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1. A method for cooling milk in an automatic milking system, which comprises a milking robot (1) for milking animals, such as cows; a milk storage tank (5) connected to said milking robot for storing milk extracted by the milking robot; and a cooling device (7) for cooling milk stored or to be stored in said milk storage tank by cooling a bottom portion (14) of said milk storage tank, said method being characterized by the steps of: measuring an amount of milk extracted by said milking robot by means of a milk flow meter; determining a cooling need for milk stored or to be stored in said milk storage tank based on said measured amount of milk; measuring a quantity indicative of a temperature of an inner surface area (16) of the bottom portion of the milk storage tank; and cooling said bottom portion of said milk storage tank in consecutive periods, such that each period of cooling (τ1, τ3) is followed by a respective period of non-cooling (τ2, τ4), wherein the duration of each period of cooling and/or non-cooling is based on said measured quantity indicative of the inner surface temperature, and said determined cooling need. 2. The method as claimed in claim 1 wherein at least one period of cooling is started while said milk storage tank is empty of milk. 3. The method as claimed in claim 1 or 2 wherein the steps of measuring an amount of milk; determining a cooling need; measuring a quantity indicative of a temperature of an inner surface area of the bottom portion of the milk storage tank; and cooling said bottom portion of said milk storage tank in consecutive periods are preceded by the step of cooling said milk storage tank by means of cooling the bottom portion thereof. 4. The method as claimed in claim 3 wherein the step of cooling said milk storage tank by means of cooling the bottom portion thereof is performed after cleaning of said milk storage tank. 5. The method as claimed in claim 1 wherein at least one period of cooling is started while milk from at most five animals has been stored in said milk storage tank. 6. The method as claimed in any of claims 1-5 wherein said quantity indicative of a temperature of an inner surface area of the bottom portion of the milk storage tank is a temperature of the cooling device. 7. The method as claimed in claim 6 wherein said cooling device includes an evaporator (8); a compressor (9); a condenser (10); and an expansion valve (12), all of which being interconnected by a connecting piping (13), such that a cooling medium can be circulated therein; and wherein said quantity indicative of a temperature of an inner surface area of the bottom portion of the milk storage tank is a temperature of the connecting piping between the expansion valve and the evaporator, of the expansion valve or of the evaporator. 8. The method as claimed in any of claims 1-5 wherein said cooling device includes an evaporator (8); a compressor (9); a condenser (10); and an expansion valve (12), all of which being interconnected by a connecting piping (13), such that a cooling medium can be circulated therein; and wherein said quantity indicative of a temperature of an inner surface area of the bottom portion of the milk storage tank is a pressure in said evaporator or in the connecting piping between the evaporator and the compressor. 9. The method as claimed in any of claims 1-8 wherein said step of cooling said milk storage tank is performed while ensuring that the temperature of said inner surface area of the bottom portion of the milk storage tank is kept above 0° C. 10. The method as claimed in any of claims 1-9 wherein said amount of milk is measured in said milking robot. 11. The method as claimed in any of claims 1-9 wherein said amount of milk is measured in or at said milk storage tank. 12. The method as claimed in any of the preceding claims comprising the step of cooling said bottom portion of said milk storage tank in response to a temperature of milk stored in said milk storage tank if a level of milk in said milk storage tank exceeds a threshold level. 13. The method as claimed in any of the preceding claims wherein the step of determining a cooling need for milk stored or to be stored in said milk storage tank includes the establishment of an expected temperature of the milk stored or to be stored in the milk store tank at the end of the step of cooling said bottom portion of said milk storage tank in consecutive periods; the temperature of the milk is established at the end of the step of cooling said bottom portion of said milk storage tank in consecutive periods; and the method for cooling milk as claimed in any of the preceding claims is repeated, wherein the duration of each period of cooling and/or non-cooling is based also on any difference between said expected and established temperatures. 14. An arrangement for cooling milk in an automatic milking system, said milking system comprising a milking robot (1) for milking animals, such as cows; a milk flow meter (2) for measuring an amount of milk extracted by said milking robot; a milk storage tank (5) connected to the milking robot for storing milk extracted by the milking robot; and a cooling device (7) for cooling milk stored or to be stored in said milk storage tank by cooling a bottom portion (14) of said milk storage tank, characterized in that said arrangement comprises: a sensor (15) for measuring a quantity indicative of a temperature of an inner surface area (16) of the bottom portion of the milk storage tank; and a controller (17) for determining a cooling need for milk stored or to be stored in said milk storage tank based on said measured amount of milk; and for controlling said cooling device to cool said bottom portion of said milk storage tank in consecutive periods, such that each period of cooling (τ1, τ3) is followed by a respective period of non-cooling (τ2, τ4), wherein the duration of each period of cooling and/or non-cooling is based on said measured quantity indicative of the inner surface area temperature, and said determined cooling need. 15. The arrangement as claimed in claim 14 wherein the controller is adapted to control said cooling device to start at least one period of cooling while said milk storage tank is empty of milk. 16. The arrangement as claimed in claim 14 wherein the controller is adapted to control said cooling device to start at least one period of cooling while milk from at most five animals has been stored in said milk storage tank. 17. The arrangement as claimed in any of claims 14-16 wherein said quantity indicative of a temperature of an inner surface area of the bottom portion of the milk storage tank is a temperature of the cooling device. 18. The arrangement as claimed in claim 17 wherein said cooling device includes an evaporator (8); a compressor (9); a condenser (10); and an expansion valve (12), all of which being interconnected by a connecting piping (13), such that a cooling medium can be circulated therein; and wherein said sensor for measuring the quantity indicative of a temperature of an inner surface area of the bottom portion of the milk storage tank is a temperature sensor (15) arranged at the connecting piping between the expansion valve and the evaporator, at the expansion valve or at the evaporator. 19. The arrangement as claimed in any of claims 14-16 wherein said cooling device includes an evaporator (8); a compressor (9); a condenser (10); and an expansion valve (12), all of which being interconnected by a connecting piping (13), such that a cooling medium can be circulated therein; and wherein said sensor for measuring said quantity indicative of a temperature of an inner surface area of the bottom portion of the milk storage tank is a pressure sensor arranged to measure the pressure in said evaporator, or in the connecting piping between the evaporator and the compressor. 20. The arrangement as claimed in any of claims 14-19 wherein said controller is adapted to control said cooling device to cool said milk storage tank while ensuring that the temperature of said inner surface area of the bottom portion of the milk storage tank is kept above 0° C. 21. The arrangement as claimed in any of claims 14-20 wherein said milk flow meter for measuring said amount of milk extracted by said milking robot is arranged in said milking robot. 22. The arrangement as claimed in any of claims 14-20 wherein said milk flow meter for measuring said amount of milk extracted by said milking robot is arranged in or at said milk storage tank. 23. The arrangement as claimed in any of claims 14-22 wherein the controller is adapted to control said cooling device to cool said bottom portion of said milk storage tank in response to a temperature of milk stored in said milk storage tank depending on a level of milk in said milk storage tank exceeding a threshold level. |
<SOH> BACKGROUND <EOH>In dairy farming animals are milked and their milk is thereafter stored in a milk storage tank for collection on a regular time basis, e.g. every second day. In order to maintain the quality of the milk, it is cooled to approximately 4° C. as quickly as possible. It is necessary to be careful during cooling of the milk since the quality is deteriorated if the milk freezes. At a dairy farm provided with an automatic milking system, the milk usually enters the milk storage tank in small amounts spread during the day and night, compared to a dairy farm without an automatic milking system, where all animals are milked together two or three times a day. The milk storage tank is usually equipped with a cooling device, which lowers the temperature of the milk to about 4° C. and maintains this temperature in a filled milk storage tank. When the milk storage tank only contains small amounts of milk, there is a considerable risk of cooling the milk too much since the cooling device, when cooling, operates at full capacity. Usually the cooling of milk in a milk storage tank is controlled in response to the temperature of the milk in the tank. The milk temperature is usually measured on the outside of the tank due to hygienic requirements, and this results in a rather slow response when the temperature of the milk within the tank is changed. Such a sluggish temperature response gives rise to problems, in particular when the volume of stored milk is small. The temperature of a small milk volume is lowered rapidly at the risk of freezing the milk. Furthermore, the milk storage tank is typically provided with a stirrer, which stirs the milk in the tank to obtain a uniform temperature of the milk. Such a stirrer is usually not able to stir the milk in the tank when the tank contains only small amounts of milk, which thus involves a further risk of freezing milk locally at the beginning of the filling of the tank with milk. If cooling, on the other hand, is omitted while there is only a small volume of milk in the milk storage tank, this milk is not cooled at once, and thus the quality of the milk is deteriorated. |
<SOH> SUMMARY OF THE INVENTION <EOH>An object of the present invention is to provide a method and an arrangement, respectively for cooling milk rapidly in an automatic milking system without risk of freezing the milk, particularly when only small amounts of milk are present in the milk storage tank. This object, among others, is attained by methods and arrangements, respectively, as claimed in the appended patent claims. By cooling in consecutive periods according to the present invention, milk is cooled without risk of being frozen, and thus a high milk quality can be safeguarded. In accordance with one preferred embodiment of the present invention the milk storage tank itself is pre-cooled, i.e. cooled before milk is entered therein. In such manner, milk transported to the milk storage tank can be cooled faster, and thus an enhanced milk quality is obtained. In accordance with another preferred embodiment of the present invention the cooling of milk may be performed firstly, when there is only a small volume of milk in the milk storage tank, as set out in the independent claims, and then, when a level of milk in said milk storage tank exceeds a threshold level, the cooling is controlled in response to the temperature of the milk in a conventional fashion. Further features and advantages of the present invention will be evident from the following description. |
Drainage and irrigation approach and structure as well as its implementation |
This Invention relates to a drainage and irrigation approach, which primarily includes the initial stage of green belt irrigation and the later stage of rainwater drainage. The drainage and irrigation structures employed in this approach consist of curb stones, edge stones, border stones, and boundary stones, which may have combinations of drainage troughs [3], rainwater ports [6] and drainage holes [4], or drainage troughs [3] and rainwater ports [6], or drainage troughs [3] and drainage holes [4]. Depending on the conditions of planted areas, the drainage and irrigation structures discussed above may be deployed along roads, landscaping on top of architectural structures, planted areas, seeded strips or barren hill slopes. |
1. A drainage and irrigation approach suitable for both rainwater and road cleaning water; the said approach consists of two stages: the initial stage of green belt irrigation and the later stage of rainwater drainage; firstly, rainwater at the beginning of a rainfall or in the event of road cleaning with debris flows through drainage holes in the drainage and irrigation structure into the planted areas, thereby providing the nutrients and water necessary for plant growth; secondly, if the rain continues falling and the rainwater rises to a certain level, clear water flows through the rainwater ports in the drainage and irrigation structure into the underground pipes and corresponding connected pipelines; the clear rainwater is then stored in underground reservoirs for storage or flows into rivers and lakes; from there it evaporates into the ecological cycle of nature. 2. The drainage and irrigation approach as described in claim 1, wherein, for the initial green belt irrigation stage, flat curb stones are laid horizontally along one side or both sides of the road, their top surface flush with the pavement surface; the curb barrier stones are laid perpendicularly to and tightly against the flat curb stones along the outer edge of the road, their top surface higher than the road pavement; on either side of the road, there is a green belt [A] on the outer side of the curb barrier stones; the surface elevation [e] of the green belt is lower than the bottom edge of the exit opening of the drainage hole in each of the said curb barrier stones so that there is an elevation drop; at the beginning of a rainfall, the rainwater flows along a drainage channel consisting of a trough in each flat curb stone and a drainage hole in each curb barrier stone. It then flows through the exit opening of the drainage hole onto the green belt [A]. 3. The drainage and irrigation approach as described in claim 1, wherein, for the later drainage stage, rainwater ports are installed at a certain interval in the row of curb barrier stones; the rainwater ports are connected to the underground drainage pipeline; the highest position of the said drainage hole in each curb barrier stone should be lower than the bottom edge [b] of the said rainwater port; when the rainwater accumulates in the green belt [A] and the water level rises to a point [a] above the rainwater port, it flows through the outlet into the underground pipeline; the drainage holes in curb barrier stones can also effectively prevent floating debris in the green belt from entering the rainwater ports so that the water flowing into the outlets is virtually filtered. 4. The drainage and irrigation approach as described in claim 1, wherein the following engineering requirements must be met: the drainage initiation point of the pavement is at Elevation b, the drainage initiation point of the rainwater port is at Elevation a, the drainage or irrigation initiation point of the curb is at Elevation C, the blocking point of the floating debris in the green belt is at Elevation d, and the water accumulation initiation point in the green belt and the seeded strips is at Elevation e. 5. The drainage and irrigation approach as described in claim 4, wherein the elevation between b and C is 6 cm; the elevation between b and d is about 12 cm; and the elevation between d and e is about 5 cm. 6. The curb stones used in a type of irrigation and drainage structure, which curb stones consist of flat curb stones and curb barrier stones having the following features: each of the said flat curb stones has a longitudinal drainage trough in its top surface and each of the said curb barrier stones has a half-circular cut-out at either end so that a drainage hole is formed when two curb barrier stones are laid end-to-end; the said longitudinal trough on the flat curb stones and the said drainage holes on the curb barrier stones are interconnected to form an unobstructed channel for water flows. 7. The curb stones as described in claim 6, wherein the said curb barrier stones have rainwater ports installed at a certain interval and the rainwater ports are connected to underground pipelines; the highest point of the drainage hole on each curb barrier stone is lower than the bottom edge of the said rainwater port. 8. The curb stones as described in claim 6, wherein the said longitudinal trough stretches on the top surface of the said flat curb stone and along the upper edge close to the said curb barrier stone; it is a trough cut along the said upper edge of the flat curb stone. 9. The curb stones as described in claim 6, wherein the said drainage hole on the curb barrier stone goes through either end of the curb barrier stone; its opening at the flat curb stone side is the water entry and on the outer side of the curb barrier stone is the water exit; the bottom edge of the entry opening is flush with the bottom of the longitudinal trough on the flat curb stone; the exit opening is somewhat lower than the entry opening to produce a horizontal elevation drop; the entry opening and the exit opening may be set level. 10. The curb stones as described in claim 8, wherein the said longitudinal trough is a half-circular trough cut along the edge of the flat curb stone close to the said curb barrier stone; additional cut-outs may or may not be made at proper positions along the trough in correspondence to the drainage holes on the curb barrier stones. 11. The curb stones as described in claim 6, wherein the half-circular drainage hole cut-out in the curb barrier stone may be a horizontal or a slanting trough; in other words, its entry and its exit may be at the same elevation or may have a proper elevation drop. 12. The edge stones used in a second type of irrigation and drainage structure, which edge stones consist of flat stones and barrier stones having the following features: Each of the said flat stones has a longitudinal trough on the side of its top surface close to the barrier stones, and the longitudinal trough connects with a drainage hole at either end of each barrier stone and several evenly spaced drainage holes in the middle section of the barrier stone to form an unobstructed channel for water flows. 13. The edge stones as described in claim 12, wherein the bottom edge of the multiple holes in the said barrier stone is flush with the bottom of the longitudinal trough in the flat stone; the drainage hole at either end of the barrier stone is flush with and connects to the bottom of the longitudinal trough; the hole is a half-circular trough at the end of each barrier stone; when two barrier stones are set end-to-end, a circular drainage hole is formed. 14. The edge stones as described in claim 12, wherein the said flat stone and the said barrier stone may form an “L”-shaped or “--|”-shaped structure. 15. The edge stones as described in claim 14, wherein the said flat stone and the said barrier stone may be a single-piece unit or two separate pieces joined together. 16. The edge stones as described in claim 15, wherein, as separate pieces to be joined together, the said flat stones are laid horizontally on one side or both sides of the road with their top surface flush with the pavement surface while the barrier stones are set perpendicularly to and tightly against the flat stones with their top surface higher than the pavement; the green belt lies on the outer side of the barrier stones; the surface elevation [e] of the green belt is lower than the bottom edge of the exit opening of the drainage hole in each of the said barrier stones so that there is an elevation drop. 17. The edge stones as described in claim 15, wherein, when the flat stones and the barrier stones form single-piece units, the edge stones may be directly laid on one side or both sides of the road; the top surface of the barrier stones is higher than the pavement surface and the top surface of the flat stones is flush with the pavement surface; the green belt lies on the outer side of the barrier stones; the surface elevation [e] of the green belt is lower than the bottom edge of the exit opening of the drainage hole in each of the said barrier stones so that there is an elevation drop. 18. The border stones used in a third type of irrigation and drainage structure, which border stones consist of flat stones and barrier stones having the following features: This structure also includes supplemental barrier stones; each of the flat stones has a stopper at either end and a trough in the middle section near one side; the trough is connected with the multiple drainage holes in the barrier stones; the top surface of the barrier stones is higher than the supplemental barrier stones, whose top surface is higher than the top surface of the flat stones; the top surface of the flat stones is flush with the upper edge of the trough and higher than the drainage holes in the barrier stones; the barrier stones and the supplemental barrier stones form an overflow relief trough between themselves. 19. The border stones as described in claim 18, wherein the flat stone, the barrier stone and the supplemental barrier stone may form a “|_|”-shaped structure. 20. The border stones as described in claim 18, wherein the said flat stone and the said supplemental barrier stone may be a single-piece “L”-shaped structure, which may then join the barrier stone to form a “|_|”-shaped structure. 21. The border stones as described in claim 18, wherein the said flat stone, barrier stone, and supplemental barrier stone may be a single-piece unit or three separate pieces joined together. 22. The border stones as described in claim 18, wherein the said supplemental barrier stones may have overflow outlets and overflow pipelines connected to those overflow outlets; they constitute the border stones used in a fourth type of drainage and irrigation structure. 23. The border stones as described in claim 18, wherein, in actual application, the said border stones are laid on one side of a green belt, which lies beyond the barrier stones; the surface elevation [e] of the green belt is lower than the bottom edge of the exit opening of the drainage hole in each of the said barrier stones so that there is an elevation drop; the rainwater flows into the trough in the flat stones and then through the drainage holes in the barrier stones onto the green belt; the supplemental barrier stones prevent the rainwater from flowing randomly. 24. The border stones as described in claim 22, wherein, in actual application, the border stones, consisting of the said flat stones, the said barrier stones and the said supplemental barrier stones, may be laid on one side of a sloping planted area, which lies outside the barrier stones; the surface elevation [e] of the planted area is lower than the bottom edge of the drainage hole in each of the said barrier stones so that there is an elevation drop; the water in the planted area flows through the drainage holes in the barrier stones into the overflow outlets in the supplemental barrier stones, and then through the connected overflow pipelines into drainage pipelines. 25. The boundary stones used in a fifth type of drainage and irrigation structure, wherein the boundary stones are U-shaped tubular structures; the tubular wall on one side has multiple drainage holes, which are connected with the water trough in the stones; the tubular stones have internal stoppers on both sides, or have no stopper. 26. The boundary stones as described in claim 25, wherein the drainage holes on the tubular wall are positioned lower than the top of the stoppers, which is a level surface and functions as the drainage initiation point. 27. The boundary stones as described in claim 25, wherein, in actual application, the U-shaped tubular boundary stones may be laid on one side of a planted area with the drainage holes on their walls opening towards the planted area; excessive rainwater in the planted area may flow through the drainage holes into the trough in the tubular boundary stones, which function like a dike to hold water around the tiered fields on the hill slopes; the boundary stones may also be used together with the border stones discussed earlier on barren hill slopes. 28. The curb stones as described in claim 6, wherein, in actual application, the said curb barrier stones may be set perpendicularly along one side of the road foundation with their rainwater ports below the top surface of the foundation; the top surface of the curb barrier stones is higher than the pavement surface; another row of curb barrier stones and flat curb stones is laid on the other side of the road; outside the curb barrier stones are planted areas; the rainwater ports in the curb barrier stones on both sides of the road are connected to underground drainage pipelines, which in turn lead into municipal sewage pipelines. 29. The drainage and irrigation structures as described in one of claims 6, 12, 18, and 25 wherein, depending on the conditions of the planted areas and the types of terrain, the said curb stones, edge stones, border stones and boundary stones may be set at one side of the planted areas, on one side or both sides of the roads, or at appropriate locations on barren hills; they may be connected to underground pipelines which lead to main drainage pipes, which in turn may be connected to drainage ditches. |
<SOH> BACKGROUND TECHNOLOGY <EOH>Roads, drainage pipeline networks, and landscaping are important urban infrastructures, which have a direct impact on the quality of the urban environment. The conventional road curb generally consists of curb barrier stones and flat curb stones. The flat curb stones are laid horizontally along one side or both sides of the road, with their upper surface flush with the road pavement. The curb barrier stones are laid perpendicularly to and tightly against the flat curb stones along the outer edge of the road, with their upper surface higher than the road pavement. To facilitate the drainage of rainwater, a curb barrier stone with a rainwater port is placed at a certain interval (generally 30-50 meters) along the road, or a special grated rainwater port is placed in its place. The said rainwater port is connected through a pipe to an underground drainage pipeline. To improve the rainwater drainage speed and efficiency, rainwater ports should be placed at the lowest positions on the road. Therefore, when necessary, longitudinal slopes are built on the road, that is, the pavement is laid with a specific sloping grade in the longitudinal direction of the road. Then, rainwater ports or special rainwater drainage devices are placed at the lowest positions on the pavement slopes. As is disclosed in a Japanese patent document (Publication No. 6-248610) and shown in FIG. 15 , a row of curb barrier stones are laid on either side of the seeded median strip of the road pavement. Each of the curb barrier stones has two evenly embedded pre-fabricated blocks, which are water pervious. The curb barrier stones are laid in such a manner that the bottoms of the pre-fabricated pervious blocks are flush with the surface of the pavement. In the event of a rainfall or road cleaning, the rainwater or the cleaning water is allowed to flow through the pores in the pre-fabricated pervious blocks into the soil in the seeded median strip (in the direction indicated by the arrow in FIG. 15 ) for irrigation purposes. Sands and other debris carried by the water are stopped and stranded between the curb barrier stones and the pavement. Consequently, regular maintenance work such as removing the debris is necessitated. Such maintenance work requires a lot of labor and expenses. Also, as is disclosed in a German patent document (DE 295 08 680 U1) and shown in FIG. 4 , a row of curb barrier stones are laid on either side of the sloping road pavement. A drainage outlet is formed between each pair of adjacent curb barrier stones. On the outer side of either row of the curb barrier stones are green barrier slopes or seeded areas. In the event of a rainfall or road cleaning, the rainwater or the cleaning water flows down the sloping pavement and through the drainage outlets in the curb barrier stones directly into the green barrier slopes or seeded areas for irrigation purposes. In a heavy rainfall, the vegetation may easily be damaged, the soil be washed away, and the water resources be wasted. Again, as is disclosed in a French patent document (N° 1.368.655) and shown in FIG. 5 , a row of curb barrier stones with pre-installed slanting pipe joints is laid along either side of the road. The pipe joints are connected to an underground pipeline. The cross section of the curb barrier stones assumes an “L” shape. The curb barrier stones are laid in such a manner that their top surface is even with the pavement surface. The bottom part of the curb barrier stone tilts upward and slants inward. The lower part of the opening of the pre-installed pipe joint is flush with the inner edge of the tilting section of the bottom part of the curb barrier stone. In the event of a rainfall or road cleaning, the rainwater or cleaning water flows through the opening of the pipe joint directly into the underground sewage pipeline. Sludge and other debris carried by the water may contaminate and clog the pipes. Debris removal will require a lot of labor and expenses. In addition, the water resources are wasted. The conventional technology as evidenced in the examples presented in the previous paragraphs has created a lot of problems in road debris removing, which has been costly and labor intensive. Road maintenance often generates a lot of dust, thus continually causing pollution. In addition, rainfalls or irrigation of the green belts and seeded strips along the roads cause soil loss, underground pipe clogging and contaminating, and contamination of downstream water resources. In short, in the event of rainfalls, the existing curb structures cause debris to be washed from green belts and surfaces of structures to the road pavements and then to underground pipelines and rivers and lakes. We are confronted not only with the issue of soil and water loss, but also with the risk of on-going contamination. |
Apparatus and method for displaying virtual endoscopy display |
An apparatus and method for displaying a three-dimensional virtual endoscopic image are provided. In the method, information on a virtual endoscopic image is input in the form of volume data expressed as a function of three-dimensional position. A two-dimensional reference image, a three-dimensional reference image, and a virtual endoscopic image are detected from the volume data. The detected images are displayed on one screen. Virtual cameras are respectively displayed on areas, in which the two and three-dimensional reference images are respectively displayed. Here, a camera display sphere and a camera display circle are defined on the basis of a current position of each virtual camera. When information regarding to one image, among the two and three-dimensional reference images and the virtual endoscopic image on one screen, is changed by a user's operation, information regarding to the other images is changed based on the information changed by the user's operation. |
1. An apparatus for displaying a three-dimensional virtual endoscopic image, the apparatus comprising: a volume data input unit, which inputs information on a virtual endoscopic image in the form of volume data expressed as a function of three-dimensional position; a two-dimensional reference image display control unit, which derives a two-dimensional reference image from the volume data and displays the two-dimensional reference image; a three-dimensional reference image display control unit, which derives a three-dimensional reference image from the volume data using parallel volume rendering and displays the three-dimensional reference image; a virtual endoscopic image display control unit, which applies position and direction information of a virtual camera used for collecting virtual endoscopic image information to a predetermined perspective ray-casting algorithm, thereby generating image information, and displays a virtual endoscopic image based on the generated image information; a user interface unit, which receives a user's operating signal; and a controller, which when the operating signal for changing image information displayed by one among the two-dimensional reference image display control unit, the three-dimensional reference image display control unit, and the virtual endoscopic image display control unit is input through the user interface unit, controls the two- and three-dimensional reference image display control units and the virtual endoscopic image display control unit to be associated with one another based on the operating signal. 2. The apparatus of claim 1, wherein the two-dimensional reference image display control unit comprises: a control signal input section, which receives a control signal for controlling a two-dimensional reference image among the operating signal input through the user interface unit; an image information receiver, which receives changed three-dimensional reference image information from the three-dimensional reference image display control unit and changed virtual endoscopic image information from the virtual endoscopic image display control unit; a painting section, which receives painting information regarding to a region of interest in the two-dimensional reference image from the control signal input section and changes two-dimensional reference image information based on the painting information; a display area control section, which changes the two-dimensional reference image information based on position change information regarding to a region of interest in the two-dimensional reference image, which is received through the control signal input section, or based on position change information regarding to a region of interest in the three-dimensional reference image or the virtual endoscopic image, which is received through the image information receiver; a camera control section, which changes virtual camera display information with respect to the two-dimensional reference image based on virtual camera display information regarding to the two-dimensional reference image, which is received through the control signal input section, or based on virtual camera display information regarding to the three-dimensional reference image and the virtual endoscopic image, which is received through the image information receiver; a display format determining section, which determines one among an axial image, a coronal image, a sagittal image, and a path MPR image as the format of the two-dimensional reference image, based on display information regarding to the two-dimensional reference image, which is received through the control signal input section; a two-dimensional image generator, which receives the changed three-dimensional reference image information and the changed virtual endoscopic image information from the image information receiver and converts the received three-dimensional reference image information and the received virtual endoscopic image information to two-dimensional image information; an image information display section, which detects a changed two-dimensional reference image based on the two-dimensional reference image information received from the painting section, the display area control section, the camera control section, the display format determining section, and the two-dimensional image generator, and displays the changed two-dimensional reference image; and an image information support section, which when the two-dimensional reference image information is changed in the painting section, the display area control section, and the camera control section in response to the control signals received from the control signal input section, transmits the changed two-dimensional reference image information to the three-dimensional reference image display control unit and the virtual endoscopic image display control unit so that the changed two-dimensional reference image information is applied to the three-dimensional reference image and the virtual endoscopic image. 3. The apparatus of claim 1, wherein the three-dimensional reference image display control unit comprises: a control signal input section, which receives a control signal for controlling a three-dimensional reference image among the operating signal input through the user interface unit; an image information receiver, which receives changed two-dimensional reference image information from the two-dimensional reference image display control unit and changed virtual endoscopic image information from the virtual endoscopic image display control unit; a painting section, which receives painting information regarding to a region of interest in the three-dimensional reference image from the control signal input section and changes three-dimensional reference image information based on the painting information; a display area control section, which changes the three-dimensional reference image information based on position change information regarding to a region of interest in the three-dimensional reference image, which is received through the control signal input section, or based on position change information regarding to a region of interest in the two-dimensional reference image or the virtual endoscopic image, which is received through the image information receiver; a camera control section, which changes virtual camera display information with respect to the three-dimensional reference image based on virtual camera display information regarding to the three-dimensional reference image, which is received through the control signal input section, or based on virtual camera display information regarding to the two-dimensional reference image and the virtual endoscopic image, which is received through the image information receiver; a three-dimensional volume rendering section, which receives the changed two-dimensional reference image information and the changed virtual endoscopic image information from the image information receiver and performs three-dimensional volume rendering based on the received two-dimensional reference image and virtual endoscopic image information; an image information display section, which detects a changed three-dimensional reference image based on the three-dimensional reference image information received from the painting section, the display area control section, the camera control section, and the three-dimensional volume rendering section, and displays the changed three-dimensional reference image; and an image information support section, which when the three-dimensional reference image information is changed in the painting section, the display area control section, and the camera control section in response to the control signals received from the control signal input section, transmits the changed three-dimensional reference image information to the two-dimensional reference image display control unit and the virtual endoscopic image display control unit so that the changed three-dimensional reference image information is applied to the two-dimensional reference image and the virtual endoscopic image. 