spacer is sufficient to dislodge any material adhering to the cutter. Typically, the spacer ring is provided with a wear surface or wear portion. More typically, the wear surface is a ring, either a complete ring or a partial ring, located at or towards one or both sides of the spacer ring. Even more typically, the wear ring is metallic or is a resilient material made from a foam material, or is a plastically deformable material. Even more typically, the foam material is a sound deadening material for reducing the noise of impact of or against the cutters. Even more typically, the wear portion is made from the same material as the spacer ring or from a material which is more durable or harder than the spacer ring. Typically, the device of the present invention is provided with a recycling assembly for returning items back to the shredder for further shredding so as to produce pieces of a more uniform size. More typically, the recycling assembly is located to one side of the shredder, preferably coaxially to one side. The recycling assembly is provided with means for transporting material to the shredder. More typically, the transporting means is a slide, baskets or similar for holding the severed material. More typically, the recycling assembly is a trommel or trommel-like device which rotates to reintroduce cut pieces into the shredder. Even more typically, the trommel is a rotating wheel-like framework having baskets for transporting cut pieces back to the receiving hopper of the shredder. Even more typically, the trommel is located axially off-set or to one end of the shredder for introducing cut pieces to one region of the counter rotating cutters, preferably the portion of the array having cutters of reduced or relatively smaller widths, such as for example, from about 40 to 70 mm wide. More typically, the trommel is provided with a slide, preferably an adjustably movable slide for collecting and/or distributing material. |
Induction of tolerance by apoptotic and/or necrotic cells |
The present invention is directed to a method of inducing tolerance to self-antigens in a subject having an autoimmune diseases. In particular, the invention provides a pharmaceutical composition and method of use thereof for the modulation of immunogical activity in an animal subject. Said modulation may be an increased tolerance to self apoptotic cells, a reduction in the tissue levels of autoantibodies associated with apoptotic cells, a reduction in the tissue levels of autoantibodies associated with an autoimmune disease, a reduction in the level of inflammation and inflammatory mediators associated with an autoimmune disease, a reduction in the level of tissue damage associated with an autoimmune disease, or a combination thereof. |
1. A method of treating a subject having an autoimmune disease, comprising the steps of: obtaining cells from the subject; inducing cell death in said cells resulting in apoptotic and/or necrotic cells; administering to the subject an amount of said apoptotic and/or necrotic cells effective to produce a modified immune response in said subject, thereby treating the subject with the autoimmune disease. 2. A method of treating a subject having an autoimmune disease, comprising the step of administering to the subject an amount of apoptotic and/or necrotic cells effective to produce a modified immune response in said subject, thereby treating the subject with the autoimmune disease. 3. The method of claim 1 whereby the step of inducing cell death is obtained by exposing said cells to an apoptosis-inducing agent or to a necrosis-inducing agent. 4. The method of claim 1 whereby the step of inducing cell death is obtained by exposing said cells to an apoptosis-inducing treatment or to a necrosis-inducing treatment. 5. The method according to any one of claims 1 to 4, whereby said modified immune response is an increased tolerance to self-apoptotic cells. 6. The method according to claim 5, whereby said modified immune response in said subject is a reduction in the tissue level of auto-antibodies associated with self-apoptotic cells. 7. The method according to claim 6, whereby said auto-antibodies are anti-nuclear antibodies, anti-single stranded DNA antibodies, anti-double stranded DNA antibodies, anti-cardiolipin antibodies, anti-phosphatidylserine antibodies, anti-2GPI antibodies, anti-Sm antibodies, anti-RNP antibodies, or anti-Ku antibodies. 8. The method according to claim 5, whereby said modified immune response in said subject is a reduction in the level of inflammatory response. 9. The method according to claim 8, whereby said inflammatory response is associated with chemokines, cytokines, eicosanoids, complement proteins, C-reactive proteins, TNF, dendritic cells or a combination thereof. 10. The method according to claim 1 or 2, whereby said autoimmune disease is associated with an immune response to self-antigens appearing on apoptotic cells. 11. The method according to claim 10, whereby said autoimmune disease is systemic or discoid lupus, erythematosis, rheumatoid arthritis, polymyositis, or vasculitis. 12. The method according to claim 1 or 2, whereby said cells are hematopoetic cells, thymocytes, splenocytes, lymphocytes, monocytes, a cultured cell line or a combination thereof. 13. The method according to claim 3, whereby said apoptosis-inducing agent is a steroid, a peptide, a protein, a sugar, a lipid, an antibody, or a combination thereof. 14. The method according to claim 13, whereby said steroid is dexamethasone. 15. The method according to claim 13, whereby said protein is perforin. 16. The method according to claim 1 or 2, whereby said composition is administered in combination with an immunosuppressing molecules. 17. A pharmaceutical composition comprising an effective amount of apoptotic and/or necrotic cells, whereby the administration of said composition to a subject suffering from an autoimmine disease produces a modified immune response in said subject. 18. The composition according to claim 17, wherein said modified immune response is a reduction in the tissue level of auto-antibodies associated with apoptotic cells in said subject. 19. The composition according to claim 18, wherein said auto-antibodies are anti-nuclear antibodies, anti-single stranded DNA antibodies, anti-double stranded DNA antibodies, anti-cardiolipin antibodies, anti-phosphatidylserine antibodies, anti-2GPI antibodies, anti-Sm antibodies, anti-RNP antibodies, anti-Ku antibodies, or a combination thereof. 20. The composition according to claim 17, wherein said modified immune response is a reduction in the level of inflammatory response. 21. The composition according to claim 20, wherein said inflammatory response is associated with chemokines, cytokines, eicosanoids, complement proteins, C-reactive proteins, TNF, dendritic cells or a combination thereof. 22. The composition according to claim 17, wherein said autoimmune disease is associated with an immune response to self-antigens appearing on apoptotic cells. 23. The composition according to claim 17, wherein said cells are from autologous origin. 24. The composition according to claim 23, wherein said cells are hematopoetic cells, thymocytes, splenocytes, lymphocytes, monocytes, or a combination thereof. 25. The composition according to claim 17, whereby said apoptotic and/or necrotic cells are obtained by contacting the cells with an apoptosis-inducing agent or a necrotic-inducing agent or by exposing the cells to an apoptosis-inducing treatment or a necrosis-inducing treatment, or a combination thereof. 26. The composition according to claim 25, wherein said apoptosis-inducing agent is a steroid, a peptide, a protein, a sugar, a lipid, an antibody, or a combination thereof. 27. The composition according to claim 26, wherein said steroid is dexamethasone. 28. The composition according to claim 26, wherein said protein is perforin. 29. The composition according to claim 17, wherein said composition is suitable for administration via an intravenous route, an intradermal route, a subdermal route, an intramuscular route, oral administration or a combination thereof. 30. The composition according to claim 17, wherein said composition is administered in combination with an immunosuppressing molecules. