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1. A telecommunication element having a color identifying coating thereon, the telecommunication element comprising: an elongated communication transmission medium; and a coating having an identifying color applied on at least a portion of the transmission medium, wherein said coating comprises a radiation-cured, crosslinked, polymeric network, and wherein the identifying color in the coating is at least in part provided by at least one chromophore moiety covalently bonded by at least one covalent bond to said polymeric network. 2. The telecommunications element of claim 1, wherein: (a) the elongated transmission medium is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon; and (b) the identifying color is thermally stable and light fast. 3. The telecommunications element of claim 1, wherein the identifying color in the coating is at least in part provided by at least one anthraquinone moiety covalently bonded by at least one covalent bond to said polymeric network. 4. The telecommunications element of claim 1, wherein the identifying color in the coating is at least in part provided by at least one methine moiety covalently bonded by at least one covalent bond to said polymeric network. 5. The telecommunications element of claim 1, wherein the identifying color in the coating is at least in part provided by at least one azo moiety covalently bonded by at least one covalent bond to said polymeric network. 6. A radiation-curable, chromophore-containing compound comprising at least one radiation-curable group and at least one chromophore moiety, wherein the radiation-curable, chromophore-containing compound comprises the reaction product of: (a) an isocyanate-containing compound comprising (i) at least one isocyanate group and (ii) the at least one chromophore moiety, and (b) a radiation-curable compound comprising (i) at least one isocyanate-reactive functional group and (ii) the at least one radiation-curable group, wherein the at least one radiation-curable group of said radiation-curable, chromophore-containing compound is covalently bonded to said radiation-curable, chromophore-containing compound by at least one covalent bond formed by reacting an isocyanate-reactive functional group (i) of said radiation-curable compound (b) with an isocyanate group (i) of said isocyanate-containing compound (a), and said isocyanate-containing compound (a) is the reaction product of: (c) a chromophore-containing compound comprising (i) a chromophore moiety and (ii) at least one isocyanate-reactive functional group; and (d) a polyisocyanate. 7. The radiation-curable, chromophore-containing compound of claim 6, wherein the at least one isocyanate-reactive functional group (i) of said radiation-curable compound (b) and the at least one isocyanate-reactive functional group (ii) of said chromophore-containing compound (c) are independently selected from the group consisting of —OH, —NH2 and —SH. 8. The radiation-curable, chromophore-containing compound of claim 6, wherein the at least one radiation-curable group (ii) of said radiation-curable compound (b) comprises ethylenic unsaturation. 9. The radiation-curable, chromophore-containing compound of claim 8, wherein the ethylenically-unsaturated, radiation-curable group (ii) of said radiation-curable compound (b) comprises a (meth)acrylate. 10. The radiation-curable, chromophore-containing compound of claim 6, wherein the at least one radiation-curable group (ii) of said radiation-curable compound (b) comprises an epoxy group. 11. The radiation-curable, chromophore-containing compound of claim 6, wherein the polyisocyanate (d) is a diisocyanate. 12. The radiation-curable, chromophore-containing compound of claim 6, wherein the at least one chromophore moiety (i) of said chromophore-containing compound (c) is a methine dye. 13. The radiation-curable, chromophore-containing compound of claim 6, wherein the at least one chromophore moiety (i) of said chromophore-containing compound (c) is an azo dye. 14. The radiation-curable, chromophore-containing compound of claim 6, wherein the at least one chromophore moiety (i) of said chromophore-containing compound (c) is an anthraquinone dye. 15. The radiation-curable, chromophore-containing compound of claim 14, wherein the anthraquinone dye has the following formula: wherein R groups R1, R2, R3, R4, R5, R6, R7 and R8 are each independently selected from the group consisting of hydrogen, amino, hydroxy, halogen, nitro, carboxylated alkali metal, sulfated alkali metal and a hydrocarbyl group optionally containing one or more heteroatoms, provided that at least two of R groups R1 through R8 have at least one isocyanate-reactive functional group selected from the group consisting of —OH, —NH2 and —SH, and further wherein adjacent R groups from among R1 through R8 can form a ring. 16. The radiation-curable, chromophore-containing compound of claim 15, wherein from 1 to 3 of said R groups R1 through R8 have the following formula: wherein R9 is hydrogen or an alkyl group having from 1 to about 12 carbon atoms, X is —CH2—, a is an integer from 1 to about 6, Y represents polymeric units of hydroxy alkylenes or alkylene oxide monomers selected from the group consisting of ethylene oxide, propylene oxide, butylene oxide, cyclohexane oxide, and glycidol, b is either 0 or 1, and Z is —OH, —NH2, or —SH group, and further wherein the remainder of said R groups R1 through R8 are selected from the group consisting of hydrogen, amino, hydroxy, halogen, nitro, carboxylated alkali metal and sulfated alkali metal. 17. The radiation-curable, chromophore-containing compound of claim 15, wherein the anthraquinone dye has the following formula: wherein R9 and R10 are independently selected from hydrogen or an alkyl group having from 1 to about 12 carbon atoms, X is —CH2—, a and a′ independently are integers from 1 to about 6, Y and Y′ are independently represent polymeric units of hydroxy alkylenes or alkylene oxide monomers selected from the group consisting of ethylene oxide, propylene oxide, butylene oxide, cyclohexane oxide, and glycidol, b and b′ are independently either 0 or 1, and Z and Z′ independently are —OH, —NH2, or —SH groups. 18. The radiation-curable, chromophore-containing compound of claim 17, wherein the anthraquinone dye has the formula: wherein n, n′, m, m′, p, and p′ independently have a value of from 0 to about 40. 19. The radiation-curable, chromophore-containing compound of claim 17, wherein the anthraquinone dye has the formula: 20. The radiation-curable, chromophore-containing compound of claim 14, wherein the anthraquinone dye is selected from the group consisting of 1,5-bis((3-hydroxy-2,2-dimethylpropyl)amino)-9,10-anthracenedione; 2,2′-((9,10-dihydro-9, 10-dioxo-1,5-anthracenediyl)bis(thio))bis-benzoic acid, 2-hydroxyethyl ester, and 1,5-bis((2,2-dimethyl-3-hydroxypropyl)amino)-4,8-bis((4-methylphenyl)thio) anthraquinone. 21. The radiation-curable, chromophore-containing compound of claim 14, wherein the anthraquinone dye is 1,5-bis((2,2-dimethyl-3-hydroxypropyl)amino)-4,8-bis((4-methylphenyl)thio) anthraquinone. 22. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a radiation-curable, chromophore-containing compound according to claim 6. 23. The radiation-curable composition of claim 22, wherein the composition further comprises TiO2. 24. The radiation-curable composition of claim 22, wherein the composition further comprises one or more of a photoinitiator, a reactive diluent, a stabilizer, and a surfactant. 25. A substrate having the radiation-curable composition of claim 22 on at least a portion thereof. 26. The substrate of claim 25, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 27. A method for producing a color-identifying, radiation-cured composition on at least a portion of a substrate, wherein the color-identifying, radiation-cured composition has at least one chromophore moiety covalently bonded to at least one other component of the color-identifying, radiation-cured composition, the method comprising the steps of: providing a substrate; providing a radiation-curable composition according to claim 22; applying the radiation-curable composition of claim 22 to at least a portion of the substrate; and subjecting the applied composition for a suitable period of time to radiation of a suitable wavelength and intensity to cause curing of the composition into the color-identifying, radiation-cured composition. 28. The method of claim 27, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 29. A dye concentrate comprising the radiation-curable, chromophore-containing compound of claim 6 and tetrahydrofurfuryl acrylate (THFA). 30. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 29. 31. The radiation-curable composition of claim 30, wherein the composition further comprises TiO2. 32. The radiation-curable composition of claim 30, wherein the composition further comprises one or more of a photoinitiator, a reactive diluent, a stabilizer, and a surfactant. 33. A substrate having the radiation-curable composition of claim 30 on at least a portion thereof. 34. The substrate of claim 33, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 35. A method for producing a color-identifying, radiation-cured composition on at least a portion of a substrate, wherein the color-identifying, radiation-cured composition has at least one chromophore moiety covalently bonded to at least one other component of the color-identifying, radiation-cured composition, the method comprising the steps of: providing a substrate; providing a radiation-curable composition according to claim 30; applying the radiation-curable composition of claim 30 to at least a portion of the substrate; and subjecting the applied composition for a suitable period of time to radiation of a suitable wavelength and intensity to cause curing of the composition into the color-identifying, radiation-cured composition. 36. The method of claim 35, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 37. The dye concentrate of claim 29, wherein the dye concentrate comprises greater than 5 wt % chromophore moiety, based on the total weight of the dye concentrate. 38. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 37. 39. The dye concentrate of claim 29, wherein the dye concentrate comprises from about 10 wt % to about 35 wt % of chromophore moiety, based on the total weight of the dye concentrate. 40. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 39. 41. A dye concentrate comprising a blend of two or more radiation-curable, chromophore-containing compounds according to claim 6 and tetrahydrofurfuryl acrylate (THFA), wherein at least the chromophore moiety of a first radiation-curable, chromophore-containing compound according to claim 6 is different from the chromophore moiety of a second radiation-curable, chromophore-containing compound according to claim 6. 42. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 41. 43. A radiation-curable, chromophore-containing compound comprising at least one radiation-curable group and at least one chromophore moiety, wherein the radiation-curable, chromophore-containing compound comprises the reaction product of: (a) a chromophore-containing compound comprising (i) at least one isocyanate-reactive functional group and (ii) the at least one chromophore moiety; and (b) a radiation-curable compound comprising (i) at least one isocyanate group and (ii) the, at least one radiation-curable group, wherein the at least one radiation-curable group of said radiation-curable, chromophore-containing compound is covalently bonded to said radiation-curable, chromophore-containing compound by at least one covalent bond formed by reacting an isocyanate group (i) of said radiation-curable compound (b) with an isocyanate-reactive functional group (i) of said chromophore-containing compound (a). 44. The radiation-curable, chromophore-containing compound of claim 43, wherein the at least one isocyanate-reactive functional group (i) of said chromophore-containing compound (a) is selected from the group consisting of —OH, —NH2 and —SH. 45. The radiation-curable, chromophore-containing compound of claim 43, wherein the at least one radiation-curable group (ii) of said radiation-curable compound (b) comprises ethylenic unsaturation. 46. The radiation-curable, chromophore-containing compound of claim 45, wherein the ethylenically-unsaturated, radiation-curable group (ii) of said radiation-curable compound (b) comprises a meth(acrylate). 47. The radiation-curable, chromophore-containing compound of claim 43, wherein the at least one radiation-curable group (ii) of said radiation-curable compound (b) comprises an epoxy group. 48. The radiation-curable, chromophore-containing compound of claim 43, wherein the at least one chromophore moiety (ii) of said chromophore-containing compound (a) is a methine dye. 49. The radiation-curable, chromophore-containing compound of claim 43, wherein the at least one chromophore moiety (ii) of said chromophore-containing compound (a) is an azo dye. 50. The radiation-curable, chromophore-containing compound of claim 43, wherein the at least one chromophore moiety (ii) of said chromophore-containing compound (a) is an anthraquinone dye. 51. The radiation-curable, chromophore-containing compound of claim 43, wherein the chromophore-containing compound (a) is an anthraquinone dye having the formula: wherein R groups R1, R2, R3, R4, R5, R6, R7 and R8 are each independently selected from the group consisting of hydrogen, amino, hydroxy, halogen, nitro, carboxylated alkali metal, sulfated alkali metal and a hydrocarbyl group optionally containing one or more heteroatoms, provided that at least two of R groups R1 through R8 have at least one isocyanate-reactive functional group selected from the group consisting of —OH, —NH2 and —SH, and further wherein adjacent R groups from among R1 through R8 can form a ring. 52. The radiation-curable, chromophore-containing compound of claim 51, wherein from 1 to 3 of said R groups R1 through R8 have the following formula: wherein R9 is hydrogen or an alkyl group having from 1 to about 12 carbon atoms, X is —CH2—, a is an integer from 1 to about 6, Y represents polymeric units of hydroxy alkylenes or alkylene oxide monomers selected from the group consisting of ethylene oxide, propylene oxide, butylene oxide, cyclohexane oxide, and glycidol, b is either 0 or 1, and Z is —OH, —NH2, or —SH group, and further wherein the remainder of said R groups R1 through R8 are selected from the group consisting of hydrogen, amino, hydroxy, halogen, nitro, carboxylated alkali metal and sulfated alkali metal. 