4. The apparatus of claim 1, wherein the virtual endoscopic image display control unit comprises: a control signal input section, which receives a control signal for controlling a virtual endoscopic image among the operating signal input through the user interface unit; an image information receiver, which receives changed two-dimensional reference image information from the two-dimensional reference image display control unit and changed three-dimensional reference image information from the three-dimensional reference image display control unit; a painting section, which receives painting information regarding to a region of interest in the virtual endoscopic image from the control signal input section and changes virtual endoscopic image information based on the painting information; a perspective ray-casting section, which applies the processing result received from the painting section and the changed two- and three-dimensional reference image information received from the image information receiver to a predetermined perspective ray-casting algorithm so as to adjust the opacity of the region of interest, and then changes the virtual endoscopic image information based on the result of the adjustment; a display area control section, which changes the virtual endoscopic image information based on position change information regarding to the region of interest in the two- or three-dimensional reference image, which is received through the image information receiver; a camera control section, which changes the virtual endoscopic image information based on position and direction information of a virtual camera, which is received through the control signal input section; a focal zone generator, which designates a region of a user's interest on the virtual endoscopic image based on a control signal transmitted through the control signal input section; an image information display section, which detects a changed virtual endoscopic image based on the virtual endoscopic image information received from the perspective ray-casting section, the display area control section, the camera control section, and the focal zone generator, and displays the changed virtual endoscopic image; and an image information support section, which when the virtual endoscopic image information is changed in the painting section, the perspective ray-casting section, the camera control section, and the focal zone generator in response to the control signals received from the control signal input section, transmits the changed virtual endoscopic image information to the two-dimensional reference image display control unit and the three-dimensional reference image display control unit so that the changed virtual endoscopic image information is applied to the two-dimensional reference image and the three-dimensional reference image. 5. A method of displaying a three-dimensional virtual endoscopic image, the method comprising: inputting information on a virtual endoscopic image in the form of volume data expressed as a function of three-dimensional position; detecting a two-dimensional reference image, a three-dimensional reference image, and a virtual endoscopic image from the volume data; displaying the two-dimensional reference image, the three-dimensional reference image, and the virtual endoscopic image on one screen; displaying virtual cameras on areas, respectively, in which the two-dimensional reference image and the three-dimensional reference image are respectively displayed, wherein a camera display sphere and a camera display circle are defined on the basis of a current position of each virtual camera; and when information regarding to one image among the two-dimensional reference image, the three-dimensional reference image, and the virtual endoscopic image, which are displayed on one screen, is changed by a user's operation, changing information regarding to the other images based on the information that has been changed by the user's operation. 6. The method of claim 5, wherein the detection comprises detecting the three-dimensional reference image from the volume data by performing parallel volume rendering. 7. The method of claim 5, wherein the detection comprises detecting virtual endoscopic image information by applying position and direction information of each virtual camera to a predetermined perspective ray-casting algorithm. 8. The method of claim 5, wherein the displaying of the virtual cameras comprises: displaying a sphere, which has a predetermined radius from the current position of the virtual camera on the two-dimensional reference image, as the camera display sphere and displaying a set of intersection points between the camera display sphere and the two-dimensional reference image as the camera display circle; and displaying a sphere, which has a predetermined radius from the current position of the virtual camera on the three-dimensional reference image, as the camera display sphere and displaying a set of intersection points between the camera display sphere and a plane, which passes the current position of the virtual camera displayed on the three-dimensional reference image and is parallel with the screen, as the camera display circle. 9. The method of claim 8, wherein the displaying of the virtual cameras comprises: assuming that there is an infinite ray in the direction of each virtual camera, detecting an intersection point between the infinite ray and a corresponding camera display sphere, and representing the direction of the virtual camera with an arrow starting from the center of the virtual camera and passing the intersection point; and setting a corresponding camera display circle and the camera to have different colors when the virtual camera faces the surface of a hemisphere in front of an image and when the virtual camera faces the surface of a hemisphere in the rear of the image. 10. The method of claim 5, wherein the change of the information comprises: when information regarding to the two-dimensional reference image is changed by the user's operation, applying changed content to the two-dimensional reference image and then applying the changed content to the virtual endoscopic image using a predetermined perspective ray-casting algorithm and to the three-dimensional reference image using three-dimensional volume rendering; when information regarding to the three-dimensional reference image is changed by the user's operation, applying changed content to the three-dimensional reference image and then applying the changed content to the virtual endoscopic image using the predetermined perspective ray-casting algorithm and to the two-dimensional reference image using two-dimensional image generation; and when information regarding to the virtual endoscopic image is changed by the user's operation, applying changed content to the virtual endoscopic image using the predetermined perspective ray-casting algorithm and then applying the changed content to the two-dimensional reference image using two-dimensional image generation and to the three-dimensional reference image using three-dimensional volume rendering. |
<SOH> BACKGROUND ART <EOH>In order to avoid giving patients discomfort provoked by an endoscope used for observing the state of the patients' internal organs, virtual endoscopy has been developed and widely and usefully used with respect to a bronchus, a blood vessel, a large intestine, a joint, and so on in a human body. In virtual endoscopy, an image similar to a real endoscopic image is generated using a series of two-dimensional images collected through CT or MRI. In other words, a virtual endoscope is noninvasive and thus allows the inside of a patient's internal organs to be observed without giving the patient pain, so recently, it is very usefully used. A real endoscope allows the inside of internal organs to be observed only in a direction in which a camera moves. Unlikely, a virtual endoscope allows a user to observe the inside of internal organ in desired directions and is thus very helpful to diagnosis. However, a conventional virtual endoscope is difficult to operate due to inefficient user interface. For example, a conventional virtual endoscopic method is composed of a two-dimensional reference image acquired using an image input apparatus and a virtual endoscopic image generated from the two-dimensional reference image. Accordingly, in the conventional virtual endoscopic method, a correlation between the reference image and the virtual endoscopic image is two-dimensionally shown, so it is difficult to express a three-dimensional correlation and detect the position of a part shown in the virtual endoscopic image or a relative relation with other structures. In the conventional virtual endoscopic method, a virtual camera is controlled and expressed in a two-dimensional reference image. Accordingly, it is difficult for a user to operate the virtual camera in a desired direction and recognize the direction of the virtual camera. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG. 1 is a schematic block diagram of an apparatus for displaying a three-dimensional virtual endoscopic image according to an embodiment of the present invention. FIG. 1A is a block diagram of a two-dimensional reference image display control unit according to the embodiment of the present invention. FIG. 1B is a block diagram of a three-dimensional reference image display control unit according to the embodiment of the present invention. FIG. 1C is a block diagram of a virtual endoscopic image display control unit according to the embodiment of the present invention. FIG. 2 is a flowchart of a method of displaying a three-dimensional virtual endoscopic image according to an embodiment of the present invention. FIG. 2A is a flowchart of a procedure of displaying virtual cameras according to the embodiment of the present invention. FIG. 2B is a flowchart of a procedure of changing a display area according to the embodiment of the present invention. FIGS. 3A through 3E show examples of a method of displaying and operating a virtual camera according to the embodiment of the present invention. FIGS. 4A through 4G show examples of a three-dimensional virtual endoscopic image according to the embodiment of the present invention. detailed-description description="Detailed Description" end="lead"? |
Modulation of gene expression associated with inflammation proliferation and neurite outgrowth using nucleic acid based technologies |
The present invention relates to nucleic acid molecules, including antisense, enzymatic nucleic acid molecules, and RNA interference molecules, such as hammerhead ribozymes, DNAzymes, allozymes, siRNA, decoys and antisense, which modulate the expression of prostaglandin D2 (PTGDS), prostaglandin D2 receptor (PTGDR), adenosine receptor, NOGO and NOGO receptor, and IKK genes, such as IKK-gamma, IKK-alpha, or IKK-beta, and PKR genes. |
1. A nucleic acid molecule that down regulates expression or inhibits function of a receptor for a neurite growth inhibitor. 2. A nucleic acid molecule of claim 1, wherein the receptor is a NOGO receptor. 3. The nucleic acid of claim 1, wherein said nucleic acid molecule is adapted for use to treat conditions selected from the group consisting of CNS injury, spinal cord injury, and cerebrovascular accident. 4. The nucleic acid molecule of claim 1 or claim 2, wherein said nucleic acid molecule is an enzymatic nucleic acid molecule having at least one binding arm. 5. The nucleic acid molecule of claim 4, wherein said enzymatic nucleic acid molecule has an endonuclease activity to cleave RNA encoded by a NOGO receptor gene. 6. The nucleic acid of claim 4, wherein the at least one binding arm of the enzymatic nucleic acid molecule comprises a sequence complementary to a sequence selected from the group consisting of SEQ ID NOs. 1-1023. 7. An enzymatic nucleic acid molecule comprising a sequence selected from the group consisting of SEQ ID NOs. 5484-7055. 8. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule is an antisense nucleic acid molecule. 9. An antisense nucleic acid molecule comprising a sequence complementary to a sequence selected from the group consisting of SEQ ID NOs. 1-1023. 10. The enzymatic nucleic acid molecule of claim 4, wherein said enzymatic nucleic acid molecule is in a hammerhead (HH) motif. 11. The enzymatic nucleic acid molecule of claim 4, wherein said enzymatic nucleic acid molecule is in a hairpin, hepatitis Delta virus, group I intron, VS nucleic acid, amberzyme, zinzyme or RNAse P nucleic acid motif. 12. The enzymatic nucleic acid molecule of claim 11, wherein said zinzyme motif comprises a sequence selected from the group consisting of SEQ ID NOs. 6030-6272. 13. The enzymatic nucleic acid molecule of claim 11, wherein said amberzyme motif comprises a sequence selected from the group consisting of SEQ ID NOs. 6630-7055. 14. The enzymatic nucleic acid molecule of claim 4, wherein said enzymatic nucleic acid molecule is in a NCH motif. 15. The enzymatic nucleic acid molecule of claim 4, wherein said enzymatic nucleic acid molecule is in a G-cleaver motif. 16. The enzymatic nucleic acid molecule of claim 4, wherein said enzymatic nucleic acid molecule is a DNAzyme. 17. The nucleic acid molecule of claim 2, wherein said nucleic acid molecule comprises between 12 and 100 bases complementary to RNA encoded by a NOGO receptor gene. 18. The nucleic acid molecule of claim 2, wherein said nucleic acid molecule comprises between 14 and 24 bases complementary to the RNA encoded by a NOGO receptor gene. 19. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule is chemically synthesized. 20. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule comprises at least one 2′-sugar modification. 21. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule comprises at least one nucleic acid base modification. 22. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule comprises at least one phosphate backbone modification. 23. A mammalian cell comprising the nucleic acid molecule of claim 1. 24. The mammalian cell of claim 23, wherein said mammalian cell is a human cell. 25. A method of reducing NOGO receptor activity in a cell, comprising the step of contacting said cell with the nucleic acid molecule of claim 2, under conditions suitable for said inhibition. 26. A method of treatment of a patient having a condition associated with levels of a NOGO receptor, comprising contacting cells of said patient with the nucleic acid molecule of claim 2, under conditions suitable for said treatment. 27. The method of claim 26 further comprising the use of one or more drug therapies under conditions suitable for said treatment. 28. A method of cleaving RNA encoded by a NOGO receptor gene comprising contacting the nucleic acid molecule of claim 2 with said RNA under conditions suitable for the cleavage of said RNA. 29. The method of claim 28, wherein said cleavage is carried out in the presence of a divalent cation. 30. The method of claim 29, wherein said divalent cation is Mg2+. 31. The nucleic acid molecule of claim 1, wherein said nucleic acid comprises a cap structure, wherein the cap structure is at the 5′-end, 3′-end, or both the 5′-end and the 3′-end. 32. The enzymatic nucleic acid molecule of claim 10, wherein said hammerhead motif comprises a sequence selected from the group consisting of SEQ ID NOs. 5484-5583. 33. The enzymatic nucleic acid molecule of claim 14, wherein said NCH motif comprises a sequence selected from the group consisting of SEQ ID NOs. 5584-6029. 34. The enzymatic nucleic acid molecule of claim 16, wherein said DNAzyme comprises a sequence selected from the group consisting of SEQ ID NOs. 6273-6629. 35. The method of claim 25, wherein said nucleic acid molecule is in a hammerhead motif. 36. The method of claim 25, wherein said nucleic acid molecule is a DNAzyme. 37. An expression vector comprising at least one nucleic acid molecule of claim 1 in a manner that allows expression of the nucleic acid molecule. 38. A mammalian cell comprising an expression vector of claim 37. 39. The mammalian cell of claim 38, wherein said mammalian cell is a human cell. 40. The expression vector of claim 37, wherein said expression vector encodes a nucleic acid molecule having a hammerhead motif. 41. The expression vector of claim 37, wherein said expression vector further comprises a sequence for an antisense nucleic acid molecule complementary to RNA encoded by a NOGO receptor gene. 42. The expression vector of claim 37, wherein said expression vector comprises a two or more of said nucleic acid molecules, which can be the same or different. 43. The expression vector of claim 42, wherein said expression vector comprises a sequence encoding an antisense nucleic acid molecule complementary to RNA encoded by a NOGO receptor gene. 44. A method for treatment of conditions selected from the group consisting of CNS injury and cerebrovascular accident comprising the step of administering to a patient the nucleic acid molecule of claim 1 under conditions suitable for said treatment. 45. The method of claim 44, wherein said treatment of CNS injury is treatment of spinal cord injury. 46. A method for treatment of conditions selected from the group consisting of CNS injury and cerebrovascular accident comprising the step of administering to a patient the antisense nucleic acid molecule of claim 9 under conditions suitable for said treatment. 47. The method of claim 44, wherein said nucleic acid molecule is in a hammerhead motif. 48. The method of claim 44, wherein said method further comprises administering to said patient one or more other therapies. 49. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule comprises at least five ribose residues, at least ten 2′-O-methyl modifications, and a 3′-end modification. 50. The nucleic acid molecule of claim 49, wherein said nucleic acid molecule further comprises phosphorothioate linkages on at least three of the 5′ terminal nucleotides. 51. The nucleic acid molecule of claim 49, wherein said 3′-end modification is 3′-3′ inverted abasic moiety. 52. The enzymatic nucleic acid molecule of claim 16, wherein said DNAzyme comprises at least ten 2′-O-methyl modifications and a 3′-end modification. 53. The enzymatic nucleic acid molecule of claim 52, wherein said DNAzyme further comprises phosphorothioate linkages on at least three of the 5′ terminal nucleotides. 54. The enzymatic nucleic acid molecule of claim 52, wherein said 3′-end modification is 3′-3′ inverted abasic moiety. 55. An enzymatic nucleic acid molecule that down regulates expression of a nucleic acid molecule encoding an IkappaB kinase (IKK) subunit. 56. An enzymatic nucleic acid molecule that down regulates expression of a nucleic acid molecule encoding protein kinase PKR. 57. An enzymatic nucleic acid molecule comprising a sequence selected from the group consisting of SEQ ID NOs. 7056-11665. 58. An enzymatic nucleic acid molecule comprising at least one binding arm wherein one or more of said binding arms comprises a sequence complementary to a sequence selected from the group consisting of SEQ ID NOs. 1024-4414. 59. An antisense nucleic acid molecule comprising a sequence complementary to a sequence selected from the group consisting of SEQ ID NOs. 1024-4414. 60. The enzymatic nucleic acid molecule of any of claims 55-58, wherein said enzymatic nucleic acid molecule is adapted to treat cancer. 61. The antisense nucleic acid molecule of claim 59, wherein said antisense nucleic acid molecule is adapted to treat cancer. 62. The enzymatic nucleic acid molecule of any of claims 55-58, wherein said enzymatic nucleic acid molecule has an endonuclease activity to cleave RNA encoded by a IKK-gamma gene or PKR gene. 63. The enzymatic nucleic acid molecule of claim 55 or claim 56, wherein said enzymatic nucleic acid molecule is in an Inozyme configuration. 64. The enzymatic nucleic acid molecule of claim 55 or claim 56, wherein said enzymatic nucleic acid molecule is in a Zinzyme configuration. 65. The enzymatic nucleic acid molecule of claim 55 or claim 56, wherein said enzymatic nucleic acid molecule is in a G-cleaver configuration. 66. The enzymatic nucleic acid molecule of claim 55 or claim 56, wherein said enzymatic nucleic acid molecule is in an Amberzyme configuration. 67. The enzymatic nucleic acid molecule of claim 55 or claim 56, wherein said enzymatic nucleic acid molecule is in a DNAzyme configuration. 68. The enzymatic nucleic acid molecule of claim 55 or claim 56, wherein said enzymatic nucleic acid molecule is in a Hammerhead configuration. 69. The enzymatic nucleic acid molecule of claim 63, wherein said Inozyme comprises a sequence complementary to a sequence selected from the group consisting of SEQ ID NOs. 1218-1721 and 3051-3549. 70. The enzymatic nucleic acid molecule of claim 63, wherein said Inozyme comprises a sequence selected from the group consisting of SEQ ID NOs. 7250-7753 and 9701-10199. 71. The enzymatic nucleic acid molecule of claim 64, wherein said Zinzyme comprises a sequence complementary to a sequence selected from the group consisting of SEQ ID NOs. 1722-1998 and 3550-3768. 72. The enzymatic nucleic acid molecule of claim 64, wherein said Zinzyme comprises a sequence selected from the group consisting of SEQ ID NOs 7754-8030 and 10200-10418. 73. The enzymatic nucleic acid molecule of claim 66, wherein said Amberzyme comprises a sequence selected from the group consisting of SEQ ID NOs 8441-9069 and 11001-11547. 74. The enzymatic nucleic acid molecule of claim 67, wherein said DNAzyme comprises a sequence selected from the group consisting of SEQ ID NOs 8031-8440 and 10419-11000. 75. The enzymatic nucleic acid molecule of claim 68, wherein said Hammerhead comprises a sequence complementary to a sequence selected from the group consisting of SEQ ID NOs. 1024-1217 and 2420-3050. 76. The enzymatic nucleic acid molecule of claim 68, wherein said Hammerhead comprises a sequence selected from the group consisting of SEQ ID NOs 7056-7249 and 9070-9700. 77. The enzymatic nucleic acid molecule of any of claims 55-58, wherein said enzymatic nucleic acid molecule comprises between 12 and 100 bases complementary to RNA encoded by an IKK-gamma gene or PKR gene. 78. The enzymatic nucleic acid molecule of any of claims 55-58, wherein said enzymatic nucleic acid molecule comprises between 14 and 24 bases complementary to RNA encoded by an IKK-gamma gene or PKR gene. 79. The enzymatic nucleic acid molecule of any of claims 55-58 wherein said enzymatic nucleic acid molecule is chemically synthesized. 80. The antisense nucleic acid molecule of claim 59, wherein said antisense nucleic acid molecule is chemically synthesized. 81. The enzymatic nucleic acid molecule of any of claims 55-58, wherein said enzymatic nucleic acid molecule comprises at least one 2′-sugar modification. 82. The antisense nucleic acid molecule of claim 59, wherein said antisense nucleic acid molecule comprises at least one 2′-sugar modification. 83. The enzymatic nucleic acid molecule of any of claims 55-58, wherein said enzymatic nucleic acid molecule comprises at least one nucleic acid base modification. 84. The antisense nucleic acid molecule of claim 59, wherein said antisense nucleic acid molecule comprises at least one nucleic acid base modification. 85. The enzymatic nucleic acid molecule of any of claims 55-58, wherein said enzymatic nucleic acid molecule comprises at least one phosphate backbone modification. 86. The antisense nucleic acid molecule of claim 59, wherein said antisense nucleic acid molecule comprises at least one phosphate backbone modification. 87. A mammalian cell including the enzymatic nucleic acid molecule of any of claims 55-58. 88. The mammalian cell of claim 87, wherein said mammalian cell is a human cell. 89. A method of down-regulating PKR activity in a cell, comprising contacting said cell with the enzymatic nucleic acid molecule of claim 56, under conditions suitable for down-regulating of PKR activity. 90. A method of treatment of a patient having a condition associated with the level of PKR, comprising contacting cells of said patient with the enzymatic nucleic acid molecule of any of claims 55-59, under conditions suitable for said treatment. 91. A method of down-regulating IKK-gamma activity in a cell, comprising contacting said cell with the enzymatic nucleic acid molecule of any of claims 55-59, under conditions suitable for down-regulating of IKK-gamma activity. 92. A method of treatment of a patient having a condition associated with the level of IKK-gamma, comprising contacting cells of said patient with the enzymatic nucleic acid molecule of any of claims 55-59, under conditions suitable for said treatment. 93. The method of claim 89 further comprising the use of one or more drug therapies under conditions suitable for said treatment. 94. The method of claim 90 further comprising the use of one or more drug therapies under conditions suitable for said treatment. 95. The method of claim 91 further comprising the use of one or more drug therapies under conditions suitable for said treatment. 96. The method of claim 92 further comprising the use of one or more drug therapies under conditions suitable for said treatment. 97. A method of cleaving RNA encoded by a PKR gene comprising contacting an enzymatic nucleic acid molecule of claim 56 with said RNA under conditions suitable for the cleavage. 98. A method of cleaving RNA encoded by an IKK-gamma gene comprising contacting an enzymatic nucleic acid molecule of claim 55 with said RNA under conditions suitable for the cleavage. 99. The method of claim 98, wherein said cleavage is carried out in the presence of a divalent cation. 100. The method of claim 99, wherein said cleavage is carried out in the presence of a divalent cation. 101. The method of claim 100, wherein said divalent cation is Mg2+. 102. The method of claim 101, wherein said divalent cation is Mg2+. 103. The enzymatic nucleic acid molecule of any of claims 55-58, wherein said enzymatic nucleic acid comprises a cap structure, wherein the cap structure is at the 5′-end, the 3′-end, or both the 5′-end and the 3′-end. 104. The antisense nucleic acid molecule of claim 59, wherein said antisense nucleic acid comprises a cap structure, wherein the cap structure is at the 5′-end, the 3′-end, or both the 5′-end and the 3′-end. 105. The enzymatic nucleic acid molecule of claim 103, wherein the cap structure at the 5′-end, 3′-end, or both the 5′-end and the 3′-end comprises a 3′,3′-linked or 5′,5′-linked deoxyabasic ribose derivative. 106. The antisense nucleic acid molecule of claim 104, wherein the cap structure at the 5′-end, 3′-end, or both the 5′-end and the 3′-end comprises a 3′,3′-linked or 5′,5′-linked deoxyabasic ribose derivative. 107. The method of claim 89, wherein said enzymatic nucleic acid molecule is in a Zinzyme configuration. 108. An expression vector comprising at least one enzymatic nucleic acid molecule of claim 55 or claim 56 in a manner that allows expression of the nucleic acid molecule. 109. A mammalian cell comprising the expression vector of claim 108. 110. The mammalian cell of claim 109, wherein said mammalian cell is a human cell. 111. The expression vector of claim 108, wherein said enzymatic nucleic acid molecule is in a hammerhead configuration. (DOES THIS MAKE SENSE?) 112. The expression vector of claim 108, wherein said expression vector further comprises a sequence for an antisense nucleic acid molecule complementary to RNA encoded by an IKK-gamma subunit gene or PKR gene. 113. The expression vector of claim 108, wherein said expression vector comprises a nucleic acid sequence encoding two or more of said enzymatic nucleic acid molecules, which can be the same or different. 114. The expression vector of claim 108, wherein said expression vector further comprises a sequence encoding an antisense nucleic acid molecule complementary to RNA encoded by an IKK-gamma gene or PKR gene. 115. A method for treatment of cancer comprising administering to a patient the enzymatic nucleic acid molecule of any of claims 55-58 under conditions suitable for said treatment. 116. The method of claim 115, wherein said cancer is breast cancer, lung cancer, prostate cancer, colorectal cancer, brain cancer, esophageal cancer, stomach cancer, bladder cancer, pancreatic cancer, cervical cancer, head and neck cancer, ovarian cancer, melanoma, lymphoma, glioma, or multidrug resistant cancer. 117. A method for treatment of cancer comprising administering to a patient the antisense nucleic acid molecule of claim 59 under conditions suitable for said treatment. 118. The method of claim 117, wherein said cancer is breast cancer, lung cancer, prostate cancer, colorectal cancer, brain cancer, esophageal cancer, stomach cancer, bladder cancer, pancreatic cancer, cervical cancer, head and neck cancer, ovarian cancer, melanoma, lymphoma, glioma, or multidrug resistant cancer. 119. The method of claim 115, wherein said enzymatic nucleic acid molecule is in a Zinzyme configuration. 120. The method of claim 115, wherein said method further comprises administering to said patient one or more other therapies. 121. The method of claim 117, wherein said method further comprises administering to said patient one or more other therapies. 122. The nucleic acid molecule of any of claims 55, 56, or 58, wherein said nucleic acid molecule comprises at least five ribose residues, at least ten 2′-O-methyl modifications, and a 3′-end modification. 123. The nucleic acid molecule of claim 122, wherein said nucleic acid molecule further comprises phosphorothioate linkages on at least three of the 5′ terminal nucleotides. 124. The nucleic acid molecule of claim 122, wherein said 3′-end modification is a 3′-3′ inverted abasic moiety. 125. The method of claim 93 wherein said other drug therapies are monoclonal antibodies, IKK-gamma or PKR-specific inhibitors, chemotherapy, or radiation therapy. 126. The method of claim 125, wherein said chemotherapy is paclitaxel, docetaxel, cisplatin, methotrexate, cyclophosphamide, doxorubin, fluorouracil carboplatin, edatrexate, gemcitabine, or vinorelbine. 127. The method of claim 94 wherein said other drug therapies are monoclonal antibodies, IKK-gamma or PKR-specific inhibitors, chemotherapy, or radiation therapy. 128. The method of claim 127, wherein said chemotherapy is paclitaxel, docetaxel, cisplatin, methotrexate, cyclophosphamide, doxorubin, fluorouracil carboplatin, edatrexate, gemcitabine, or vinorelbine. 129. The method of claim 95 wherein said other drug therapies are monoclonal antibodies, IKK-gamma or PKR-specific inhibitors, chemotherapy, or radiation therapy. 130. The method of claim 129, wherein said chemotherapy is paclitaxel, docetaxel, cisplatin, methotrexate, cyclophosphamide, doxorubin, fluorouracil carboplatin, edatrexate, gemcitabine, or vinorelbine. 131. The method of claim 96 wherein said other drug therapies are monoclonal antibodies, IKK-gamma or PKR-specific inhibitors, chemotherapy, or radiation therapy. 132. The method of claim 131, wherein said chemotherapy is paclitaxel, docetaxel, cisplatin, methotrexate, cyclophosphamide, doxorubin, fluorouracil carboplatin, edatrexate, gemcitabine, or vinorelbine. 133. The method of claim 120, wherein said other therapies are monoclonal antibodies, IKK-gamma or PKR-specific inhibitors, chemotherapy, or radiation therapy. 134. The method of claim 133, wherein said chemotherapy is paclitaxel, docetaxel, cisplatin, methotrexate, cyclophosphamide, doxorubin, fluorouracil carboplatin, edatrexate, gemcitabine, or vinorelbine. 135. The method of claim 121, wherein said other therapies are monoclonal antibodies, IKK-gamma or PKR-specific inhibitors, chemotherapy, or radiation therapy. 136. The method of claim 135, wherein said chemotherapy is paclitaxel, docetaxel, cisplatin, methotrexate, cyclophosphamide, doxorubin, fluorouracil carboplatin, edatrexate, gemcitabine, or vinorelbine. 137. A method for treatment of an inflammatory disease comprising the step of administering to a patient the enzymatic nucleic acid molecule of any of claims 55-58 under conditions suitable for said treatment. 138. The method of claim 137, wherein said inflammatory disease is rheumatoid arthritis, restenosis, asthma, Crohn's disease, diabetes, obesity, autoimmune disease, lupus, multiple sclerosis, transplant/graft rejection, gene therapy applications, ischemia/reperfusion injury, glomerulonephritis, sepsis, allergic airway inflammation, inflammatory bowel disease, or infection. 139. A method for treatment of an inflammatory disease comprising the step of administering to a patient the antisense nucleic acid molecule of claim 59 under conditions suitable for said treatment. 140. The method of claim 139, wherein said inflammatory disease is rheumatoid arthritis, restenosis, asthma, Crohn's disease, diabetes, obesity, autoimmune disease, lupus, multiple sclerosis, transplant/graft rejection, gene therapy applications, ischemia/reperfusion injury (CNS and myocardial), glomerulonephritis, sepsis, allergic airway inflammation, inflammatory bowel disease, or infection. 141. The method of claim 137, wherein said enzymatic nucleic acid molecule is in a Zinzyme configuration. 142. The method of claim 137, wherein said method further comprises administering to said patient one or more other therapies. 143. The method of claim 139, wherein said method further comprises administering to said patient one or more other therapies. 144. A pharmaceutical composition comprising an enzymatic nucleic acid molecule of any of claims 55-58 in a pharmaceutically acceptable carrier. 145. A pharmaceutical composition comprising an antisense nucleic acid molecule of claim 59 in a pharmaceutically acceptable carrier. 146. The enzymatic nucleic acid molecule of claim 55, wherein said subunit of IKK-is IKK-gamma. 147. The enzymatic nucleic acid molecule of claim 55, wherein said subunit of IKK-is IKK-alpha. 148. The enzymatic nucleic acid molecule of claim 55, wherein said subunit of IKK-is IKK-beta. 149. A method of administering to a cell an enzymatic nucleic acid molecule of any of claims 55-57 comprising contacting said cell with the enzymatic nucleic acid molecule under conditions suitable for said administration. 150. The method of claim 149, wherein said cell is a mammalian cell. 151. The method of claim 149, wherein said cell is a human cell. 152. The method of claim 149, wherein said administration is in the presence of a delivery reagent. 153. The method of claim 152, wherein said delivery reagent is a lipid. 154. The method of claim 153, wherein said lipid is a cationic lipid. 155. The method of claim 153, wherein said lipid is a phospholipid. 156. The method of claim 152, wherein said delivery reagent is a liposome. 157. A nucleic acid molecule that down regulates expression of a prostaglandin D2 receptor (PTGDR) gene. 158. The nucleic acid molecule of claim 157, wherein said nucleic acid molecule is an enzymatic nucleic acid molecule. 159. The nucleic acid molecule of claim 157, wherein said nucleic acid molecule is an antisense nucleic acid molecule. 160. The enzymatic nucleic acid molecule of claim 158, wherein said enzymatic nucleic acid molecule comprises a sequence selected from the group of sequences consisting of SEQ ID NOs: 11666-13262. 161. The enzymatic nucleic acid molecule of claim 158, wherein said enzymatic nucleic acid molecule comprises at least one binding arm wherein the at least one binding arm comprises a sequence complementary to a sequence selected from the group of sequences consisting of SEQ ID NOs: 4415-5483. 162. The antisense nucleic acid molecule of claim 159, wherein said antisense nucleic acid molecule comprises a sequence complementary to a sequence selected from the group of sequences consisting of SEQ ID NOs: 4415-5483. 163. The nucleic acid molecule of claim 157, wherein said nucleic acid molecule is adapted to treat asthma. 164. The enzymatic nucleic acid molecule of any of claims 158, 160 or 161, wherein said enzymatic nucleic acid molecule has an endonuclease activity to cleave RNA encoded by a PTGDR gene. 165. The enzymatic nucleic acid molecule of claim 157, wherein said enzymatic nucleic acid molecule is in a hammerhead configuration. 166. The enzymatic nucleic acid molecule of claim 157, wherein said enzymatic nucleic acid molecule is in an Inozyme configuration. 167. The enzymatic nucleic acid molecule of claim 157, wherein said enzymatic nucleic acid molecule is in a Zinzyme configuration. 168. The enzymatic nucleic acid molecule of claim 157, wherein said enzymatic nucleic acid molecule is in a DNAzyme configuration. 169. The enzymatic nucleic acid molecule of claim 157, wherein said enzymatic nucleic acid molecule is in a G-cleaver configuration. 170. The enzymatic nucleic acid molecule of claim 157, wherein said enzymatic nucleic acid molecule is in an Amberzyme configuration. 171. The enzymatic nucleic acid molecule of claim 165, wherein said hammerhead configuration comprises a sequence complementary to a sequence selected from the group of sequences consisting of SEQ ID NOs: 4415-4641. 172. The enzymatic nucleic acid molecule of claim 165, wherein said hammerhead configuration comprises a sequence selected from the group of sequences consisting of SEQ ID NOs: 11666-11892. 173. The enzymatic nucleic acid molecule of claim 166, wherein said Inozyme configuration comprises a sequence complementary to a sequence selected from the group of sequences consisting of SEQ ID NOs: 4642-5017. 174. The enzymatic nucleic acid molecule of claim 166, wherein said Inozyme configuration comprises a sequence selected from the group of sequences consisting of SEQ ID NOs: 11893-12268. 175. The enzymatic nucleic acid molecule of claim 167, wherein said Zinzyme configuration comprises a sequence complementary to a sequence selected from the group of sequences consisting of SEQ ID NOs: 5018-5248. 176. The enzymatic nucleic acid molecule of claim 167, wherein said Zinzyme configuration comprises a sequence selected from the group of sequences consisting of SEQ ID NOs: 12269-12499. 177. The enzymatic nucleic acid molecule of claim 168, wherein said DNAzyme configuration comprises a sequence complementary to a sequence selected from the group of sequences consisting of SEQ ID NOs: 4415-5294. 178. The enzymatic nucleic acid molecule of claim 168, wherein said DNAzyme configuration comprises a sequence selected from the group of sequences consisting of SEQ ID NOs: 12500-12842. 179. The enzymatic nucleic acid molecule of claim 170, wherein said Amberzyme configuration comprises a sequence complementary to a sequence selected from the group of sequences consisting of SEQ ID NOs: 5018-5248, and 5295-5483. 180. The enzymatic nucleic acid molecule of claim 170, wherein said Amberzyme configuration comprises a sequence selected from the group of sequences consisting of SEQ ID NOs: 12843-13262. 181. The enzymatic nucleic acid molecule of any of claims 158, 160 or 161, wherein said enzymatic nucleic acid molecule comprises between 8 and 100 bases complementary to a RNA molecule encoded by a PTGDR gene. 182. The enzymatic nucleic acid molecule of any of claims 158, 160 or 161, wherein said enzymatic nucleic acid molecule comprises between 14 and 24 bases complementary to a RNA molecule encoded by a PTGDR gene. 183. The enzymatic nucleic acid molecule of any of claims 158, 160 or 161, wherein said enzymatic nucleic acid molecule is chemically synthesized. 184. The antisense nucleic acid molecule of claim 159, wherein said antisense nucleic acid molecule is chemically synthesized. 185. The enzymatic nucleic acid molecule of any of claims 158, 160 or 161, wherein said enzymatic nucleic acid molecule comprises at least one 2′-sugar modification. 