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The immune system of animals is a complex and multivarious network comprising cells, antibodies, solid and non-solid organs, and chemical messenger molecules which allow for communication between these structures. A hallmark of a healthy immune system is the ability to recognize bacteria, viruses, and other foreign bodies and to effectively attack such pathogens while continuing to distinguish between the foreign bodies and the molecules, cells, tissues and organs comprising the individual organism. When this aspect of an animal's immune system is deficient the result is a state of disease, often one in which the immune system attacks one or more specific molecules or cells leading to tissue and organ damage. Since the immune system destroys bodies recognized as foreign, often known as antigens, through a complex process known as inflammation, of which many different types exist, the immediate and chronic types of tissue damage in autoimmune and inflammatory diseases are frequently the result of one or more types of inflammation. In autoimmune disease an immune response directed against one or more components of the animal's own tissues or cells results in damage to one or more organs or tissues. In mammals, particularly in humans, many clinically different types of autoimmune disease occur, including subtypes of particular autoimmune disease. Although each type of autoimmune disease is associated with a spectrum of clinical symptoms and aberrant laboratory parameteres, signs and symptoms of autoimmune diseases frequently overlap so that one or more are diagnosed in the same patient. The vast majority cases in which one or more autoimmune disease has been diagnosed are characterized by the presence in the affected subject of autoantibodies. The autoantibodies are directed to one or more molecular or cellular targets, known as antigens, within the animal. Such autoantibodies are present at tissue levels, which are often ten to one hundred times the normal level in healthy individuals and give rise to a significant proportion of the organ and tissue damage associated with the particular autoimmune disease. For example, in the autoimmune disease myasthenia gravis, autoantibodies against a receptor in meuromuscular junction are associated with muscle weakness, while in systemic lupus erythematosus, anti-dsDNA antibodies are associated with nephritis in human patients and can cause nephritis upon injection to normal mice. The tissue and organ damage is attributed to the presence of autoantibodies and to the inflammation, which arises to due inflammatory immune responses, set off by autoantibodies. Thus the signs and symptoms of disease are due to autoantibodies, the autoimmune inflammatory response, or a combination thereof. Autoimmune diseases include rheumatoid arthritis, graft versus host disease, systemic lupus erythromatosus (SLE), scleroderma, multiple sclerosis, diabetes, organ rejection, inflammatory bowel disease, psoriasis, and other afflictions. It is becoming increasingly apparent that many vascular disorders, including atherosclerotic forms of such disorders, have an autoimmune component, and a number of patients with vascular disease have circulating autoantibodies. Autoimmune diseases may be divided into two general types, namely systemic autoimmune diseases (exemplified by lupus and scleroderma), and organ specific (exemplified by multiple sclerosis, diabetes and atherosclerosis, in which latter case the vasculature is regarded as a specific organ). One important autoimmune disease is lupus and this disease is a model for enraveling the physiology and developing inventive treatments for autoimmune disease in general. It has long been appreciated that DNA and histones are major autoantigens in systemic lupus erythematosis (SLE). However, only recently has evidence been provided that the DNA-histone complex, i.e., nucleosomes, are the preferred targets of autoantibodies in SLE. The question then arises as to how nucleosomes and several other intracellular antigen targets can be immunogenic in SLE. During apoptosis, the membrane of cells undergoing apoptosis form cytoplasmic blebs, some of which are shed as apoptotic bodies. It was recently demonstrated that exposure of kertinocytes to high frequency light induces apoptosis, and that the cell surface expression of Ro and La, but also of nucleosomes and ribosomes, can be explained by translocation of certain intracellular particles to the apoptotic surface blebs. Significantly, another translocation which occurs during apoptosis is that of phosphatidyl serine (PS), an acidic phospholipid that normally resides on the inside of the cell, but flips to the outside of the cell membrane when the cell undergoes apoptosis. PS like, cardiolipin, is a major autoantigen for anti-phospholipid (aPL) antibodes in SLE. Taken together, these findings provide a unifying hypothesis to explain antigen selection in SLE, e.g., that SLE patients are responding to the exposure of intracellular proteins translocated to the cell surface during apoptosis. Thus, SLE patients form an immune response to apoptotic material. Although there may be many possible explanations to explain this observation, any explanation must take into account that in SLE patients the uptake of apoptotic cells by macrophages in vitro is reduced. Furthermore, brief, limited administration of syngeneic apoptotic cells to normal strains of mice leads to induction of autoantibodies and glomerular depositions. In addition, it has been shown that the complement system is important in clearance of uptake of apoptotic cells, suggesting the novel hypothesis disclosed herein for the reason why greater than 90% of patients homozygous for C1q and greater than 70% of C4 deficiency patients develop SLE. This novel understanding of the pathogenesis of SLE may suggest a different approach to the treatment of SLE. Manipulation of the immune system to prevent a deleterious response has been the goal of immunologists for many years in transplantation biology and autoimmune diseases. Traditionally, the main effort was to induce immunosuppression and the current therapy for a classical systemic autoimmune disease such as SLE is drug treatment with corticosteroids, azathyoprine, cyclopohosphamide, and cyclosporine, all of which are administered with the aim suppressing the immune system. Immunosuppression was an important step in ameliorating the 5-years survival rate of SLE patients in the last three decades but it is far from the ideal treatment since no cure is achieved and patients suffer from very serious side effects leading to high rates of morbidity and being the main cause of premature mortality. In that regard, even the newly developed biologics currently under toxicity and efficacy evaluation, such as anti-CD40 ligand, and CTLA4 lg, are non-specific for the autoimmune B and T cell clones and, if successful for autoimmunity, will suppress probably the entire immune system. In addition to fighting infections, the immune system has other roles in maintaining the normal state of health and function of the animal. Throughout the life span of an animal, tissues become reshaped with areas of cells being removed. This is accomplished by the cells' undergoing a process called programmed cell death or apoptosis, the apoptotic cells disintegrating and being phagocytosed while not becoming disrupted. In many organs, for example, a certain percentage of the cells die off every day while different branches of the immune system are typically called in to remove the dead cells and parts thereof to make room for the new cells which are born to replace them. Were it not for the cellular debri removing cells of the immune system, often known as macrophages, tissue and organ growth would be impossible due to a lack for space. In fact, the process of apoptosis is considered to be particularly important in the development and maintenance of the immune system itself, where the immune cells which recognize or attack other normal cells of the animal are destroyed and removed by this process. Thus, while apoptosis is a process used by the immune system in protecting the organism, it is also used to maintain tolerance to self antigens and therefore allowing the immune system to fulfill its role in distinguishing the animal's own cells from those of non-self invaders. Immature dendritic cells (IDC) engulf apoptotic cells and are able to acquire antigens found in the dying cells. IDC that capture apoptotic macrophages infected by killed influenza-virus, mature and activate lymphocytes to mount virus-specific CTL responses in the presence of conditioned media. However, in the absence of infection and conditioned media, IDC do not mature following uptake of apoptotic cells and as a consequence are less able to efficiently present acquired antigens. Furthermore, it has been suggested that following interaction with apoptotic material, IDC may have a role in maintaining peripheral tolerance to self-antigens that are permanently created at different sites. In support of this, autoimmunity or lupus like disease has been observed in mice and human deficient in receptors important for uptake of apoptotic cells such as ABC1 cassette transporter, Mer, and complement deficiencies. Clearance via specific receptors may dictate specific immune response or tolerance as demonstrated by TGF-β and IL-10 secretion by macrophages following uptake by macrophages. So, cytokines, chemokines, eicosanoids, and additional materials found in the milieu of the interaction, may polarize the immune response. Thus, the aim of the present Invention is to induce tolerance to self antigens in a subject having an autoimmune disease, mainly antigens related to apoptotic and/or necrotic cells. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is directed to a method of inducing tolerance to self-antigens in a subject having an autoimmune disease. According to the present invention, a lack of tolerance to apoptotic and/or necrotic cells is an important aspect in the development of autoimmune disease and an important target for therapy. In particular, the invention provides a pharmaceutical composition and method of use thereof for the modulation of immunogical activity in an animal subject wherein said modulation is an increased tolerance to apoptotic and/or necrotic cells, a reduction in the tissue levels of autoantibodies associated with apoptotic and/or necrotic cells, a reduction in the tissue levels of autoantibodies associated with an autoimmune disease, a reduction in the level of inflammation and inflammatory mediators associated with an autoimmune disease, a reduction in the level of tissue damage associated with an autoimmune disease, or a combination thereof. A composition for treating autoimmune diseases according to the present invention should contain antigens, i.e. apoptotic and/or necrotic cells, or fragments thereof, i.e. blebs of apoptotc cells, membrane fragments, and peptides, that upon administration, interact with the immune system of the animal to produce enhanced tolerance to self antigens. In addition, the antigens or fragment thereof should be present in a form which can be recognized by the subject's immune system when the composition is administered to the subject. The desirable antigens may be present on intact apoptotic cells or necrotic cells on fragments thereof. |
Superior molecular vaccine based on self-replicating rna, suicidal dna or naked dna vector, that links antigen with polypeptide that promotes antigen presentation |
Improved molecular vaccines comprise nucleic acid vectors that encode a fusion polypeptide that includes polypeptide or peptide physically linked to an antigen. The linked polypeptide is one that (a) promotes processing of the expressed fusion polypeptide via the MHC class I pathway and/or (b) promotes development or activity of antigen presenting cells, primarily dendritic cells. These vaccines employ one of several types of nucleic acid vectors, each with its own relative advantages: naked DNA plasmids, self-replicating RNA replicons and suicidal DNA-based on viral RNA replicons. Administration of such a vaccine results in enhance immune responses, primarily those mediated by CD8+ cytotoxic T lymphocytes, directed against the immunizing antigen part of the fusion polypeptide. Such vaccines are useful against tumor antigens, viral antigens and antigens of other pathogenic microorganisms and can be used in the prevention or treatment of diseases that include cancer and infections. |
1. A nucleic acid molecule encoding a fusion polypeptide useful as an immunogenic composition, which molecule comprises: (a) a first nucleic acid sequence encoding a first polypeptide or peptide that (i) promotes antigen processing via the MHC class I pathway (MHC-I-PP) and/or (ii) promotes development or activity of an antigen presenting cell (APC); (b) optionally, fused in frame with the first nucleic acid sequence, a linker nucleic acid sequence encoding a linker peptide; and (c) a second nucleic acid sequence that is linked in frame to said first nucleic acid sequence or to said linker nucleic acid sequence and that encodes an antigenic polypeptide or peptide. 2. The nucleic acid molecule of claim 4, wherein the antigenic peptide comprises an epitope that binds to a MHC class I protein. 3. The nucleic acid molecule of claim 2, wherein said epitope is between about 8 amino acid residues and about 11 amino acid residues in length. 4. The nucleic acid molecule of claim 1 wherein the first polypeptide or peptide is Hsp70, an active C-terminal domain thereof, a functional derivative of Hsp70 or of said C-terminal domain, Flt3 ligand (FL), the extracellular domain of FL, or a functional derivative of FL or of said extracellular domain. 5. (canceled) 6. The nucleic acid molecule of claim 4, wherein the first polypeptide is SEQ ID NO:4 or the full length sequence of Hsp70 a set forth in GENBANK Z95324 AL123456 and encoded by nucleotides 10633-12510 of the Mycobacterium tuberculosis genome. 7. The nucleic acid molecule of claim 4 wherein the first polypeptide consists essentially of said HSP-70 C-terminal domain having a sequence from about residue 517 to about the C-terminal amino acid residue of: (i) SEQ ID NO:4, or (ii) the full length native sequence of Hsp70 a set forth in GENBANK Z95324 AL123456 and encoded by nucleotides 10633-12510 of the Mycobacterium tuberculosis genome. 8. The nucleic acid molecule of claim 4, wherein the first polypeptide is FL. 9. The nucleic acid molecule of claim 4, wherein the first polypeptide consists essentially of the sequence SEQ ID NO:10. 10. The nucleic acid molecule of claim 4 wherein the antigen is one which is present on, or/cross-reactive with an epitope of, a pathogenic organism, cell, or virus. 11. The nucleic acid molecule of claim 10, wherein the virus is a human papilloma virus. 12. The nucleic acid molecule of claim 11, wherein the antigen is the E7 or the E6 polypeptide of HPV-16 or an antigenic fragment thereof. 13. (canceled) 14. (canceled) 15. The nucleic acid molecule of claim 10, wherein the pathogenic cell is a tumor cell, and wherein the antigen is a tumor-specific or tumor-associated antigen, or any antigenic epitope thereof. 16. (canceled) 17. The nucleic acid molecule of claim 15, wherein the antigen comprises the HER-2/neu protein or a peptide thereof, mutant p53 or a melanoma-associated antigens selected from the group consisting of MAGE-1, MAGE-3, MART-1/Melan-A, tyrosinase, gp75, gp100, BAGE, GAGE-1, GAGE-2, GnT-V, and p15. 18. The nucleic acid molecule of claim 4 operatively linked to a promoter. 19. The nucleic acid molecule of claim 18, wherein the promoter is one which is expressed in an APC. 20. The nucleic acid molecule of claim 19, wherein the APC is a dendritic cell. 21. An isolated nucleic acid molecule that, under stringent hybridization conditions, hybridizes with a nucleic acid of claim 4, said hybridization occurring simultaneously with: (i) at least part of said first nucleic acid sequence and at least part of said second nucleic acid sequence, (ii) at least part of said first nucleic acid sequence and part of said linker nucleic acid sequence, (iii) at least part of said second nucleic acid sequence and part of said linker nucleic acid sequence, or (iv) at least part of said first nucleic acid sequence, at least part of said first nucleic acid sequence and said linker nucleic acid sequence. 22. An expression vector comprising the nucleic acid molecule of claim 4 operatively linked to (a) a promoter; and (b) optionally, additional regulatory sequences that regulate expression of said nucleic acid in a eukaryotic cell. 23. The expression vector of claim 22 which is a viral vector or a plasmid. 24. The expression vector of claim 22 wherein said plasmid is pcDNA3 which is derived from the sequence of SEQ ID NO:15, into which is inserted the nucleic acid sequences encoding said first or said second polypeptides. 25. The expression vector of claim 22 which is a self-replicating RNA replicon. 26. The expression vector of claim 25, wherein the self-replicating RNA replicon is a Sindbis virus self-replicating RNA replicon. 27. The expression vector of claim 26, wherein the replicon is SINrep5 which is derived from the sequence of SEQ ID NO:16 into which is inserted, the nucleic acid sequences encoding said first or said second polypeptides. 28. The expression vector of claim 22 which is a suicidal DNA vector. 29. The expression vector of claim 28 wherein said suicidal DNA vector is an alphavirus DNA vector. 30. The expression vector of claim 29 wherein said alphavirus is Semliki Forest virus (SFV). 31. The expression vector of claim 30 wherein said SFV vector is pSCA1. 32. The expression vector of claim 28 wherein the suicidal DNA is derived from the sequence of SEQ ID NO:17 into which is inserted the nucleic acid sequences encoding said first or said second polypeptides. 33. The expression vector of claim 22 wherein the first encoded polypeptide or peptide is Hsp70, an active C-terminal domain thereof, a functional derivative of Hsp70 or of said C-terminal domain, is FL, the extracellular domain of FL, or a functional derivative of FL or of said extracellular domain. 34. The expression vector of claim 33 wherein the first encoded polypeptide consists essentially of the sequence SEQ ID NO:4 or the full length sequence of Hsp70 as set forth in GENBANK Z95324 AL123456 and encoded by nucleotides 10633-12510 of the Mycobacterium tuberculosis genome. 35. The expression vector of claim 33 wherein the first polypeptide consists essentially of said C-terminal domain having a sequence from about residue 517 to about the C-terminal amino acid residue of: (i) SEQ ID NO:4, or (ii) the full length native sequence of Hsp70 a set forth in GENBANK Z95324 AL123456 and encoded by nucleotides 10633-12510 of the Mycobacterium tuberculosis genome. 