53. The radiation-curable, chromophore-containing compound of claim 51, wherein the anthraquinone dye has the formula: wherein R9 and R10 are independently selected from hydrogen or an alkyl group having from 1 to about 12 carbon atoms, X is —CH2—, a and a′ independently are integers from 1 to about 6, Y and Y′ are independently represent polymeric units of hydroxy alkylenes or alkylene oxide monomers selected from the group consisting of ethylene oxide, propylene oxide, butylene oxide, cyclohexane oxide, and glycidol, b and b′ are independently either 0 or 1, and Z and Z′ independently are —OH, —NH2, or —SH groups. 54. The radiation-curable, chromophore-containing compound of claim 53, wherein the anthraquinone dye has the formula: wherein n, n′, m, m′, p, and p′ independently have a value of from 0 to about 40. 55. The radiation-curable, chromophore-containing compound of claim 53, wherein the anthraquinone dye has the formula: 56. The radiation-curable, chromophore-containing compound of claim 43, wherein the chromophore-containing compound (a) is selected from the group consisting of 1,5-bis((3-hydroxy-2,2-dimethylpropyl)amino)-9,10-anthracenedione; 2,2′-((9,10-dihydro-9,10-dioxo-1,5-anthracenediyl)bis(thio))bis-benzoic acid, 2-hydroxyethyl ester; and 1,5-bis((2,2-dimethyl-3-hydroxypropyl)amino)-4,8-bis((4-methylphenyl)thio) anthraquinone. 57. The radiation-curable, chromophore-containing compound of claim 56, wherein the chromophore-containing compound (a) is 1,5-bis((2,2-dimethyl-3-hydroxypropyl)amino)-4,8-bis((4-methylphenyl)thio) anthraquinone. 58. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a radiation-curable, chromophore-containing compound according to claim 43. 59. The radiation-curable composition of claim 58, wherein the composition further comprises TiO2. 60. The radiation-curable composition of claim 58, wherein the composition further comprises one or more of a photoinitiator, a reactive diluent, a stabilizer, and a surfactant. 61. A substrate having the radiation-curable composition of claim 58 on at least a portion thereof. 62. The substrate of claim 61, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 63. A method for producing a color-identifying, radiation-cured composition on at least a portion of a substrate, wherein the color-identifying, radiation-cured composition has at least one chromophore moiety covalently bonded to at least one other component of the color-identifying, radiation-cured composition, the method comprising the steps of: providing a substrate; providing a radiation-curable composition according to claim 58; applying the radiation-curable composition of claim 58 to at least a portion of the substrate; and subjecting the applied composition for a suitable period of time to radiation of a suitable wavelength and intensity to cause curing of the composition into the color-identifying, radiation-cured composition. 64. The method of claim 63, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 65. A dye concentrate comprising the radiation-curable, chromophore-containing compound of claim 43 and tetrahydrofurfuryl acrylate (THFA). 66. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 65. 67. The radiation-curable composition of claim 66, wherein the composition further comprises TiO2. 68. The radiation-curable composition of claim 66, wherein the composition further comprises one or more of a photoinitiator, a reactive diluent, a stabilizer, and a surfactant. 69. A substrate having the radiation-curable composition of claim 66 on at least a portion thereof. 70. The substrate of claim 69, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 71. A method for producing a color-identifying, radiation-cured composition on at least a portion of a substrate, wherein the color-identifying, radiation-cured composition has at least one chromophore moiety covalently bonded to at least one other component of the color-identifying, radiation-cured composition, the method comprising the steps of: providing a substrate; providing a radiation-curable composition according to claim 66; applying the radiation-curable composition of claim 66 to at least a portion of the substrate; and subjecting the applied composition for a suitable period of time to radiation of a suitable wavelength and intensity to cause curing of the composition into the color-identifying, radiation-cured composition. 72. The method of claim 71, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 73. The dye concentrate of claim 65, wherein the dye concentrate comprises greater than 5 wt % chromophore moiety, based on the total weight of the dye concentrate. 74. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 73. 75. The dye concentrate of claim 65, wherein the dye concentrate comprises from about 10 wt % to about 35 wt % of chromophore moiety, based on the total weight of the dye concentrate. 76. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 75. 77. A dye concentrate comprising a blend of two or more radiation-curable, chromophore-containing compounds according to claim 43 and tetrahydrofurfuryl acrylate (THFA), wherein at least the chromophore moiety of a first radiation-curable, chromophore-containing compound according to claim 43 is different from the chromophore moiety of a second radiation-curable, chromophore-containing compound according to claim 43. 78. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 77. 79. A radiation-curable, chromophore-containing compound comprising at least one radiation-curable group and at least one chromophore moiety. 80. The radiation-curable, chromophore-containing compound of claim 79, wherein the at least one radiation-curable group comprises ethylenic unsaturation. 81. The radiation-curable, chromophore-containing compound of claim 80, wherein the ethylenically-unsaturated, radiation-curable group comprises a meth(acrylate). 82. The radiation-curable, chromophore-containing compound of claim 79, wherein the at least one radiation-curable group comprises an epoxy group. 83. The radiation-curable, chromophore-containing compound of claim 79, wherein the at least one chromophore moiety is a methine dye, and the methine dye comprises a methine core group with at least one substituent comprising the at least one radiation-curable group. 84. The radiation-curable, chromophore-containing compound of claim 79, wherein the at least one chromophore moiety is an azo dye, and the azo dye comprises an azo core group with at least one substituent comprising the at least one radiation-curable group. 