186. The antisense nucleic acid molecule of claim 159, wherein said antisense nucleic acid molecule comprises at least one 2′-sugar modification. 187. The enzymatic nucleic acid molecule of any of claims 158, 160 or 161, wherein said enzymatic nucleic acid molecule comprises at least one nucleic acid base modification. 188. The antisense nucleic acid molecule of claim 159, wherein said antisense nucleic acid molecule comprises at least one nucleic acid base modification. 189. The enzymatic nucleic acid molecule of any of claims 158, 160 or 161, wherein said enzymatic nucleic acid molecule comprises at least one phosphate backbone modification. 190. The antisense nucleic acid molecule of claim 159, wherein said antisense nucleic acid molecule comprises at least one phosphate backbone modification. 191. A mammalian cell comprising the enzymatic nucleic acid molecule of any of claims 158, 160 or 161. 192. The mammalian cell of claim 191, wherein said mammalian cell is a human cell. 193. A method of reducing PTGDR activity in a cell, comprising contacting said cell with the enzymatic nucleic acid molecule of any of claims 158, 160 or 161, under conditions suitable for said reduction. 194. A method of reducing PTGDR activity in a cell, comprising contacting said cell with the antisense nucleic acid molecule of claim 159 under conditions suitable for said reduction. 195. A method of treatment of a patient having a condition associated with the level of PTGDR, comprising contacting cells of said patient with the enzymatic nucleic acid molecule of any of claims 158, 160 or 161, under conditions suitable for said treatment. 196. A method of treatment of a patient having a condition associated with the level of PTGDR, comprising contacting cells of said patient with the antisense nucleic acid molecule of claim 159, under conditions suitable for said treatment. 197. The method of claim 193 further comprising the use of one or more drug therapies under conditions suitable for said treatment. 198. The method of claim 194 further comprising the use of one or more drug therapies under conditions suitable for said treatment. 199. The method of claim 195 further comprising the use of one or more drug therapies under conditions suitable for said treatment. 200. The method of claim 196 further comprising the use of one or more drug therapies under conditions suitable for said treatment. 201. A method of cleaving a RNA molecule encoded by a PTGDR gene comprising contacting the enzymatic nucleic acid molecule of any of claims 158, 160 or 161 with said RNA molecule encoded by a PTGDR gene under conditions suitable for the cleavage. 202. The method of claim 201, wherein said cleavage is carried out in the presence of a divalent cation. 203. The method of claim 202, wherein said divalent cation is Mg2+. 204. The enzymatic nucleic acid molecule of any of claims 158, 160 or 161, wherein said enzymatic nucleic acid comprises a cap structure, wherein the cap structure is at the 5′-end, 3′-end, or both the 5′-end and the 3′-end. 205. The antisense nucleic acid molecule of claim 159, wherein said antisense nucleic acid comprises a cap structure, wherein the cap structure is at the 5′-end, 3′-end, or both the 5′-end and the 3′-end. 206. The enzymatic nucleic acid molecule of claim 204, wherein the cap structure at the 5′-end, 3′-end, or both the 5′-end and the 3′-end comprises a 3′,3′-linked or 5′,5′-linked deoxyabasic ribose derivative. 207. The antisense nucleic acid molecule of claim 205, wherein the cap structure at the 5′-end, 3′-end, or both the 5′-end and the 3′-end comprises a 3′,3′-linked or 5′,5′-linked deoxyabasic ribose derivative. 208. The method of claim 193, wherein said enzymatic nucleic acid molecule is in a Zinzyme configuration. 209. An expression vector comprising a nucleic acid molecule encoding at least one enzymatic nucleic acid molecule of claim 158 or claim 160 in a manner that allows expression of the nucleic acid molecule. 210. A mammalian cell comprising the expression vector of claim 209. 211. The mammalian cell of claim 210, wherein said mammalian cell is a human cell. 212. The expression vector of claim 209, wherein said enzymatic nucleic acid molecule is in a hammerhead configuration. 213. The expression vector of claim 209, wherein said expression vector further comprises a sequence for an antisense nucleic acid molecule complementary to a RNA molecule encoded by a PTGDR gene. 214. The expression vector of claim 209, wherein said expression vector comprises a nucleic acid sequence encoding two or more of said enzymatic nucleic acid molecules, which can be the same or different. 215. The expression vector of claim 214, wherein said expression vector further comprises a sequence encoding an antisense nucleic acid molecule complementary to a RNA molecule encoded by a PTGDR gene. 216. A method for treatment of an allergic condition comprising the step of administering to a patient the enzymatic nucleic acid molecule of any of claims 156-159 under conditions suitable for said treatment. 217. The method of claim 216, wherein said allergic condition is asthma, allergic rhinitis, or atopic dermatitis. 218. A method for treatment of an allergic condition comprising administering to a patient the antisense nucleic acid molecule of claim 159 under conditions suitable for said treatment. 219. The method of claim 218, wherein said allergic condition is asthma, allergic rhinitis, or atopic dermatitis. 220. The method of claim 216, wherein said enzymatic nucleic acid molecule is in a Zinzyme configuration. 221. The method of claim 216, wherein said method further comprises administering to said patient one or more other treatment therapies. 222. The method of claim 218, wherein said method further comprises administering to said patient one or more other treatment therapies. 223. The enzymatic nucleic acid molecule of any of claims 158, 160 or 161, wherein said enzymatic nucleic acid molecule comprises at least five ribose residues, at least ten 2′-O-methyl modifications, and a 3′-end modification. 224. The enzymatic nucleic acid molecule of claim 223, wherein said enzymatic nucleic acid molecule further comprises phosphorothioate linkages on at least three of the 5′ terminal nucleotides. 225. The nucleic acid molecule of claim 223, wherein said 3′-end modification is a 3′-3′ inverted abasic moiety. 226. The method of claim 197 wherein said other drug therapies are bronchodilators, anti-inflammatories, adenosine inhibitors, or adenosine A1 receptor inhibitors. 227. A pharmaceutical composition comprising an enzymatic nucleic acid molecule of any of claims 158, 160 or 161. 228. A pharmaceutical composition comprising an antisense nucleic acid molecule of claim 159. 229. A method of administering to a mammal the nucleic acid molecule of claim 157, comprising contacting said mammal with the nucleic acid molecule under conditions suitable for said administration. 230. The method of claim 229, wherein said mammal is a human. 231. The method of claim 229 wherein said administration is in the presence of a delivery reagent. 232. The method of claim 231, wherein said delivery reagent is a lipid. 233. The method of claim 232, wherein said lipid is a cationic lipid. 234. The method of claim 232, wherein said lipid is a phospholipid. 235. The method of claim 231, wherein said delivery reagent is a liposome. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The present invention provides compounds, compositions, and methods for the study, diagnosis, and treatment of conditions relating to the expression of NOGO and NOGO receptor genes. In particular, the invention provides nucleic acid molecules that are used to modulate the expression of NOGO and NOGO receptor gene products. The present invention further relates to therapeutic compositions and methods for the treatment or diagnosis of diseases or conditions related to IKK gamma (IKKG) and PKR levels, such as cancer, inflammatory, and autoimmune diseases and/or disorders. In addition, the present invention also relates to therapeutic compositions and methods for the treatment or diagnosis of diseases or conditions related to allergic response. Specifically, the invention provides compositions and methods for the treatment of diseases or conditions related to levels of factors involved in allergic conditions such as asthma, for example prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS) and adenosine A1 receptor (ADORA1). The discussions that follow are not meant to be complete and are provided only to assist understanding the invention that follows. The summary is not an admission that any of the work described below is prior art to the claimed invention. The following is a brief description of the current understanding of NOGO and NOGO receptors. The ceased growth of neurons following development has severe implications for lesions of the central nervous system (CNS) caused by neurodegenerative disorders and traumatic accidents. Although CNS neurons have the capacity to rearrange their axonal and dendritic foci in the developed brain, the regeneration of severed CNS axons spanning distance does not exist. Axonal growth following CNS injury is limited by the local tissue environment rather than intrinsic factors, as indicated by transplantation experiments (Richardson et al., 1980 , Nature, 284, 264-265). Non-neuronal glial cells of the CNS, including oligodendrocytes and astrocytes, have been shown to inhibit the axonal growth of dorsal root ganglion neurons in culture (Schwab and Thoenen, 1985 , J. Neurosci., 5, 2415-2423). Cultured dorsal root ganglion cells can extend their axons across glial cells from the peripheral nervous system, (ie; Schwann cells), but are inhibited by oligodendrocytes and myelin of the CNS (Schwab and Caroni, 1988 , J. Neurosci., 8, 2381-2393). The non-conducive properties of CNS tissue in adult vertebrates is thought to result from the existence of inhibitory factors rather than the lack of growth factors. The identification of proteins with neurite outgrowth inhibitory or repulsive properties include NI-35, NI-250 (Caroni and Schwab, 1988 , Neuron, 1, 85-96), myelin-associated glycoprotein (Genbank Accession No M29273), tenascin-R (Genbank Accession No X98085), and NG-2 (Genbank Accession No X61945). Monoclonal antibodies (mAb IN-1) raised against NI-35/250 have been shown to partially neutralize the growth inhibitory effect of CNS myelin and oligodendrocytes. IN-1 treatment in vivo has resulted in long distance fiber regeneration in lesioned adult mammalian CNS tissue (Weibel et al., 1994 , Brain Res., 642, 259-266). Additionally, IN-1 treatment in vivo has resulted in the recovery of specific reflex and locomotor functions after spinal cord injury in adult rats (Bregman et al., 1995 , Nature, 378, 498-501). Recently, the cloning of NOGO-A (Genbank Accession No AJ242961), the rat complementary DNA encoding NI-220/250 has been reported (Chen et al., 2000 , Nature, 403, 434-439). The NOGO gene encodes at least three major protein products (NOGO-A, NOGO-B, and NOGO-C) resulting from both alternative promoter usage and alternative splicing. Recombinant NOGO-A inhibits neurite outgrowth from dorsal root ganglia and the spreading of 3T3 firboblasts. Monoclonal antibody IN-1 recognizes NOGO-A and neutralizes NOGO-A inhibition of neuronal growth in vitro. Evidence supports the proposal that NOGO-A is the previously described rat NI-250 since NOGO-A contains all six peptide sequences obtained from purified bNI-220, the bovine equivalent of rat NI-250 (Chen et al supra). Prinjha et al., 2000 , Nature, 403, 383-384, report the cloning of the human NOGO gene which encodes three different NOGO isoforms that are potent inhibitors of neurite outgrowth. Using oligonucleotide primers to amplify and clone overlapping regions of the open reading frame of NOGO cDNA, Phrinjha et al., supra identified three forms of cDNA clone corresponding to the three protein isoforms. The longest ORF of 1,192 amino acids corresponds to NOGO-A (Accession No. AJ251383). An intermediate-length splice variant that lacks residues 186-1,004 corresponds to NOGO-B (Accession No. AJ251384). The shortest splice variant, NOGO-C (Accession No. AJ251385), appears to be the previously described rat vp20 (Accession No. AF051335) and foocen-s (Accession No. AF132048), and also lacks residues 186-1,004. According to Prinjha et al., supra, the NOGO amino-terminal region shows no significant homology to any known protein, while the carboxy-terminal tail shares homology with neuroendocrine-specific proteins and other members of the reticulon gene family. In addition, the carboxy-terminal tail contains a consensus sequence that may serve as an endoplasmic-reticulum retention region. Based on the NOGO protein sequence, Prinjha et al., supra, postulate NOGO to be a membrane associated protein comprising a putative large extracellular domain of 1,024 residues with seven predicted N-linked glycosylation sites, two or three transmembrane domains, and a short carboxy-terminal region of 43 residues. Grandpre et al., 2000 , Nature , also report the identification of NOGO as a potent inhibitor of axon regeneration. The 4.1 kilobase NOGO human cDNA clone identified by Grandpre et al., supra, KIAA0886, is homologous to a cDNA derived from a previous effort to sequence random high molecular-weight brain derived cDNAs (Nagase et al., 1998 , DNA Res., 31, 355-364). This cDNA clone encodes a protein that matches all six of the peptide sequences derived from bovine NOGO. Grandpre et al., supra demonstrate that NOGO expression is predominantly associated with the CNS and not the peripheral nervous system (PNS). Cellular localization of NOGO protein appears to be predominantly reticluar in origin, however, NOGO is found on the surface of some oligodentrocytes. An active domain of NOGO has been identified, defined as residues 31-55 of a hydrophilic 66-residue lumenal/extracellular domain. A synthetic fragment corresponding to this sequence exhibits growth-cone collapsing and outgrowth inhibiting activities (Grandpre et al., supra). A receptor for the NOGO-A extracellular domain (NOGO-66) is described in Fournier et al., 2001, Nature, 409, 341-346. Fournier et al., have shown that isolated NOGO-66 inhibits axonal extension but does not alter non-neuronal cell morphology. The receptor identified has a high affinity for soluble NOGO-66, and is expressed as a glycophosphatidylinostitol-linked protein on the surface of CNS neurons. Furthermore, the expression of the NOGO-66 receptor in neurons that are NOGO insensitive results in NOGO dependent inhibition of axonal growth in these cells. Cleavage of the NOGO-66 receptor and other glycophosphatidylinostitol-linked proteins from axonal surfaces renders neurons insensitive to NOGO-66 inhibition. As such, disruption of the interaction between NOGO-66 and the NOGO-66 receptor provides the possibility of treating a wide variety of neurological diseases, injuries, and conditions. Hauswirth and Flannery, International PCT Publication No. WO 98/48027, describe materials and methods for the specific expression of proteins in retinal photoreceptor cells consisting of an adeno-associated viral vector contacting a rod or cone-opsin promoter. In addition, ribozymes which degrade mutant mRNA are described for use in the treatment of retinitis pigmentosa. Fechteler et al., Interanational PCT Publication No. WO 00/03004 describe ribozymes targeting presenilin-2 RNA for the use in treating neurodegenerative diseases such as Alzheimer's disease. Eldadah et al., 2000 , J. Neurosci., 20, 179-186, describe the protection of cerebellar granule cells from apoptosis induced by serum-potassium deprivation from ribozyme mediated inhibition of caspase-3. Seidman et al., 1999 , Antisense Nucleic Acid Drug Dev., 9, 333-340, describe in general terms, the use of antisense and ribozyme constructs for treatment of neurodegenerative diseases. Denman et al., 1994 , Nucleic Acids Research, 22, 2375-82, describe the ribozyme mediated degradation of beta-amyloid peptide precursor mRNA in COS-7 cells. Schwab and Chen, International PCT publication No. WO 00/31235, describe NOGO proteins and inhibitors of NOGO. Blatt et al., International PCT publication No. WO 01/59103, describe nucleic acid molecules for modulating expression of NOGO genes. The following is a brief description of the physiological role of nuclear factor kappa B (NFKB), IKK kinases, and protein kinase PKR. Nuclear factor kappa B (NFKB) is a multiunit transcription factor which regulates the expression of genes involved in a number of physiologic and pathologic processes. NFKB is a key component of the TNF signaling pathway. These processes include, but are not limited to: apoptosis, immune, inflammatory and acute phase responses. The REL-A gene product (a.k.a. RelA or p65), and p50 subunits of NFKB, have been implicated in the induction of inflammatory responses and cellular transformation. NFKB exists in the cytoplasm as an inactive heterodimer of the p50 and p65 subunits. NFKB is complexed with an inhibitory protein complex, IkappaB (IKK complex), until activated by the appropriate stimuli. NFKB activation can occur following stimulation of a variety of cell types by inflammatory mediators, for example TNF and IL-1, and reactive oxygen intermediates. In response to induction, NFKB can stimulate production of pro-inflammatory cytokines such as TNF-alpha, IL-1-beta, IL-6 and iNOS, thereby perpetuating a positive feedback loop. NFKB appears to play a role in a number of disease processes including: ischemia/reperfusion injury (CNS and myocardial), glomerulonephritis, sepsis, allergic airway inflammation, inflammatory bowel disease, infection, arthritis, and cancer. The nuclear DNA-binding protein, NFKB, was first identified as a factor that binds and activates the immunoglobulin kappa light chain enhancer in B cells. NFKB now is known to activate transcription of a variety of other cellular genes (e.g., cytokines, adhesion proteins, oncogenes and viral proteins) in response to a variety of stimuli (e.g., phorbol esters, mitogens, cytokines and oxidative stress). In addition, molecular and biochemical characterization of NFKB has shown that the activity is due to a homodimer or heterodimer of a family of DNA binding subunits. Each subunit bears a stretch of 300 amino acids that is homologous to the oncogene, v-rel. The activity first described as NFKB is a heterodimer of p49 or p50 with p65. The p49 and p50 subunits of NFKB (encoded by the NF-kappa B2 or NF kappa B1 genes, respectively) are generated from the precursors NFKB1 (p105) or NFKB2 (p100). The p65 subunit of NFKB (now termed REL-A) is encoded by the rel-A locus. The roles of each specific transcription-activating complex now are being elucidated in cells (Perkins, et al., 1992 , Proc. Natl. Acad. Sci USA, 89, 1529-1533). For instance, the heterodimer of NFKB1 and Rel A (p50/p65) activates transcription of the promoter for the adhesion molecule, VCAM-1, while NFKB2/RelA heterodimers (p49/p65) actually inhibit transcription (Shu, et al., 1993 , Mol. Cell. Biol., 13, 6283-6289). Conversely, heterodimers of NFKB2/RelA (p49/p65) act with Tat-I to activate transcription of the HIV genome, while NFKB1/RelA (p50/p65) heterodimers have little effect (Liu et al., 1992 , J. Virol., 66, 3883-3887). Similarly, blocking rel A gene expression with antisense oligonucleotides specifically blocks embryonic stem cell adhesion; blocking NFKB1 gene expression with antisense oligonucleotides had no effect on cellular adhesion (Narayanan et al., 1993 , Mol. Cell. Biol., 13, 3802-3810). Thus, the promiscuous role initially assigned to NFKB in transcriptional activation (Lenardo, and Baltimore, 1989 , Cell, 58, 227-229) represents the sum of the activities of the rel family of DNA-binding proteins. This conclusion is supported by recent transgenic “knock-out” mice of individual members of the rel family. Such “knock-outs” show few developmental defects, suggesting that essential transcriptional activation functions can be performed by more than one member of the rel family. A number of specific inhibitors of NFKB function in cells exist, including treatment with phosphorothioate antisense oliogonucleotide, treatment with double-stranded NFKB binding sites, and over expression of the natural inhibitor MAD-3 (an Ikappa-B family member). These agents have been used to show that NFKB is required for induction of a number of molecules involved in cancer and/or inflammation, as described below. NFkB is required for phorbol ester-mediated induction of IL-6 (Kitajima, et al., 1992 , Science, 258, 1792-5) and IL-8 (Kunsch and Rosen, 1993 , Mol. Cell. Biol., 13, 6137-46). NFkB is required for induction of the adhesion molecules ICAM-1 (Eck, et al., 1993 , Mol. Cell. Biol., 13, 6530-6536), VCAM-1 (Shu et al., supra), and E-selectin (Read, et al., 1994 , J. Exp. Med., 179, 503-512) on endothelial cells. NFkB is involved in the induction of the integrin subunit, CD18, and other adhesive properties of leukocytes (Eck et al., 1993 supra). HER2/Neu overexpression induces NFKB via a PI3-kinase/Akt pathway involving calpain-mediated degradation of IKB-alpha. Breast cancer has been shown to typify the aberrant expression of NFKB/REL factors (Pianetti et al., 2001 , Oncogene, 20, 1287-1299; Sovak et al., 1999 , J. Clin. Invest., 100, 2952-2960). Inhibition of NFKB activity has been shown to induce apoptosis in murine hepatocytes (Bellas et al., 1997 , Am. J. Pathol., 151, 891-896). NFKB has been shown to regulate cyclooxygenase-2 expression and cell proliferation in human gastric cancer cells (Joo Weon et al., 2001 , Laboratory Investigation, 81, 349-360). The above studies suggest that NFKB is integrally involved in the induction of cytokines and adhesion molecules by inflammatory mediators and is involved in the transformation of cancerous cells. Two reported studies point to another connection between NFKB and inflammation: glucocorticoids can exert their anti-inflammatory effects by inhibiting NFKB. The glucocorticoid receptor and p65 both act at NFKB binding sites in the ICAM-1 promoter (van de Stolpe, et al., 1994 , J. Biol. Chem., 269, 6185-6192). Glucocorticoid receptor inhibits NFKB-mediated induction of IL-6 (Ray and Prefontaine, 1994 Proc. Natl. Acad. Sci USA, 91, 752-756). Conversely, overexpression of p65 inhibits glucocorticoid induction of the mouse mammary tumor virus promoter. Finally, protein cross-linking and co-immunoprecipitation experiments demonstrated direct physical interaction between p65 and the glucocorticoid receptor. The IKK complex that sequesters NFKB in the cytoplasm comprises IkappaB (IκB) proteins (IκB-alpha, IκB-beta, IκKB-epsilon, p105, and p100). The phosphorylation of IκB proteins results in the release of NFKB from the IκB complex which is transported to the nucleus via the unmasking of nuclear translocation signals. Phosphorylation marks IkB proteins for ubiquitination and degradation via the proteosome pathway. Most NFKB inducing stimuli initiate activation of an IκB kinase (IKK) complex that contains two catalytic subunits, IKK-alpha (IKK1) and IKK-beta (IKK2), that phosphorylate IκB-alpha and IκB-beta, with IKK-beta playing a predominant role in pro-inflammatory signaling. In addition to the two kinases, the IKK complex contains regulatory subunits, including IKK-gamma (NEMO/IKKAP1). IKK-gamma is a protein that is critical for the assembly of the IKK complex. IKK-gamma directly binds to IKK-beta and is required for activation of NFKB, for example by TNF-alpha, IL-1-beta, lipopolysaccharide, phorbol 12-myristate 13-acetate, the human T-cell lymphotrophic virus (HTLV-1), or double stranded RNA. Genomic rearrangements in IKK-gamma have been shown to impair NFKB activation and result in incontinentia pigmenti. Additional proteins that associate with the IKK complex include, MEK kinase (MEKK1), NFKB inducing kinase (NIK), receptor interacting protein (RIP), protein kinase CK2, and IKK-associated protein (IKAP), which appears to be associated with the IκB Kinase (IKK) complex, but does not appear to be an integral component of the tripartate IKK complex as does IKK-gamma (Krappmann et al., 2001 , J. Biol. Chem., 275, 29779-87). The RNA-dependent protein kinase PKR is a signal transducer for NFKB and IFN regulatory factor-1. PKR is required for activation of NFKB by IFN-gamma via a STAT-1 independent pathway (Amitabha et al., 2001 , J. Immunol., 166, 6170-6180). The induction of NFKB by PKR takes place though phosphorylation of IκB-alpha, and appears not to require the catalytic activity of PKR, thereby proceeding independently of the dsRNA-binding properties of PKR (Ishii et al., 2001 , Oncogene, 20, 1900-1912). PKR also plays an important role in the regulation of protein synthesis by modulating the activity of eukaryotic initiation factor 2 (eIF-2-alpha) through interferon induction. Kamiya, JP 2000253884, describes specific antisense oligonucleotides for inhibiting IκB-kinase subunit expression. Krappmann et al., 2001 , J. Biol. Chem ., describe specific antisense oligonucleotides to IKK-gamma. The following is a description of molecular targets involved in diseases or conditions related to allergic response. Asthma is a chronic inflammatory disorder of the lungs characterized by airflow obstruction, bronchial hyper-responsiveness, and airway inflammation. T-lymphocytes that produce TH2 cytokines and eosinophilic leukocytes infiltrate the airways. In the airway and in bronchial alveolar lavage (BAL) fluid of individuals with asthma, high concentrations of TH2 cytokines, interleukin-4 (14), IL-5, and IL-13, are present along with increased levels of adenosine. In contrast to normal individuals, asthmatics respond to adenosine challenge with marked airway obstruction. Upon allergen challenge, mast cells are activated by cross-linked IgE-allergen complexes. Large amounts of prostaglandin D2 (PGD2), the major cyclooxygenase product of arachidonic acid are released. PGD2 is generated from PGH2 via the activity of prostaglandin D2 synthetase (PTGDS). PGD2 receptors and adenosine A1 receptors are present in the lungs and airway along with various other tissues in response to allergic stimuli (Howarth, 1997 , Allergy, 52, 12). The significance of PGD2 as a mediator of allergic asthma has been established with the development of mice deficient in the PGD2 receptor (DP). DP is a heterotrimeric GTP-binding protein-coupled, rhodopsin-type receptor specific for PGD2 (Hirata et al., 1994 , PNAS USA., 91, 11192). These mice fail to develop airway hyperreactivity and have greatly reduced eosinophil infiltration and cytokine accumulation in response to allergens. Upon allergen challenge mice deficient in the prostaglandin D2 (PGD2) receptor (DP) did not develop airway hyperactivity. Cytokine, lymphocyte and eosinophil accumulation in the lungs were greatly reduced (Matsuoka et al., 2000 , Science, 287, 2013). The DP −/− mice exhibited no behavioral, anatomic, or histological abnormalities. Primary immune response is not affected by DP disruption. Asthma affects more than 100 million people worldwide and more than 17 million Americans (5% of the population). Since 1980 the incidence has more than doubled and deaths have tripled (5,000 deaths in 1995). Annual asthma-related healthcare costs in the US alone were estimated to exceed $14.5 billion in 2000. Current therapies such as inhalant anti-inflammatories and bronchodilators can be used to treat symptoms, however, these therapies do not prevent or cure asthma. Sandberg et al., 2001 , Prog. Respir. Res., 31, 370-373, describes ribozyme therapy for asthma and COPD. Sullivan et al., International U.S. Pat. No. 5,616,488, describes ribozymes targeting interleukin-5 for treatment and diagnosis of asthma and other inflammatory disorders. Stinchcomb et al., International PCT Publication No. WO 95/23225, describes ribozymes and methods for inhibiting the expression of disease related genes including genes associated with asthma. Nyce, International PCT Publication Nos. WO 00/62736, WO 00/09525, WO 99/13886, WO 98/23294, WO 96/40266 and U.S. Pat. No. 6,025,339 describe specific antisense oligonucleotides targeting certain mRNAs encoding particular adenosine receptors. |
<SOH> SUMMARY OF THE INVENTION <EOH>The invention features novel nucleic acid-based molecules, for example, enzymatic nucleic acid molecules, allozymes, antisense nucleic acids, 2-5A antisense chimeras, triplex forming oligonucleotides, decoy RNA, dsRNA, siRNA, aptamers, and antisense nucleic acids containing RNA cleaving chemical groups, and methods to modulate gene expression; for example, gene(s) encoding prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), and adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3; gene(s) encoding NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors; and genes encoding an IkappaB kinase (IKK) subunit or protein kinase PKR. In one embodiment, the instant invention features nucleic-acid based techniques to inhibit the expression of NOGO-A (Accession No. AJ251383), NOGO-B (Accession No. AJ251384), and/or NOGO-C (Accession No. AJ251385), NOGO-66 receptor (Accession No AF283463, Fournier et al., 2001, Nature, 409, 341-346), NI-35, NI-220, and/or NI-250, myelin-associated glycoprotein (Genbank Accession No M29273), tenascin-R (Genbank Accession No X98085), and NG-2 (Genbank Accession No X61945). The description below of the various aspects and embodiments is provided with reference to the exemplary NOGO-A and NOGO-66 receptor genes. However, the various aspects and embodiments are also directed to other genes which express NOGOA-like inhibitor proteins and other receptors involved in neurite outgrowth inhibition. Those additional genes can be analyzed for target sites using the methods described for NOGO and the NOGO-66 receptor, referred to alternatively as NOGO receptor. Thus, the inhibition and the effects of such inhibition of the other genes can be performed as described herein. The invention features one or more enzymatic nucleic acid-based molecules and methods that independently or in combination modulate the expression of gene(s) encoding a member of the IκB kinase IKK complex or PKR. In particular embodiments, the invention features nucleic acid-based molecules and methods that modulate the expression of a member of the IκB kinase IKK complex, for example IKK-alpha (IKK1), IKK-beta (IKK2), or IKK-gamma (IKKγ) and/or a protein kinase PKR protein, such as IKK-alpha (IKK1) gene (Genbank Accession No. NM — 001278); IKK-beta (IKK2) gene, for example (Genbank Accession No. AF080158), IKK-gamma (IKKγ) gene, for example (Genbank Accession No. NM — 003639), and protein kinase PKR gene, for example (Genbank Accession No. NM — 002759). The description below of the various aspects and embodiments is provided with reference to the exemplary IKK-gamma and PKR genes. IKK-gamma is also known as NEMO/IKKAP1. However, the various aspects and embodiments are also directed to other genes which encode other subunits of the IKK complex, such as IKK-alpha (IKK1) or IKK-beta (IKK2). Those additional genes can be analyzed for target sites using the methods described for IKK-gamma or PKR. Thus, the inhibition and the effects of such inhibition of the other genes can be performed as described herein. In one embodiment, an enzymatic nucleic acid molecule of the invention is in a hammerhead, Inozyme, Zinzyme, DNAzyme, Amberzyme, or G-cleaver configuration. In another embodiment, a nucleic acid molecule of the invention comprises between 8 and 100 bases complementary to the RNA of the target gene. In another embodiment, a nucleic acid molecule of the invention comprises between 14 and 24 bases complementary to a RNA molecule of the target gene. In one embodiment, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups of the invention is chemically synthesized. In another embodiment, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups of the invention comprises at least one 2′-sugar modification. In another embodiment, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups of the invention comprises at least one nucleic acid base modification. In another embodiment, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups of the invention comprises at least one phosphate backbone modification. In one embodiment, the invention features a mammalian cell, for example a human cell, including the nucleic acid molecule of the invention. In another embodiment, the invention features a method of reducing target gene expression or activity in a cell, comprising contacting the cell with a nucleic acid molecule of the invention, such as an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups, under conditions suitable for the reduction. In yet another embodiment, the invention features a method of treatment of a patient having a condition associated with the level of a target gene, such prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR, comprising contacting cells of the patient with an enzymatic nucleic acid molecule of the invention, under conditions suitable for the treatment. In another embodiment, a method of treatment of a patient having a condition associated with the level of a target gene, such prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR is featured, wherein the method further comprises the use of one or more drug therapies under conditions suitable for the treatment. In another embodiment, the invention features a method of cleaving a RNA molecule of a target gene, such prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR gene, comprising contacting an enzymatic nucleic acid molecule of the invention with a RNA molecule of the corresponding gene under conditions suitable for the cleavage, for example, wherein the cleavage is carried out in the presence of a divalent cation, such as Mg 2+ . In one embodiment, a nucleic acid molecule of the invention comprises a cap structure, for example a 3′,3′-linked or 5′,5′-linked deoxyabasic ribose derivative, wherein the cap structure is at the 5′-end, or 3′-end, or both the 5′-end and the 3′-end of the enzymatic nucleic acid molecule. In one embodiment, the invention features an expression vector comprising a nucleic acid sequence encoding at least one nucleic acid molecule of the invention, in a manner which allows expression of the nucleic acid molecule. In another embodiment, the invention features a mammalian cell, for example, a human cell, including an expression vector of the invention. In yet another embodiment, the expression vector of the invention further comprises a sequence for an antisense nucleic acid molecule complementary to a RNA molecule of a target gene, such prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR gene. In one embodiment, an expression vector of the invention comprises a nucleic acid sequence encoding two or more nucleic acid molecules, such as enzymatic nucleic acid molecules, antisense, aptamers, decoys, siRNA, or 2-5A chimeras which can be the same or different. In one embodiment, the method of treatment features an enzymatic nucleic acid molecule of the invention comprises at least five ribose residues, at least ten 2′-O-methyl modifications, and a 3′-end modification, such as a 3′-3′ inverted abasic moiety. In another embodiment, an enzymatic nucleic acid molecule or antisense nucleic acid molecule of the invention further comprises phosphorothioate linkages on at least three of the 5′ terminal nucleotides. In another embodiment, the invention features a method of administering to a mammal, for example a human, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acid containing RNA cleaving chemical groups of the invention, comprising contacting the mammal with the nucleic acid molecule under conditions suitable for the administration, for example, in the presence of a delivery reagent such as a lipid, cationic lipid, phospholipid, or liposome. In yet another embodiment, the invention features a method of administering to a mammal an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acid containing RNA cleaving chemical groups of the invention in conjunction with a therapeutic agent, comprising contacting the mammal, for example a human, with the nucleic acid molecule and the therapeutic agent under conditions suitable for the administration. In one embodiment, the invention features the use of an enzymatic nucleic acid molecule, which can be in a hammerhead, NCH, G-cleaver, Amberzyme, Zinzyme, and/or DNAzyme motif, to down-regulate the expression of a a target gene, such as prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR gene. By “inhibit”, “down-regulate”, or “reduce”, it is meant that the expression of the gene, or level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or activity of one or more proteins or protein subunits, such as prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR subunits, is reduced below that observed in the absence of the nucleic acid molecules of the invention. In one embodiment, inhibition, down-regulation or reduction with an enzymatic nucleic acid molecule is below that level observed in the presence of an enzymatically inactive or attenuated molecule that is able to bind to the same site on the target RNA molecule, but is unable to cleave that RNA molecule. In another embodiment, inhibition, down-regulation, or reduction with antisense oligonucleotides is below that level observed in the presence of, for example, an oligonucleotide with scrambled sequence or with mismatches. In another embodiment, inhibition, down-regulation, or reduction of the target gene with a nucleic acid molecule of the instant invention is greater in the presence of the nucleic acid molecule than in its absence. By “up-regulate” is meant that the expression of a gene, or level of RNA molecules or equivalent RNA molecules encoding one or more proteins, protein subunits, or activity of one or more proteins or protein subunits, such as a target gene, such as prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR subunits, is greater than that observed in the absence of the nucleic acid molecules of the invention. For example, the expression of a gene, such as prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR gene, can be increased in order to treat, prevent, ameliorate, or modulate a pathological condition caused or exacerbated by an absence or low level of gene expression. By “modulate” is meant that the expression of the gene, or level of RNA molecules or equivalent RNA molecules encoding one or more protein subunits, or activity of one or more protein subunits is up-regulated or down-regulated, such that the expression, level, or activity is greater than or less than that observed in the absence of a nucleic acid molecule of the invention. By “enzymatic nucleic acid molecule” it is meant a nucleic acid molecule that has complementarity in a substrate binding region to a specified gene target, and also has an enzymatic activity that is active to specifically cleave target a RNA molecule. That is, the enzymatic nucleic acid molecule is able to intermolecularly cleave a RNA molecule and thereby inactivate a target RNA molecule. These complementary regions allow sufficient hybridization of an enzymatic nucleic acid molecule to a target RNA molecule and thus permit cleavage. One hundred percent complementarity is preferred, but complementarity as low as 50-75% can also be useful in this invention (see for example Werner and Uhlenbeck, 1995 , Nucleic Acids Research, 23, 2092-2096; Hammann et al., 1999 , Antisense and Nucleic Acid Drug Dev., 9, 25-31). The nucleic acids can be modified at the base, sugar, and/or phosphate groups. The term enzymatic nucleic acid is used interchangeably with phrases such as ribozymes, catalytic RNA, enzymatic RNA, catalytic DNA, aptazyme or aptamer-binding ribozyme, regulatable ribozyme, catalytic oligonucleotides, nucleozyme, DNAzyme, RNA enzyme, endoribonuclease, endonuclease, minizyme, leadzyme, oligozyme or DNA enzyme. All of these terminologies describe nucleic acid