36. The expression vector of an of claims 33, that is a self-replicating RNA replicon that also encodes HPV protein E7, and has the nucleotide sequence SEQ ID NO:19. 37. The expression vector of claim 33, that is a suicidal DNA vector which also encodes HPV protein E7, and has the nucleotide sequence of SEQ ID NO:20. 38. (canceled) 39. The expression vector of claim 33 wherein the first encoded polypeptide consists essentially of the extracellular domain of FL having a sequence SEQ ID NO:10. 40. The expression vector of claim 39 that comprises a naked DNA plasmid pcDNA3 that includes the coding sequence for HPV protein E7 and the FL extracellular domain, and has the nucleotide sequence SEQ ID NO:21. 41. A cell which has been modified to comprise and express the expression vector of claim 22. 42. (canceled) 43. The cell of claim 41 which is an APC. 44. The cell of claim 43, wherein the APC is a dendritic cell, a keratinocyte, a macrophage, a monocyte, a B lymphocyte, a microglial cell, an astrocyte, or an activated endothelial cell. 45. A particle comprising the expression vector of claim 22. 46. The particle of claim 45 which comprises a material that is suitable for introduction into a cell or an animal by particle bombardment. 47. The particle of claim 46, wherein the material is gold. 48. A fusion or chimeric polypeptide comprising (a) a first polypeptide or peptide that (i) promotes processing via the MHC class I pathway and/or (ii) promotes development or activity of an APC; and (b) a second polypeptide comprising an antigenic peptide or polypeptide. 49. The fusion or chimeric polypeptide of claim 53, wherein the antigenic peptide or polypeptide comprises an epitope that binds to, and is presented on the cell surface by, MHC class I proteins. 50. The fusion or chimeric polypeptide of claim 53 wherein the first polypeptide and the antigenic polypeptide or peptide are linked by a chemical linker. 51. The fusion polypeptide of claim 53 wherein the first polypeptide is N-terminal to the second polypeptide. 52. The fusion polypeptide of claim 53 wherein the second polypeptide is N-terminal to the first polypeptide. 53. The fusion or chimeric polypeptide of claim 48 wherein the first polypeptide or peptide is Hsp70, an active C-terminal domain thereof, a functional derivative of Hsp70 or of said C-terminal domain, FL, the extracellular domain of FL, or a functional derivative of FL or of said extracellular domain. 54. (canceled) 55. The fusion polypeptide of claim 53, wherein the first polypeptide is SEQ ID NO:4 or the full length sequence of Hsp70 a set forth in GENBANK Z95324 AL123456 and encoded by nucleotides 10633-12510 of the Mycobacterium tuberculosis genome. 56. The fusion polypeptide of claim 53, wherein the first polypeptide consists essentially of the sequence SEQ ID NO:10. 57. The fusion polypeptide encoded by the nucleic acid molecule of claim 4. 58. The fusion polypeptide encoded by the expression vector of claim 22. 59. A pharmaceutical composition capable of inducing or enhancing an antigen-specific immune response, comprising: (a) pharmaceutically and immunologically acceptable excipient in combination with; the nucleic acid molecule claim 4; 60. A method of inducing or enhancing an antigen specific immune response in a subject comprising administering to the subject an effective amount of the pharmaceutical composition of claims 78, thereby inducing or enhancing said response. 61-62. (canceled) 63. The method of claim 60 wherein said subject is a human. 64. A method of inducing or enhancing an antigen specific immune response in cells or in a subject comprising contacting said cells with, or administering to said subject, an effective amount of the pharmaceutical composition of claim 78, thereby inducing or enhancing said response. 65. The method of claim 64, comprising contacting said cells ex vivo with said composition. 66. The method of claim 65 wherein said cells comprise APCs. 67. The method of claim 66, wherein said APCs are dendritic cells. 68. (canceled) 68. (canceled) 69. The method of claim 65, further comprising a step of administering said cells, which were contacted with the composition ex vivo, to (i) a histocompatible subject or (ii) the subject from which said cells were obtained. 70-73. (canceled) 74. A method of increasing the numbers or lytic activity of CD8+ CTLs specific for a selected antigen in a subject, comprising administering to said subject an effective amount of the pharmaceutical composition of claim 78, wherein (i) said expression vector encodes said selected antigen, and (ii) said selected antigen comprises an epitope that binds to, and is presented on the cell surface by, MHC class I proteins, thereby increasing the numbers or activity of said CTLs. 75. A method of inhibiting growth or preventing re-growth of a tumor in a subject, comprising administering to said subject an effective amount of a pharmaceutical composition of claim 78, wherein said expression vector encodes one or more tumor-associated or tumor-specific epitopes present on said tumor in said subject, thereby inhibiting said growth or preventing said re-growth. 76-77. (canceled) 78. A pharmaceutical composition capable of inducing or enhancing an antigen-specific immune response, comprising: (a) pharmaceutically and immunologically acceptable excipient in combination with; (b) the expression vector of claim 22. 79. A pharmaceutical composition capable of inducing or enhancing an antigen-specific immune response, comprising: (a) pharmaceutically and immunologically acceptable excipient in combination with; (b) the cell of claim 41. 80. A pharmaceutical composition capable of inducing or enhancing an antigen-specific immune response, comprising: (a) pharmaceutically and immunologically acceptable excipient in combination with; (b) the particle of claim 45. 81. A pharmaceutical composition capable of inducing or enhancing an antigen-specific immune response, comprising: (a) pharmaceutically and immunologically acceptable excipient in combination with; (b) the fusion or chimeric polypeptide of claim 48. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention The present invention in the fields of molecular biology, immunology and medicine relates to chimeric nucleic acids encoding fusion proteins and their use as vaccines to enhance immune responses, primarily cytotoxic T lymphocyte (CTL) responses to specific antigens such as tumor antigens. The fusion proteins comprise an antigenic polypeptide fused to a protein that promotes processing via the MHC class I pathway and/or promotes development or activity of antigen presenting cells (APCs), primarily dendritic cells (DCs). Preparation of the foregoing nucleic acid constructs as naked DNA plasmids, self-replicating RNA replicons and suicidal DNA-based viral RNA replicons confer various advantages on these molecular vaccines. 2. Description of the Background Art Antigen-specific cancer immunotherapy has emerged as a promising approach because it is capable of engendering specific immunity against neoplastic cells while sparing normal cells. Increasing evidence suggests that professional antigen-presenting cells (APCs), particularly dendritic cells (DCs), are central players in this process. An effective vaccine strategy includes targeting the tumor antigen to professional APCs that in turn activate antigen-specific T cells (for review, see (Chen, CH et al., J Biomed Sci. 5:231-52, 1998.). Recently, DNA vaccines have become attractive as an approach for generating antigen-specific immunotherapy (for review, see (Robinson, H L Vaccine 15:785-778, 1997; Robinson, H L et al., Semin Immunol. 9:271-83, 1997; Pardoll, D M et al., Immunity. 3:165-9, 1995; Donnelly, J J et al., Annu Rev Immunol. 15: 617-48, 1997). The advantages of naked DNA include purity, ease of preparation and stability. In addition, DNA-based vaccines can be prepared inexpensively and rapidly in large-scale. Furthermore, multiple DNA vaccines can be administered simultaneously. However, naked DNA vaccines raise concerns such as potential integration into the host genome and cell transformation. Because they do not have the intrinsic ability to amplify in vivo as do viral vaccines, DNA vaccines may be more limited in their potency. The present inventors conceived that a directing a DNA vaccine encoding an antigen (in the form of a fusion protein) to cells which activate immune responses, such as DCs, would enhance the vaccine's potency. Others demonstrated that linking DNA encoding the cytokine GM-CSF gene to DNA encoding an HIV or hepatitis C antigen enhanced the potency of DNA vaccines (Lee, A H et al., Vaccine 17: 473-9, 1999; Lee, S W et al., J Virol. 