85. The radiation-curable, chromophore-containing compound of claim 79, wherein the at least one chromophore moiety is an anthraquinone dye, and the anthraquinone dye comprises an anthraquinone core group with at least one substituent comprising the at least one radiation-curable group. 86. The radiation-curable, chromophore-containing compound of claim 85, wherein the anthraquinone dye has the formula: wherein R groups R1, R2, R3, R4, R5, R6, R7 and R8 are each independently selected from the group consisting of hydrogen, amino, hydroxy, halogen, nitro, carboxylated alkali metal, sulfated alkali metal and a hydrocarbyl group optionally containing one or more heteroatoms, provided that at least one of R groups R11 through R18 comprises at least one ethylenically unsaturated radiation-curable group. 87. The radiation-curable, chromophore-containing compound of claim 86, wherein one or two of said R groups R11 through R18 have a (meth)acrylic functionality and at least four of said R groups R11 through R18 are hydrogen. 88. The radiation-curable, chromophore-containing compound of claim 85, wherein the anthraquinone dye has one of the following formulas: wherein R29, R30, R31, R32, R33, and R34 are the same or different and are independently hydrogen or a C1 to C6 alkyl optionally substituted with one or more substituents selected from the group consisting of —OH, —NH2, —SH, —NO2, —CN and halogen. 89. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a radiation-curable, chromophore-containing compound according to claim 79. 90. The radiation-curable composition of claim 89, wherein the composition further comprises TiO2. 91. The radiation-curable composition of claim 89, wherein the composition further comprises one or more of a photoinitiator, a reactive diluent, a stabilizer, and a surfactant. 92. A substrate having the radiation-curable composition of claim 89 on at least a portion thereof. 93. The substrate of claim 92, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 94. A method for producing a color-identifying, radiation-cured composition on at least a portion of a substrate, wherein the color-identifying, radiation-cured composition has at least one chromophore moiety covalently bonded to at least one other component of the color-identifying, radiation-cured composition, the method comprising the steps of: providing a substrate; providing a radiation-curable composition according to claim 89; applying the radiation-curable composition of claim 89 to at least a portion of the substrate; and subjecting the applied composition for a suitable period of time to radiation of a suitable wavelength and intensity to cause curing of the composition into the color-identifying, radiation-cured composition. 95. The method of claim 94, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 96. A dye concentrate comprising the radiation-curable, chromophore-containing compound of claim 79 and tetrahydrofurfuryl acrylate (THFA). 97. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 96. 98. The radiation-curable composition of claim 97, wherein the composition further comprises TiO2. 99. The radiation-curable composition of claim 97, wherein the composition further comprises one or more of a photoinitiator, a reactive diluent, a stabilizer, and a surfactant. 100. A substrate having the radiation-curable composition of claim 97 on at least a portion thereof. 101. The substrate of claim 100, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 102. A method for producing a color-identifying, radiation-cured composition on at least a portion of a substrate, wherein the color-identifying, radiation-cured composition has at least one chromophore moiety covalently bonded to at least one other component of the color-identifying, radiation-cured composition, the method comprising the steps of: providing a substrate; providing a radiation-curable composition according to claim 97; applying the radiation-curable composition of claim 97 to at least a portion of the substrate; and subjecting the applied composition for a suitable period of time to radiation of a suitable wavelength and intensity to cause curing of the composition into the color-identifying, radiation-cured composition. 103. The method of claim 102, wherein the substrate is selected from the group consisting of (i) an optical fiber having a core and a cladding surrounding the core, (ii) an optical fiber having a core, a cladding surrounding the core, and one or more polymeric coatings on the cladding, (iii) a plurality of optical fibers arranged in an array, and (iv) an optical fiber ribbon. 104. The dye concentrate of claim 96, wherein the dye concentrate comprises greater than 5 wt % chromophore moiety, based on the total weight of the dye concentrate. 105. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 104. 106. The dye concentrate of claim 96, wherein the dye concentrate comprises from about 10 wt % to about 35 wt % of chromophore moiety, based on the total weight of the dye concentrate. 107. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 106. 108. A dye concentrate comprising a blend of two or more radiation-curable, chromophore-containing compounds according to claim 79 and tetrahydrofurfuryl acrylate (THFA), wherein at least the chromophore moiety of a first radiation-curable, chromophore-containing compound according to claim 79 is different from the chromophore moiety of a second radiation-curable, chromophore-containing compound according to claim 79. 109. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 108. 110. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and two or more of (b) a radiation-curable, chromophore-containing compound according to claim 6; (c) a radiation-curable, chromophore-containing compound according to claim 43; and (d) a radiation-curable, chromophore-containing compound according to claim 79. 111. A dye concentrate comprising tetrahydrofurfuryl acrylate (THFA) and two or more of a radiation-curable, chromophore-containing compound according to claim 6, a radiation-curable, chromophore-containing compound according to claim 43, and a radiation-curable, chromophore-containing compound according to claim 79. 112. A radiation-curable composition, comprising: (a) a non-chromophore-containing, radiation-curable oligomer; and (b) a dye concentrate according to claim 111. 113. A colored oligomer for providing color to a coating on a communications element, said colored oligomer comprising the reaction product of: (a) an isocyanate end capped oligomer, and (b) a radiation curable monomer having both (i) a reactive functionality which is reactive with isocyanate and (ii) ethylenic unsaturation, wherein said colored oligomer is end capped with radiation curable groups by covalent linkages formed by reacting said reactive functionality (i) of said radiation curable monomer (b) with an isocyanate moiety of said isocyanate end capped oligomer (a), and said isocyanate end capped oligomer (a) is the reaction product of: (c) at least one polyfunctional compound having at least two isocyanate reactive groups; and (d) at least one polyisocyanate, said polyfunctional compound (c) comprising at least one dye having at least two isocyanate reactive functionalities. 