molecules with enzymatic activity. The specific enzymatic nucleic acid molecules described in the instant application are not limiting in the invention and those skilled in the art will recognize that all that is important in an enzymatic nucleic acid molecule of this invention is that it has a specific substrate binding site that is complementary to one or more of the target nucleic acid regions, and that it have nucleotide sequences within or surrounding that substrate binding site which impart a nucleic acid cleaving and/or ligation activity to the molecule (Cech et al., U.S. Pat. No. 4,987,071; Cech et al., 1988, 260 JAMA 3030). By “nucleic acid molecule” as used herein is meant a molecule having nucleotides. The nucleic acid can be single, double, or multiple stranded and can comprise modified or unmodified nucleotides or non-nucleotides or various mixtures and combinations thereof. Exemplary nucleic acid molecules of the invention include enzymatic nucleic acid molecules, allozymes, antisense nucleic acids, 2-5A antisense chimeras, triplex forming oligonucleotides, decoy RNA, dsRNA, siRNA, aptamers, and/or antisense nucleic acids containing RNA cleaving chemical groups. By “enzymatic portion” or “catalytic domain” is meant that portion/region of the enzymatic nucleic acid molecule essential for cleavage of a nucleic acid substrate (for example see FIGS. 1-4 ). By “substrate binding arm” or “substrate binding domain” is meant that portion/region of a enzymatic nucleic acid that is able to interact, for example via complementarity (i.e., able to base-pair with), with a portion of its substrate. Such complementarity can be 100%, but can be less if desired. For example, as few as 10 bases out of 14 can be base-paired (see for example Werner and Uhlenbeck, 1995 , Nucleic Acids Research, 23, 2092-2096; Hammann et al., 1999 , Antisense and Nucleic Acid Drug Dev., 9, 25-31). Examples of such arms are shown generally in FIGS. 1-4 . That is, these arms contain sequences within an enzymatic nucleic acid that are intended to bring enzymatic nucleic acid and target RNA together through complementary base-pairing interactions. The enzymatic nucleic acid of the invention can have binding arms that are contiguous or non-contiguous and can be of varying lengths. The length of the binding arm(s) can be greater than or equal to four nucleotides and of sufficient length to stably interact with a target RNA; in one embodiment they can be 12-100 nucleotides; in another embodiment they can be 14-24 nucleotides long (see for example Werner and Uhlenbeck, supra; Hamman et al., supra; Hampel et al., EP0360257; Berzal-Herranze et al., 1993 , EMBO J., 12, 2567-73) or between 8 and 14 nucleotides long. If two binding arms are chosen, the design is such that the length of the binding arms are symmetrical (i.e., each of the binding arms is of the same length; e.g., four and four, five and five nucleotides, or six and six nucleotides, or seven and seven nucleotides long) or asymmetrical (i.e., the binding arms are of different length; e.g., three and five, six and three nucleotides; three and six nucleotides long; four and five nucleotides long; four and six nucleotides long; four and seven nucleotides long; and the like). By “Inozyme” or “NCH” motif or configuration is meant, an enzymatic nucleic acid molecule comprising a motif as is generally described as NCH Rz in FIG. 1 . Inozymes possess endonuclease activity to cleave RNA substrates having a cleavage triplet NCH/, where N is a nucleotide, C is cytidine and H is adenosine, uridine or cytidine, and/represents the cleavage site. H is used interchangeably with X. Inozymes can also possess endonuclease activity to cleave RNA substrates having a cleavage triplet NCN/, where N is a nucleotide, C is cytidine, and / represents the cleavage site. “I” r in FIG. 1 represents an Inosine nucleotide, including a ribo-Inosine or xylo-Inosine nucleoside. By “G-cleaver” motif or configuration is meant, an enzymatic nucleic acid molecule comprising a motif as is generally described as G-cleaver Rz in FIG. 1 . G-cleavers possess endonuclease activity to cleave RNA substrates having a cleavage triplet NYN/, where N is a nucleotide, Y is uridine or cytidine and / represents the cleavage site. G-cleavers can be chemically modified as is generally shown in FIG. 1 . By “amberzyme” motif or configuration is meant, an enzymatic nucleic acid molecule comprising a motif as is generally described in FIG. 2 . Amberzymes possess endonuclease activity to cleave RNA substrates having a cleavage triplet NG/N, where N is a nucleotide, G is guanosine, and / represents the cleavage site. Amberzymes can be chemically modified to increase nuclease stability through substitutions as are generally shown in FIG. 2 . In addition, differing nucleoside and/or non-nucleoside linkers can be used to substitute the 5′-gaaa-3′ loops shown in the figure. Amberzymes represent a non-limiting example of an enzymatic nucleic acid molecule that does not require a ribonucleotide (2′-OH) group within its own nucleic acid sequence for activity. By “zinzyme” motif or configuration is meant, an enzymatic nucleic acid molecule comprising a motif as is generally described in FIG. 3 . Zinzymes possess endonuclease activity to cleave RNA substrates having a cleavage triplet including but not limited to YG/Y, where Y is uridine or cytidine, and G is guanosine and / represents the cleavage site. Zinzymes can be chemically modified to increase nuclease stability through substitutions as are generally shown in FIG. 3 , including substituting 2′-O-methyl guanosine nucleotides for guanosine nucleotides. In addition, differing nucleotide and/or non-nucleotide linkers can be used to substitute the 5′-gaaa-2′ loop shown in the figure. Zinzymes represent a non-limiting example of an enzymatic nucleic acid molecule that does not require a ribonucleotide (2′-OH) group within its own nucleic acid sequence for activity. By ‘DNAzyme’ is meant, an enzymatic nucleic acid molecule that does not require the presence of a 2′-OH group within its own nucleic acid sequence for activity. In particular embodiments the enzymatic nucleic acid molecule can have an attached linker or linkers or other attached or associated groups, moieties, or chains containing one or more nucleotides with 2′-OH groups. DNAzymes can be synthesized chemically or expressed endogenously in vivo, by means of a single stranded DNA vector or equivalent thereof. An example of a DNAzyme is shown in FIG. 4 and is generally reviewed in Usman et al., U.S. Pat. No. 6,159,714; Chartrand et al., 1995 , NAR 23, 4092; Breaker et al., 1995 , Chem. Bio. 2, 655; Santoro et al., 1997 , PNAS 94, 4262; Breaker, 1999 , Nature Biotechnology, 17, 422-423; and Santoro et. al., 2000 , J. Am. Chem. Soc., 122, 2433-39. The “10-23” DNAzyme motif is one particular type of DNAzyme that was evolved using in vitro selection (see Santoro et al., supra). Additional DNAzyme motifs can be selected for using techniques similar to those described in these references, and hence, are within the scope of the present invention. By “sufficient length” is meant an oligonucleotide of greater than or equal to 3 nucleotides that is of a length great enough to provide the intended function under the expected condition. For example, for binding arms of enzymatic nucleic acid “sufficient length” means that the binding arm sequence is long enough to provide stable binding to a target site under the expected binding conditions. The binding arms are not so long as to prevent useful turnover of the nucleic acid molecule. By “stably interact” is meant interaction of the oligonucleotides with target nucleic acid (e.g., by forming hydrogen bonds with complementary nucleotides in the target under physiological conditions) that is sufficient to the intended purpose (e.g., cleavage of target RNA by an enzyme). By “equivalent” or “related” RNA to NOGO is meant to include those naturally occurring RNA molecules having homology (partial or complete) to NOGO-A, NOGO-B, NOGO-C and/or NOGO receptor proteins or encoding for proteins with similar function as NOGO or NOGO receptor proteins in various organisms, including human, rodent, primate, rabbit, pig, protozoans, fungi, plants, and other microorganisms and parasites. The equivalent RNA sequence also includes in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like. By “equivalent” or “related” RNA to IKK-gamma is meant to include those naturally occurring RNA molecules having homology (partial or complete) to IKK-gamma proteins or encoding for proteins with similar function as IKK-gamma proteins in various organisms, including human, rodent, primate, rabbit, pig, protozoans, fungi, plants, and other microorganisms and parasites. The equivalent RNA sequence also includes in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like. By “equivalent” or “related” RNA to PKR is meant to include those naturally occurring RNA molecules having homology (partial or complete) to PKR proteins or encoding for proteins with similar function as PKR proteins in various organisms, including human, rodent, primate, rabbit, pig, protozoans, fungi, plants, and other microorganisms and parasites. The equivalent RNA sequence also includes in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like. By “equivalent” or “related” RNA to PTGDS is meant to include RNA molecules having homology (partial or complete) to RNA molecules encoding PTGDS proteins or encoding proteins with similar function as PTGDS proteins in various organisms, including human, rodent, primate, rabbit, pig, plants, protozoans, fungi, and other microorganisms and parasites. The equivalent RNA sequence can also include in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like. By “equivalent” or “related” RNA to PTGDR is meant to include RNA molecules having homology (partial or complete) to RNA molecules encoding PTGDR proteins or encoding proteins with similar function as PTGDR proteins in various organisms, including human, rodent, primate, rabbit, pig, plants, protozoans, fungi, and other microorganisms and parasites. The equivalent RNA sequence can also include in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like. By “equivalent” or “related” RNA to ADORA1 is meant to include RNA molecules having homology (partial or complete) to RNA molecule encoding ADORA1 proteins or encoding proteins with similar function as ADORA1 proteins in various organisms, including human, rodent, primate, rabbit, pig, plants, protozoans, fungi, and other microorganisms and parasites. The equivalent RNA sequence can also include in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like. By “homology” is meant the nucleotide sequence of two or more nucleic acid molecules is partially or completely identical. By “antisense nucleic acid”, it is meant a non-enzymatic nucleic acid molecule that binds to target RNA by means of RNA-RNA or RNA-DNA or RNA-PNA (protein nucleic acid; Egholm et al., 1993 Nature 365, 566) interactions and alters the activity of the target RNA (for a review, see Stein and Cheng, 1993 Science 261, 1004 and Woolf et al., U.S. Pat. No. 5,849,902). Typically, antisense molecules are complementary to a target sequence along a single contiguous sequence of the antisense molecule. However, in certain embodiments, an antisense molecule can bind to substrate such that the substrate molecule forms a loop, and/or an antisense molecule can bind such that the antisense molecule forms a loop. Thus, the antisense molecule can be complementary to two (or even more) non-contiguous substrate sequences or two (or even more) non-contiguous sequence portions of an antisense molecule can be complementary to a target sequence or both. For a review of current antisense strategies, see Schmajuk et al., 1999 , J. Biol. Chem., 274, 21783-21789, Delihas et al., 1997 , Nature, 15, 751-753, Stein et al., 1997 , Antisense N. A. Drug Dev., 7, 151, Crooke, 2000 , Methods Enzymol., 313, 3-45; Crooke, 1998 , Biotech. Genet. Eng. Rev., 15, 121-157, Crooke, 1997 , Ad. Pharmacol., 40, 1-49. In addition, antisense DNA can be used to target RNA by means of DNA-RNA interactions, thereby activating RNase H, which digests the target RNA in the duplex. The antisense oligonucleotides can comprise one or more RNAse H activating region, which is capable of activating RNAse H cleavage of a target RNA. Antisense DNA can be synthesized chemically or expressed via the use of a single stranded DNA expression vector or equivalent thereof. By “RNase H activating region” is meant a region (generally greater than or equal to 4-25 nucleotides in length, and in one embodiment from 5-11 nucleotides in length) of a nucleic acid molecule capable of binding to a target RNA to form a non-covalent complex that is recognized by cellular RNase H enzyme (see for example Arrow et al., U.S. Pat. No. 5,849,902; Arrow et al., U.S. Pat. No. 5,989,912). The RNase H enzyme binds to the nucleic acid molecule-target RNA complex and cleaves the target RNA sequence. The RNase H activating region comprises, for example, phosphodiester, phosphorothioate (at least four of the nucleotides are phosphorothiote substitutions; and in another embodiment, 4-11 of the nucleotides are phosphorothiote substitutions); phosphorodithioate, 5′-thiophosphate, or methylphosphonate backbone chemistry or a combination thereof. In addition to one or more backbone chemistries described above, the RNase H activating region can also comprise a variety of sugar chemistries. For example, the RNase H activating region can comprise deoxyribose, arabino, fluoroarabino or a combination thereof, nucleotide sugar chemistry. Those skilled in the art will recognize that the foregoing are non-limiting examples and that any combination of phosphate, sugar and base chemistry of a nucleic acid that supports the activity of RNase H enzyme is within the scope of the definition of the RNase H activating region and the instant invention. By “2-5A antisense chimera” is meant an antisense oligonucleotide containing a 5′-phosphorylated 2′-5′-linked adenylate residue. These chimeras bind to target RNA in a sequence-specific manner and activate a cellular 2-5A-dependent ribonuclease which, in turn, cleaves the target RNA (Torrence et al., 1993 Proc. Natl. Acad. Sci. USA 90, 1300; Silverman et al., 2000 , Methods Enzymol., 313, 522-533; Player and Torrence, 1998 , Pharmacol. Ther., 78, 55-113). By “aptamer” or “nucleic acid aptamer” as used herein is meant a nucleic acid molecule that binds specifically to a target molecule wherein the nucleic acid molecule has sequence that is distinct from sequence recognized by the target molecule in its natural setting. Alternately, an aptamer can be a nucleic acid molecule that binds to a target molecule where the target molecule does not naturally bind to a nucleic acid. The target molecule can be any molecule of interest. For example, the aptamer can be used to bind to a ligand-binding domain of a protein, thereby preventing interaction of the naturally occurring ligand with the protein. This is a non-limiting example and those in the art will recognize that other embodiments can be readily generated using techniques generally known in the art, see for example Gold et al., 1995 , Annu. Rev. Biochem., 64, 763; Brody and Gold, 2000 , J. Biotechnol., 74, 5; Sun, 2000 , Curr. Opin. Mol. Ther., 2, 100; Kusser, 2000 , J. Biotechnol., 74, 27; Hermann and Patel, 2000 , Science, 287, 820; and Jayasena, 1999 , Clinical Chemistry, 45, 1628. By “triplex forming oligonucleotides” is meant an oligonucleotide that can bind to a double-stranded DNA in a sequence-specific manner to form a triple-strand helix. Formation of such triple helix structure has been shown to inhibit transcription of the targeted gene (Duval-Valentin et al., 1992 Proc. Natl. Acad. Sci. USA 89, 504; Fox, 2000 , Curr. Med. Chem., 7, 17-37; Praseuth et al., 2000 , Biochim. Biophys. Acta, 1489, 181-206). By “gene” it is meant a nucleic acid that encodes an RNA, for example, nucleic acid sequences including but not limited to structural genes encoding a polypeptide. “Complementarity” refers to the ability of a nucleic acid to form hydrogen bond(s) with another RNA molecule by either traditional Watson-Crick or other non-traditional types. In reference to the nucleic molecules of the present invention, the binding free energy for a nucleic acid molecule with its target or complementary sequence is sufficient to allow the relevant function of the nucleic acid to proceed, e.g., enzymatic nucleic acid cleavage, antisense or triple helix inhibition. Determination of binding free energies for nucleic acid molecules is well known in the art (see, e.g., Turner et al., 1987 , CSH Symp. Quant. Biol. LII pp. 123-133; Frier et al., 1986 , Proc. Nat. Acad. Sci. USA 83:9373-9377; Turner et al., 1987 , J. Am. Chem. Soc. 109:3783-3785). A percent complementarity indicates the percentage of contiguous residues in a nucleic acid molecule that can form hydrogen bonds (e.g., Watson-Crick base pairing) with a second nucleic acid sequence (e.g., 5, 6, 7, 8, 9, 10 out of 10 being 50%, 60%, 70%, 80%, 90%, and 100% complementary). “Perfectly complementary” means that all the contiguous residues of a nucleic acid sequence will hydrogen bond with the same number of contiguous residues in a second nucleic acid sequence. By “RNA” is meant a molecule comprising at least one ribonucleotide residue. By “ribonucleotide” or “2′-OH” is meant a nucleotide with a hydroxyl group at the 2′ position of a β-D-ribo-furanose moiety. By “decoy RNA” is meant an RNA molecule or aptamer that is designed to preferentially bind to a predetermined ligand. Such binding can result in the inhibition or activation of a target molecule. The decoy RNA or aptamer can compete with a naturally occurring binding target for the binding of a specific ligand. For example, it has been shown that over-expression of HIV trans-activation response (TAR) RNA can act as a “decoy” and efficiently binds HIV tat protein, thereby preventing it from binding to TAR sequences encoded in the HIV RNA (Sullenger et al., 1990, Cell, 63, 601-608). This is but a specific example and those in the art will recognize that other embodiments can be readily generated using techniques generally known in the art, see for example Gold et al., 1995 , Annu. Rev. Biochem., 64, 763; Brody and Gold, 2000 , J. Biotechnol., 74, 5; Sun, 2000 , Curr. Opin. Mol. Ther., 2, 100; Kusser, 2000 , J. Biotechnol., 74, 27; Hermann and Patel, 2000 , Science, 287, 820; and Jayasena, 1999 , Clinical Chemistry, 45, 1628. Similarly, a decoy RNA can be designed to bind to a D2 receptor and block the binding of PTGDS or a decoy RNA can be designed to bind to PTGDS and prevent interaction with the D2 receptor. The term “short interfering RNA” or “siRNA” as used herein refers to a double stranded nucleic acid molecule capable of RNA interference “RNAi”, see for example Bass, 2001 , Nature, 411, 428-429; Elbashir et al., 2001 , Nature, 411, 494-498; and Kreutzer et al., International PCT Publication No. WO 00/44895; Zernicka-Goetz et al., International PCT Publication No. WO 01/36646; Fire, International PCT Publication No. WO 99/32619; Plaetinck et al., International PCT Publication No. WO 00/01846; Mello and Fire, International PCT Publication No. WO 01/29058; Deschamps-Depaillette, International PCT Publication No. WO 99/07409; and Li et al., International PCT Publication No. WO 00/44914. As used herein, siRNA molecules need not be limited to those molecules containing only RNA, but further encompasses chemically modified nucleotides and non-nucleotides. The term “allozyme” as used herein refers to an allosteric enzymatic nucleic acid molecule, see, e.g., George et al., U.S. Pat. Nos. 5,834,186 and 5,741,679, Shih et al., U.S. Pat. No. 5,589,332, Nathan et al., U.S. Pat. No. 5,871,914, Nathan and Ellington, International PCT publication No. WO 00/24931, Breaker et al., International PCT Publication Nos. WO 00/26226 and 98/27104, and Sullenger et al., International PCT publication No. WO 99/29842. The term “2-5A chimera” as used herein refers to an oligonucleotide containing a 5′-phosphorylated 2′-5′-linked adenylate residue. These chimeras bind to target RNA in a sequence-specific manner and activate a cellular 2-5A-dependent ribonuclease which, in turn, cleaves the target RNA (Torrence et al., 1993 Proc. Natl. Acad. Sci. USA 90, 1300; Silverman et al., 2000 , Methods Enzymol., 313, 522-533; Player and Torrence, 1998 , Pharmacol. Ther., 78, 55-113). The term “triplex forming oligonucleotides” as used herein refers to an oligonucleotide that can bind to a double-stranded DNA in a sequence-specific manner to form a triple-strand helix. Formation of such triple helix structure has been shown to inhibit transcription of the targeted gene (Duval-Valentin et al., 1992 Proc. Natl. Acad. Sci. USA 89, 504; Fox, 2000 , Curr. Med. Chem., 7, 17-37; Praseuth et. al., 2000 , Biochim. Biophys. Acta, 1489, 181-206). Several varieties of naturally-occurring enzymatic RNAs are known presently. Each can catalyze the hydrolysis of RNA phosphodiester bonds in trans (and thus can cleave other RNA molecules) under physiological conditions. Table I summarizes some of the characteristics of these ribozymes. In general, enzymatic nucleic acids act by first binding to a target RNA. Such binding occurs through the target binding portion of a enzymatic nucleic acid that is held in close proximity to an enzymatic portion of the molecule that acts to cleave the target RNA. Thus, the enzymatic nucleic acid first recognizes and then binds a target RNA through complementary base-pairing, and once bound to the correct site, acts enzymatically to cut the target RNA. Strategic cleavage of such a target RNA will destroy its ability to direct synthesis of an encoded protein. After an enzymatic nucleic acid has bound and cleaved its RNA target, it is released from that RNA to search for another target and can repeatedly bind and cleave new targets. Thus, a single ribozyme molecule is able to cleave many molecules of target RNA. In addition, the ribozyme is a highly specific inhibitor of gene expression, with the specificity of inhibition depending not only on the base-pairing mechanism of binding to the target RNA, but also on the mechanism of target RNA cleavage. Single mismatches, or base-substitutions, near the site of cleavage can completely eliminate catalytic activity of a ribozyme. In one embodiment of the inventions described herein, the enzymatic nucleic acid molecule is formed in a hammerhead or hairpin motif, but can also be formed in the motif of a hepatitis delta virus, group I intron, group II intron or RNase P RNA (in association with an RNA guide sequence), Neurospora VS RNA, DNAzymes, NCH cleaving motifs, or G-cleavers. Examples of such hammerhead motifs are described by Dreyfus, supra, Rossi et al., 1992 , AIDS Research and Human Retroviruses 8, 183; of hairpin motifs by Hampel et al., EP0360257, Hampel and Tritz, 1989 Biochemistry 28, 4929, Feldstein et al., 1989 , Gene 82, 53, Haseloff and Gerlach, 1989 , Gene, 82, 43, and Hampel et al., 1990 Nucleic Acids Res. 18, 299; Chowrira & McSwiggen, U.S. Pat. No. 5,631,359; of the hepatitis delta virus motif is described by Perrotta and Been, 1992 Biochemistry 31, 16; of the RNase P motif by Guerrier-Takada et al., 1983 Cell 35, 849; Forster and Altman, 1990 , Science 249, 783; Li and Altman, 1996 , Nucleic Acids Res. 24, 835; Neurospora VS RNA ribozyme motif is described by Collins (Saville and Collins, 1990 Cell 61, 685-696; Saville and Collins, 1991 Proc. Natl. Acad. Sci. USA 88, 8826-8830; Collins and Olive, 1993 Biochemistry 32, 2795-2799; Guo and Collins, 1995 , EMBO. J. 14, 363); Group II introns are described by Griffin et al., 1995 , Chem. Biol. 2, 761; Michels and Pyle, 1995 , Biochemistry 34, 2965; Pyle et al., International PCT Publication No. WO 96/22689; of the Group I intron by Cech et al., U.S. Pat. No. 4,987,071 and of DNAzymes by Usman et al., International PCT Publication No. WO 95/11304; Chartrand et al., 1995 , NAR 23, 4092; Breaker et al., 1995 , Chem. Bio. 2, 655; Santoro et al., 1997 , PNAS 94, 4262, and Beigelman et al., International PCT publication No. WO 99/55857. NCH cleaving motifs are described in Ludwig & Sproat, International PCT Publication No. WO 98/58058; and G-cleavers are described in Kore et al., 1998 , Nucleic Acids Research 26, 4116-4120 and Eckstein et al., International PCT Publication No. WO 99/16871. Additional motifs such as the Aptazyme (Breaker et al., WO 98/43993), Amberzyme (Class I motif; FIG. 2 ; Beigelman et al., U.S. Ser. No. 09/301,511) and Zinzyme ( FIG. 3 ) (Beigelman et al., U.S. Ser. No. 09/301,511), all included by reference herein including drawings, can also be used in the present invention. These specific motifs or configurations are not limiting in the invention and those skilled in the art will recognize that all that is important in an enzymatic nucleic acid molecule of this invention is that it has a specific substrate binding site which is complementary to one or more of the target gene RNA regions, and that it have nucleotide sequences within or surrounding that substrate binding site which impart an RNA cleaving activity to the molecule (Cech et al., U.S. Pat. No. 4,987,071). In one embodiment of the present invention, a nucleic acid molecule of the instant invention can be between 12 and 100 nucleotides in length. Exemplary enzymatic nucleic acid molecules of the invention are shown in Tables III-XXIII. For example, enzymatic nucleic acid molecules of the invention can be between 15 and 50 nucleotides in length, and in another embodiment between 25 and 40 nucleotides in length, e.g., 34, 36, or 38 nucleotides in length (for example see Jarvis et al., 1996 , J. Biol. Chem., 271, 29107-29112). Exemplary DNAzymes of the invention are can between 15 and 40 nucleotides in length, and in one embodiment, between 25 and 35 nucleotides in length, e.g., 29, 30, 31, or 32 nucleotides in length (see, e.g., Santoro et al., 1998 , Biochemistry, 37, 13330-13342; Chartrand et al., 1995, Nucleic Acids Research, 23, 4092-4096). Exemplary antisense molecules of the invention can be between 15 and 75 nucleotides in length, and in one embodiment between 20 and 35 nucleotides in length, e.g., 25, 26, 27, or 28 nucleotides in length (see for example Woolf et al., 1992 , PNAS., 89, 7305-7309; Milner et al., 1997 , Nature Biotechnology, 15, 537-541). Exemplary triplex forming oligonucleotide molecules of the invention are between 10 and 40 nucleotides in length, and in one embodiment are between 12 and 25 nucleotides in length, e.g., 18, 19, 20, or 21 nucleotides in length (see for example Maher et al., 1990 , Biochemistry, 29, 8820-8826; Strobel and Dervan, 1990 , Science, 249, 73-75). Those skilled in the art will recognize that all that is required is for the nucleic acid molecule to be of length and conformation sufficient and suitable for the nucleic acid molecule to catalyze a reaction contemplated herein. The length of the nucleic acid molecules of the instant invention are not limiting within the general limits stated. In one embodiment, a nucleic acid molecule that modulates, for example, down-regulates, the expression of a target gene comprises between 8 and 100 bases complementary to a RNA molecule of prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR. In another embodiment, a nucleic acid molecule that modulates the expression of a target gene comprises between 14 and 24 bases complementary to a RNA molecule of prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR. The invention provides a method for producing a class of nucleic acid-based gene modulating agents that exhibit a high degree of specificity for the RNA of a desired target. For example, the enzymatic nucleic acid molecule is can be targeted to a highly conserved sequence region of target RNAs encoding prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR (e.g., prostaglandin D2 receptor (PTGDR)), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR genes) such that specific treatment of a disease or condition can be provided with either one or several nucleic acid molecules of the invention. Such nucleic acid molecules can be delivered exogenously to specific tissue or cellular targets as required. Alternatively, the nucleic acid molecules (e.g., ribozymes, antisense, aptamers, and/or siRNA) can be expressed from DNA and/or RNA vectors that are delivered to specific cells. As used in herein “cell” is used in its usual biological sense, and does not refer to an entire multicellular organism. The cell can, for example, be in vitro, e.g., in cell culture, or present in a multicellular organism, including, e.g., birds, plants and mammals such as humans, cows, sheep, apes, monkeys, swine, dogs, and cats. The cell may be prokaryotic (e.g., bacterial cell) or eukaryotic (e.g., mammalian or plant cell). By “NOGO proteins” is meant, a protein, protein receptor or a mutant protein derivative thereof, comprising neuronal inhibitor activity, preferably CNS neuronal growth inhibitor activity. By “IKK-gamma proteins” is meant, a peptide or protein comprising a IKK-gamma or NEMO/IKKAP1 component of the IKK complex, for example a regulatory IKK subunit involved in the assembly of the high molecular weight IKK complex and/or induction of NFKB. By “PKR proteins” is meant, a peptide or protein comprising a protein kinase PKR activity, for example the activation of NFKB. By “PTGDR proteins” is meant, a protein receptor or a mutant protein or peptide derivative thereof, having prostaglandin D2 receptor activity, for example, having the ability to bind prostaglandin D2 and/or having GTP-binding protein coupled activity. By “PTGDS proteins” is meant, a prostaglandin synthetase protein or a mutant protein or peptide derivative thereof, having prostaglandin D2 synthetase activity, for example, having the ability to convert PGH2 to PGD2. By “highly conserved sequence region” is meant, a nucleotide sequence of one or more regions in a target gene does not vary significantly from one generation to the other or from one biological system to the other. The nucleic acid-based inhibitors of the invention can be added directly, or can be complexed with cationic lipids, packaged within liposomes, or otherwise delivered to target cells or tissues, for example by pulmonary delivery of an aerosol formulation with an inhaler or nebulizer. The nucleic acid or nucleic acid complexes can be locally administered to relevant tissues ex vivo, or in vivo through inhalation, injection or infusion pump, with or without their incorporation in biopolymers. In preferred embodiments, the enzymatic nucleic acid inhibitors comprise sequences that are complementary to the substrate sequences in Tables III to XXIII. Examples of such enzymatic nucleic acid molecules also are shown in Tables III to XXIII. Examples of such enzymatic nucleic acid molecules consist essentially of sequences defined in these tables. In another embodiment, the invention features antisense nucleic acid molecules, siRNA and 2-5A chimeras including sequences complementary to the substrate sequences shown in Tables III to XXIII. Such nucleic acid molecules can include sequences as shown for the binding arms of the enzymatic nucleic acid molecules in Tables III to XXIII. Similarly, triplex molecules can be provided targeted to the corresponding DNA target regions, and containing the DNA equivalent of a target sequence or a sequence complementary to the specified target (substrate) sequence. Typically, antisense molecules are complementary to a target sequence along a single contiguous sequence of the antisense molecule. However, in certain embodiments, an antisense molecule can bind to substrate such that the substrate molecule forms a loop, and/or an antisense molecule can bind such that the antisense molecule forms a loop. Thus, the antisense molecule can be complementary to two (or even more) non-contiguous substrate sequences or two (or even more) non-contiguous sequence portions of an antisense molecule can be complementary to a target sequence or both. By “consists essentially of” is meant that the active nucleic acid molecule of the invention, for example, an enzymatic nucleic acid molecule, contains an enzymatic center or core equivalent to those in the examples, and binding arms able to bind RNA such that cleavage at the target site occurs. Other sequences can be present that do not interfere with such cleavage. Thus, a core region can, for example, include one or more loop, stem-loop structure, or linker which does not prevent enzymatic activity. Thus, the underlined regions in the sequences in Tables III, IV, VIII, IX, XIII, XIV, XIX, and XX can be such a loop, stem-loop, nucleotide linker, and/or non-nucleotide linker and can be represented generally as sequence “X”. For example, a core sequence for a hammerhead enzymatic nucleic acid can comprise a conserved sequence, such as 5′-CUGAUGAG-3′ and 5′-CGAA-3′ connected by “X”, where X is 5′- GCCGUUAGGC -3′ (SEQ ID NO: 13274), or any other Stem II region known in the art, or a nucleotide and/or non-nucleotide linker. Similarly, for other nucleic acid molecules of the instant invention, such as Inozyme, G-cleaver, amberzyme, zinzyme, DNAzyme, antisense, 2-5A antisense, triplex forming nucleic acid, siRNA and decoy nucleic acids, other sequences or non-nucleotide linkers can be present that do not interfere with the function of the nucleic acid molecule. Sequence X can be a linker of ≧2 nucleotides in length, including 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 26, 30, where the nucleotides can be internally base-paired to form a stem of ≧2 base pairs. Alternatively or in addition, sequence X can be a non-nucleotide linker. In yet another embodiment, the nucleotide linker X can be a nucleic acid aptamer, such as an ATP aptamer, HIV Rev aptamer (RRE), HIV Tat aptamer (TAR) and others (for a review see Gold et al., 1995 , Annu. Rev. Biochem., 64, 763; and Szostak & Ellington, 1993, in The RNA World , ed. Gesteland and Atkins, pp. 511, CSH Laboratory Press). A “nucleic acid aptamer” as used herein is meant to indicate a nucleic acid sequence capable of interacting with a ligand. The ligand can be any natural or a synthetic molecule, including but not limited to a resin, metabolites, nucleosides, nucleotides, drugs, toxins, transition state analogs, peptides, lipids, proteins, amino acids, nucleic acid molecules, hormones, carbohydrates, receptors, cells, viruses, bacteria and others. In yet another embodiment, the non-nucleotide linker X is as defined herein. The term “non-nucleotide” as used herein include either abasic nucleotide, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, or polyhydrocarbon compounds. Specific examples include those described by Seela and Kaiser, Nucleic Acids Res. 1990, 18:6353 and Nucleic Acids Res. 1987, 15:3113; Cload and Schepartz, J. Am. Chem. Soc. 1991, 113:6324; Richardson and Schepartz, J. Am. Chem. Soc. 1991, 113:5109; Ma et al., Nucleic Acids Res. 1993, 21:2585 and Biochemistry 1993, 32:1751; Durand et al., Nucleic Acids Res. 1990, 18:6353; McCurdy et al., Nucleosides & Nucleotides 1991, 10:287; Jschke et al., Tetrahedron Lett. 1993, 34:301; Ono et al., Biochemistry 1991, 30:9914; Arnold et al., International Publication No. WO 89/02439; Usman et al., International Publication No. WO 95/06731; Dudycz et al., International Publication No. WO 95/11910 and Ferentz and Verdine, J. Am. Chem. Soc. 1991, 113:4000, all hereby incorporated by reference herein. A “non-nucleotide” further means any group or compound that can be incorporated into a nucleic acid chain in the place of one or more nucleotide units, including either sugar and/or phosphate substitutions, and allows the remaining bases to exhibit their enzymatic activity. The group or compound can be abasic in that it does not contain a commonly recognized nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine. Thus, in a preferred embodiment, the invention features an enzymatic nucleic acid molecule having one or more non-nucleotide moieties, and having enzymatic activity to cleave an RNA or DNA molecule. In another aspect of the invention, nucleic acid molecules that interact with target RNA molecules and down-regulate target genes (e.g., prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR gene) activity are expressed from transcription units inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. Enzymatic nucleic acid molecule or antisense expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. The recombinant vectors capable of expressing the enzymatic nucleic acid molecules or antisense can be delivered as described above, and persist in target cells. Alternatively, viral vectors can be used that provide for transient expression of enzymatic nucleic acid molecules or antisense. Such vectors can be repeatedly administered as necessary. Once expressed, the enzymatic nucleic acid molecules or antisense bind to the target RNA and down-regulate its function or expression. Delivery of enzymatic nucleic acid molecule or antisense expressing vectors can be systemic, such as by intravenous or intramuscular administration, by administration to target cells ex-planted from the patient followed by reintroduction into the patient, or by any other means that would allow for introduction into the desired target cell. Antisense DNA can be expressed via the use of a single stranded DNA intracellular expression vector. By “vectors” is meant any nucleic acid- and/or viral-based technique used to deliver a desired nucleic acid. By “patient” is meant an organism, which is a donor or recipient of explanted cells, or the cells themselves. “Patient” also refers to an organism to which the nucleic acid molecules of the invention can be administered. A patient can be a mammal or mammalian cells. In one embodiment, a patient is a human or human cells. By “enhanced enzymatic activity” is meant to include activity measured in cells and/or in vivo where the activity is a reflection of both the catalytic activity and the stability of the nucleic acid molecules of the invention. In this invention, the product of these properties can be increased in vivo compared to an all RNA enzymatic nucleic acid or all DNA enzyme. In some cases, the activity or stability of the nucleic acid molecule can be decreased (i.e., less than ten-fold), but the overall activity of the nucleic acid molecule is enhanced, in vivo. The nucleic acid molecules of the instant invention, individually, or in combination or in conjunction with other drugs, can be used to treat diseases or conditions discussed above. For example, to treat a disease or condition associated with the levels of prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR, the patient can be treated, or other appropriate cells can be treated, as is evident to those skilled in the art, individually or in combination with one or more drugs under conditions suitable for the treatment. In a further embodiment, the described molecules, such as antisense or enzymatic nucleic acid molecules, can be used in combination with other known treatments to treat conditions or diseases discussed above. For example, the described molecules can be used in combination with one or more known therapeutic agents to treat allergic diseases or conditions, including but not limited to asthma, allergic rhinitis, atopic dermatitis, and/or other allergic or inflammatory diseases and conditions which respond to the modulation of prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), adenosine receptors (AR) such as adenosine receptor A1 (ADORA1), A2a, A2b, and A3, NOGO-A, NOGO-B, NOGO-C, NI-35, NI-220, NI-250, myelin-associated glycoprotein, tenascin-R, NG-2 and/or their corresponding receptors, an IkappaB kinase (IKK) subunit and/or protein kinase PKR expression. By “comprising” is meant including, but not limited to, whatever follows the word “comprising”. Thus, use of the term “comprising” indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present. By “consisting of” is meant including, and limited to, whatever follows the phrase “consisting of”. Thus, the phrase “consisting of” indicates that the listed elements are required or mandatory, and that no other elements may be present. Other features and advantages of the invention will be apparent from the following description of the preferred embodiments thereof, and from the claims. |