72: 8430- 6, 1998). The chigmeric GM-CSF/antigen is believed to act as an immunostimulatory signal to DCs, inducing their differentiation from an immature form (Banchereau, J et al., Nature 392: 245-52, 1998). Since DCs and their precursors express high levels of GM-CSF receptors, the chimeric GM-CSF/antigen should target and concentrate the linked antigen to the DCs and further improve the vaccine's potency. Use of self-replicating RNA vaccines (RNA replicons) has also been identified as an important strategy in nucleic acid vaccine development. RNA replicon vaccines may be derived from alphavirus vectors, such as Sindbis virus (Hariharan, M J et al., 1998 . J Virol 72:950-8.), Semliki Forest virus (Berglund, P M et al., 1997 . AIDS Res Hum Retroviruses 13:1487-95; Ying, H T et al., 1999 . Nat Med 5:823-7) or Venezuelan equine encephalitis virus (Pushko, P M et al., 1997 . Virology 239:389-401). These self-replicating and self-limiting vaccines may be administered as either (1) RNA or (2) DNA which is then transcribed into RNA replicons in cells transfected in vitro or in vivo (Berglund, P C et al., 1998 . Nat Biotechnol 16:562-5; Leitner, W W et al., 2000 . Cancer Res 60:51-5). Self-replicating RNA infects a diverse range of cell types and allows the expression of a linked antigen of interest at high levels (Huang, H V 1996 . Curr Opin Biotechnol 7:531-5) Because viral replication is toxic to infected host cells, such self-replicating RNA preparations eventually causes lysis of the transfected cells (Frolov, I et al., 1996 . J Virol 70:1182-90). These vectors cannot integrate into the host genome, and therefore do not raise concerns of associated with naked DNA vaccines. This is particularly important for vaccine development where target proteins are potentially oncogenic, such as human papillomavirus (HPV) E6 and E7 proteins. The present inventors and their colleagues recently demonstrated that linkage of HPV-16 E7 antigen to Mtb heat shock protein 70 (Hsp70) leads to the enhancement of DNA vaccine potency (Chen, CH et al., 2000 . Cancer Research 60:1035-1042). (See also co-pending patent applications U.S. Ser. No. 09/501,097, filed 9 Feb. 2000; and U.S. Ser. No. 099/421,608, filed 20 Oct. 1999, all of which are incorporated by reference in their entirety.) Immunization with HSP complexes isolated from tumor or virus-infected cells induced potent anti-tumor immunity (Janetzki, S et al., 1998 . J Immunother 21:269-76) or antiviral immunity (Heikema, A E et al, Immunol Lett 57:69-74). In addition, immunogenic HSP-peptide complexes dould be reconstituted in vitro by mixing the peptides with HSPs (Ciupitu, AM et al., 1998 . J Exp Med 187:685-91). Furthermore, HSP-based protein vaccines have been created by fusing antigens to HSPs (Suzue, K et al., 1996 . J Immunol 156:873-9). The results of these investigations point to HSPs a attractive candidates for use in immunotherapy. However, prior to the present inventors' work, HSP vaccines were all peptide/protein-based vaccines or, in more recent cases, were in the form of naked DNA. To date, there have been no reports of HSPs incorporated into self-replicating RNA vaccines. Another molecule that stimulates growth of DC precursors and can help in generating large numbers of DCs in vivo is Flt3-ligand (“FL”) (Maraskovsky, E et al., J Exp Med 184: 1953-62, 1996, Shurin, M R et al., Cell Lmmunol. 179: 174-84, 1997). FL has emerged as an important molecule in the development of tumor vaccines that augment numbers and action of DCs in vivo. Flt3, a murine tyrosine kinase receptor, first described in 1991 (Rosnet, O et al., Oncogene. 6: 1641-50, 1991), was found to be a member of the type III receptor kinase family which includes -kit and c-fms (for review, see (Lyman, S D Curr Opin Hematol. 5:192-6, 1998). In hematopoietic tissues, the Flt3expression is restricted to the CD34+ progenitor population. Flt3 has been used to identify and subsequently clone the corresponding ligand, Flt3-ligand or “FL” (Lyman, S D et al., Cell. 75: 1157-67, 1993; Hannum, C et al., Nature. 368: 643-8, 1994). The predominant form of FL is synthesized as a transmembrane protein from which the soluble form is believed to be generated by proteolytic cleavage. The soluble form of FL (the extracellular domain or “ECD”) is functionally similar to intact FL (Lyman, S D et al., Cell. 75: 1157-67, 1993). These proteins function by binding to and activating unique tyrosine kinase receptors. Expression of the Flt3 receptor is primarily restricted, among hematopoietic cells, to the most primitive progenitor cells, including DC precursors. The soluble ECD of FL induced strong anti-tumor effects against several murine model tumors including fibrosarcoma (Lynch, D H et al., Nat Med. 3: 625-31, 1997), breast cancer (Chen, K et al Cancer Res. 57: 3511-6, 1997; Braun, S E et al., Hum Gene Ther. 10: 2141-51, 1999), liver cancer (Peron, J M et al., J Immunol. 161: 6164-70, 1998), lung cancer (Chakravarty, P K et al., Cancer Res. 59: 6028-32, 1999), melanoma and lymphoma (Esche, C et al., Cancer Res. 58: 380-3, 1998). There is a need in the art for improved molecular vaccines, such as nucleic acid vaccines, that combine potency and safety. The present invention helps meet this need by its design of novel fusion or chimeric polypeptides and nucleic acids coding therefor, that link the antigen with specialized polypeptides that promote antigen presentation by various mechanisms and that exploit delivery of these constructs by various nucleic acid vectors. |
<SOH> SUMMARY OF THE INVENTION <EOH>Self-replicating RNA vaccines (RNA replicons) have emerged as an important strategy for nucleic acid vaccine development. The present inventors evaluated the effect of linking HPV type 16 (HPV-16) E7 as a model antigen to Mycobacterium tuberculosis (Mtb) heat shock protein 70 (Hsp70) on the potency of antigen-specific immunity generated by a Sindbis virus self-replicating RNA vector, SINrep5. The results indicated that this RNA replicon vaccine containing E7/Hsp70 fusion genes generated significantly greater E7-specific T cell-mediated immunity than vaccines comprising wild type E7 DNA. HPV-16 E7 was selected as a model antigen for vaccine development because .HPVs, particularly HPV-16, are associated with most cervical cancers. HPV oncogenic proteins, E6 and E7, are co-expressed in most HPV-containing cervical cancers and are important in the induction and maintenance of cell transformation. Therefore, vaccines targeting E6 or E7 provide an opportunity to prevent and treat HPV-associated cervical malignancies. HPV-16 E7 is a well-characterized cytoplasmic/nuclear protein that is more conserved than E6 in HPV-associated cancer cells; E7 has been tested in a variety of HPV vaccines. Furthermore, in vitro studies demonstrated that E7 antigen from apoptotic cells that have been transfected with E7/Hsp70 RNA replicons is taken up by bone marrow-derived dendritic cells (DC's) and presented more efficiently through the MHC class I pathway compared to antigen from than apoptotic cells transfected by wild-type E7 RNA replicons. Importantly, the fusion of Hsp70 to E7 converted a less effective vaccine into one with significant potency against E7-expressing tumors. This antitumor effect involved NK cells and CD8 + T cells. Thus, fusion of a nucleic acid sequence encoding Hsp70 to nucleic acid encoding an antigen of interest in the form of a self-replicating RNA vaccine greatly enhances the potency of this vaccine. Naked DNA vaccines represent an attractive approach for generating antigen-specific immunity because of their stability and simplicity of delivery. Concerns with DNA vaccines include potential integration into the host genome, cell transformation, and limited potency. The use of DNA-based alphaviral RNA replicons (“suicidal DNA vectors”), as disclosed herein, may alleviate concerns surrounding DNA integration or cell transformation since suicidal DNA vectors eventually cause lysis of the cells they transfect. To further improve the potency of suicidal DNA vaccines, the present inventors linked Hsp70 to E7 (as a model antigen) using DNA-based Semliki Forest virus (SFV) RNA vector, pSCA1. This suicidal DNA vaccine containing E7/Hsp70 fusion DNA produced a significantly greater E7-specific T cell-mediated immune response in mice than did vaccines containing the wild type E7 DNA alone. Importantly, this fusion converted a less effective vaccine into one with significant therapeutic potency against established E7-expressing metastatic tumors. The antitumor effect was dependent upon CD8+ T cells. Thus, linkage of Hsp70 to an antigen enhances the potency of a suicidal DNA vaccine. Flt3 (fms-like tyrosine kinase 3)-ligand is an important cytokine in the development and differentiation of professional APCs, particularly DCs. A recombinant chimeric or fusion polypeptide molecule comprising the extracellular domain (ECD) of Flt3-ligand (FL) linked to an antigen targets the antigen to DCs and their precursors. Using HPV-16 E7 as a model antigen, the present inventors linked FL to E7 and caused stimulation of an antigen-specific immune response by a naked DNA vaccine administered intradermally via gene gun. Vaccines that included DNA encoding a chimeric FL-E7 fusion polypeptide dramatically increased the frequency of E7-specific CD8 + T cells when compared to vaccines of only E7 DNA. Cells transfected in vitro with FL-E7 DNA presented E7 via the MHC class I pathway more efficiently than did cells transfected