114. The colored oligomer of claim 113, wherein said dye is an anthraquinone dye and said anthraquinone dye has the following formula: wherein R groups R1, R2, R3, R4, R5, R6, R7 and R8 are each independently selected from the group consisting of hydrogen, amino, hydroxy, halogen, nitro, carboxylated alkali metal, sulfated alkali metal and a hydrocarbyl group optionally containing one or more heteroatoms, provided that at least two of R groups R1 through R8 have at least one isocyanate reactive functionality selected from the group consisting of —OH, —NH2 and —SH, and further wherein adjacent R groups from among R1 through R8 can form a ring. 115. The colored oligomer of claim 114, wherein from 1 to 3 of said R groups R1 through R8 have the following formula: wherein R9 is hydrogen or an alkyl group having from 1 to about 12 carbon atoms, X is —CH2—, a is an integer from 1 to about 6, Y represents polymeric units of hydroxy alkylenes or alkylene oxide monomers selected from the group consisting of ethylene oxide, propylene oxide, butylene oxide, cyclohexane oxide, and glycidol, b is either 0 or 1, and Z is a reactive —OH, —NH2, or —SH group, and further wherein the remainder of said R groups R1 through R8 are selected from the group consisting of hydrogen, amino, hydroxy, halogen, nitro, carboxylated alkali metal and sulfated alkali metal. 116. The colored oligomer of claim 114, wherein said anthraquinone dye has the following formula: wherein R9 and R10 are independently selected from hydrogen or an alkyl group having from 1 to about 12 carbon atoms, X is —CH2—, a and a′ independently are integers from 1 to about 6, Y and Y′ are independently represent polymeric units of hydroxy alkylenes or alkylene oxide monomers selected from the group consisting of ethylene oxide, propylene oxide, butylene oxide, cyclohexane oxide, and glycidol, b and b′ are independently either 0 or 1, and Z and Z′ independently are reactive —OH, —NH2, or —SH groups. 117. The colored oligomer of claim 116, wherein said isocyanate end capped oligomer (a) is a urethane oligomer and said anthraquinone dye has the following formula: wherein n, n′, m, m′, p, and p′ independently have a value of from 0 to about 40. 118. The colored oligomer of claim 116, wherein said anthraquinone dye has the formula 119. The colored oligomer of claim 114, wherein said anthraquinone dye is selected from the group consisting of 1,5-bis((3-hydroxy-2,2-dimethylpropyl)amino)-9,10-anthracenedione; 2,2′-((9,10-dihydro-9,10-dioxo-1,5-anthracenediyl)bis(thio))bis-benzoic acid, 2-hydroxyethyl ester; and 1,5-bis((2,2-dimethyl-3-hydroxypropyl)amino)-4,8-bis((4-methylphenyl)thio) anthraquinone. 120. The colored oligomer of claim 114, wherein said anthraquinone dye is 1,5-bis((2,2-dimethyl-3-hydroxypropyl)amino)-4,8-bis((4-methylphenyl)thio) anthraquinone. 121. The colored oligomer of claim 113 wherein a (meth)acrylic group represents the ethylenic unsaturation (ii) in the radiation curable monomer (b). 122. A photocurable resin composition for forming a colored, cured coating on an optical fiber, said resin composition comprising: (e) at least one (meth)acrylate end capped urethane oligomer; (f) at least one photoinitiator; (g) at least one reactive diluent; and (h) at least one colored oligomer according to claim 113. 123. An optical fiber comprising a colored, cured coating, said colored, cured coating having been formed from the photocurable resin composition of claim 122. 124. A reactive anthraquinone dye for providing color to a coating on an optical fiber, said reactive anthraquinone dye comprising an anthraquinone core group with at least one substituent comprising a radiation curable group. 125. The reactive anthraquinone dye of claim 124, wherein said radiation curable group is an ethylenically unsaturated group or an epoxy group. 126. The reactive anthraquinone dye of claim 124, wherein said radiation curable group is a (meth)acrylic group. 127. The reactive anthraquinone dye of claim 124, wherein said reactive anthraquinone dye has the following formula: wherein R groups R1, R2, R3, R4, R5, R6, R7 and R8 are each independently selected from the group consisting of hydrogen, amino, hydroxy, halogen, nitro, carboxylated alkali metal, sulfated alkali metal and a hydrocarbyl group optionally containing one or more heteroatoms, provided that at least one of R groups R11 through R18 have at least one ethylenically unsaturated radiation curable functionality. 128. The reactive anthraquinone dye of claim 127, wherein one or two of said R groups R11 through R18 have a (meth)acrylic functionality and at least four of said R groups R11 through R18 are hydrogen. 129. The reactive anthraquinone dye of claim 127, wherein the reactive anthraquinone dye has one of the following formulas: wherein R29, R30, R31, R32, R33, and R34 are the same or different and are independently hydrogen or a C1 to C6 alkyl optionally substituted with one or more substituents selected from the group consisting of —OH, —NH2, —SH, —NO2, —CN and halogen. 130. A photocurable resin composition for forming a colored, cured coating on an optical fiber, said resin composition comprising: (a) at least one (meth)acrylate end capped urethane oligomer; (b) at least one photoinitiator; (c) at least one reactive diluent; and (d) at least one reactive anthraquinone dye according to claim 127. 131. An optical fiber comprising a colored, cured coating, said colored, cured coating having been formed from the photocurable resin composition of claim 130. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention The present invention relates to colored, radiation-curable compositions for use in a wide variety of applications, including colored, radiation-curable coating compositions for telecommunications elements, such as optical fibers and optical fiber ribbons, and colored, radiation-curable ink compositions for other applications, such as textile, electronics, and printing applications. More particularly, the present invention relates to colored, radiation-curable compositions for producing a cured, colored coating on a substrate, such as a telecommunications element, the cured compositions having an identifying color provided by chromophore-containing compounds that are covalently bonded to other components within the cured composition. 