Glazing laminates |
There is provided a glazing laminate comprising a polyvinyl butyral core layer having polyvinyl butyral/IR dye layers on each side. The dye containing layers are solvent cast from methanol onto the core layer. The dyes in each layer are different comprising BN 18 type PVB containing either 2 mg Epoline III-125 or 2 mg of Epolite III-57 dyes per grams of resin and dissolved in 10 ml of analytical grade of methanol for casting. The polymer is sandwiched between two glass layers under pressure and heat treatment at 135-140° C. An overall concentration of the dyes of ˜0.1 g/m2 results in an energy absorption over 65%. The heating effect of absorption is reduced by conduction through the polymer to the laminate edges, internal reflection, spherical reradiation and the relative insulative effect of the glass layers. |
1. A laminated glazing element including an optically clear laminated sheet including at least two polymer layers having dispersed therein respective dyes, said layers having between them at least two dyes selected to absorb IR radiation of different or overlapping ranges of frequencies, said ranges including frequencies corresponding to IR intensity peaks in incident solar radiation, said polymer layers being sandwiched between a UV absorbent outer support layer and an insulative inner support layer, said glazing element having at least one edge portion associated with a heat sink selected to disperse heat from said laminated sheet. 2. The laminated glazing element according to claim 1, wherein said polymer layers are selected to have higher thermal conductivity than said inner support layer. 3. The laminated glazing element according to claim 1, wherein said heat sink is configured to disperse heat to either the interior or exterior of a glazed structure from the at least one edge portion. 4. A The laminated glazing element according to claim 1, wherein said heat sink is configured to selectively disperse the heat to the interior or exterior of the glazed structure. 5. The laminated glazing element according to claim 1, wherein said heat sink is a thermal mass configured to radiate and/or convect heat. 6. The laminated glazing element according to claim 1, wherein the IR dyes are selected from aminium, bisammonium and bisimmonium salt dyes. 7. The laminated glazing sheet according to claim 1, wherein said polymer layers are selected from polyvinyl butyral (PVB) polymers. 8. The laminated glazing element according to claim 1, wherein said polymer layers are selected from polyvinyl butryal (PVB) polymers. 9. The laminated glazing element according to claim 1, wherein the inner support layer has a range of colouring dyes. 10. The laminated glazing element according to claim 6, wherein one or more of the IR dyes have graduated wavelengths from 900 to 1700 nanometers. 11. The laminated glazing sheet according to any of the preceding claims, excluding claim 7, wherein said polymers layers are selected from methyl acrylate polymers. 12. The laminated glazing sheet according to any of the preceding claims 1 to 10, excluding claim 7, wherein said polymer layers are selected from polycarbonate polymers. 13. The laminated glazing sheet according to claim 7, wherein a plolyvinyl butral layer is complimented by a liquid polycarbonate polymer containing selected dyes. 14. A laminated glazing sheet including a polymer layer comprising a polyvinyl butyral having a hydroxyl content of at least 18 wt % and at least one aminium salt IR absorbing dye, said at least one dye being selected to absorb IR radiation of different or overlapping ranges of frequencies, said ranges including frequencies corresponding to IR intensity peaks in incident solar radiation, and at least one UV absorbent outer support layer, wherein said polymer layer is formed by forming a mixture of methanolic solution of said polyvinyl butyral having a hydroxyl content of at least 18 wt % and a saturated methanolic solution of said at least one dye, and drying said mixture. 15. A method of forming a glazing laminate including the steps of: (a) solvent casting a polyvinyl butyral film; (b) solvent casting a polyvinyl butyral/dye film to each surface of said polyvinyl butyral film, said polyvinyl butyral/dye films collectively including at least two dyes selected to absorb IR radiation of different or overlapping ranges of frequencies, said ranges including frequencies corresponding to IR intensity peaks in incident solar radiation; and (c) laminating the multilayer film of step (b) between glass sheets under heat and pressure. 16. The laminated glazing element according to claim 7, wherein one or more of the IR dyes have graduated wavelengths from 900 to 1700 nanometers. 17. The laminated glazing element according to claim 8, wherein one or more of the IR dyes have graduated wavelengths from 900 to 1700 nanometers. 18. The laminated glazing element according to claim 9, wherein one or more of the IR dyes have graduated wavelengths from 900 to 1700 nanometers. |
<SOH> BACKGROUND OF THE INVENTION <EOH>This invention relates to glazing laminates. This invention has particular but not exclusive application to glazing laminates for glazing in construction, and for illustrative purposes reference will be made to such application. However, it is to be understood that this invention could be used in other applications, such as automotive glass and the like. A significant contributor to the cost of running commercial buildings or maintenance of comfort in residences is the energy required to provide control of the internal climate of the building. In buildings in hot climates having windows that are exposed to a relatively high flux of solar radiation, or buildings with large areas exposed to solar radiation, a major cost is air cooling by airconditioning. The conventional means of controlling heat passage into a building is to reduce the flux of solar radiation through the glazing by tinting to reduce a broad spectrum of radiation passing through the glass, with or without the provision of an outer partially reflective layer. By these means, a reduction of the total flux effects a partial reduction of the heat transmitted to the interior of the building. Clear window glass has a natural absorption maximum in the UV spectrum. Whilst the UV part of the solar spectrum at the Earth's surface is of high energy, the proportion of the total EM flux in the UV at the earth's surface is quite small due to upper atmospheric absorption. The visible spectrum is not substantially absorbed by clear window glass, but is quite strongly absorbed by tinted glass. The absorbed energy is converted to heat, which is in part spherically radiated in the infrared, and otherwise dispersed by convective transfer and conduction. The infrared component of the incident light is generally transmitted directly (with refraction) to the interior of the building in addition to the internally directed IR component arising from absorbed visible/UV radiation. In terms of energy density, the IR/visible bands contribute the greatest part if the incident energy density transmitted to the interior of a building through the windows or glass walls. In fact, energy density reaches a maximum at a wavelength of about 600 nm, with about 90% of the energy of solar radiation incident at the earth's surface being of wavelengths between 500 nm and 1750 nm, with several distinct maxima. Where tinting and/or partially reflective films is used, this generally reduces the visible flux and requires stronger lighting, which carries its own heat burden. In the case of tinting alone, the heat loading transmitted is only reduced by the proportion of IR reradiation back out of the building. International patent publication WO97/44690 discloses an optical element comprising a transparent layers comprising one or more passive layers and one or more active layers wherein said passive layers facilitate the transmission of electromagnetic radiation in a substantially unaltered form and the at least one active layers include an active material dispersed through the active layer and having the capacity to intercept electromagnetic radiation of a wavelength or range of wavelengths and redirect some of the energy of the intercepted radiation into the interior of the optical element, the layers being in face to face relationship and being optically coupled to each other. The described embodiments use a luminophore as the active material to absorb IR in the active layer and to spherically reemit IR by luminescent decay of the luminophore from the excited state. The basic concept of causing dissipation of IR by absorbance dye in a polymer film was shown to work, but no practical polymer film was produced. |
<SOH> SUMMARY OF THE INVENTION <EOH>This invention in one aspect resides broadly in a laminated glazing sheet including at least one polymer layer having dispersed therein at least two dyes selected to absorb IR radiation of different or overlapping ranges of frequencies, said ranges including frequencies corresponding to IR intensity peaks in incident solar radiation, and at least one UV absorbent outer support layer. The at least two dyes may be dispersed in one, both or more of the polymer layers depending on the number of layers, choice of polymers and compatibility of the dyes in polyemer dispersion in practice. The polymer layer may include a tint such as by means of a dye absorbing in the visible band. A practical difficulty experienced with such constructs is that suitable dyes tend to be barely compatible with the polymer matrices in which they are dispersed, and may be reactive with the IR dyes. The problems range from lack of homogeneity in monolayer films, films changed colour and destroyed the IR properties, and when laminated in two layers to keep the dyes separate, the panels were blotchy, patchy, uneven in colour and torn. Accordingly, in a further aspect this invention resides in a laminated glazing sheet including at least two polymer layers having dispersed therein respective dyes, said layers having between them at least two dyes selected to absorb IR radiation of different or overlapping ranges of frequencies, said ranges including frequencies corresponding to IR intensity peaks in incident solar radiation, at least one other of said layers including a dye absorbing in the visible spectrum, a polymer interlayer between adjacent dye bearing ones of said polymer layers, and at least one UV absorbent outer support layer. The absorbed IR radiation will be for the most part dispersed in the laminate as heat. In order to minimize the heat transmitted to the interior of the building, the heat is preferably dispersed from the laminate to reduce heat build-up and the black body effect to the interior of a glazed structure. Accordingly, in a yet further aspect the present invention resides broadly in a laminated glazing element including an optically clear laminated sheet including at least two polymer layers having dispersed therein respective dyes, said layers having between them at least two dyes selected to absorb IR radiation of different or overlapping ranges of frequencies, said ranges including frequencies corresponding to IR intensity peaks in incident solar radiation, said polymer layers being sandwiched between a UV absorbent outer support layer and an insulative inner support layer, said glazing element having at least one edge portion associated with a heat sink selected to disperse heat from said laminated sheet. The inner support layer is of an insulative material to reduce heat transfer to the interior of a glazed structure by convection. The at least one polymer layer may be selected from polymers that have a higher then usual thermal conductivity, and that the heat sink be configured to disperse heat to the outside of the glazed structure from the at least one edge thereof. For example, the heat sink may be a thermal mass being configured to either or both of radiate and convect heat to the exterior. Alternatively, the heat sink may be configured to selectively disperse the heat to the interior or exterior of the glazed structure. By this means, in winter the filtered IR may be used to heat the building, whilst in summer the trapped heat is disposed to the exterior. In one embodiment the heat sink is the atmosphere per se, rendered useful by installing the laminate having the at least one edge exposed. The relative refractive indices of the inner and outer support layers and the at least one polymer layer may be selected to maximize the amount of black body radiation which is internally reflected to the edges of the laminate. |
Metal sheet coated with thermoplastic resin and can obtained therefrom |
A thermoplastic-resin-coated metal sheet in which the thermoplastic resin has been applied to the metal sheet without through an adhesive primer and which, even after having been formed into a can, is excellent in impact resistance at low temperatures and resistance to corrosion by acid contents; and a can obtained from the coated metal sheet. The thermoplastic-resin-coated metal sheet is obtained by coating a metal sheet with a thermoplastic resin composition comprising a blend of a polyester resin with a polyolefin resin or polyolefin elastomer. This metal sheet is formed into a can through drawing/ironing with thickness reduction. |
1. A thermoplastic resin-coated metal sheet in which a thermoplastic resin composition comprising a blend of polybutylene terephthalate and a polyolefin ingredient comprising at least one member selected from the group consisting of polyolefin resins and polyolefin elastomers is coated in a substantially non-orientation state on at least one surface of a metal sheet. 2. A thermoplastic resin-coated metal sheet according to claim 1, wherein a polyolefin resin is used as the polyolefin ingredient. 3. A thermoplastic resin-coated metal sheet according to claim 1, wherein a polyolefin elastomer is used as the polyolefin ingredient. 4. A thermoplastic resin-coated metal sheet according to claim 1, wherein a polyolefin resin and a polyolefin elastomer are used as the polyolefin ingredient. 5. A thermoplastic resin-coated metal sheet according to any one of claim 1, wherein the ratio of a melt index of the polyolefin ingredient upon heat melting (VPOL) to that of the polybutylene terephthalate upon heat melting (VPES), that is, VPOL/VPES is 1.2 or less in the thermoplastic resin composition. 6. A thermoplastic resin-coated metal sheet according to claim 1, wherein a polybutylene terephthalate layer formed by coating the polybutylene terephthalate in a substantially non-orientation state is formed on the coating layer of the thermoplastic resin composition. 7. A thermoplastic resin-coated metal sheet according to claim 1, wherein a polybutylene terephthalate layer coating in which the polybutylene terephthalate is in a substantially non oriented state is formed on the surface of the metal sheet, and a coating layer of the thermoplastic resin composition is formed on the polyester resin layer. 8. A thermoplastic resin-coated metal sheet according to claim 1 wherein the polyolefin resin is a resin comprising one or more of 1-alkene polymer resins having a number or carbon atoms of 2 to 8. 9. A thermoplastic resin-coated metal sheet according to claim 8, wherein the 1-alkene polymer resin is one of polyethylene, polypropylene, and an ethylene-propylene copolymer. 10. A thermoplastic resin-coated metal sheet according to claim 8, wherein the polyolefin resin is a polyolefin resin polymerized by means of a metallocene catalyst. 11. A thermoplastic resin-coated metal sheet according to claim 1, wherein at least a portion of the polyolefin resin is a modified polyolefin resin modified with one of maleic acid anhydride, acrylic acid, acrylic acid ester and diglycidyl methacrylate. 12. A thermoplastic resin-coated metal sheet according to claim 1 wherein the polyolefin elastomer is an in-plant produced ethylene-propylene copolymerized elastomer having a melt flow rate (MFR, 230° C.) of 0.4 to 30 g/10 min). 13-15. (canceled) 16. A thermoplastic resin-coated metal sheet according to claims 1 to 12, wherein the thermoplastic resin composition contains 1 to 30% by weight of a polyolefin ingredient. 17. A thermoplastic resin-coated metal sheet according to claim 1, wherein the thermoplastic resin composition contains 70 to 95% by weight of the polybutylene terephthalate. 18. A thermoplastic resin-coated metal sheet according to claim 1, wherein the thermoplastic resin composition is heat melted and directly extruded from a T-die to a metal sheet for coating. 19. A thermoplastic resin-coated metal sheet according to claim 1, wherein the thermoplastic resin composition is heat melted, extruded from a T-die onto a casting roll, cooled to solidify into a film and then the film is hot press bonded on a metal sheet. 20. A thermoplastic resin-coated metal sheet according to claim 18, wherein, wherein the polyolefin ingredient in the coating layer comprising a thermoplastic resin composition is present being dispersed with a size of 1 to 10 μm in the extruding direction and 0.1 to 2 μm in the direction perpendicular to the extruding direction. 21. A thermoplastic resin-coated metal sheet according to claim 1, wherein the metal sheet is one of electrolytic chromic acid treated steel sheet, tin-plated sheet and aluminum alloy sheet. 22. A can using a thermoplastic resin-coated metal sheet according to claim 1. |
<SOH> BACKGROUND ART <EOH>In recent years, cans formed by coating a polyester resin on a metal sheet and improved with thickness reduction ratio for side wall portions by applying severe forming such as thickness-reducing drawing or thickness-reducing ironing have been used as cans mainly for drink applications. In a case of forming a polyester resin coated metal sheet by applying severe such as wall reducing drawing or wall reducing ironing, it is necessary to coat in a non-orientation state capable of attaining excellent fabricability such that the resin is not peeled upon forming or cracks are not formed to the resin. However, when applying thickness-reducing drawing or thickness-reducing ironing is applied to a resin-coated metal sheet with a non-orientation state of a polyester resin, then applying coating printing to the outer surface of the can and heating for baking, since the upper portion of the can is thermally set in a state where it is molecularly oriented in the direction of a height of the can by the fabrication, elongation of the not oriented resin in the circumferential direction of the can is extremely small to become brittle. Particularly, cracks tend to be formed to the resin layer by mere collision between cans to each other, particularly, at low temperature. Further, since the portion for the can bottom scarcely undergoes the forming, the resin crystals are grown into a coarse state and brittled upon heating for baking after the coating printing also tending to cause cracks upon receiving impacts particularly at low temperature. That is, a polyester resin-coated metal sheet in a non-orientation state is poor in the impact resistance, particularly, the impact resistance at low temperature after formed into the can. Poor impact resistance after forming can be improved by interposing an adhesive primer between the polyester resin and the metal sheet upon coating the polyester resin on the metal sheet. However, a method of interposing the adhesive primer may possibly cause undesired effects on the environment due to evaporation of organic solvents and, further, requires surplus steps of coating and drying to increase the cost. Further, in a case where acidic contents at pH of 5 or less are filled, the underlying metal sheet may sometimes be corroded in the upper portion of the can undergoing high degree of fabrication after lapse of a long time. The present invention intends to provide a thermoplastic resin-coated metal sheet in which a resin layer, after applying severe forming to a thermoplastic resin-coated metal sheet where the resin layer after coating to the metal sheet is in a non-oriented state, is excellent in impact resistance, particularly, impact resistance at low temperature, coated to the metal sheet without interposing an adhesive primer and is excellent in corrosion resistance to acidic contents even in a case where it is formed into a can, as well as a can using the same. |
Diagnosis and treatment of blood disorders |
Based on the discovery of the nucleotide and amino acid differences which distinguish the Gova and Govb allelic forms of the membrane glycoprotein CD109, and which comprise the biallelic Gov platelet alloantigen system, compositions and methods are provided for determining the Gov genotype and phenotype of individuals. Also provided, on the basis of this discovery, are compositions and methods for treating disorders associated with Gov alloantigen incompatibility, such as the bleeding disorders post-transfusion purpura, post-transfusion platelet refractoriness, and neonatal alloimmune thrombocytopenia. The two allelic forms of CD109 differ by a single amino acid. The Gova allelic form has Tyr at amino acid position 703 in the CD109 sequence. The Govb allelic form has Ser at the same position. This amino acid difference is due to a single change, from A for the Gova allele to C for the Govb allele, in the CD109 gene. |
1. An oligonucleotide comprising a sequence which binds specifically to (i) a region of CD109 nucleic acid that includes a single nucleotide polymorphism that is distinctive of a Gova allele and/or (ii) a region of CD109 nucleic acid that includes a single nucleotide polymorphism that is distinctive of a Govb allele. 2. The oligonucleotide of claim 1, comprising 8 to 50 nucleotides. 3. The oligonucleotide of claim 1, wherein the nucleic acid specifically binds to one of (i) or (ii) under high stringency hybridization conditions. 4. The oligonucleotide of claim 2, wherein the stringent hybridization conditions comprise 0.1×SSC, 0.1% SDS at 65° C. 5. The oligonucleotide of claim 1, wherein the CD109 nucleic acid comprises genomic DNA, cDNA, or RNA corresponding to the Gova allele of the CD109 gene or locus, or comprises genomic DNA, cDNA, or RNA corresponding to the Govb allele of the CD109 gene or locus. 6. The oligonucleotide of claim 5, wherein the Gova allele comprises an A at a position corresponding to position 2108 of SEQ ID NO:1 and corresponding to position 954 of SEQ ID No:5. 7. The oligonucleotide of claim 5, wherein the Govb allele comprises a C at a position corresponding to position 2108 of SEQ ID NO:3 and corresponding to position 954 of SEQ ID NO:5. 8. The oligonucleotide of claim 1, comprising a sequence complementary to the Gova allele or to the Govb allele. 9. The oligonucleotide of claim 6, comprising a sequence selected from the group consisting of: (a) 8-50 nucleotides of SEQ ID NO:1; (b) a sequence that is complementary to a sequence specified in (a); and (c) a sequence having at least 70% sequence identity to a sequence in (a) or (b), wherein the sequence having identity is capable of hybridization to CD109 under high stringency hybridization conditions. 10. The oligonucleotide of claim 7, comprising a sequence selected from the group consisting of: (a) 8-50 nucleotides of SEQ ID NO:3; (b) a sequence that is complementary to a sequence specified in (a); and (c) a sequence having at least 70% sequence identity to a sequence in (a) or (b), wherein the sequence having identity is capable of hybridization to CD109 under high stringency hybridization conditions. 11. A oligonucleotide comprising all or part of any one of SEQ ID NO:6-SEQ ID NO:14 or a complement thereof. 12. The oligonucleotide of claim 11, comprising 8 to 50 nucleic acids. 13. The oligonucleotide of claim 1, wherein the nucleic acid is capable of use as a probe in a hybridization assay. 14. The oligonucleotide of claim 1, wherein the nucleic acid sequence is detectably labelled. 15. The oligonucleotide of claim 14, wherein the detectable label comprises: (a) a fluorogenic dye; and/or (b) a biotinylation modification; and/or (c) a radiolabel. 16. The oligonucleotide of claim 1, wherein the sequence comprises DNA, a DNA analog, RNA or an RNA analog. 17. The oligonucleotide of claim 1, wherein the oligonucleotide is attached to a substrate. 18. The oligonucleotide of claim 1, wherein the oligonucleotide is capable of use as a primer that will specifically bind proximate to, and/or cause elongation through, a CD109 sequence, including the single nucleotide polymorphism distinctive of the Gova or Govb alleles. 19. A Gov genotyping kit comprising a detection agent for detecting the presence of a Gov allele-specific target sequence in a CD109 nucleic acid derived from a subject. 20. The kit of claim 19, wherein the detection agent comprises a nucleic acid and/or a restriction enzyme. 21. The kit of claim 19, further comprising a container. 22. The kit of claim 21, wherein the container comprises a biological sample container for housing the detection agent. 23. The kit of claim 19, further comprising a plate having a plurality of wells and having bound thereto probes having a nucleic acid sequence which specifically binds to a CD109 sequence including a Gova or a Govb allele target sequence. 24. The kit of claim 20, wherein the restriction enzyme is selected from the group consisting of Bst2UI, BstNI, BstOI, EcoRII, MaeIII, MspR91, MvaI, ScrFI or an isoschizomer thereof. 25. The kit of claim 19, further comprising an amplification agent for amplifying the nucleic acid. 26. The kit of claim 25, wherein the amplification agent amplifies a region of CD109 platelet, T cell, or endothelial cell mRNA including the single nucleotide polymorphism distinctive of a Gova or Govb allele. 27. The kit of claim 25, wherein the amplification agent comprises a primer set including first and second primers, wherein the first primer is a nucleic acid that will specifically bind proximate to, and/or cause elongation through, CD109 sequence that includes the single nucleotide polymorphism distinctive of a Gova allele and the second primer is a nucleic acid that will specifically bind proximate to, and/or cause elongation through, CD109 sequence that includes the single nucleotide polymorphism distinctive of a Govb allele. 28. The kit of claim 19, wherein the nucleic acid is obtained by amplification with all or part of the nucleic acid of any one of SEQ ID NO:6- SEQ ID NO:14 or the complement thereof. 29. The kit of claim 19, further comprising all or part of a CD109 gene, a CD109-encoding mRNA, or a CD109 cDNA made from a CD109-encoding mRNA. 30. The kit of claim 19, comprising the oligonucleotide of any of claims 1 to 18. 31. The kit of claim 19, for detecting that the subject has or is at risk of a disease, disorder or abnormal physical state. 32. The kit of claim 31, wherein the disease, disorder or abnormal physical state comprises a blood disease, disorder or abnormal physical state. 33. The kit of claim 32, wherein the blood disease, disorder or abnormal physical state comprises bleeding of the subject, or increased risk of bleeding, due to destruction of blood platelets. 34. The kit of claim 33, wherein the blood disease, disorder or abnormal physical state comprises post-transfusion purpura (“PTP”), post-transfusion platelet refractoriness (“PR”) or neonatal alloimmune thrombocytopenia (“NAIT”). 35. The kit of claim 33, wherein the nucleic acid is obtained from mRNA from human platelets, T cells, endothelial cells, or human genomic DNA. 36. A method of Gov alloantigen genotyping a subject comprising: (a) providing a CD109 nucleic acid sample derived from the subject; and (b) detecting a region of CD109 nucleic acid that includes a single nucleotide polymorphism distinctive of a Gova or a Govb allele. 37. The method of claim 36 comprising determining whether the subject is homozygous or heterozygous for the Gov alleles. 38. The method of claim 37, wherein the subject is a human and the Gov genotype is used to determine that the subject has, or is at risk of a disease, disorder or abnormal physical state. 39. The method of claim 38, wherein the disease, disorder or abnormal physical state comprises a blood disease, disorder or abnormal physical state. 40. The method of claim 39, wherein the blood disease, disorder or abnormal physical state comprises bleeding of the subject, or increased risk of bleeding, due to destruction of blood platelets. 41. The method of claim 40, wherein the blood disease, disorder or abnormal physical state comprises post-transfusion purpura (“PTP”), post-transfusion platelet refractoriness (“PR”) or neonatal alloimmune thrombocytopenia (“NAIT”). 42. The method of claim 41, wherein the nucleic acid is obtained by amplifying the nucleic acid from the subject. 43. The method of claim 42, wherein the nucleic acid is obtained by amplification with all or part of the oligonucleotide of any of claims 1 to 18. 44. The method of claim 41, wherein the nucleic acid is obtained from mRNA from human platelets, T cells, endothelial cells, or human genomic DNA. 45. The method of claim 36, wherein the detection step comprises determining the nucleotide sequence of the CD109 nucleic acid. 46. The method of clam 36, wherein the detection step comprises contacting the nucleic acid with the oligonucleotide of any of claims 1 to 18 under high stringency conditions. 47. The method of claim 46, wherein the oligonucleotide will selectively hybridize to (i) a region of CD109 nucleic acid that includes a single polymorphism distinctive of a Gova allele or (ii) a region of CD109 nucleic acid that includes a single polymorphism distinctive of a Govb allele. 48. The method of claim 36, wherein the detecting step comprises: (a) performing a restriction endonuclease digestion of the nucleic acid, thereby providing a nucleic acid digest; and (b) contacting the digest with the oligonucleotide. 49. The method of claim 47, wherein the hybridization occurs either during or subsequent to PCR amplification and the analysis is by “Real-Time” PCR analysis, or fluorimetric analysis. 50. The method of claim 36, wherein the detection step comprises: (a) incubation of the amplified nucleic acid with a restriction endonuclease under conditions whereby the DNA will be cleaved if the nucleic acid comprises a recognition site for the enzyme; and (b) determining whether the nucleic acid contains a recognition site for the restriction enzyme characteristic of cDNA made from mRNA encoding a Gova or Govb allele of CD109. 51. The method of claim 50, wherein the restriction enzyme is selected from the group consisting of Bst2UI, BstNI, BstOI, EcoRII, MaeIII, MspR91, MvaI, ScrFI or an isoschizomer thereof. 52. The method of claim 50, wherein the determination step includes size analysis of the nucleic acid. 53. The method of claim 50, wherein the amplified nucleic acid is analyzed by electrophoretic mobility and the mobility of the amplified nucleic acid is compared to the characteristic mobility of amplified nucleic acid fragments corresponding to the Gova or Govb alleles of CD109. 54. A method of amplifying CD109 mRNA comprising amplifying the mRNA by PCR using an oligonucleotide of claim 1. 55. A Gova specific antibody. 56. The antibody of claim 55, that recognizes specifically a Gova allele-specific CD109 epitope corresponding to the polypeptide encoded by a CD109 nucleic acid containing an A at the position corresponding to position 2108 of SEQ ID NO: and position 954 of SEQ ID NO:5., and containing the amino acid Tyrosine at the position corresponding to position 703 of the CD109 protein encoded by SEQ ID NO:1. 57. A Govb specific antibody. 58. The antibody of claim 57, that recognizes specifically a Govb allele-specific CD109 epitope corresponding to the polypeptide encoded by a CD109 nucleic acid containing a C at the position corresponding to position 2108 of SEQ ID NO:3 and position 954 of SEQ ID NO:5., and containing the amino acid Serine at the position corresponding to position 703 of the CD109 protein encoded by SEQ ID NO:3. 59. The antibody of claim 55, further comprising a monoclonal antibody or a polyclonal antibody. 60. The antibody of claim 55, further comprising a detectable label. 61. An immunogenic composition comprising a Gov specific antibody of claim 55. 62. A method of Gov alloantigen phenotyping a subject, comprising: (a) providing a CD109 polypeptide sample derived from the subject; and (b) detecting the presence of a Gova or a Govb antigen in the CD109 polypeptide. 63. The method of claim 62, wherein the CD109 is membrane bound CD109 or isolated CD109. 64. The method of claim 62, wherein the detection step comprises contacting the polypeptide sample with the antibody of claim 55. 65. A diagnostic kit for Gov alloantigen phenotyping a subject, comprising a Gova antibody and/or a Govb antibody of claim 55. 66. The kit of claim 65, further comprising a container. 67. An isolated polypeptide containing Gova allele-specific amino acid sequence and which is specifically reactive with a Gova antibody. 68. An isolated polypeptide containing Govb allele-specific amino acid sequence and which is specifically reactive with a Govb antibody. 69. The isolated polypeptide of claim 67, comprising between 4 and 100 amino acids. 70. The isolated polypeptide of claim 67, comprising a full-length CD109 polypeptide, or a fragment of a CD109 polypeptide. 71. An isolated CD109 polypeptide fragment, comprising a Gova or a Govb antigen. 72. The polypeptide fragment of claim 71, comprising all of, or a fragment of, the protein encoded by SEQ ID NO:1., and in which the amino acid corresponding to position 703 of the protein encoded by SEQ ID NO:1 is a Tyrosine. 73. The polypeptide fragment of claim 72, comprising all of, or a fragment of, the protein encoded by SEQ ID NO:3., and in which the amino acid corresponding to position 703 of the protein encoded by SEQ ID NO:3 is a Serine. 74. The polypeptide fragment of claim 72, comprising between 4 and 100 amino acids. 75. The polypeptide fragment of claim 74, comprising between 7 and 50 amino acids. 76. The polypeptide fragment of claim 67, in which the polypeptide is purified from native CD109, is synthetic, or is prepared by recombinant means. 77. The polypeptide fragment of claim 67, in which the polypeptide is bound to a substrate. 78. A fusion compound comprising the polypeptide of any of claim 67 connected to an immunogenic carrier. 79. The fusion compound of claim 78, wherein the immunogenic carrier comprises a proteinaceous carrier. 80. The fusion compound of claim 79, wherein the immunogenic carrier comprises a detectable label. 81. A Gova or Govb specific antibody recognizing the fusion compound of claim 80. 82. An immunogenic composition comprising the polypeptide, polypeptide fragment or fusion compound of claim 67. 83. A method of producing a Gova or Govb specific antibody, comprising contacting an animal with the immunogenic composition of claim 82, so that the animal produces antibodies against the immunogenic composition. 84. The method of claim 83, wherein the animal is a bird or a mammal. 85. A method of screening an antibody producing culture to determine whether the culture produces Gova or Govb specific antibody, comprising: (a) contacting a polypeptide of the invention with the culture; and (b) detecting Gova or Govb specific antibody. 86. The method of claim 85, wherein the polypeptide comprises a detectable label. 87. The method of claim 86, wherein the polypeptide is attached to a substrate. 88. A method of purifying a Gov allele-specific antibody from a sample, comprising: (a) contacting a Gov allele-specific antibody with a polypeptide of the invention comprising a Gova or Govb antigen, so that an antibody:polypeptide complex is formed; (b) separating the complex from the sample; and (c) next separating the antibody from the polypeptide. 89. The method of claim 88, wherein the polypeptide is bound to a substrate. 90. The method of claim 89, wherein the polypeptide comprises a detectable label. 91. A method of purifying a Gov polypeptide from a sample, comprising: (a) contacting a Gov allele-specific antibody with a polypeptide of the invention containing a Gova or Govb-specific epitope, so that an antibody: polypeptide complex is formed; (b) separating the complex from the sample; and (c) next separating the antibody from the polypeptide. 92. The method of claim 91, wherein the antibody is bound to a substrate. 93. The method of claim 92, wherein the antibody comprises a detectable label. 94. A method of screening a subject sample to determine whether the sample contains Gova or Govb-specific antibodies, comprising: (a) contacting a polypeptide of the invention with the sample; and (b) detecting the presence or absence of Gova or Govb specific antibody. 95. The method of claim 94, wherein the polypeptide comprises a detectable label. 96. The method of claim 95, wherein the polypeptide is attached to a substrate. 97. The method of claim 96, wherein the subject comprises a mother of a fetus or a newborn infant, or the fetus or newborn infant itself, and the presence of Gova or Govb-specific antibody indicates that the fetus or infant has, or is at risk of NAIT. 98. The method of claim 97, wherein the presence of Gova or Govb specific antibody indicates that the subject has, or is at risk of a blood disease, disorder or abnormal physical state. 99. The method of claim 98, wherein the blood disease, disorder or abnormal physical state comprises bleeding of the subject, or increased risk of bleeding, due to destruction of blood platelets. 100. The method of claim 99, wherein the blood disease, disorder or abnormal physical state comprises post-transfusion purpura (“PTP”), post-transfusion platelet refractoriness (“PR”) or neonatal alloimmune thrombocytopenia (NAIT). 101. The method of claim 100, wherein the sample comprises human serum or plasma. 102. A diagnostic kit for detection of Gova or Govb specific antibody, comprising a polypeptide of claim 67. 103. The kit of claim 102, further comprising a container. 104. A method of determining Gov antibody specificity, comprising: (a) contacting an antibody with a first polypeptide comprising a Gova antigen and a second polypeptide comprising a Govb antigen; and (b) determining whether the antibody binds to either or both of the first and second polypeptide. 105. A method of blocking Gova antibody binding to an antigen, comprising: contacting the antibody with a polypeptide of the invention comprising a Gova antigen so that an antibody:polypeptide complex is formed. 106. The method of claim 105, wherein the polypeptide comprises a detectable label. 107. The method of claim 106, wherein the polypeptide is bound to a substrate. 108. A method of blocking Govb antibody binding to an antigen, comprising: contacting the antibody with a polypeptide of the invention comprising a Govb antigen so that an antibody:polypeptide complex is formed. 109. The method of claim 108, wherein the polypeptide comprises a detectable label. 110. The method of claim 109, wherein the polypeptide is bound to a substrate. 111. A pharmaceutical composition comprising the polypeptide of claim 67. 112. A method of immunizing a subject so that the subject will produce anti-idiotypic antibodies, comprising administering to the subject the immunogenic composition of claim 82. 113. A method of blocking Gova or Govb specific antibodies from binding to CD109 in a subject, comprising: administering to the subject polypeptides of the invention capable of binding to Gova and/or Govb specific antibodies. 114. The method of claim 113, wherein the polypeptide comprises a detectable label. 115. The method of claim 114, wherein the binding of the polypeptide to the Gova or Govb specific antibody prevents alloimmune cell destruction by the antibody. 116. The method of claim 115, wherein the binding of the polypeptide to the Gova or Govb-specific antibody depletes the antibody. 117. The method of claim 116, wherein the subject has or is at risk of a blood disease, disorder or abnormal physical state. 118. The method of claim 117, wherein the blood disease, disorder or abnormal physical state comprises bleeding of the subject, or increased risk of bleeding, due to alloimmune destruction of blood platelets. 119. The method of claim 118, wherein the blood disease, disorder or abnormal physical state comprises post-transfusion purpura (“PTP”), post-transfusion platelet refractoriness (“PR”) or neonatal alloimmune thrombocytopenia (“NAIT”). |