with wild-type E7 DNA. Furthermore, bone marrow-derived DCs pulsed with lysates of cells that had been transfected to express an FL-E7 fusion protein presented E7 (via the MHC class I pathway) more efficiently than did DCs pulsed with lysates of cells expresssing (after transfection) E7 protein alone. More importantly, this fusion construct rendered a less effective vaccine highly potent in inducing a therapeutic response against established E7-expressing metastatic tumors. The FL-E7 fusion vaccines mainly targeted CD8 + T cells as anti-tumor effects were completely independent of CD4+ T cells. Thus, fusion of DNA encoding the ECD of FL to DNA encoding an antigen markedly enhances the potency of a DNA vaccine acting via CD8-dependent pathways. In one embodiment, the antigen (e.g., the MHC class I-binding peptide epitope) is derived from a pathogen, e.g., it comprises a peptide expressed by a pathogen. The pathogen can be a virus, such as, e.g., a papilloma virus, a herpesvirus, a retrovirus (e.g., an immunodeficiency virus, such as HIV-1), an adenovirus, and the like. The papilloma virus can be a human papilloma virus; for example, the antigen (e.g., the Class I-binding peptide) can be derived from an HPV-16 E7 polypeptide. In one embodiment, the HPV-16 E7 polypeptide is substantially non-oncogenic, i.e., it does not bind retinoblastoma polypeptide (pRB) or binds pRB with such low affinity that the HPV-16 E7 polypeptide is effectively non-oncogenic when expressed or delivered in vivo. In alternative embodiments, the pathogen is a bacteria, such as Bordetella pertussis; Ehrlichia chaffeensis; Staphylococcus aureus; Toxoplasma gondii; Legionella pneumophila; Brucella suis; Salmonella enterica; Mycobacterium avium; Mycobacterium tuberculosis; Listeria monocytogenes; Chlamydia trachomatis; Chlamydia pneumoniae; Rickettsia rickettsii ; or, a fungi, such as, e.g., Paracoccidioides brasiliensis ; or other pathogen, e.g., Plasmodium falciparum. In another embodiment, the MHC class I-binding peptide epitope is derived from a tumor cell. The tumor cell-derived peptide epitope can comprise a tumor associated antigen, e.g., a tumor specific antigen, such as, e.g., a HER-2/neu antigen. In one embodiment, the isolated or recombinant nucleic acid molecule is operatively linked to a promoter, such as, e.g., a constitutive, an inducible or a tissue-specific promoter. The promoter can be expressed in any cell, including cells of the immune system, including, e.g., antigen presenting cells (APCs), e.g., in a constitutive, an inducible or a tissue-specific manner. In alternative embodiments, the APCs are dendritic cells, keratinocytes, astrocytes, monocytes, macrophages, B lymphocytes, a microglial cell, or activated endothelial cells, and the like. The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims. All publications, patents, patent applications, GenBank sequences and ATCC deposits, cited herein are hereby expressly incorporated by reference for all purposes. |
Burner unit and method for operation thereof |
Arranged upstream of each swirl-stabilized premix burner (1) of a burner system there is respectively an adjustable distributing device (5) with control valves (V1, . . . ,V8) and/or on/off valves (V″1, . . . , V″16), by means of which various axial mass flow distributions of the fuel introduced can be set. Preferably, those which have particularly favorable values with respect to characteristics such as NOx emission and maximum amplitudes of pulsations occurring are chosen. For this purpose, Pareto solutions are determined with respect to the said characteristics, in that a distributing device (5) is represented by a tree structure with distributing parameters, and values for the distributing parameters on the basis of which the distributing device (5) is set by means of a control unit are iteratively generated in a data-processing system by an evolutionary algorithm. On the basis of the values determined by a measuring unit, solutions which are especially favorable with respect to the characteristics mentioned, especially Pareto-optimal, are selected. The distributing devices are then set during operation on the basis of the solutions of this type corresponding to respective requirements. |
1. A burner system for use with a fuel source and a fuel-source feed line connected to the fuel source, the system comprising: at least one swirl-stabilized premix burner having a plurality of inlet openings; and a distributing device by which the inlet openings in the premix burner can be connected to the fuel source, the distributing device comprising a system of valves or restrictors for being positioned between the fuel-source feed line and the inlet openings of the premix burner, for setting the mass flow distribution between the inlet openings. 2. The burner system as claimed in claim 1, wherein the distributing device includes a fuel input, wherein the system of valves or restrictors comprises at least one control valve, and wherein at least one of said plurality of inlet openings is fluidly preceded by said at least one control valve for establishing the mass flow, the at least one control valve being connected directly to the fuel input of the distributing device. 3. The burner system as claimed in claim 1, wherein the distributing device further comprises: a tree structure for branching from the feed line; and multiway valves connected to the tree structure. 4. The burner system as claimed in claim 3, wherein the multiway valves each comprises a three-way valve. 5. The burner system as claimed in claim 1, wherein the distributing device comprises on/off valves. 6. The burner system as claimed in claim 5, wherein the on/off valves are each arranged directly upstream of at least one burner inlet opening. 7. A method for operating a burner system as claimed in claim 1, the method comprising: determining determination variable comprising a vector from a determination set which is a subset of an n-dimensional domain; setting the mass flow distribution with the valves or restrictors so that a target variable comprising a vector from a target set which is a subset of an m-dimensional domain, assumes a specific value. 8. The method as claimed in claim 7, further comprising: selecting a determination variable depending on the mode of operation, a requirement for the target variable, or both, the determination variable selected from a plurality of determination variables, the determination variable being a basis for setting the mass flow distribution. 9. The method as claimed in claim 7, wherein the determination variable corresponds at least approximately to a Pareto solution, wherein for every solution in which one component of the target variable has a more favorable value, at least one other component has a less favorable value. 10. The method as claimed in claim 7, wherein the components of the determination variable correspond to the mass flows to inlet openings or groups of inlet openings of the at least one premix burner, or are variables from which the mass flows can be derived. 11. The method as claimed in claim 7, wherein the concentration of at least one pollutant forms a component of the target variable. 12. The method as claimed in claim 7, wherein a measure of the pulsations occurring in the burner system forms a component of the target variable. 13. The burner system as claimed in claim 1, further comprising: a fuel source; and a fuel-source feed line fluidly connected between the fuel source and the distributing device. 14. The method as claimed in claim 11, wherein the at least one pollutant comprises NOx concentration in exhaust gas. 15. The method as claimed in claim 12, wherein the measure comprises a maximum amplitude of the pulsations. |
<SOH> TECHNICAL FIELD <EOH>The invention relates to a burner system according to the precharacterizing clause of claim 1 . Burner systems of this type are used in particular in gas turbines. It also relates to a method for operating a system of this type. |
<SOH> SUMMARY OF THE INVENTION <EOH>The invention is based on the object of stabilizing the combustion by the simplest possible means and to achieving the most favorable possible results with regard to specific characteristics of the combustion, especially reduction of the pulsations and pollutant emission, in particular the NO x emission. It was found that the way in which the combustion proceeds is influenced strongly by the mass flow distribution of the fuel introduced into the premix burners. In the case of the burner system according to the invention, the distribution of the mass flow can be set. Therefore, especially on the basis of the operating method according to the invention, it can be set such that the way in which the combustion proceeds is in each case optimized to a great extent with respect to specific requirements such as low pollutant emission and low pulsations, or changing weightings of a number of such requirements. |
Method for the production of a burner unit |