2. Description of Related Art For many years now, optical fibers have been used as a reliable transmission medium in telecommunications cables. Typically, an optical fiber comprises a core, a cladding and one or more coatings applied over the cladding. One purpose of the coatings is to protect the surface of the optical fiber from mechanical scratches and abrasions typically caused by subsequent handling and use. Another purpose of the coatings is to protect the glass from exposure to moisture. The coating or coatings may also have some influence over the fiber's optical characteristics because the coatings are physically responsive to external mechanical forces and temperature. The coating compositions applied to the optical fiber are typically liquid, radiation-curable compositions. Typically, the coating compositions are cured on the optical fiber by exposing the coating composition to ultraviolet radiation, electron beam radiation or ionizing radiation for a predetermined period of time deemed suitable for effective curing. Telecommunications cables containing optical fibers come in a variety of configurations. In some cables, the optical fibers are held loosely inside a buffer tube. In other cables, the optical fibers are arranged in a planar array to form an optical fiber ribbon. The planar array is typically encapsulated by one or more radiation-curable matrix material layers. The radiation-curable matrix layers are cured by exposing the matrix material to ultraviolet radiation, electron beam radiation, ionizing radiation or infrared radiation for a predetermined period of time deemed suitable for effective curing. In a telecommunications cable containing multiple optical fibers, the optical fibers may be distinguished from each other by the use of a color coating layer which has been applied over a coated optical fiber. Colors in the color coating layer are usually obtained by dispersing colored pigment particles in a suitable liquid carrier and applying the liquid carrier over the coating. Alternatively, the optical fibers may be distinguished from each other by the use of a so-called colored primary coating, which is a colored coating applied directly onto an optical fiber. Unfortunately, the use of pigment particles to provide color in color coatings for optical fibers has presented manufacturing and performance problems. For example, the pigment particles and the liquid carrier tend to gradually separate into two distinct phases. As a result, pigmented color coatings have a relatively short shelf life. In addition, the phase separation in a pigmented coloring system is further complicated by the concurrent agglomeration of pigment particles. Undesirably, the presence of pigment particle agglomerates in a color coating on a coated optical fiber can induce micro-bending which results in transmission losses. Typically, a relatively high concentration of pigment material is required to achieve an opaque or translucent ultraviolet radiation-curable color coating. Unfortunately, the required high concentration inhibits the transmission of incident ultraviolet radiation which is necessary to cure the color coating material because the pigments refract, reflect and scatter the incident radiation. The inhibition of the ultraviolet radiation results in a reduction in processing speed of the optical fiber along a manufacturing line and thereby increases production costs. Also, the slow cure speed of pigmented color coatings causes the processing and the cure of these materials to be sensitive to minor changes in the thickness of the color coatings. The use of dyes to provide color in color coatings has been considered as an alternative to pigment-based color coatings. Dyes have the advantage over pigments of faster curing because the dyes do not scatter the curing radiation, although some dyes may absorb light which could slow curing. Dyes, however, are generally not preferred because they diffuse (bleed) out into common cable filling compounds resulting in a color loss. In an effort to reduce the bleeding, U.S. Pat. No. 5,074,643 teaches the use of a polymeric dye in a color coating. The polymeric dyes are macromolecular chromophore-containing molecules that become entrapped in the cross-linked coating network. While the entrapment results in a slowing of the bleeding process, the dyes nevertheless still bleed. Over time, even with the entrapped polymeric dyes, the color imparted to the fibers is likely to be lost and if the color is lost from the fibers, then identification of each of the fibers becomes extremely difficult and time-consuming in the field during fiber-splicing. The color imparted to the fibers will be lost over time if the dyes themselves lack stability. In particular, the dyes should have sufficient thermal stability and light fastness to impart the desired color for an extended period of time. If a telecommunications cable has many optical fiber ribbons, it is generally desirable to distinguish one optical fiber ribbon from another by coloring each of the optical fiber ribbons. Typically, color in a colored optical fiber ribbon is obtained in the same way as color is obtained in a color-coated optical fiber. The optical fiber ribbon matrix composition is either provided with pigments or a polymer dye is used. The same problems mentioned above with respect to colored optical fibers apply to colored optical fiber ribbons. It is desirable to provide a composition that can provide a durable cured color coating that can be used for color coding a telecommunications element, such as an optical fiber, where the coating has the ability to withstand the conditions in a typical operational environment that such elements are typically found. It would also be desirable to provide a composition that can provide a durable cured ink, dye, coating, colorant, etc. that can be used for substrates in other fields, such as textiles, electronics, or printing, where the ink, dye, coating, colorant, etc. has the ability to withstand the conditions in a typical operational environment that such substrates are typically found. The present invention provides such a composition. |
<SOH> SUMMARY OF THE INVENTION <EOH>In one aspect of the present invention, there is provided a chromophore-containing compound, wherein the chromophore-containing compound comprises one or more functional groups that are capable of being reacted to covalently bond the chromophore-containing compound with any another molecule or series of molecules in a radiation-curable composition, such that the chromophore-containing compound is incorporated via covalent bonding into the radiation-cured composition. In one embodiment of the first aspect of the invention, the chromophore-containing compound includes, as one or more functional groups, one or more radiation-curable groups. The chromophore-containing compound comprising one or more radiation-curable groups becomes covalently bonded by one or more covalent bonds to other constituents of a radiation-curable composition during the curing step. In a second embodiment of the first aspect of the invention, the chromophore-containing compound is a colored oligomer. For example, there may be provided a chromophore-containing compound comprising a chromophore covalently bonded to or within an oligomeric backbone, such that the chromophore of the chromophore-containing compound is attached to the remainder of the oligomer by one or more covalent bonds. The colored oligomer may include radiation-curable groups that become covalently bonded to other constituents of a radiation-curable composition during the curing step. In another aspect of the present invention, there is provided a radiation-curable composition comprising a chromophore-containing compound, wherein the chromophore-containing compound comprises one or more functional groups that are capable of reacting to covalently bond the