<SOH> BACKGROUND OF THE INVENTION <EOH>Among the disorders, which the invention concerns, are those involving abnormal and excessive bleeding due to destruction of blood platelets (“platelets”). These disorders include, but are not restricted to, post-transfusion purpura (“PTP”) and post-transfusion platelet refractoriness (“PTPR”), which are suffered by some persons who receive blood, platelets, leukocyte concentrates, or plasma from other persons by transfusion or the like. The disorders also include one that is suffered by fetuses and newborns and is known as “neonatal alloimmune thrombocytopenia” (“NATP”). This disorder can cause death of fetuses and serious birth defects or death of newborns. NATP is estimated to affect about 1 in 1000 newborns. In NATP, fetal platelets, which enter the mother's blood stream, induce production in the mother of antibodies directed against fetal platelets. These maternal antibodies then pass with the mother's blood into the fetus and mediate destruction of platelets in the fetus. A mother, whose fetus or newborn suffers from NATP, is at increased risk of suffering PTP or PTPR. When platelets from a first human (a “donor”) are introduced into the blood system of a second human (a “recipient”) by transfusion, through the placenta (in the case of fetal blood entering the mother), or the like, the recipient may mount an immune response against the platelets from the donor. Such an immune response is referred to as an “alloimmune” response, because it involves antibodies reacting-against antigens of a different individual of the same species. The alloimmune response to platelets is due to an immune response of the recipient against “alloantigens” (antigens of the same species as that mounting the immune response) on platelets from the donor. These alloantigens are found on membrane glycoproteins that occur in the cell membranes, which define the outer surfaces of platelets (“platelet membranes”). In this invention, the glycoprotein is anchored to the membrane in an atypical manner through an anchor consisting of glycosylphosphatidylinositol (GPI), which anchors an extracellular domain or segment of the glycoprotein exposed to the outside of the platelet. It is thought that alloantibodies, which are generated in an alloimmune response against platelet alloantigens, interact with the extracellular domains of the alloantigens. The platelet alloantigens that a person has are determined by the person's genetics. A donor, because of his or her genetics, may have a platelet alloantigen, which a recipient, who receives blood, platelets, leukocytes or plasma from the donor, does not have, because of the recipient's genetics. In such a situation, the immune system of the recipient may recognize the donor's alloantigen as “non-self,” and raise an immune response against, the platelet alloantigen, which the donor has but the recipient does not. Membrane glycoprotein alloantigens have been characterised for both human red blood cells and human platelets. It is noteworthy, however, that they also occur on other cell types, such as leukocytes and endothelial cells, where they may also occasion various disorders through alloimmune responses. Recognised classes of red blood cell and platelet alloantigens have been described, over the past 30 years, based on observations of antibody reactions occurring when blood recipients have been exposed to blood from donors. A recent review of human platelet alloantigen systems is provided by Ouwehand, W., and Navarrete, C., in Molecular Haematology , Provan, D. and Gribben, J. eds. Blackwell (1999). Several biallelic platelet alloantigen “systems” have been characterised. In each of these systems, there are two alloantigens, each of which is provided by one of two alleles of the gene comprising the system. Because each gene occurs twice in the normal human genome, a person can be homozygous for one or the other of the two alloantigens, or heterozygous for the two alloantigens, comprising a biallelic system. The alloantigens described to date occur on glycoprotein molecules which may exist in various forms (transmembrane, GPI-linked and soluble, for example). In such a case, the alloantigens are found on each of the variant forms of the glycoprotein. For all of the biallelic platelet alloantigen systems that have been characterised at the level of protein and gene sequences, it has been found in all cases, except for one, that the difference between the two alleles is based on a single nucleotide polymorphism in the relevant gene. One biallelic system of human platelet alloantigens is the Gov a /Gov b biallelic system associated with CD109, a membrane glycoprotein which occurs on platelets and various other cell types, including leukocytes and endothelial cells. Each Gov allele corresponds to one CD109 glycoprotein (Sutherland, D. R. et al, 1991; Smith et al., 1995; Berry, J. et al., 2000), consistent with the known tissue distribution of CD109. The frequencies for the Gov alleles are 0.4 for Gov a and 0.6 for Gov b in the Caucasian population. Thus, in this population, 40.7% are heterozygous for the Gov alleles, and will not mount an alloimmune response due to Gov incompatibility (not possessing the Gov alloantigen found on platelets received from another). In contrast, 19.8% of Caucasians are homozygous for the Gov a allele and thus may mount an immune response due to Gov alloantigen incompatibility against platelets received from anyone in the 80.5% of the Caucasian population that is not homozygous for the Gov a allele, while 39.8% are homozygous for the Gov b allele and thus may mount an immune response due to Gov alloantigen incompatibility against platelets received from anyone in the 60.2% of the Caucasian population that is not homozygous for the Gov b allele. As indicated above, alloimmunization based on Gov incompatibility (the introduction into the blood stream of donor platelets bearing a Gov alloantigen not carried by the recipient) can result in bleeding disorders due to platelet destruction, including NATP, PTPR, and PTP. The location of the Gov antigens within the CD109 molecule, and the nature of the CD109 polymorphism which underlies the Gov a /Gov b alloantigen (both at the protein and at the gene level), have not heretofore been known. Furthermore, it has not heretofore been possible to generate non-human antibody (polyclonal or monoclonal), as from a rat, mouse, goat, chicken, or the like, with specificity for the Gov a alloantigen but not the Gov b alloantigen (or vice-versa) sufficient for use in an immunoassay, for typing for Gov phenotype using platelets or CD109 molecules. Previously developed technology, involving gene-specific amplification of platelet RNA-derived cDNA, followed by the determination of the nucleotide sequence of the amplified DNA, has been applied successfully to the elucidation of the molecular basis of other biallelic platelet alloantigen systems (Newman et al., J. Clin. Invest. 82,739-744 (1988); Newman et al., J. Clin. Invest. 83, 1778-1781 (1989)(P1A or HPA-1 system); Lyman et al., Blood 75, 2343-2348 (1990)(Bak or HPA-3system); Kuijpers et al., J. Clin. Invest. 89, 381-384 (1992)(HPA-2 or Ko system); Wang et al., J. Clin. Invest. 90, 2038-2043 (1992)(Pen system). With one exception, it has been found in each case that a single amino acid difference at a single position differentiates the amino acid sequences of the two alleles, and that this difference arises from a single allele-specific nucleotide substitution in the coding region of the mRNA and gene. There remains a need to elucidate the molecular basis of the biallelic Gov platelet alloantigen system. |
<SOH> SUMMARY OF THE INVENTION <EOH>The Gov a/Gov b CD109 Single Nucleotide Polymorphism We have now discovered that a single amino acid difference in the CD109 glycoprotein distinguishes the Gov a and Gov b allelic forms. The two alleles differ at amino acid position 703 of the full-length 1445 amino acid CD109 molecule, with the Gov a allele [SEQ ID NO:2] containing a Tyr at this position, while the Gov b allele [SEQ ID NO:4] contains Ser. Further, we have discovered that this difference in amino acid sequence between the allelic forms of CD109 is due to a single nucleotide polymorphism at position 2108 of the coding portion of full-length mRNA encoding CD109, or of the corresponding coding strand of the cDNA corresponding to this mRNA. Specifically, the Gov a allele [SEQ ID NO:1] contains adenine at position 2108, the second nucleotide of the codon encoding the amino acid at position 703 of the full-length CD109 protein, while the Gov b allele contains cytosine at position 2108, as shown in SEQ ID NO:3 The Gov a /Gov b single nucleotide polymorphism of CD109, lies at position 2108 in SEQ ID NO:1. SEQ ID NO:1 is the cDNA sequence encoding the full-length 1445 amino acid CD109 precursor encoding the Gov a alleleln the Gov b allele form [SEQ ID NO:3], C occurs at position 2108, rather than A. The ATG at the 5′-end of the sequence in SEQ ID NO:1 corresponds to the translation start of the full-length precursor form (including leader peptide) of CD109. The triplet corresponding to the N-terminal amino acid of the mature CD109 protein is at positions 64-66 in SEQ ID NO:1. The Gov a /Gov b single nucleotide polymorphism of CD109, lies at position 954 in SEQ ID NO:5. SEQ ID NO:5 is the genomic DNA sequence of human CD109 exon 19 and the contiguous introns, introns 18 and 19. The Gov a /Gov b single nucleotide polymorphism of CD109 is found within CD109 exon 19, and specifically is located at position 3 of CD109 exon 19. The sequence presented in SEQ ID NO:5 contains A at position 954, and thus corresponds to the Gov a allele. The corresponding Gov b sequence contains C at position 954 of SEQ ID NO:5 (nucleotide position 3 of exon 19). In view of this discovery, it will be readily apparent to the skilled what the present invention provides: Gov allele-specific oligonucleotides and polynucleotides: Based on the discovery, the present invention provides oligonucleotides and polynucleotides (seems repetitive), including (but not limited to) probes which can be used to determine whether a person is homozygous for one or the other of the Gov alleles, or heterozygous for these alleles, thereby to determine that person's Gov genotype, and by extension, their Gov phenotype (i.e., the Gov alloantigen(s) which their cells express). Further, the invention provides methods of using such oligonucleotides, and test kits to facilitate their use, in such Gov genotype and phenotype determinations. These oligonucleotides of the invention can be used to determine whether, in the CD109 gene, or in the mRNA encoding CD109, the internal nucleotide (nucleotide 2108) of the codon (in CD109 gene or in the mRNA encoding CD109) which corresponds to the amino acid at position 703 in the sequence of full-length CD109 is adenine or cytosine. Such probes will typically be cDNA but may be genomic DNA, mRNA or RNA, and may be labelled for detection. The oligonucleotides of the invention can be used as probes to detect nucleic acid molecules according to techniques known in the art (for example, see U.S. Pat. Nos. 5,792,851 and 5,851,788). For example, an oligonucleotide of the invention may be converted to a probe by being end-labelled using digoxigenin-11-deoxyuridine triphosphate. Such probes may be detected immunologically using alkaline-phosphate-conjugated polyclonal sheep antidigoxigenin F(ab) fragments and nitro blue tetrazolium with 5-bromo-4-chloro-3-indoyl phosphate as chromogenic substrate. Gov allele-specific antibodies: Still further, based on the discovery, which underlies the invention, of the molecular basis for the Gov a /Gov b alloantigen system, the invention provides non-human polyclonal and monoclonal antibodies, which can be used to distinguish one Gov allelic form of CD109 from the other, whether the CD109 is part of a complex embedded in or isolated from a membrane or is isolated. These antibodies of the invention, which are preferably provided in an aqueous buffer solution, and the immunoassays of the invention which employ such antibodies, are useful for determining whether a person has one or both of the Gov alloantigens and for Gov phenotyping. Methods of using the antibodies of the invention in the immunoassays of the invention, and in such determinations, are also encompassed by the invention. The invention also provides test kits to facilitate carrying out such immunoassays and determinations. Gov allele-specific peptides and polypeptides: Again, based on the discovery that underlies the invention, of the molecular basis for the Gov a /Gov b alloantigen system, the invention provides peptides or polypeptides, which are useful for various purposes. These peptides or polypeptides are typically between 4 and 100, and more typically between 7 and 50, amino acids in length, and have amino acid sequences identical or having sequence identity to those of segments of the CD109 sequences, that include the amino acid at position 703 of full-length mature CD109. This amino acid (position 703) corresponds the triplet at positions 2107-2109 in the CD109 cDNA sequence presented in SEQ ID NO:1, or in the corresponding sequence for the CD109 cDNA that encodes the Gov b allelic form [SEQ ID NO:3]. These peptides or polypeptides may be synthetic, may be purified from native CD109 or may be prepared by recombinant means. For guidance, one may consult the following U.S. Pat. Nos. 5,840,537, 5,850,025, 5,858,719, 5,710,018, 5,792,851, 5,851,788, 5,759,788, 5,840,530, 5,789,202, 5,871,983, 5,821,096, 5,876,991, 5,422,108, 5,612,191, 5,804,693, 5,847,258, 5,880,328, 5,767,369, 5,756,684, 5,750,652, 5,824,864, 5,763,211, 5,767,375, 5,750,848, 5,859,337, 5,563,246, 5,346,815, and WO9713843. Many of these patents also provide guidance with respect to experimental assays, probes and antibodies, methods, transformation of host cells, which are described below. These patents, like all other patents, publications (such as articles and database publications) in this application, are incorporated by reference in their entirety. Gov allele-specific peptides and polypeptides as antigens and immunogens, and Gov allele-specific polyclonal and monoclonal antibodies: These peptides or polypeptides are useful as antigens (usually coupled to a larger, immunogenic carrier [proteinaceous or otherwise], as known in the art) for making the polyclonal or monoclonal antibodies of the invention. The peptides or polypeptides are also useful in screening monoclonal antibody-producing cultures (hybridoma cultures/ E. coli cultures or so-called V gene phage antibodies) to identify those that produce monoclonal antibodies of the invention. The invention also encompasses immunogenic compositions which comprise a peptide, polypeptide or fusion compound of the invention and which are immunogenic in a bird, including, without limitation, a chicken, or a mammal, such as, a mouse, rat, goat, rabbit, guinea pig, sheep or human. The compositions may include an immunogenicity-imparting “carrier” which may be but is not necessarily a protein as known in the art, that is immunogenic in a bird or mammal, coupled to at least one peptide or polypeptide of the invention, which has an amino acid sequence that is the same as that of a segment of the sequence for CD109, that includes the amino acid at position 703 of the full length CD109 molecule. The present invention also provides methods of using the peptides, polypeptides and immunogenic compositions of the invention for making antibodies of the invention, and methods of using the peptides and polypeptides of the invention in screening monoclonal antibody-producing hybridoma cultures or bacterial clones for those that produce monoclonal antibodies or fragments thereof of the invention. Therapeutic and diagnostic application of Gov allele-specific peptides, polypeptides, and antibodies: These peptides or polypeptides, as well as antibodies, which are specific for the Gov a [SEQ ID NO:2] or Gov b [SEQ ID NO:4], but not both, allelic forms of CD109 in the platelet membrane, and which can be produced by a mammal (including an human) immunized with the peptides or polypeptides, which themselves happen to be immunogenic, or the immunogenic compositions of the invention, are also useful both therapeutically and diagnostically. The invention also provides the methods of using the peptides and polypeptides of the invention, and antibodies made using the peptides that are immunogenic and the immunogenic compositions of the invention, in therapeutic and diagnostic applications. The Gov allele-specific peptides or polypeptides can also be used diagnostically to detect the presence of Gov a or Gov b specific antibodies in human plasma or serum samples, using methods that are readily apparent to those skilled in the art. Such analyses would be useful in the investigation of cases of acquired alloimmune thrombocytopenia, including PTP, PTPR, and NATP. In the latter case, this approach could also be used to detect the presence of Gov allele-specific antibodies in the mother of the affected fetus or newborn. The presence of Gov allele-specific antibodies can also be detected using platelets of known Gov phenotype. However, this approach has numerous technical disadvantages that are eliminated by the use of Gov allele-specific peptides or polypeptides for Gov allele-specific antibody detection. Administration to a person, who is suffering from, or at risk for, for example, PTP or PTPR, or a mother at risk for passing NATP-causing alloantibodies to her fetus, of one of the peptides or polypeptides, that would be bound by the anti-Gov alloantibodies in such a person, would inhibit the binding of the alloantibodies to the person's (or the fetus's platelets and thereby inhibit the platelet destruction and abnormal bleeding associated with the disorders. Alternatively, administration to such a person of antibodies (particularly human antibodies), which are produced using a peptide or polypeptide of the invention, which is immunogenic by itself, or an immunogenic composition of the invention, and which are specific for the Gov allelic form of the CD109 on the person's platelets which is associated with the PTP or PTPR, from which the person is suffering or may suffer, would induce the production of anti-idiotypic antibodies, which, in turn, would inhibit the platelet-destructive effects of the anti-Gov alloantibodies, which are generated by the person's own immune system and which are causing or threatening to cause the PTP, PTPR or NATP. These therapeutic applications of peptides and polypeptides of the invention would be especially useful in treating NATP in a newborn, because the alloantibody giving rise to NATP in the newborn is not continuously produced by the immune system of the newborn, but rather is acquired passively, and therefore in limited, non-replenished quantity, by the newborn from its mother. Thus, in accordance with one aspect of the present invention, an oligonucleotideprobe is provided that hybridizes to a portion of the CD109 gene, or a portion of CD109-encoding mRNA or cDNA prepared from such mRNA, which portion includes a nucleotide corresponding to the internal nucleotide of the codon for the amino acid at position 703 of the full-length CD109 molecule, and that is capable of distinguishing one Gov allele from the other through the ability to hybridize under stringent conditions to the portion in question only when the nucleotide in question is A (or dA), when the probe is to detect the Gov a allele, or C (or dC), when the probe is to detect the Gov b allele. The nucleotide in question is at position 2108 of the coding region of the CD109 cDNA sequence and lies at position 2108 in SEQ ID NO:1. The cDNA sequence has A at this position, and so is the sequence corresponding to the Gov a allele. The nucleotide in question lies at position 954 of the sequence presented as SEQ ID NO:5 and contains an A in this position, and thus also corresponds to Gov a allele. The Gov allele-specific oligonucleotide hybridization probes of the invention may comprise genomic DNA, cDNA, or RNA, although preferably it is DNA. Such oligonucleotide probes can be synthesised by automated synthesis and will preferably contain about 10-30 bases, although as understood in the oligonucleotide probe hybridization assay art, as few as 8 and as many as about 50 nucleotides may be useful, depending on the position within the probe where the potential mismatch with the target is located, the extent to which a label on the probe might interfere with hybridization, and the physical conditions (e.g., temperature, pH, ionic strength) under which the hybridization of probe with target is carried out. In accordance with another aspect of the present invention, a test kit for Gov alloantigen typing is provided comprising: (a) means for amplifying nucleic acid that comprises at least a portion of a CD109 gene, a CD109-encoding mRNA, or a CD109 cDNA made from such RNA, wherein the portion includes a nucleotide (nucleotide 2108 in SEQ ID NO:1, or nucleotide 954 in SEQ ID NO:5) corresponding to the internal nucleotide of the codon encoding amino acid 703 of the full length CD109 protein. (b) an oligonucleotide probe of the invention, that distinguishes one Gov allele from the other. The “means for amplifying” will, as the skilled will readily understand, depend on the amplification method to be used. Thus, for example, these means might include suitable primers, a suitable DNA polymerase, and the four 2′-deoxyribonucleoside triphosphates (dA, dC, dG, dT), if amplification is to be by the PCR method. To cite another example, if the amplification is to be by a method relying on transcription, such as the 3SR method, the means will include two primers, at least one of which, when made double-stranded, will provide a promoter, an RNA polymerase capable of transcribing from that promoter, a reverse transcriptase to function in primer-initiated, DNA-directed and RNA-directed, DNA polymerization and possibly also in RNAse H degradation of RNA to free DNA strands from RNA/RNA hybrids, the four ribonucleoside triphosphates (A, C, G and U), and the four 2′-deoxyribonucleoside triphosphates. In another example, if the amplification is by the ligase chain reaction, the means will include two oligonucleotides (DNAs) and a suitable DNA ligase that will join the two if a target, to which both can hybridize adjacent to one another in ligatable orientation, is present. The oligonucleotide probes of the invention will preferably be labelled. The label may be any of the various labels available in the art for such probes, including, but not limited to 32 P; 35 S; biotin (to which a signal generating moiety, bound to or complexed with avidin can be complexed); a fluorescent moiety; an enzyme such as alkaline phosphatase (which is capable of catalysing a chromogenic reaction); digoxigenin, as described above; or the like. As indicated in the examples, RFLP analysis can be employed, using BstNI (or isoschizomers thereof), in analysing cDNA or genomic DNA (with or without amplification) to determine Gov genotype. As indicated further in the examples, electrophoretic SSCP analysis may be used to determine Gov genotype. And as indicated in the examples, the hybridization studies outlined above may use fluorescent probes, and may be directly coupled to the DNA amplification step, as in “Real-Time PCR” or related methods. There has also been provided, in accordance with another aspect of the present invention, a method of typing for Gov allele-specific target sequence in a CD 109 nucleic acid derived from a subject, comprising the steps of, (a) obtaining, by a target nucleic acid amplification process applied to mRNA from human platelets, endothelial cells, or T cells, an assayable quantity of amplified nucleic acid with a sequence that is that of a subsequence (or the complement of a subsequence) of the mRNA that encodes a CD109 said subsequence including the nucleotide at the position in the mRNA corresponding to position 2108 in SEQ ID NO:1 or to nucleotide 954 in SEQ ID NO:5; and (b) analyzing (e.g., in a nucleic acid probe hybridization assay employing an oligonucleotide probe or probes according to the invention) the amplified nucleic acid obtained in step (a) to determine the base or bases at the position in the amplified nucleic acid that corresponds to position 2108 in SEQ ID NO:1 or to nucleotide 954 in SEQ ID NO:5. It is noteworthy that, if the product of the amplification is double-stranded DNA, analysis for Gov genotype can be carried out by a RFLP (restriction fragment length polymorphism) analysis comprising exposing the amplified DNA to the restriction endonuclease BstNI (or isoschizomer thereof) under conditions whereby the DNA will be cleaved if it includes a site for cleavage by that enzyme. Such DNA, prepared from mRNA encoding the Gov b alloantigen, containing a C rather than an A at the position corresponding to nucleotide 2108 in SEQ ID NO:1 (or to nucleotide 954 in SEQ ID NO:5), includes a recognition site for that endonuclease, while such DNA prepared from mRNA encoding the Gov a alloantigen, does not If the analysis, by whatever method, of the amplified nucleic acid reveals that there is only an A (or dA) at the position corresponding to position 12108, the platelets (and blood from which they came) have only the Gov a alloantigen, and the individual from whom the platelets came, is homozygous for Gov a . Alternatively, if the analysis of the amplified nucleic acid reveals that there is only a C (or dC) at the position corresponding to position 2108, the platelets (and blood from which they came) have only the Gov b alloantigen and the individual, from whom the platelets came, is homozygous for the Gov b allele. Finally, if the analysis indicates that there is either an A (or dA) or a C (or dC) at that position, the platelets (and blood from which they came) have both Gov alloantigens, and the individual from whom the platelets came, is heterozygous for Gov alloantigen. In one application of the typing methods of the invention, the methods are applied to two individuals to determine whether blood or platelets from one would provoke an alloimmune response, and possibly PTP or PTPR, in the other. The typing method can be applied with a man and a woman, who are contemplating conceiving or have conceived a child together, to determine the risk that the child would be at risk for NATP and the risk that the woman would be at increased risk for PTP or PTPR. If the woman were heterozygous for the Gov alloantigens there would be, due to Gov alloantigen incompatibility, no risk of NATP and no increased risk for the woman of PTP or PTPR. If, however, the woman were homozygous for one of the Gov alloantigens, there would be, due to Gov alloantigen incompatibility, risk of NATP in a child and increased risk of PTP or PTPR for the woman, unless the man is homozygous for the same Gov alloantigen as is the woman. In accordance with yet another aspect of the present invention, a method of typing an individual for Gov alloantigen is provided that comprises analyzing the genomic DNA of the individual to determine the Gov alloantigen(s) of the individual. Applications of this method are substantially the same as those of the method of the invention for typing for Gov alloantigen that begins with platelet, endothelial cell, or T cell mRNA. This method of the invention, entailing analysis of genomic DNA, can be carried out in substantially the same way as outlined above for analysis of mRNA, namely first amplifying the genomic DNA and then analyzing to product of the amplification to ascertain whether there is only dA, only dC, or both dA and dC, at the position in the coding region of the genomic DNA corresponding to position 2108 in SEQ ID NO:1, or to nucleotide 954 in SEQ ID NO:5. In accordance with a further aspect of the present invention, a test kit for Gov alloantigen typing is provided comprising a non-human antibody (or antibodies) that distinguishes the two allelic forms of CD109. The antibody (or antibodies) of the kit may be polyclonal, or preferably monoclonal, and in addition to its (their) specificity for either but not both Gov alloantigens (on the surface of platelets or separated therefrom) or the CD109 subunit of one but not both of such alloantigens, typically will recognise a polypeptide molecule encoded by a nucleotide sequence encoding at least amino acid 703 of a CD109 polypeptide (the amino acid at the position corresponding to nucleotides 2107-2109 in SEQ ID NO:1, or to nucleotides 953-955 in SEQ ID NO:5). detailed-description description="Detailed Description" end="lead"? |
Alkylated c-sugars and uses thereof |
The present invention relates to alkylated C-sugars which are new carbohydrate derivatives based on the “C-Sugar” platform. These alkylated C-sugars are converted from hydrophilic, hydrogen-bonded saccaride derivatives and are very stable, highly soluble and relatively low molecular weight structures. These alkylated C-sugars can serve as effective bioconjugates and pharmaceutical carriers. The alkylated C-sugar have the formula wherein n=0, 1, 2, 3, 4, Rx=aryl, alkyl, or halogen-substituted aryl or alkyl, Y=(CH2)mNH2,(CH2)mCO2H,(CH2)mOH,(CH2)mCH, or (CH2)mCl, and m=0, 1, 2 or 3. |
1. An alkylated C-sugar of the formula wherein n=0, 1, 2, 3 or 4, Rx=aryl, alkyl, or halogen-substituted aryl or alkyl, 2. An alkylated C-sugar of the formula wherein X═CO2H, CHO, OH, NH2, Cl, Br or CH2OH, and R=alkyl or aryl. 3. A method of making the alkylated C-sugar according to claim 1, comprising alkylating a C-sugar to eliminate hydroxyl groups and obtain ether linkages. 4. A method of making the alkylated C-sugar according to claim 2, comprising alkylating a C-sugar to eliminate hydroxyl groups and obtain ether linkages. 5. The method according to claim 3, wherein the C-sugar to be alkylated is derived from mannose, glucose or galactose. 6. The method according to claim 4, wherein the C-sugar to be alkylated is derived from mannose, glucose or galactose. 7. The method according to claim 3, wherein the alkylated C-sugar is structured to comprise a free carboxylic acid, amine or other functional groups which facilitate coupling of said alkylated C-sugar to a drug or peptide. 8. The method according to claim 4, wherein the alkylated C-sugar is structured to comprise a free carboxylic acid, amine or other functional groups which facilitate coupling of said alkylated C-sugar to a drug or peptide. 9. A composition comprising a drug and the alkylated C-sugar according to claim 1. 10. A composition comprising a drug and the alkylated C-sugar according to claim 2. 11. The composition according to claim 9, wherein the alkylated C-sugar serves as a bioconjugate. 12. The composition according to claim 10, wherein the alkylated C-sugar serves as a bioconjugate. 13. The composition according to claim 9, wherein the drug is a peptide or polypeptide. 14. The composition according to claim 10, wherein the drug is a peptide or polypeptide. 15. The composition according to claim 9, wherein the alkylated C-sugar is coupled to said drug. 16. The composition according to claim 10, wherein the alkylated C-sugar is coupled to said drug. 17. A method of making the composition according to claim 9, comprising coupling said alkylated C-sugar to said drug. 18. A method of making the composition according to claim 10, comprising coupling said alkylated C-sugar to said drug. 19. A bioconjugate comprising the alkylated C-sugar according to claim 1. 20. A bioconjugate comprising the alkylated C-sugar according to claim 2. 21. A glycopeptide comprising the alkylated C-sugar according to claim 1, said alkylated C-sugar being coupled to the N-terminal, C-terminal and/or side chain of a peptide or polypeptide. 22. A glycopeptide comprising the alkylated C-sugar according to claim 2, said alkylated C-sugar being coupled to the N-terminal, C-terminal and/or side chain of a peptide or polypeptide. 23. A method of making the glycopeptide according to claim 21, comprising coupling said alkylated C-sugar to the N-terminal, C-terminal and/or side chain of the peptide or polypeptide. 24. A method of making the glycopeptide according to claim 22, comprising coupling said alkylated C-sugar to the N-terminal, C-terminal and/or side chain of the peptide or polypeptide. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention The present invention relates to alkylated C-sugars which are new carbohydrate derivatives based on the “C-Sugar” platform. These alkylated C-sugars are converted from hydrophilic, hydrogen-bonded saccharide derivatives and are very stable, highly soluble and relatively low molecular weight structures. These alkylated C-sugars can serve as effective bioconjugates and pharmaceutical carriers. 2. Description of the Related Art Advances in the synthetic chemistry of oligosaccharides, the most structurally diverse biopolymers, have been driven principally by the growing understanding of their role as encoders of biological information. Glycoconjugates on cell surfaces have a major role in biological phenomena such as the immune response, intercellular recognition, cellular adhesion, intracellular targeting, cell growth regulation, metastasis, and inflammation (Dwek, 1996; Kahane, 1996; Garegg, 1993). Glycosylation of proteins affects a variety of properties such as folding, packing, proteolytic resistance, conformational stability, quaternary structure, and water structure (Dwek, 1996; St. Hilaire, 1998). For these reasons, interest is strong in the creation and biological evaluation of oligosaccharide and glycopeptide libraries (Armstrong, 1996; Seeberger, 1996; Sofia, 1998). While cell surface carbohydrates are, typically, oligosaccharides, recognition by a protein typically involves a monosaccharide or disaccharide unit of the oligosaccharide. Consequently, monoglycosylated amino acids (and analogs) and a few glycosyl-glycosylated amino acids have been the building blocks of glycopeptide libraries. More interest has focused on the glycopeptide libraries, principally because cell surface carbohydrates, while they bind to proteins with high specificity, typically have weak binding constants (and limited bioavailability due to their biological instability). With glycopeptide libraries, the principal aim has been to create glycomimetics targeted for lectins, enzymes, and other cell surface receptors which bind carbohydrates (St. Hilaire et al., 1998; Sutherlin et al., 1996). For example, considerable interest has focused on E-selectin and its ligand, the sialyl Lewis X antigen because of their role in inflammation. Glycomimetics of sialyl Lewis X, a tetrasaccharide, have been found with binding potencies higher than the natural compound (Bertozzi, 1992) and glycosylation of peptides is known to enhance binding to this receptor (St. Hilaire, 1998). Various methods are available for developing small molecule therapeutics ranging from structure-based combinatorial techniques (Ellman, 1996; Gallop, 1994), computer screening approaches (Li et al., 1997), and various peptidomimetic strategies (Hruby, 1993; Spatola, 1983; Sawyer, 1997). Once a lead is identified, the developmental emphasis is directed toward optimizing potency, enhancing receptor or target specificity, and ensuring bioavailability with growing emphasis on nuclear-based therapies. It is clear that entry into the nucleus through membrane barriers becomes a formal obstacle. Various workers have reported success with pro-drugs, other bioconjugates (Niidome, 1999; Prouillat, 1998), and tactics designed to mask difficulties such as desolvation penalties by using, for example, N-methyl amides to enhance the solubility of polyamide species such as peptides (Conradi, 1992). More recently, there has been renewed interest in stabilized glycopeptides as drug conjugates. Currently, polyethylene glycol (PEG) is used as an effective bioconjugate because of its global solubility, enhanced stability through the ester linkage, and an optimum blend of non-reactivity and non-immunogenicity (Greenwald, 2000, Zalipsky, 1995). This knowledge and the benefits of glycoconjugation has led the present inventors to research C-sugars as bioconjugates and determine their effect when coupled to peptides and drugs. Through their research, the present inventors believe that the alkylated C-sugars of the present invention are particularly effective as bioconjugates. The presently claimed alkylated C-sugars possess some of the beneficial attributes of PEG and additionally provides stereochemical versatility via five stereocenters. The alkylated C-sugars can be easily and covalently linked to either a backbone or side chain peptide or pseudopeptide functionality or a drug. Thus, the invention has been completed based on this understanding and research. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is directed to alkylated C-sugars of the following formula: wherein n=0, 1, 2, 3 or 4, R x =aryl, alkyl, or halogen-substituted aryl or alkyl, The present invention is also directed to compositions comprising a drug and the alkylated C-sugars, as well as methods of making and using said alkylated C-sugars and compositions. detailed-description description="Detailed Description" end="lead"? |
Circuit arrangement for switching a current of and off without the occurrence of overcurrent |
There is provided a circuit arrangement for switching a current on and off without overcurrent. The circuit arrangement includes a current source, a load associated with the current source, a switching transistor for switching the current source on and off, where the switching transistor has a parasitic capacitance between a control electrode and a first output electrode, and a shorting device between the control electrode and the first output electrode of the switching transistor for switching off the current source. The current can be used, for example, to drive a laser diode. |