In the case of swirl-stabilized premix burners (1), an axial mass flow distribution of the fuel introduced which has especially favorable values with respect to characteristics such as NOx emission and maximum amplitudes of pulsations occurring is used. For this purpose, Pareto solutions are determined with respect to the said characteristics, in that a distributing device (5) with control valves is represented by a tree structure with distributing parameters, and values for the distributing parameters on the basis of which the distributing device (5) is set by means of a control unit (10) are iteratively generated in a data-processing system (9) by an evolutionary algorithm. On the basis of the values determined by a measuring unit (11), solutions which are especially favorable with respect to the characteristics mentioned, espectially Pareto-optimal, are selected. The distributing devices or the premix burners of the burner system are then formed in a way corresponding to such a solution. |
1. A method for producing a burner system having a fuel source, at least one swirl-stabilized premix burner having a plurality of inlet openings, and a distributing device, by which the plurality of inlet openings in the premix burner are connected to the fuel source, the method comprising: determining a desired mass flow distribution into the at least one premix burner; generating determination variables fixing the mass flow distribution, comprising vectors from a determination set which is a subset of an n-dimensional domain, one after the other with a data-processing system; setting a mass flow distribution in a test setup with at least one premix burner and at least one adjustable distributing device, the mass flow distribution being set on the basis of the determination variable, and measuring a target variable, comprising a vector from a target set which is a subset of an m-dimensional domain, on the test setup; and selecting a determination variable is selected on the basis of the target variables, wherein the at least one premix burner or the at least one distributing device of the burner system is configured such that the mass flow distribution corresponds to that which is fixed by the selected determination variable. 2. The method as claimed in claim 1, further comprising: forming the components of the determination variables at least partly by the distributing parameters of the branching points of a tree structure, by which tree structure the distribution of the mass flow between inlet openings or groups of inlet openings of the at least one premix burner is determined. 3. The method as claimed in claim 1, further comprising: determining Pareto solutions, wherein for every solution in which one component of the target variable has a more favorable value, at least one other component has a less favorable value, at least approximately with the data-processing system; and selecting a determination variable from among the Pareto solutions. 4. The method as claimed in claim 3, wherein determining Pareto solutions comprises determining starting variables serving as a set of determination variables are determined and, further comprising: carrying out iteration steps with the data processing system until a terminating criterion is satisfied including determining a new set of determination variables from a set of determination variables by generating from the set of determination variables a set of test variables respectively lying in the determination set, from which set of test variables the new set of determination variables is selected in each case on the basis of the target variables which were measured for the mass flow distribution fixed by the determination variables. 5. The method as claimed in claim 4, wherein generation of the test variables from the set of determination variables comprises random mutation or recombination of the determination variables using the data-processing system. 6. The method as claimed in claim 1, wherein the concentration of at least one pollutant, forms a component of the target variable. 7. The method as claimed in claim 1, wherein a measure of the pulsations occurring in the burner system forms a component of the target variable. 8. The method as claimed in claim 1, wherein the inlet openings are provided at least partly axially in succession. 9. The method as claimed in claim 1, further comprising: dimensioning the inlet openings at least partially to achieve the desired mass flow distribution. 10. The method as claimed in claim 1, wherein the distributing device comprises restrictors, diverters, or both, to achieve the desired mass flow distribution. 11. The method as claimed in claim 6, wherein the at least one pollutant comprises NOx concentration in an exhaust gas. 12. The method as claimed in claim 7, wherein the measure of the pulsations comprises pulsation maximum amplitude. |
<SOH> TECHNICAL FIELD <EOH>The invention relates to a method for producing a burner system according to the precharacterizing clause of claim 1 . Burner systems of this type are used in particular in gas turbines. |
<SOH> SUMMARY OF THE INVENTION <EOH>The invention is based on the object of providing a method for producing burner systems of the generic type which are of a simple construction and in which the combustion proceeds favorably, in particular with regard to the reduction of pulsations and low emission of pollutants, especially NO x . It was found that the way in which the combustion proceeds is influenced strongly by the mass flow distribution of the fuel introduced into the premix burners. According to the invention, the burner systems are formed in such a way that the fuel is introduced into the premix burners with a specific mass flow distribution, which ensures favorable characteristics of the combustion, especially with regard to pulsations and pollutant emission. |
Fabricating a semiconductor device using fully cured bisbenzocyclobutene |
The present invention is a method of fabricating a semiconductor device using a fully cured BCB layer and removing the same using wet etching. The first step is selecting a substrate. The second step of the method is producing an oxide layer or other coating on the substrate. The third step is applying a BCB layer on the oxide layer. The fourth step is fully curing the BCB layer. The fifth step is processing the device. The sixth step is stressing the substrate, preferably causing the substrate to warp. The seventh step is wet etching the BCB layer. The eighth step is removing the BCB layer. The ninth step is removing the oxide layer. |
1. A method of fabricating semiconductor device comprising the steps of: a) selecting a substrate having a front side and a back side; b) applying a BCB layer on the front side of the substrate; c) fully curing the BCB layer; d) stressing the substrate; e) wet etching the BCB layer; and f) removing the BCB layer from the substrate. 2. The method of claim 1, further including the step of producing an oxide layer or any other suitable layer on the front of the substrate between the BCB layer and the substrate. 3. The method of claim 2, further including the step of removing the oxide layer from the substrate. 4. The method of claim 3, further including the step of processing the semiconductor using a process selected from the group of processes composed of grinding, vapor deposition, defining patterns in the substrate, any other suitable semiconductor fabrication technique, and any combination thereof. 5. The method of claim 4, wherein the step of fully curing the BCB layer further includes the steps of: a) baking the BCB layer until the BCB layer is partially cured; and b) baking the BCB layer until the BCB layer is fully cured. 6. The method of claim 5, wherein the step of baking the BCB layer until partially cured is performed at approximately 80 degrees Celsius and the step of baking the BCB layer until fully cured is performed at approximately 250 degrees Celsius. 7. The method of claim 6, further including the step of bonding said semiconductor device to a second substrate when said BCB layer is in a partially cured state. 8. The method of claim 5, wherein the step of wet etching the BCB layer is performed using a chemical etch selected from the group of chemical etches composed of hydrogen fluoride, nitric acid, sulfuric acid and phosphoric acid or any combination thereof. 9. The method of claim 8, wherein the step of stressing the substrate is performed by a process selected from the group of processes composed of mechanically grinding the substrate from the back side, etching the back side of the substrate, polishing the back side of the substrate, depositing a material film on the front or back side of the substrate, depositing an oxide layer on the front or back side of the substrate, patterning material layers on the front or back side of the substrate, depositing multiple BCB layers on the front or back side of the substrate, depositing multiple oxide layers on the front or back side of the substrate, any other suitable process or any combination thereof. 10. The method of claim 9, wherein the step of applying a BCB layer is performed by spin coating. 