chromophore-containing compound with any another molecule or series of molecules in the radiation-curable composition. Upon being subjected to the appropriate level of radiation, the radiation-curable composition provides a radiation-cured composition having a chromophore-containing compound covalently bonded to other molecules or series of molecules within the radiation-cured composition. In yet another aspect of the invention, there is provided a color concentrate, or masterbatch, comprising the chromophore-containing compound, wherein the color concentrate or masterbatch is a vehicle for the delivery of the chromophore-containing compound to the particularly desired application or embodiment. There is also provided a method for manufacturing such a color concentrate or masterbatch. In a further aspect of the present invention, there is provided a telecommunication element having a color-identifying coating thereon, the telecommunication element comprising an elongated communication transmission medium and a coating having an identifying color applied on at least a portion of the transmission medium, wherein the coating comprises a radiation-cured, crosslinked polymeric network, and wherein the identifying color in the coating is provided by at least one chromophore molecule covalently bonded by at least one covalent bond to the polymeric network. The color of a telecommunication element prepared in this manner according to the invention does not bleed in the presence of cable-filling compounds. For example, the telecommunication element may be an optical fiber ribbon, and the ribbon has a colored matrix or a colored coating applied over an uncolored matrix, wherein the color of the colored matrix or colored coating does not bleed in the presence of cable-filling compounds. It is a still further aspect of the present invention to provide: a method for producing a colored, radiation-curable composition having at least one chromophore-containing compound, wherein the chromophore-containing compound comprises one or more functional groups that are capable of being reacted to covalently bond the chromophore-containing compound with any another molecule or series of molecules in the radiation-curable composition; and a method of providing at least a portion of a substrate with a radiation-cured composition having at least one chromophore-containing compound, wherein the chromophore-containing compound is covalently bonded to other molecules or series of molecules within the radiation-cured composition. The method may comprise the steps of: providing a substrate, such as, for example, a transmission medium; providing a colored, radiation-curable composition comprising a chromophore-containing compound; applying the radiation-curable composition to at least a portion of the substrate; and exposing the radiation-curable composition for a suitable period of time to radiation of a suitable wavelength and intensity to cause curing of the composition into a radiation-cured, crosslinked polymeric network, wherein the identifying color in the coating is provided by at least one chromophore molecule covalently bonded by at least one covalent bond to the polymeric network. The invention will be more fully understood when reference is made to the following detailed description taken in conjunction with the accompanying drawings. |
Protease deficient caulobacter host cells |
Caulobacter optimized for use in expression of heterologous peptides as part of a hybrid Caulobacter S-layer protein are provided as well as methods for producing such Caulobacter. Caulobacter of this invention are deficient in a native Caulobacter protease that cleaves hybrid S-layer proteins. Also provided are Caulobacter libraries and methods for sorting such libraries using cell sorting technoloey such as FACS. |
1. An isolated Caulobacter deficient in a protease native to Caulobacter that cleaves hybrid Caulobacter S-layer protein monomers. 2. The isolated Caulobacter of claim 1, wherein said protease comprises a metalloprotease active site. 3. The isolated Caulobacter of claim 1 or 2, wherein said protease comprises one or more RTX sequences. 4. The isolated Caulobacter of claim 1, wherein said protease comprises a C-terminal portion having a first sequence of about 150 to about 250 amino acids, the first sequence having 30% or more sequence identity to a second sequence of about 150 to about 250 amino acids, the second sequence being an N-terminal portion of a Caulobacter S-layer protein monomer. 5. The isolated Caulobacter of claim 4, wherein the first and second sequences independently have about 190 to about 230 amino acids. 6. The isolated Caulobacter of claim 4, wherein the second sequence is amino acids 23 to 242 of SEQ ID NO: 1. 7. The isolated Caulobacter of claim 6, wherein the first sequence is about 200 amino acids in length. 8. The isolated Caulobacter of claim 1, wherein said protease comprises an amino acid sequence having about 75% or more sequence identity to SEQ ID NO: 3. 9. The isolated Caulobacter of claim 8, wherein the identity to SEQ ID NO: 3 is about 80% or more. 10. The isolated Caulobacter of claim 8, wherein the identity to SEQ ID NO: 3 is about 85% or more. 11. The isolated Caulobacter of claim 8, wherein the identity to SEQ ID NO: 3 is about 90% or more. 12. The isolated Caulobacter of claim 8, wherein the identity to SEQ ID NO: 3 is about 95% or more. 13. The isolated Caulobacter of claim 1, wherein said protease comprises an amino acid sequence substantially identical to SEQ ID NO: 3 or a sequence in which one or more amino acids of SEQ ID NO: 3 are conservatively substituted. 14. The isolated Caulobacter of claim 1, wherein said protease comprises SEQ ID NO: 3. 15. The isolated Caulobacter according to claim 1, wherein the isolated Caulobacter is deficient in the protease as a result of a deletion, substitution, or addition of nucleotide in a gene encoding the protease. 16. The isolated Caulobacter of claim 1, wherein the isolated Caulobacter is incapable of secreting an S-layer protein monomer encoded by a gene native to the Caulobacter. 17. The isolated Caulobacter of claim 16, wherein the isolated Caulobacter comprises a recombinant DNA capable of expression of an S-layer protein monomer. 18. The isolated Caulobacter of claim 17, wherein the S-layer monomer expressed by the recombinant DNA is a hybrid S-layer protein monomer. 19. The isolated Caulobacter of claim 18, wherein the hybrid S-layer protein monomer is secreted by the isolated Caulobacter, is capable of attachment to a cell surface of the Caulobacter, and comprises one or more peptides heterologous to Caulobacter S-layer protein monomer. 20. A library of Caulobacter in which members of the library are different isolated Caulobacter according to claim 19, wherein the members differ by amino acid sequence of the one or more heterologous peptides. 21. A method of sorting a library of claim 20 according to ability for a heterologous peptide to bind to a target, comprising exposing the library to the target, and separating members of the library that bind to the target from members which do not bind to the target. 