1. A circuit arrangements for switching a current on and off without overcurrent, comprising: a current source; a load associated with the current source; a switching transistor for switching the current source on and off, wherein the switching transistor has a parasitic capacitance between a control electrode and a first output electrode, which is not at constant potential; and a shorting device between the control electrode and the first output electrode of the switching transistor for switching off the current source. 2. The circuit arrangement as claimed in claim 1, wherein the shorting device comprises a transistor having: a control electrode for having a switch-on/switch-off signal applied to it; an output electrode connected to the first output electrode; and an output electrode connected to the control electrode of the switching transistor. 3. The circuit arrangement as claimed in claim 1, wherein the shorting device has an associated pull-up device. 4. The circuit arrangement as claimed in claim 1, wherein one or more of the current source, the switching transistor, the shorting device and the pull-up device are in a form selected from the group consisting of a bipolar transistor and a field-effect transistor. 5. The circuit arrangement as claimed in claim 1, wherein the load is a laser diode. 6. The circuit arrangement as claimed in claim 1, wherein the current is switched off by the switching transistor if a constant voltage applied to a control electrode of the current source is lower than a sum of threshold voltages of the current source and the switching transistor. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Circuit arrangements for switching a current driving a load on and off are known. FIG. 1 shows such a circuit arrangement, which uses a transistor 20 ′ as a current source or current sink. The control electrode of the transistor 20 ′ is connected to a DC voltage source 80 ′. The current source 20 ′ is connected in series with a load 40 ′, for example with a laser diode. A switching transistor 50 ′, which is likewise connected in series with the current source 20 ′, is used as a switch for the current source. The switching transistor 50 ′, which is in the form of a field-effect transistor, for example, is connected by means of its gate electrode 51 ′ to a driving device which comprises the transistors 75 ′ and 70 ′, for example. The input 1 of the driving device 70 ′, 75 ′ has a switch-on/switch-off signal P applied to it, as illustrated in FIG. 2 , for example. Between the gate 51 ′ and the drain electrode 52 ′ of the switching transistor 50 ′ there is a manufacture-dependent parasitic capacitance 60 ′, which is also known as the Miller capacitance. As will be explained later, this parasitic capacitance is the cause of overcurrents that are brought about when the current source 20 ′ is switched off. The gate/source voltage GS of the switching transistor 50 ′ controls the state of the switching transistor. If this gate/source voltage GS is lower than the threshold voltage of the switching transistor 50 ′, the current IL is interrupted by the switching transistor 50 ′. The gate/source voltage GS is S controlled by means of the driving device 70 ′, 75 ′. However, when the switching transistor 50 ′ and hence the current source 20 ′ are disconnected, an overcurrent, also called a current spike, is produced which has its origins in the parasitic capacitance 60 ′ of the switching transistor 50 ′. This is because a falling edge of the gate/source voltage GS on the switching transistor 50 ′ (the profile of said gate/source voltage being shown in FIG. 4 ) causes the parasitic capacitance 60 ′ to transfer the sudden voltage change on the control electrode 51 ′ to the output electrode 52 ′ of the switching transistor 50 ′ and to drive the transistor 20 ′ at a higher level. This causes an overcurrent through the load 40 ′, as shown in FIG. 3 at the time t a . The overcurrent flows until the parasitic capacitance's charge has been reversed. The time t a characterizes the time at which the switch-off signal is applied to the input 1 of the driving device 70 ′, 75 ′. In the case of certain applications, such current spikes can result in the load 40 ′ being destroyed, as can be the case with a laser diode, for example. |
<SOH> SUMMARY OF THE INVENTION <EOH>The invention is now based on the object of improving the initially described circuit arrangement for switching a current on and off such that overcurrents can be avoided when a current source is switched off. The invention solves this technical problem with the features of claim 1 . Accordingly, a circuit arrangement for switching a current on and off without overcurrent is provided. The circuit arrangement has a current source, a load associated with the current source and a switching transistor for switching the current source on and off. Depending on manufacture, the switching transistor contains a parasitic capacitance between the control electrode and a first output electrode, which causes charge-reversal currents when the current source is switched off. A shorting device, connected between the control electrode and the first output electrode of the switching device, for switching off the current source can be used to prevent the charge-reversal currents from resulting in overcurrents in the load. Expediently, the shorting device is in the form of a transistor whose control electrode can have a switch-on/switch-off signal applied to it. One output electrode of the shorting device is connected to the first output electrode of the switching transistor, and the other output electrode of the shorting device is connected to the control electrode of the switching transistor. In this way, the shorting device shorts the switching transistor and hence switchs off the current source. In order to be able to shorten the input and output cycles, a pull-up device is connected in parallel with the shorting device and ensures that the potential on the switching transistor's control electrode can be raised at the switch-on time. The current source, the switching transistor, the shorting device and/or the pull-up device can be in the form of bipolar transistors, field-effect transistors, particularly MOS transistors, or combinations of these technologies. The circuit arrangement is suitable particularly for driving a laser diode as a load. |
Novel genes for conditional cell ablation |
A Novel deacetylase genes for use in conditional cell ablation are described. These genes are particularly useful for the production of transgenic plants with plant parts which can be destroyed upon treatment with N-acetyl-PPT. |
1. A DNA molecule encoding a protein or polypepetide having deacetylase activity, wherein said protein has at least 70% to 75% sequence indentity with the sequence of SEQ ID No. 1. 2. The DNA molecule of claim 1, wherein said protein or polypeptide comprises the sequence of SEQ ID No. 1. 3. The DNA molecule of claim 1, comprising a nucleotide sequence which has at least 70% to 75% sequence identity to the sequence of SEQ ID No. 2. 4. The DNA molecule of claim 3, comprising the nucleotide sequence of SEQ ID No. 2. 5. The DNA molecule of claim 1, which can be isolated from Comamonas acidovorans, deposited as DSM 11070. 6. A chimeric gene comprising: a) the DNA molecule of any one of claims 1 to 5, b) a plant-expressible promoter; said DNA molecule being in the same transcriptional unit and under the control of said plant-expressible promoter. 7. A plant having plant parts which can be destroyed upon induction, said plant comprising, stably integrated in its genome, a chimeric gene, comprising a) The DNA molecule of any one of claims 1 to 5, b) a plant-expressible promoter; said DNA molecule being in the same transcriptional unit and under the control of said tissue-specific, plant-expressible promoter; wherein said plant is characterized by the fact that, upon treatment with N-acetyl-PPT, plant parts are destroyed. 8. The plant of claim 7, wherein said plant-expressible promoter is a tissue-specific promoter. 9. The plant of claim 8, wherein said tissue-specific promoter is a stamen selective promoter; 10. A plant which is inducibly male-sterile, said plant comprising, stably integrated into its genome, a foreign DNA comprising a chimeric gene comprising: a) the DNA molecule of any one of claims 1 to 5; and b) a promoter directing stamen-selective expression; said DNA being in the same transcriptional unit and under the control of said stamen-selective promoter. 11. The plant of claim 10, wherein said stamen-selective promoter is selected from the group of: the TA29 promoter, the T72 promoter, the El promoter, or the CA55 promoter. 12. The plant of claim 10, wherein said foreign DNA further comprises: a herbicide resistance gene comprising a DNA encoding an RNA, protein or polypeptide that confers herbicide resistance on said plant. 13. A process for producing a plant with selectively destroyable plant tissues or parts, said process comprising: i) introducing into a plant cell or tissue a foreign DNA comprising a chimeric gene comprising: a) The DNA molecule of any one of claims 1 to 5, b) plant-expressible promoter; said DNA molecule being in the same transcriptional unit and under the control of said plant-expressible promoter. ii) regenerating said plant from said plant cell. 14. The process of claim 13, wherein said plant-expressible promoter is a tissue-specific promoter. 15. The process of claim 14, wherein said tissue-specific promoter is a male or female organ selective promoter and said plants are inducibly male or female sterile. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Deacetylases are capable of deacetylating acetylated toxins, such as N-acetyl-phosphinothricin (N-Ac-PTC or N-Ac-PPT), intracellularly, whereupon the cytotoxic activity of the toxins is restored (formation of PTC or PTC). U.S. Pat. No. 5,650,310, U.S. Pat. No. 5,668,297, U.S. Pat. No. 5,767,370, and U.S. Pat. No. 5,767,371, describe deacetylase genes, for the production of phosphinothricin or phosphinothricyl-alanyl-alanine, as well as processes for their isolation and use. Furthermore deacetylase genes are disclosed which are isolated from Streptomyces viridochromogenes (dea) and from E. coli (ArgE). The use of such genes for the production of plants with selectively destroyable plant parts is described. More specifically, a method is disclosed for the production of conditionally male-sterile plants. This is achieved by introducing a deacetylase gene, under the control of a tapetum-specific promoter into the genome of a plant. Upon application of N-Ac-PPT to the plant, in the tapetum cells, where the deacetylase is expressed, N-Ac-PPT is converted into PPT, which is toxic to the cells. The cytotoxic activity in the tapetum cells inhibits the development of microspores and renders the plant male-sterile. WO 98/27201 describes novel genes encoding amino acid deacetylases isolated from Stenotrophomonas sp. and from Comamonas acidovorans and their use in the production of tranagenic plants. WO 98/13504 describes the use of deacetylase genes for the production of female-sterile plants. WO 98/39462 describes a method of hybrid seed production using conditional female sterility, whereby a plant is made conditionally female-sterile by transformation with the ArgE gene linked to a female-preferential promoter. EP 98116492 describes the use of the deac system for the modification of plant development. Deacetylase genes placed under control of a meristem specific promoter can be used for inducible modification of developmental characteristics of the plant such as flowering and bolting to improve yield, or influence general plant architecture. EP 98116493 describes the use of the deac system to obtain pathogen control in plants. Deacetylase genes placed under control of a pathogen-inducible promoter can be used to obtain plants which, upon treatment with N-acetyl-PPT become tolerant to pathogens, through selective destruction of pathogen-infected tissue. However, the foregoing documents fail to teach or suggest the present invention. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention relates to DNA molecules encoding a protein having the biological activity of a deacetylase. More specifically the protein having the biological activity of a deacetylase is a protein having an amino acid sequence with at least 70% to 79%, more preferably 80% to 85%, especially preferably 86% to 90%, most preferably 91% to 95%, sequence identity, more specifically which is 96% to 100% identical to the sequence of SEQ ID No. 1 or a biologically active fragment or variant thereof. Especially preferably the DNA molecule of the invention encodes a protein having the amino acid sequence of SEQ ID No. 1 or a biologically active fragment thereof Alternately or additionally, the DNA molecule of the present invention has a nucleotide sequence which has a sequence identity of at least 70% to 79%, more preferably 80% to 85%, especially preferably 86% to 90%, most preferably 91% to 95%, sequence identity, more specifically which is 96% to 100% identical to the sequence of SEQ ID No. 2. Especially preferably the DNA molecule of the invention comprises the nucleotide sequence of SEQ ID No. 2. Additionally or alternatively, the invention relates to a DNA molecule encoding a deacetylase which can be isolated from Comamonas acidovorans , deposited as DSM 11070, which comprises the sequence of SEQ ID No. 2. The invention further relates to a protein or polypeptide having the biological activity of a deacetylase, which is encoded by a DNA molecule, such as those described above. The invention further relates to a chimeric gene comprising the DNA molecule of the present invention placed in the same transcriptional unit and under the control of a plant expressible promoter, preferably a tissue-specific promoter. The invention further relates to a plant having inducibly destroyable plant parts, which plant comprises, stably integrated in its genome, a chimeric gene comprising the DNA molecule of the invention, placed under control of a plant-expressible promoter, preferably a tissue-specific promoter, the plant of the invention being characterized by the fact that, upon treatment with N-acetyl-PPT, the plant tissues in which the promoter directs expression, are destroyed. The invention further relates to an inducibly male or female-sterile plant, which plant comprises, stably integrated in its genome, the DNA molecule of the invention, placed under control of a male- or female reproductive organ selective promoter, the inducibly male- or female-sterile plant of the invention being characterized by the fact that, upon treatment with N-acetyl-PPT, specific plant tissues of the male- or female reproductive organ are destroyed. The invention further relates to a process for producing a plant with inducibly destroyable plant tissues or parts, which process comprises: introducing into a plant cell or tissue a foreign DNA comprising a chimeric gene comprising a) the DNA molecule of the invention, encoding a protein or polypeptide having the biological activity of a deacetylase, under the control of b) a plant-expressible promoter, which is preferably tissue-specific; regenerating the plant cell or tissue into a plant optionally, inducing the destruction of the plant tissue or parts by treatment of the plant with N-acetyl-PPT. detailed-description description="Detailed Description" end="lead"? |
Bushing with controlled elastic absorption of radial stress |
Bushing, in particular of the kind with controlled elastic absorption of radial stress, includes a cylindrical tubular element (1) out of self-lubricating material onto which a ring (2) is freely fitted out of an elastically buckling material, wherein the internal surface of the ring out of elastically buckling material is associated to an annular reinforcing fitting (3) out of a material that does not give radially and which gives the bushing the required radial stiffness and limits the longitudinal deformations of the ring (2). |
1. A bushing, in particular of the kind with controlled elastic absorption of radial stress, comprising a cylindrical tubular element (1) out of self-lubricating material onto which a ring (2) is freely fitted out of a elastically buckling material, characterized in that the internal surface of said ring out of elastically buckling material is associated to an annular reinforcing fitting (3) out of a material that does not give radially. 2. A bushing according to claim 1, characterized in that the external surface of said ring (2) out of elastically buckling material comprises bulge ribbings (4). 3. A bushing according to claim 1, characterized in that said bulge ribbings (4) are placed at regular distances one from the other. 4. A bushing according to claim 1, characterized in that said bulge ribbings (4) develop in a longitudinal direction substantially parallel to the axis of said bushing. 5. A bushing according to claim 1, characterized in that said bulge ribbings (4) have an annular shape. 6. A bushing according to claim 1, characterized in that said ring (2) is out of rubber. 7. A bushing according to claim 1, characterized in that said annular reinforcing insert (3) is fixed to said ring (2) by means of vulcanization. 8. A bushing according to claim 1, characterized in that said element (1) and said ring (2) are respectively provided with stop collars (5, 6). 9. A bushing according to claim 1, characterized in that it comprises a plane metal washer that may be placed against said stop collar (6). 10. A vehicle characterized in that it comprises at least one bushing according to claim 1. |
Method and compositions for evaluating risk of developing type 2 diabetes in people of chinese descent |
Methods and compositions for identifying mutations and polymorphisms in mutant genes encoding gene product involved in insulin secretion, for example, hepatocyte nuclear factor-1∝, glucokinase, amylin and mitochondrial DNA are disclosed. Specifically, a microchip comprising a combination of at least two different mutant genes wherein each gene comprises at least one mutation indicative of a predisposition for type-2 diabetes in a member of a Chinese population is disclosed. A kit comprising the microchip, an isolated nucleic acid, primers and probes which are specifically used to screen or identify the mutations in genes of hepatocyte nuclear factor-1∝, glucokinase, amylin and mitochondrial DNA are also disclosed. |
1. A microchip comprising: a combination of at least two different mutant nucleic acid sequences of a wild-type nucleic acid sequence, wherein each wild-type nucleic acid sequence encodes a protein involved in insulin secretion, wherein said gene comprises at least one mutation indicative of a predisposition for type 2 diabetes in a member of a Chinese population. 2. The microchip according to claim 1, wherein said nucleic acid sequences comprise nucleic acid selected from the group consisting of genomic DNA, complementary DNA and messenger RNA. 3. The microchip according to claim 1, wherein said type 2 diabetes is maturity onset diabetes of the young. 4. The microchip according to claim 1, wherein said microchip further comprises a genetic marker that uniquely identifies a member of a Chinese population. 5. A microchip comprising: a combination of at least two different nucleic acid sequences, wherein each nucleic acid sequence encodes a gene product involved in insulin secretion wherein said gene comprises at least one mutation indicative of a predisposition for type 2 diabetes in a human subject of a Chinese population, wherein said gene product is selected from the group consisting of a glucokinase, a hepatocyte nuclear factor 1α, an amylin and a mitochondrial tRNA (Leu) (UUR). 6. A microchip comprising: at least one each of a combination of different nucleic acid sequences, wherein each nucleic acid sequence encodes a protein selected from the group consisting of glucokinase, hepatocyte nuclear factor la, amylin and mitochondrial tRNA(Leu)(UTR), wherein said glucokinase gene comprises at least one mutation selected from the group consisting of V101M, I110T, A119D, Q239R, and G385V, and said hepatocyte nuclear factor la gene comprises at least one mutation selected from the group consisting of G20R, A116V, IVS2nt-G→A, R203H, S432C, and I618M, and said amylin gene comprises the mutation S20G, and said mitochondrial tRNA(Leu)(UUR) gene comprises the mutation A3243G. 7. A microchip comprising at least one nucleic acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7 and SEQ ID NO:10. 8. A microassay system comprising a microchip according to claim 1. 9. A kit comprising a microchip according to claim 1. 10. A nucleic acid primer comprised of SEQ ID NO: 34. 11. A nucleic acid primer comprised of SEQ ID NO: 35. 12. A nucleic acid primer comprised of SEQ ID NO: 36. 13. A nucleic acid primer comprised of SEQ ID NO: 37. 14. A nucleic acid probe that specifically anneals to a nucleic acid encoding a mutant gene of a wild-type gene involved in insulin secretion, wherein said mutant gene comprises at least one mutation indicative of increased risk for type 2 diabetes in a human subject of a Chinese population, and wherein said nucleic acid probe does not bind to said wild-type gene. 15. An isolated nucleic acid encoding a mutant gene of a wild-type gene that encodes a protein involved in the secretion of insulin, wherein said mutant gene comprises at least one mutation associated with increased risk for type 2 diabetes in a subject of a Chinese population. 16. The isolated nucleic acid according to claim 15, wherein said mutation is a single nucleotide polymorphism. 17. The isolated nucleic acid according to claim 15, wherein said mutation is selected from the group consisting of a missense, a nonsense, an insertion and a deletion mutation. 18. The isolated nucleic acid according to claim 15, wherein said wild-type gene encodes hepatocyte nuclear factor la, and said mutation is A116V. 19. The isolated nucleic acid according to claim 15, wherein said wild-type gene encodes glucokinase, and said mutation is selected from the group consisting of V101M and Q239R. 20. An isolated nucleic acid encoding a mutant gene of a wild-type gene that encodes a protein involved in the secretion of insulin, wherein said mutant gene is selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 7 and SEQ ID NO: 10. 21. An isolated amino acid sequence encoded by a mutant gene of a wild-type gene encoding a protein involved in the secretion of insulin, wherein said mutant gene comprises at least on mutation associated with increased risk for type 2 diabetes in a member of a Chinese population. 22. An antibody that specifically binds a protein encoded by a mutant gene of a wild type gene encoding a protein involved in the secretion of insulin, wherein said mutant gene comprises at least on mutation associated with increased risk for type 2 diabetes in a member of a Chinese population, and wherein said antibody does not bind to a protein encoded by said wild-type gene. 23. A method of determining a genetic predisposition of a member of a Chinese population to develop type 2 diabetes, said method comprising the step of: contacting a sample comprising nucleic acid from said member with a combination of at least two nucleic acid sequences, wherein each nucleic acid sequence encodes a mutant gene of a wild-type gene encoding a protein involved in insulin secretion, wherein each mutant gene comprises at least one mutation indicative of a predisposition of a member of a Chinese population to develop type 2 diabetes, whereby identification of at least one of said mutations in said sample is indicative of a genetic predisposition for type 2 diabetes in said member of a Chinese population. 24. A method for detecting an increased risk of an individual of a Chinese population with decreased insulin secretory function to develop type 2 diabetes, said method comprising the step of: contacting a sample comprising nucleic acid from said individual with a combination of at least two different nucleic acid sequences, wherein each nucleic acid sequence encodes a mutant gene of a wild-type gene encoding a protein involved in insulin secretion, wherein each mutant gene comprises at least one mutation indicative of a predisposition of a member of a Chinese population to develop type 2 diabetes, wherein identification of at least one of said mutations in said sample is indicative of an increased risk for type 2 diabetes in said individual of a Chinese population. 25. The method according to claim 23, wherein said combination of at least two different nucleic acid sequences are attached to a microchip. 26. The method according to claim 23, wherein said nucleic acid sample is obtained from bodily fluid or tissue. 27. The method according to claim 23, wherein said wild-type gene encodes a gene product selected from the group consisting of hepatocyte nuclear factor 1α, glucokinase, amylin and mitochondrial tRNA(Leu)(UUR). 28. A method of determining a genetic predisposition of a member of a Chinese population to develop type 2 diabetes, said method comprising the step of: contacting a sample comprising nucleic acid from said member with a combination of at least two different nucleic acid sequences selected from the group consisting of G20R, A116V, IVS2nt-G→A, R203H, S432C, and I618M of hepatocyte nuclear factor 1α; V101M, I110T, A119D, Q239R, and G385V of glucokinase; S20G of amylin, and A3243G of mitochondrial tRNA(Leu)(UUR), wherein each nucleic acid sequence encodes a mutant gene of a wild-type gene encoding a protein involved in insulin secretion, wherein each mutant gene comprises at least one mutation indicative of a predisposition of a member of a Chinese population to develop type 2 diabetes, and wherein said identification of one of said mutations in said sample is indicative of a genetic predisposition for type 2 diabetes in said member of a Chinese population. 29. A method for detecting an increased risk of an individual of a Chinese population with decreased insulin secretory function to develop type 2 diabetes, said method comprising the step of: contacting a sample from said individual with a combination of at least two different nucleic acid sequences selected from the group consisting of G20R, A116V, IVS2nt-G→A, R203H, S432C, and I618M of hepatocyte nuclear factor 1α;V101M, I110T, A119D, Q239R, and G385V of glucokinase; S20G of amylin, and A3243G of mitochondrial tRNA(Leu)(UUR), wherein each nucleic acid sequence encodes a mutant gene of a wild-type gene encoding a protein involved in insulin secretion, wherein each mutant gene comprises at least one mutation indicative of a predisposition of an individual of a Chinese population to develop type 2 diabetes, and wherein the identification of at least one of said mutations in said sample is indicative of an increased risk for type 2 diabetes in said individual of a Chinese population. 30. A method for screening for genetic mutations in an individual of a Chinese population diagnosed with type 2 diabetes, said method comprising the steps of: contacting a sample from said individual with a combination of at least two different nucleic acid sequences, wherein each nucleic acid sequence encodes a mutant gene of a wild-type gene encoding a protein involved in insulin secretion, wherein each mutant gene comprises at least one mutation indicative of a predisposition of a member of a Chinese population to develop type 2 diabetes, and wherein identification of at least one of said mutations in said sample is indicative of an etiology of said type 2 diabetes in said individual of a Chinese population. 31. The method according to claim 30, wherein said individual has been diagnosed with maturity onset diabetes of the young. 32. The method according to claim 30, wherein said individual has at least one primary family member that has been diagnosed with maturity onset diabetes of the young. 33. The method according to claim 30, wherein said mutation is selected from the group consisting of a missense, a nonsense, an insertion and a deletion mutation. 34. A method for screening for genetic mutations indicative of increased risk of an individual of a Chinese population to develop type 2 diabetes, said method comprising the steps of: contacting a sample from said individual with a combination of at least two different nucleic acid sequences selected from the group consisting of G20R, A116V, IVS2nt-G→A, R203H, S432C, and I618M of hepatocyte nuclear factor 1α;V101M, I110T, A119D, Q239R, and G385V of glucokinase; S20G of amylin, and A3243G of mitochondrial tRNA(Leu)(UTR), wherein each nucleic acid sequence encodes a mutant gene of a wild-type gene encoding a protein involved in insulin secretion, wherein each mutant gene comprises at least one mutation indicative of a predisposition of an individual of a Chinese population to develop type 2 diabetes. 35. A method for screening for a genetic predisposition to develop type 2 diabetes in an individual of a Chinese population having at least one primary family member that has been diagnosed with type 2 diabetes, said method comprising the steps of: contacting a sample comprising nucleic acid from said individual with a combination of at least two different nucleic acid sequences, wherein each nucleic acid sequence encodes a mutant gene of a wild-type gene encoding a protein involved in insulin secretion, wherein each mutant gene comprises at least one mutation indicative of a predisposition of a member of a Chinese population to develop type 2 diabetes, and wherein identification of at least one of said mutations in said sample is indicative of a genetic predisposition to develop type 2 diabetes in said individual of a Chinese population. |
<SOH> BACKGROUND <EOH>Although people of Chinese ancestry account for >20% of the world's population (Chan, et al(1997) 20: 1785), very little is known about the genetic factors that contribute to the development of diabetes in this population. The prevalence of diabetes amongst Chinese people varies from <1% in some rural areas in mainland China to 6-12% in Hong Kong, Singapore, and Taiwan (Chan, et al (1997), supra). Hong Kong can be regarded as a paradigm of future China. The prevalence of diabetes mellitus is reaching epidemic proportions amongst Hong Kong Chinese, with type 2 diabetes being the predominant form in pateints with early-or late-onset of disease (Chan and Cockram (1997) Diabetes Care 20: 1785). Type 2 diabetes mellitus is a heterogeneous disease that is caused by both genetic and environmental factors. The age-adjusted prevalence of diabetes in the Chinese population has increased from 7.7% in 1990 (Cockram, et al (1993) Diabetes Res and Clin Practice 21: 67) to 8.9% in 1995 (Cockram and Chan (1999) In: Diabetes in the New Millennium, Pot Still Press, Sydney, pp. 11-22). In a population-based study conducted in 1995, the crude prevalence of diabetes mellitus was 9.6%, rising from 1.7% in those aged under 40 years to 25% in those older than 60 years (Janus (1997) Clin Exp Pharmacol Physiol 24: 987). There is a high prevalence of obesity (43%) and positive family history of diabetes (50%) in Chinese patients presenting with acute or early onset diabetes (Chan, et al (1993) Postgrad Med J 69: 204; Ko, et al (1998) 35: 761). These findings indicate that genetic factors, in addition to environmental factors, can be an important cause of early onset diabetes in this population. Because type 2 diabetes is an insidious disease, it is estimated that as many as half of the individuals in Hong Kong that would be considered diabetic remain undiagnosed. Most patients are finally diagnosed only when presenting with overt symptoms that often are the consequence of advanced disease. Clinic as well as population-based studies reveal that about 17% of diabetic patients in Hong Kong are diagnosed before age 35 years (Chan, et al (1993) Postgrad Med 69: 204; Janus (1996) The Hong Kong cardovascular risk factor prevalence study 1995-1996 Dept of Clin Biochem, Queen Mary Hospital of Hong Kong, Hong Kong, 1997). Due to their anticipated long duration of disease, it is important to classify and characterize the nature of diabetes in these young patients to facilitate early diagnosis and appropriate treatments. Current methods of diagnosing type 2 diabetes generally involve assessing phenotypic parameters, such as measuring fasting serum glucose levels by administering an oral glucose tolerance test (OGTT) to determine impaired glucose tolerance (IGT) or impaired fasting glucose (IFG). Phenotypic assessments of persons suspected of having type 2 diabetes are important, but they are limited in that patients generally receive a diagnosis only after presentation with overt symptoms. Furthermore, because the common symptoms of type 2 diabetes are a consequence of a combination heterogenous genetic and environmental causes, the therapies provided are general with regard to the disease rather than targetted to the specific etiology of the individual patient. Numerous studies have attempted to correlate the increased risk for development of type 2 diabetes with a mutation of a specific gene, but the results of these studies repeatedly demonstrate that no one mutated gene can be attributed as the major cause of type 2 diabetes, emphasizing the heterogeneous nature of this disease. Furthermore, a mutation in a particular gene that correlates with increased risk for developing type 2 diabetes in individuals of one ethnic population is not relevant to individuals of a second ethnic population, wherein the risk for type 2 diabetes in individuals of the second ethnic population will correlate with a different mutation or a mutation in a completely different gene. It is therefore of interest to identify additional genetic mutations and polymorphisms that are indicative of an increased risk for developing type 2 diabetes in people of Chinese ancestry, and to develop methods that can be effectively employed to prophylactically identify asymptomatic Chinese individuals with a genetic predisposition for type 2 diabetes. Relevant Literature Maturity-onset diabetes of the young (MODY) is a monogenic form of diabetes characterized by autosomal dominant inheritance, early onset (usually before 25 years of age) and a primary defect in pancreatic β-cell function (Fajans (1990) Diabetes Care 13: 49; Chan, et al (1990) Diabetic Med 7: 211; Byrne, et al (1996) Diabetes 45: 1503). This form of diabetes can result from mutations in at least five different genes including those encoding the glycolytic enzyme glucokinase (Froguel, et al (1993) New Engl J Med 328: 697), the liver-enriched transcription factors expressed in the pancreatic β-cell, which are hepatocyte nuclear factors HNF-1α (Yamagata, et al (1996) Nature 384: 455), HNF-1β (Horikawa, et al (1997) Nature Genet 17: 384), and HNF-4α (Yamagata, et al (1996) Nature 384: 458), and insulin promoter factor-1 (IPF-1) (Stoffers, et al (1997) Nature Genet 17: 138). Some mutations and polymorphisms in the glucokinase and HNF-1α genes that are associated with the genetic predisposition of a Chinese individual to develop type 2 diabetes mellitus have been initially identified in Ng, et al ( Diabetic Medicine 1999, 16: 956, herein incorporated by reference), but this manuscript does not disclose how these mutations and polymorphisms might be used to identify Chinese individuals with increased risk of developing type 2 diabetes. U.S. Pat. No. 5,541,060 discloses the results of screening a cohort of sixteen French families having MODY and the identification of several missense mutations in the glucokinase gene, however none of the mutations identified are relevant to individuals of Chinese descent. U.S. Pat. No. 5,800,998 discloses a point mutation at nucleotide 414 of human HNF 1α, but this single point mutation is not associated with a genetic predisposition of a Chinese individual to develop type 2 diabetes. Major susceptibility loci for non-insulin dependent diabetes have been identified through genome scans of individuals in Mexican-American (Hanis, et al (1996) Nature Genet 13: 161) and Finnish (Mahtani, et al (1996) Nature Genet 14: 90) populations, but not in individuals of a Chinese population. Specific microsatellite regions of genomic DNA can be correlated with major susceptibility loci that closely associate with the increased risk of a Chinese subject to develop type 2 diabetes. For instance, Le Stunff, et al ( Nature Genet. (2000) 26: 444) have reported that particular alleles of the insulin gene variable number of tandem repeat (VNTR) locus are associated with obesity and type 2 diabetes. Also, microsatellite polymorphisms flanking the glucokinase have been associated with type 2 diabetes in a Taiwanese population (Wu, et al (1995) Diabetes Res Clin Pract 30: 21). |
<SOH> SUMMARY OF THE INVENTION <EOH>Compositions and methods are provided, wherein a unique combination of genetic markers indicative of a genetic predisposition for developing type 2 diabetes in members of a Chinese population is described. The invention is exemplified by a combination of mutated gene sequences from wild-type genes that are involved in insulin secretory function, including hepatocyte nuclear factor 1α (HNF-1α), glucokinase, amylin and mitochondrial DNA. The combination of representative mutations include G20R, A116V, IVS2nt→GA, R203H, S432C and 1618M of HNF-1α; V101M, 1110T, A119D, Q239R and G385V of glucokinase; S20G of amylin; and A3243G of mitochondrial tRNA Leu(UUR) . The combination of the mutated genes of interest will be most efficiently used for screening individuals at increased risk by attaching them to a microchip. Embodiments of methods for determining or detecting the genetic predisposition of a Chinese individual to develop type 2 diabetes include obtaining a sample containing genomic nucleic acid from a Chinese patient, such as a tissue biopsy or a blood sample, and contacting that sample with a representative combination of at least two mutated genes of interest, then subjecting the sample DNA together with the patient's DNA to hybridization conditions stringent enough to detect nucleotide differences of at least one base pair. Alternatively, particular genes of interest from the genomic DNA of a Chinese individual at risk are screened using PCR primer pairs and PCR-RFLP techniques to identify the presence or absence of a mutation known to be associated with type 2 diabetes. The methods further encompass screening the genomic DNA of Chinese individuals who have been diagnosed with type 2 diabetes or who have a primary family member with type 2 diabetes for additional associative mutations in identified genes or for mutations correlative with the predisposition of a member of a Chinese population to develop type 2 diabetes in additional candidate genes, such as those associated with diabetic kidney disease and obesity. The invention further provides for nucleic acid primers and probes that are specifically used to identify mutations, for instance by PCR or hybridization, of wild-type genes involved in insulin secretion that are associated with an increased risk of a Chinese subject to develop type 2 diabetes. Additionally, proteins translated from genes carrying at least one mutation associated with increased risk of a Chinese individual to develop type 2 diabetes find use in functional diagnostic assays and in the production of diagnostic antibodies that bind to the mutant but not the wild-type protein. The prophylactic detection of mutations and polymorphisms that are indicative of a genetic predisposition of a Chinese individual to develop type 2 diabetes finds application in providing clinicians with information that allows for early detection and therapy initiation before the onset of overt symptoms or complications, and that enables clinicians to administer specifically targetted therapies that address the etiology of an individual's disease. |
Propellant powder charge for barrel weapon |
The present invention relates to a method for producing propellant powder charges intended for heavy barrel weapons, such as canons and howitzers, and with a high degree of filling and a high energy content, and also to propellant powder charges produced in accordance with this method. The charges according to the invention are distinguished by the fact that 70 to 95% of their weight consists of a more coarsely grained powder and 30 to 5% consists of a granular powder with a smaller grain size. These two powders have the same of different chemical composition, and one or more of them can be surface-inhibited. |