11. The method of claim 10, wherein the step of removing the BCB layer is performed by a dump rinse tank. 12. The method of claim 11, wherein the step of removing the oxide layer is performed by an acid etch. 13. The method of claim 12, wherein the step of producing an oxide layer is composed of producing a silicon oxide layer. 14. The method of claim 13, wherein the step of selecting a substrate is composed of selecting a silicon substrate. 15. The method of claim 1, further including the step of processing the semiconductor using a process selected from the group of processes composed of grinding, vapor deposition, defining patterns in the substrate, any other suitable semiconductor fabrication technique, and any combination thereof. 16. The method of claim 1, wherein the step of curing the BCB layer further includes the steps of: a) baking the BCB layer until the BCB layer is partially cured; and b) baking the BCB layer until the BCB layer is fully cured. 17. The method of claim 16, wherein the step of baking the BCB layer until partially cured is performed at approximately 80 degrees Celsius and the step of baking the BCB layer until fully cured is performed at approximately 250 degrees Celsius. 18. The method of claim 17, further including the step of bonding said semiconductor device to a second substrate when said BCB layer is in a partially cured state. 19. The method of claim 1, wherein the step of wet etching the BCB layer is performed using a chemical etch selected from the group of chemical etches composed of hydrogen fluoride, nitric acid, sulfuric acid and phosphoric acid or any combination thereof. 20. The method of claim 1, wherein the step of stressing the substrate is performed by a process selected from the group of processes composed of mechanically grinding the substrate from the back side, etching the back side of the substrate, polishing the back side of the substrate, depositing a material film on the front or back side of the substrate, depositing an oxide layer on the front or back side of the substrate, patterning material layers on the front or back side of the substrate, depositing multiple BCB layers on the front or back side of the substrate, depositing multiple oxide layers on the front or back side of the substrate, any other suitable process or any combination thereof. 21. The method of claim 1, wherein the step of applying a BCB layer is performed by spin coating. 22. The method of claim 1, wherein the step of removing the BCB layer is performed by a dump rinse tank. 23. The method of claim 3, wherein the step of removing the oxide layer is performed by an acid etch. 24. The method of claim 2, wherein the step of producing an oxide layer is composed of producing a silicon oxide layer. 25. The method of claim 1, wherein the step of selecting a substrate is composed of selecting a silicon substrate. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Bisbenzocyclobutene (BCB) was developed for use as a dielectric in semiconductor devices, specifically multi-chip module (MCM) applications. BCB is especially well suited for these applications because it adheres well to semiconductor materials such as silicon, silicon oxide, silicon nitride, aluminum, copper and polyimide. It is also used because it has a service temperature in excess of 300 degrees Celsius. These properties make BCB additionally desirable as a masking material, a protection layer and a wafer bonding material. Despite the advantages of BCB, industry has avoided using BCB in many situations due to a significant practical problem related to BCB processing. The primary problem with the material is that it is resistant to wet chemical etching, particularly when the BCB has been fully cured. Because the BCB resists wet chemical etching, processing of BCB must be done by dry etching or plasma treatments. As is known by those skilled in the art, both of these processes commonly result in significant damage to the device leading to increased waste and excessive cost to the manufacturer. Consequently, many manufacturers have refused to incorporate BCB into semiconductor devices or have increased the cost of devices requiring the use of BCB to account for their own increased costs. U.S. Pat. No. 5,879,572 entitled “METHOD OF PROTECTING SILICON WAFERS DURING WET CHEMICAL ETCHING,” discloses a process for micromachining silicon wafers using BCB as an etch mask. In this process, a film of partially cured BCB is applied over the metallization and circuitry on the wafer. The BCB protects these objects during wet chemical etching of the substrate. After etching of the substrate, the BCB is completely removed by dry etching or, because the BCB is only partially cured, a limited number of wet etch techniques. The present invention does not involve the application of a partially cured BCB protective layer. U.S. Pat. No. 5,879,572 is hereby incorporated by reference into the specification of the present invention. U.S. Pat. No. 6,204,081 entitled “METHOD OF MANUFACTURING A SUBSTRATE OF A LIQUID CRSTAL DISPLAY DEVICE,” discloses a process for etching a substrate using a light developable BCB protection layer. A mask is placed over the BCB layer, exposing only certain portions of the layer to a light source. The light source removes the exposed portions of the light developable BCB layer. The present invention does not use a light source to remove a BCB layer. U.S. Pat. No. 6,204,081 is hereby incorporated by reference into the specification of the present invention. U.S. Pat. No. 6,284,149 entitled “HIGH-DENSITY PLASMA ETCHING OF CARBON-BASED LOW-K MATERIALS IN AN INTEGRATED CIRCUIT,” discloses a plasma etching process for a BCB dielectric layer. A hard mask is placed over an upper BCB layer, the BCB layer being etched by a gas of oxygen, a fluorocarbon and nitrogen. The present invention does not use a gas to etch BCB. U.S. Pat. No. 6,284,149 is hereby incorporated by reference into the specification of the present invention. U.S. Pat. No. 6,514,872 entitled “METHOD OF MANUFACTURING A SEMICONDUCTOR DEVICE,” discloses a semiconductor having a BCB film applied over an insulating layer. The BCB film is fully cured and dry etched in an inductive coupled plasma reactive furnace. The present invention does not involve dry etching of a fully cured BCB layer. U.S. Pat. No. 6,514,872 is hereby incorporated by reference into the specification of the present invention. As can be seen from the prior art, the difficulty with the use of BCB is that once it is fully cured wet etching techniques are ineffective. Because applications requiring precision processing often necessitate wet etching of the dielectric layer, and do not allow the substitution of partially cured BCB that may allow some degree of wet etching, BCB has been abandoned for many devices where it would otherwise be the ideal material. An example of such an application is the use of BCB as a wafer bonding material. It is therefore desirable in the art to have a method for effectively processing fully cured BCB with a wet etch process. |
<SOH> SUMMARY OF THE INVENTION <EOH>It is an object of the present invention to fabricate a semiconductor device by wet etching a fully cured BCB layer. It is a further object of the present invention to fabricate a semiconductor device by wet etching and subsequently removing a fully cured BCB layer. It is another object of the present invention to fabricate a semiconductor device by wet etching and subsequent removing a fully cured BCB layer without damage to the device surface. The present invention is a method of wet etching a semiconductor device having at least one BCB layer. The first step of the method is selecting a substrate. The substrate is preferably a silicon wafer. The second step of the method is coating the front surface of the substrate with an oxide layer. The third step of the method is applying a BCB layer on top of the oxide layer. The BCB layer is preferably applied by spin coating. The fourth step of the method is baking the device until the BCB layer is fully cured. The fifth step of the method is processing the device. Processing may, for example, involve grinding, etching, polishing or performing other techniques on the back side of the substrate, depositing layers or defining patterns on the front side of the substrate. The sixth step of the method is to stress the substrate, preferably causing the substrate to warp. The stress is preferably created by mechanically grinding the substrate such that the substrate becomes thinner. The seventh step of the method is breaking the bonds within the BCB layer and between the BCB layer and the oxide layer. This is a process commonly referred to as wet etching. The wet etch is preferably performed using a chemical mixture of hydrofluoric acid, nitric acid, phosphoric acid and sulfuric acid. The eighth step of the method is removing the BCB layer, preferably using a standard dump rinse tank. The ninth step of the method is removing the oxide layer, preferably using an acid etch. |
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