1-7. (Cancelled). 8. A method for producing progressive propellant powder charges suitable for use in barrel weapons having a high degree of filling or loading density and thus also a high energy content at which the charge is produced, the method comprising: mixing at least two types of granular powder of different grain size, of which at least one of the at least two powder types with a largest grain size and a larger number of internal priming channels has a progressive burn characteristic, wherein, as subsidiary components of an actual charge, granular powders are chosen having size, geometric shape, and quantities adapted to give a smallest possible empty spaces between the powder grains. 9. The method of claim 1, wherein a starting material chosen for the charge comprises at least two types of powder whose mutual progressivity has been optimized for a particular purpose. 10. The method of claim 1, wherein a starting material chosen for the charge comprises granular powder having a same or different chemical composition, but with different grain size and a different number of internal priming channels. 11. The method of claim 1, wherein, when choosing a starting material, powder types are chosen of which one or more are surface-inhibited. 12. The method of claim 1, wherein, of the powder types included in a finished charge, all the types are introduced contemporaneously with each other and, at the same time a casing or cartridge to which the powder is added and in which the charge is to be stored prior to use is subjected to vibrations which improve a degree of packing of the charge. 13. Propellant powder charge suitable for use in barrel weapons and having a high degree of filling or loading density and with a high energy content, the power charge comprising: at least two different types of granular powder of which at least one type has a progressive burn characteristic and in which one or more of these powders is surface-inhibited, wherein burn characteristics of each of these different powders are adapted to one another while their mutual grain sizes and quantities of each powder included in a charge are adapted to one another such that an empty space remaining between the powder grains is minimized. 14. The propellant powder charge of claim 13, wherein between 70-95% of a powder charge weight comprises relatively coarsely grained powder, and wherein 30-5% of the weight comprises a relatively more finely grained powder, wherein these two powders have a same or different chemical composition and at least one of the two powders is progressive and one or more of the two powders is surface-inhibited. 15. The propellant powder charge of claim 14, wherein between 75 to 85% of of a powder charge weight comprises relatively coarsely grained powder. 16. The propellant powder charge of claim 14, wherein the coarsely grained powder is either 19-hole or 37-hole powder cut into short sections. 17. The propellant powder charge of claim 14, wherein the finely grained powder is either 1-hole or 7-hole powder. |
Method for pumping fluids |
The production rate, useful life and operating efficiency of electric submergible pumping systems (ESP) is improved by operating the system so that the motor (220, 320) is cooled by two passes of production. Production fluids remove waste heat from the motor (220, 320) by passing the fluid in contact with the exterior of the motor (220, 320). The improved cooling permits the motor to be operated at a lower temperature improving life and efficiency, and/or to be operated at higher power at a similar temperature. Additionally, oversized and excess equipment is not required, further improving performance and economics for the user. In another aspect, the invention is a method of pumping fluids using a motor having vortex generators (366) on its exterior surfaces or on the interior surfaces of a surrounding shroud. |
1. A method comprising pumping a fluid with a submergible pumping system including a pump and an electrical motor, wherein the electrical motor has a hollow rotor shaft and said pumping system is adapted to pump at least a portion of said fluid through said hollow rotor shaft, and wherein during operation at least a portion of said fluid passes in fluid contact with exterior portions of the electrical motor and removes heat therefrom. 2. The method of claim 1, wherein the pumping system is submerged in said fluid such that production fluids entering the pumping system pass in fluid contact with exterior portions of the electrical motor and remove heat therefrom before entering the pumping system, and said liquid production fluids then pass through the hollow rotor shaft of the electrical motor as the fluid is pumped, and remove additional heat from the motor. 3. A method for producing fluids from a well having a casing that contains perforations through which production fluids enter the well, comprising (a) positioning an electrical submergible pumping system including an electric motor within said casing such that the electric motor is at or below at least some of said perforations, and (b) pumping production fluids from the well with said pumping system, wherein (1) the pumping system includes a pump, an electrical motor having a hollow rotor shaft and production fluid intakes located below said electrical motor, said pumping system being adapted to pump said production fluids through said hollow rotor shaft of the electrical motor, (2) production fluids entering the well through at least some of the perforations in the casing pass in fluid contact with exterior portions of the electrical motor and remove heat therefrom before entering the production fluid intakes, and (3) production fluids pass through the hollow rotor shaft of the electrical motor as the fluid travels to the wellhead, and remove additional heat from the motor. 4. The method of claim 3 wherein the production fluids pass through the hollow rotor shaft of the electrical motor under conditions of turbulent flow. 5. The method of claim 4 wherein the electrical motor is located between the wellhead and the pump. 6. The method of claim 5 wherein production fluids entering the well through the perforations in the casing travel downward in fluid contact with exterior portions of the electrical motor to an intake, and the liquid production fluids are subsequently pumped upwardly through the hollow rotor shaft of the electrical motor and then to the wellhead. 7. The method of claim 6, wherein the flow of production fluid through the hollow rotor shaft is characterized by a Reynolds number of at least 2300. 8. The method of claim 7, wherein the motor contains a motor fluid at a positive pressure to the exterior of the motor, and has seals which allow motor fluid to leak into the production fluid. 9. The method of claim 7, wherein conditions are selected such that heat transfer from at least one of the hollow rotor shaft and the exterior surfaces of the motor to the production fluid is characterized by a Nusselt number of at least 10. 10. The method of claim 7 wherein conditions are selected so as to generate a Brinkmann number of less than 2. 11. The method of claim 7 wherein conditions are selected so as to generate a Rossby number of more than 0.5. 12. The method of claim 4 wherein the pump is located between the wellhead and the electric motor. 13. The method of claim 4 wherein the electrical submergible pumping system contains two pumps, one of which is located above the electric motor and one of which is located below the electric motor. 14. The method of claim 1 wherein (a) the electrical motor has a hollow rotor shaft and the pumping system has a fluid intake below the electrical motor and in liquid communication with said hollow rotor shaft; (b) the pumping system has at least one outlet located at or below the electrical motor, said outlet being in liquid communication with the fluid intake, (c) the pumping system is submerged in a well having a well casing; (d) the cross-section that the electrical motor is such that the electrical motor fits within the well casing and a space is defined between the well casing and the electrical motor; and (e) said pumping system is adapted so that during operation a portion of the fluid entering the fluid intake is pumped through the hollow rotor shaft of the electrical motor, and a portion of the fluid entering the fluid intake is pumped through the outlet and upwardly in fluid contact with the outside of the electrical motor. 15. A method of pumping fluids, comprising (I) positioning a pumping system in a well with the pump being located between the electrical motor and the wellhead; wherein (a) electrical motor has a hollow rotor shaft which is in liquid communication with the pump, (e) the cross-section of the electrical motor is such that the electrical motor fits within the well casing and a space is defined between the well casing and the electrical motor; (f) the pumping system has a first fluid intake in liquid communication with said hollow rotor shaft; (g) the pumping system has a second fluid intake above the motor in liquid communication with the space defined between the well casing and the electrical motor, with the at least one outlet located at or below the electrical motor, said outlet being in liquid communication with the fluid intake; and (II) operating said pumping system under conditions such that during operation a portion of the fluids enter said first fluid intake and is pumped through the hollow rotor shaft of the electrical motor, and a second portion of the fluids are pumped through the space defined between the well casing and the electrical motor and in fluid contact with the exterior of the electrical motor to enter the second fluid intake, and said first and second portions of the fluids are then pumped to the wellhead. 16. A mechanism for providing motive force, the mechanism comprising (I) a power unit including: (a) a housing having an exterior wall; (b) a hollow rotor shaft having a longitudinal axis and opposite ends, the hollow rotor shaft being rotatably mounted within the housing for rotation of the hollow rotor shaft relative to the housing, substantially about the longitudinal axis of the hollow rotor shaft; and (c) a drive system mounted within said housing connected to the hollow rotor shaft for causing rotation of the hollow rotor shaft relative to the housing, wherein the drive system includes a plurality of magnets mounted within the housing, located around the hollow rotor shaft, wherein the magnets create magnetic forces for causing the hollow rotor shaft to rotate relative to the housing; and (II) a pumping unit located below the power unit, wherein (i) the pumping unit includes a longitudinal hollow drive shaft that is in fluid communication with the hollow rotor shaft in the power unit, the hollow drive shaft being rotated substantially about its longitudinal axis when the hollow rotor shaft of the power unit is rotated; (ii) the hollow drive shaft of the pumping unit has a shaft inlet proximate to its bottom portion for allowing fluids being pumped to enter the hollow drive shaft; (iii) at least one impeller mounted to the exterior of the hollow drive shaft of the pumping unit such that when the hollow drive shaft is rotated, the impeller pumps fluids downwardly toward the inlet in the hollow drive shaft; and (iv) the pumping unit has fluid intakes proximate to a topmost portion thereof through which fluids being pumped enter the pumping unit; the fluid intakes, hollow drive shaft, shaft inlet and impellers being adapted such that when the hollow drive shaft is rotated about its longitudinal axis, fluids are pumped into the pumping unit through said fluid intakes, downwardly within the pumping unit to the shaft inlet, and then through the hollow drive shaft of the pumping unit and through the hollow rotor shaft of the power unit. 17. A method of removing a mixture of gaseous and liquid fluids from a well having at least one point where a mixture of gaseous and liquid fluids enter the well, comprising (I) positioning a pumping system in the well above the point or points where the mixture of gaseous and liquid fluids enter the well, wherein: (a) the pumping system includes at least one power unit, at least one pumping unit, and at least one intake through which fluids being pumped enter the pumping system; (b) the power unit includes a hollow rotor shaft in fluid communication said intake, and the pumping system is adapted such that fluids entering the intake are pumped through the hollow rotor shaft of the motor; (c) the power unit and pumping unit are smaller in diameter than the well; (d) the intake is located below at least a portion of the pumping system and is of smaller diameter than at least that portion of the pumping system above the intake; (e) the intake is adapted such that liquid fluids being pumped by the pumping system are caused to flow downwardly through a section of the intake to enter the pumping system; and (II) operating the pumping system such that (i) the gaseous fluids and the liquid fluids separate at a point proximate to the intake and below that portion of the pumping system above the intake, (ii) the gaseous fluids bypass the pumping system and rise to the surface of the well, and (iii) the liquid fluids being pumped flow upwardly to the intake, then downwardly through a section of the intake and then through the hollow rotor shaft of the motor and out of the well. 18. A method of pumping fluids with a submergible pumping system including a pump and an electrical motor, wherein the pumping system is submerged in production fluids such that said fluids flow across vortex generators adapted to impart streamwise vorticity to the production fluids as they pass in fluid contact with the exterior portions of the electrical motor. 19. The method of claim 18 wherein the vortex generators are attached to the exterior portions of the electric motor. 20. The method of claim 18 wherein the vortex generators are attached to the interior surface of a motor shroud. |
Receptacle for a programmable, electronic processing device |
A mount for a programmable electronic processing device is disclosed. It is divided into at least a first functional unit (7) and a second functional unit (8) which represent separate, particularly monolithic integrated, components. The first functional unit (7) defines all input and output interfaces of the processing device with respect to their electrical properties. All essential connections of the second functional unit (8) are accessible from outside only via the first functional unit (7). To this end, the first functional unit (7) includes matching circuits which serve to electrically adapt the connections of the second functional unit (8) to the external conditions. |
1. A mount containing a programmable electronic processing device, particularly a microprocessor, which is divided into at least a first functional unit (7) and a second functional unit (8) which represent separate, particularly monolithic integrated, components, characterized in that that the first functional unit (7) defines all input and output interfaces of the processing device with respect to their electrical properties, all essential connections of the second functional unit (8) are accessible from outside only via the first functional unit (7), with direct accesses being normally, and first functional unit (7) comprises matching circuits which serve to electrically adapt the connections of the second functional unit (8) to the external conditions. 2. A mount as claimed in claim 1, characterized in that at first the mount is bare or contains only a portion of the programmable electronic processing device, and that the mount so designed that the remaining portion or the remaining portions of the processing device can be incorporated later. 3. A mount as claimed in claim 2, characterized in that the portion contained in the mount is regarded as forming part of the first functional unit (7). 4. A mount as claimed in claim 1, characterized in that in at least one of the matching circuits, a change of a voltage level takes place. 5. A mount as claimed in claim 4, characterized in that at least one supply terminal of the second functional unit (8) is fed via the first functional unit (7), in which case the matching circuit includes a voltage regulator. 6. A mount as claimed in claim 1, characterized in that in at least one of the matching circuits, a change of a current level takes place. 7. A mount as claimed in claim 1, characterized in that in at least one of the matching circuits, a conversion from a serial data stream to a parallel data stream or from a parallel data stream to a serial data stream takes place. 8. A mount as claimed in claim 1, characterized in that in at least one of the matching circuits, incoming or outgoing data are temporarily stored. 9. A mount as claimed in claim 1, characterized in that at least one of the matching circuits is programmable in terms of its electrical properties. 10. A mount as claimed in claim 9, characterized in that for at least one of the matching circuits, a measurement mode is programmable. 11. A mount as claimed in claim 10, characterized in that in the measurement mode for at least one of the matching circuits, a direct signal path from an external connection of the mount to a connection of the second functional unit (8) can be switched through by means of the program, with the indirect signal path via the matching circuit being off. |
Noise cancellation |
A noise cancellation circuit has a first and second input for a first signal having a signal element and a noise element, and a second signal comprising at least a smaller amplitude of the said signal element, a first inverter arrangement for producing an inverted signal output that is an inverted form of one of the first and second signals, a first adder for adding the other signal and the inverted signal to produce an intermediate signal, an intermediate inverter arrangement for inverting the intermediate signal to produce an inverted intermediate signal, and a second adder for adding the other signal, the inverted signal and the inverted intermediate signal to produce an output. |
1. A noise cancellation circuit comprising: a first input for a first signal having a signal element and a noise element; a second input for a second signal comprising at least a smaller amplitude of the said signal element; a first inverter arrangement for producing an inverted signal output that is an inverted form of one of the first and second signals; a first adder for adding the other signal and the inverted signal to produce an intermediate signal; an intermediate inverter arrangement for inverting the intermediate signal to produce an inverted intermediate signal; and a second adder for adding the other signal, the inverted signal and the inverted intermediate signal to produce an output. 2. A circuit as claimed in claim 1 in which the first inverter arrangement comprises a first inverter having an output of a first inverted signal which is operably connected with the first adder, and a second inverter having an output of a second inverted signal which is operably connected with the second adder. 3. A circuit as claimed in claim 1, including means for balancing the amplitude of the noise in the first signal, the inverted signal and the inverted intermediate signal, such that it is substantially cancelled in the output. 4. A circuit as claimed in claim 3 in which the means for balancing include at least one variable gain amplifier. 5. A circuit as claimed in claim 4 in which the first inverter includes the variable gain amplifier. 6. A circuit as claimed in claim 4 in which the intermediate inverter includes the variable gain amplifier. 7. A circuit as claimed in claim 4, in which the output of the second adder is connected to the variable gain amplifier. 8. A circuit as claimed in claim 1 in which the intermediate inverter attenuates the intermediate signal. 9. A circuit as claimed in claim 1, including a first transducer operably connected with the first input, and a second transducer operably connected with the second input, the second transducer being constructed and/or arranged to receive at least an attenuated amplitude of the signal element relative to the signal element received by the first transducer. 10. A circuit as claimed in claim 9 in which at least one of the first and second transducers is] constructed and arranged to inhibit the reception of the signal element by the second transducer. 11. A circuit as claimed in claim 9 in which the transducers are microphones. 12. A circuit as claimed in claim 9 in which the transducers are microphones and in which the second microphone is arranged with a baffle to reception of the signal element. 13. A circuit as claimed in claim 12 in which the microphones are directional, the first microphone being arranged in front of the second microphone along the major direction of reception of signal thereby. 14. A circuit as claimed in claim 13 in which the receiving faces of the microphones are spaced by a distance in the range 0.2 mm to 2.5 mm, preferably 0.625 mm. 15. A circuit as claimed in claim 1 in which the first inverter arrangement is arranged to invert the second signal from the second input to produce the inverted signal. 16. A circuit as claimed in claim 1 in which the first and second signals are low pass filtered to attenuate higher frequency noise. 17. A method of noise reduction comprising: comparing a first signal having a signal element and a noise element and a second signal having at least a smaller amplitude of the said signal element to produce an intermediate signal; and subtracting the intermediate signal from a comparison of the first signal and the second signal to produce an output in which the noise is reduced. 18. A microphone arrangement for a noise cancellation circuit, comprising a first microphone arranged to receive a signal element and a noise element, a second microphone arranged with a baffle such that it receives at least a smaller amplitude of signal element relative to the first microphone. 19. A microphone arrangement as claimed in claim 18 in which the second microphone is arranged in relation to the first microphone such that the first microphone acts as a baffle to receipt of the signal element by the second microphone. 20. A circuit as claimed in claim 9 in which the transducers are microphones and in which the second microphone is arranged with a baffle to reception of the signal element and in which the second microphone is arranged at a greater distance from the signal source than the first microphone. 21. A circuit as claimed in claim 9 in which the transducers are microphones and in which the second microphone is arranged at a greater distance from the signal source than the first microphone. |
Method for determining synthetic term senses using reference text |
A method for determining term senses by identifying terms having multiple “senses” or meanings. For a given source term and trigger term, a list of important terms having high relevance to a combination of the source term and trigger term is created. The list of important terms is determined to be a “sense” for the source term in accordance with the present invention. A sense is assigned to a given term of a given document by determining similarity of the document to one or more senses and assigning a sense as a function of the degree of similarity. A sense-indicative index is created to treat multiple occurrences of a term distinctly, as a function of each respective assigned sense. Accordingly, the index may be used for sense-relevant information retrieval when a sense of a query term is discernible or specified. |
1. A method for determining a sense of a source term of a document, the method comprising the steps of: (a) identifying a trigger term relating to said source term; (b) identifying a plurality of important terms relating to a combination of said source term and said trigger term; and (c) establishing a sense of said source term, said sense comprising said plurality of important terms. 2. The method of claim 1, wherein step (c) comprises the step of: (c1) storing said plurality of important terms in association with said source term. 3. The method of claim 1, wherein step (a) comprises the step of: (a1) searching a group of documents for terms important to said source term. 4. The method of claim 1, wherein step (a) comprises the steps of: (a1) determining a reference frequency for each of a plurality of terms of a reference text, said reference frequency comprising a frequency of occurrence within said reference text; (a2) identifying a sample text comprising documents comprising said source term; (a3) determining a sample frequency for each of a plurality of terms of said sample text, said sample frequency comprising a frequency of occurrence within said sample text; (a4) for each of said plurality of terms of said sample text, comparing a respective sample frequency to a respective reference frequency to determine importance as a function of said respective frequencies by calculating a difference between said respective sample frequency and said respective reference frequency; (a5) assigning an importance score to each of said plurality of terms of said sample text, said importance score being determined as a function of said difference; and (a6) defining a plurality of trigger terms to comprise each of said plurality of terms of said sample text having a respective importance score exceeding a threshold. 5. The method of claim 1, wherein step (b) comprises the step of: (b1) searching a group of documents for terms important to said source term. 6. The method of claim 1, wherein step (b) comprises the steps of: (b1) determining a reference frequency for each of a plurality of terms of a reference text, said reference frequency comprising a frequency of occurrence within said reference text; (b2) identifying a sample text comprising documents comprising said source term; (b3) determining a sample frequency for each of a plurality of terms of said sample text, said sample frequency comprising a frequency of occurrence within said sample text; (b4) for each of said plurality of terms of said sample text, comparing a respective sample frequency to a respective reference frequency to determine importance as a function of said respective frequencies by calculating a difference between said respective sample frequency and said respective reference frequency; (b5) assigning an importance score to each of said plurality of terms of said sample text, said importance score being determined as a function of said difference; and (b6) defining said plurality of important terms to comprise each of said plurality of terms having a respective importance score exceeding a threshold. 7. A method for determining senses of a source term, the method comprising the steps of: (a) identifying a plurality of trigger terms relating to said source term; (b) for one of said plurality of trigger terms, identifying a plurality of important terms relating to a combination of said source term and said one of said plurality of trigger terms, said plurality of important terms comprising a sense of said source term; (c) removing from said plurality of trigger terms all of said plurality of important terms, if any, to define a reduced plurality of trigger terms; and (d) for one of said reduced plurality of trigger terms, identifying a next 11 plurality of important terms relating to a combination of said source term and said one of said reduced plurality of trigger terms, said next plurality of important terms comprising a next sense of said source term. 8. The method of claim 7, wherein step (a) comprises the steps of: (a1) determining a reference frequency for each of a plurality of terms of a reference text, said reference frequency comprising a frequency of occurrence within said reference text; (a2) identifying a sample text comprising documents comprising said source term; (a3) determining a sample frequency for each of a plurality of terms of said sample text, said sample frequency comprising a frequency of occurrence within said sample text; (a4) for each of said plurality of terms of said sample text, comparing a respective sample frequency to a respective reference frequency to determine importance as a function of said respective frequencies by calculating a difference between said respective sample frequency and said respective reference frequency; (a5) assigning an importance score to each of said plurality of terms of said sample text, said importance score being determined as a function of said difference; and (a6) defining said plurality of trigger terms to comprise each of said plurality of terms having a respective importance score exceeding a threshold. 9. A method for assigning a sense to a document's term for facilitating sense-relevant retrieval of said document by an information retrieval system, the method comprising the steps of: identifying a term of said document; identifying a first sense for said term, said first sense comprising terms relating to said term; comparing said first sense to said document to determine similarity; assigning said first sense to said term if said first sense and said document are sufficiently similar. 10. The method of claim 9, further comprising the steps of: identifying a next sense for said term if said first sense and said document are not sufficiently similar, said sense comprising terms relating to said term; comparing said next sense to said document to determine similarity; and assigning said next sense to said term if said next sense and said document are sufficiently similar. 11. The method of claim 9, wherein said comparing step is performed using a cosine measure technique. 12. The method of claim 9, wherein said assigning step comprises storing data associating said term with said first sense. 13. The method of claim 9, wherein said identifying step comprises referencing a memory storing a plurality of senses, each of said plurality of senses comprising a plurality of terms. 14. A method for assigning a sense to a document's term for facilitating sense-relevant retrieval of said document by an information retrieval system, the method comprising the steps of: identifying a term of said document; identifying a plurality of senses for said term, each of said plurality of senses 6 comprising a plurality of terms; comparing each of said plurality of senses to said document to determine similarity, assigning to said term a sense of said plurality of senses determined to have the greatest similarity. 15. The method of claim 14, wherein said comparing step comprises the step of generating a similarity score for each of said plurality of senses, and wherein said assigning step comprises the step of assigning to said term said sense of said plurality of senses determined to have the highest similarity score. 16. A method for preparing a group of documents for sense-relevant retrieval by an information retrieval system, the method comprising the steps of: (a) creating an index for said group of documents, said index associating each of a plurality of term identifiers with a corresponding set of document identifiers, each of said plurality of term identifiers being associated with a term, each of said set of document identifiers being associated with a document; (b) for each of said term identifiers, identifying a sense corresponding to a respective term and a respective document associated with a respective one of said corresponding set of document identifiers, said sense comprising a plurality of important terms; (c) for each of said term identifiers, creating at least one sensed term identifier, each said sensed term identifier corresponding to a respective term identifier and a corresponding sense; and (d) creating a sensed index for said group of documents, said sensed index associating each of said sensed term identifiers with a corresponding set of document identifiers, each of said sensed term identifiers being associated with a respective term and a respective sense. 17. An information processing system for determining a sense of a source term of a document, the system comprising: a central processing unit (CPU) for executing programs; a memory operatively connected to said CPU; a first program stored in said memory and executable by said CPU for identifying a trigger term relating to said source term; and a second program stored in said memory and executable by said CPU for identifying a plurality of important terms relating to a combination of said source term and said trigger term, said sense comprising said plurality of important terms. 18. An information processing system for assigning a sense to a document's term for facilitating sense-relevant retrieval of the document by an information retrieval system, the system comprising: a central processing unit (CPU) for executing programs; a memory operatively connected to said CPU; a first program stored in said memory and executable by said CPU for identifying a term of said document; a second program stored in said memory and executable by said CPU for identifying a first sense for said term, said sense comprising terms relating to said term; a third program stored in said memory and executable by said CPU for comparing 11 said first sense to said document to determine similarity; and a fourth program stored in said memory and executable by said CPU for assigning said first sense to said term if said first sense and said document are sufficiently similar. 19. An information processing system for preparing a group of documents for sense-relevant retrieval by an information retrieval system, the system comprising: a central processing unit (CPU) for executing programs; a memory operatively connected to said CPU; a first program stored in said memory and executable by said CPU for creating an index for said group of documents, said index associating each of a plurality of term identifiers with a corresponding set of document identifiers, each of said plurality of term identifiers being associated with a term, each of said set of document identifiers being associated with a document; a second program stored in said memory and executable by said CPU for identifying, for each of said term identifiers, a sense corresponding to a respective term and a respective document associated with a respective one of said corresponding set of document identifiers, said sense comprising a plurality of important terms; a third program stored in said memory and executable by said CPU for creating, for each of said term identifiers, at least one sensed term identifier, each said sensed term identifier corresponding to a respective term identifier and a corresponding sense; and a fourth program stored in said memory and executable by said CPU for creating a sensed index for said group of documents, said sensed index associating each of said sensed term identifiers with a corresponding set of document identifiers, each of said sensed term identifiers being associated with a respective term and a respective sense. 20. An information processing system for facilitating sense-relevant retrieval of a document by an information retrieval system, the system comprising: a central processing unit (CPU) for executing programs; a memory operatively connected to said CPU; a document stored in said memory, said document comprising a term; and data stored in said memory associating said term with a sense comprising a plurality of terms. 21. The information processing system of claim 20, further comprising: data stored in said memory identifying said plurality of terms. 22. The information processing system of claim 21, further comprising: a first program stored in said memory and executable by said CPU for comparing said plurality of terms to a search query. 23. The information processing system of claim 22, further comprising: a second program stored in said memory and executable by said CPU for identifying said document as a relevant search result for said search query if said first program determines sufficient similarity between said plurality of terms and said search query. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention The present invention relates generally to computerized systems for processing, searching and retrieving information. In particular, the present invention relates to techniques for determining a “sense” for a term that may be used, for example, to provide more accurate search results by seeking to match not only search terms but also the search term's sense. 2. Description of the Related Art Computerized text-based information retrieval systems are now in widespread use in database, intranet and internet (e.g., World Wide Web) applications. Such systems typically search a large group of documents according to search query terms provided by a user to identify a subset of documents of the group of documents that are likely of interest to the user based on the search query terms. Since a very large number of search results is typically obtained, it is burdensome to the user to identify the relatively few documents that are actually of interest to the user. This problem is exacerbated by the fact that many document terms and search terms have multiple contextual meanings. As used herein, “term” is used broadly and may include, for example, a word, a stemmed word, a character n-gram, phrase, or any other grouping that may be used to characterize text. Term meanings are typically not accounted for as part of the automated search process. For example, when a term has more than one “sense”, documents are often returned as search results when there is a term match, but not necessarily a sense match. For illustrative purposes, consider a Web-based search engine operated by a user to search the Web for documents relating to the search term “jaguar”. The term “jaguar” has multiple senses, including an automotive sense, an animal sense, and a sports sense. As a result, when the search engine retrieves documents that contain the word “jaguar”, some will likely relate to Jaguar cars, others will likely relate to jaguar animals, and yet others will likely relate to a Jaguars football team. This is undesirable because a user must review an unnecessarily large set of search results including all such senses of the search term, although the user may be interested only in documents in which the term is used in one of the term's senses. What is needed is a method for determining a term's sense for facilitating automated sense-relevant information retrieval. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is directed toward a method and apparatus for determining synthetic term senses for facilitating automated sense-relevant information processing, such as automated sense-relevant information retrieval. Conceptually, the present invention can be embodied in two principal aspects, which may be combined. The first aspect may involve using reference text, e.g. a group of documents, to determine senses of terms. In effect, this establishes a “library” of senses for terms in the large body of documents. This may be performed before any search queries are processed, and/or may be ongoing as new documents are added to the collection. In contrast to using a dictionary to identify a “sense”, this method is advantageous because it provides “synthetic senses”—some of which likely cannot be found in a dictionary. Additionally, this method excludes dictionary senses that do not appear in the reference text, and therefore are of little relevance when searching reference text. The senses are “synthetic” in that they are based on document collection statistics, e.g. term frequencies, rather than dictionary definitions. The second aspect involves assigning an appropriate sense to a term in a given document or query. In effect, this provides an indication of the terms' senses for each document in the large body of documents. For example, the term's sense for a given document may be used for sense-relevant information retrieval, cross-language translations, etc. Once term senses have been determined and/or assigned in accordance with the present invention, sense-relevant automated information retrieval may be performed using known information retrieval and indexing techniques by using appropriate term senses in the index and in the query instead of unsensed terms, i.e. terms to which senses have not been assigned. In addition to sense-relevant automated information retrieval, the present invention may be used to provide senses for use in the context of a thesaurus, a cross-language dictionary, or a document index. |
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