22. The method of claim 21, wherein the target is labelled and presence of the label bound to a member is indicative of binding of the target to the member. 23. The method of claim 22, wherein the label is fluorescent and the separation is performed by a flow cytometer. 24. A method of producing a Caulobacter optimized for use as a host cell for expression of a hybrid Caulobacter S-layer protein monomer, comprising: i) providing a Caulobacter which exhibits protease activity whereby hybrid S-layer protein monomers expressed by the Caulobacter are cleaved; ii) mutating the Caulobacter; and iii) selecting and culturing a mutated Caulobacter which does not exhibit said activity. 25. The method of claim 24, wherein the Caulobacter is mutated by introduction into the Caulobacter of a nucleic acid capable of homologous recombination with all or part of SEQ ID NO: 2. 26. The method of claim 24, wherein the Caulobacter is mutated by transposon insertion. 27. The method of claim 24, wherein the Caulobacter is mutated by one or more of radiation and a chemical mutagen. 28. The method of claim 24, wherein the Caulobacter which exhibits protease activity expresses a protein comprising an amino acid sequence having about 75% or more sequence identity to SEQ ID NO: 3, and the Caulobacter is mutated so as to result in insertion, deletion, or substitution of at least one amino acid in said amino acid sequence. 29. A mutated Caulobacter or descendents thereof, produced by the method of claim 24. 30. A library of mutated Caulobacter or descendents thereof of claim 29, wherein members of the library express and secrete different hybrid Caulobacter S-layer protein monomers. 31. A method of sorting a library of claim 30 according to ability for the different hybrid S-layer protein monomers to bind a target, comprising exposing the library to the target, and separating members of the library that bind to the target from members which do not bind to the target. 32. The method of claim 31, wherein the target is labelled and presence of the label bound to a member is indicative of binding of the target to the member. 33. The method of claim 32, wherein the label is fluorescent and the separation is performed by a flow cytometer. 34. A method of sorting members of a Caulobacter library according to ability of S-layer on a Caulobacter to bind to a target, wherein members of the library express and secrete different hybrid S-layer protein monomers, comprising: i) exposing the library to a fluorescent labelled target; and ii) segregating members of the library with a flow cytometer according to whether the labelled target is bound to a member or is not bound to a member. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The gram-negative bacterium Caulobacter elaborates a paracrystalline protein surface layer (S-layer) which covers the surface of its outer membrane (Smit, et al., 1981; 1992). The S-layer protein monomer is secreted by a Type I secretion mechanism relying upon a C-terminal secretion signal which remains attached to the rest of the protein during the secretion process (Gilchrist, et al., 1992; Bingle, et al. 1997a; and, Awram and Smit 1998). Once the protein monomer is secreted, the S-layer forms by a process of self-assembly as a hexagonal array of about 40,000 interlinked protein monomers (Nomellini, et al., 1997). Anchoring of the S-layer protein to the cell surface is dependent on a smooth lipopolysaccharide (LPS) molecule and Ca +2 ions (Walker, et al, 1994) and involves the N-terminal portion of the S-layer monomer (Bingle, et al, 1997 a,b). The abundance, cell-surface location and geometrical packing of the S-layer protein as well as the inherent properties of Caulobacter (ease of genetic manipulation, simple growth requirements, non-pathogenic nature, and biofilm-forming characteristics) have led to the exploitation of the Caulobacter /S-layer system for biotechnology development (Smit, et al., 2000). Through use of gene fusions encoding the C-terminal secretion signal of the S-layer monomer linked to sequences encoding a heterologous peptide, the Caulobacter /S-layer system is capable of secreting large quantities of the heterologous peptide (PCT patent applications published under No. WO 97/34000 and WO 00/49153; and, Bingle, et al., 2000). Such Caulobacter expression systems are available commercially under the trade name PurePro™ (Invitrogen, Carlsbad, Calif.). The PureProm™ system is designed for expression of “C-terminal hybrid proteins” in which a heterologous peptide is linked to the S-layer monomer C-terminal secretion signal and does not form an S-layer on the host. The Caulobacter /S-layer system is also used for expression of heterologous peptides inserted into sites within a full-length (or nearly full-length) S-layer monomer resulting in production of what is termed herein “full-length hybrid protein”. In such cases, a sufficient portion of the N-terminal region of the S-layer monomer is present to provide anchoring of the hybrid protein to the cell surface. The C-terminal secretion signal is present to permit secretion of the hybrid protein. Various preferred sites for insertion of heterologous peptides into the S-layer protein have been reported (WO 97/34000; and, Bingle, et al., 1997(b)). The hybrid S-layer protein so produced is capable of self-assembly on the cell surface resulting in an array of hybrid S-layer protein monomers forming as an S-layer on the cell surface. In this manner, heterologous peptides may be expressed and presented on the cell surface. This technology has particular use in expression and presentation of antigens. A phenomenon observed during the development of the Caulobacter /S-layer system was the apparent proteolytic cleavage of various hybrid proteins. This was initially observed for both C-terminal hybrid proteins and full-length hybrid proteins. No obvious site specificity was associated with this phenomenon despite monitoring the degradation of numerous modified S-layer proteins comprising different heterologous peptides of varying lengths inserted at different places in the S-layer protein monomer (Bingle, et al. 1997a,b) it has now been reported (Simion, B. et al. 2001) that smaller proteins seen contaminating preparations of C-terminal hybrid proteins are not the result of proteolytic activity but rather the result of internal translation initiation following Met residues within the heterologous or “passenger” portion of the hybrid protein. Nevertheless, the cleavage phenomenon still places limitations on the use of Caulobacter as an expression system for full length hybrid proteins, particularly when the “passenger” peptide is unknown or uncharacterized, since it would then be difficult to know if this phenomenon has affected the hybrid product. The use of a biological system to express and display a panel or library of different peptides to assess the ability of the peptides to bind to a chosen target, has become a powerful tool for investigating interaction of cellular components (see U.S. Pat. Nos. 5,223,409 & 5,571,698). In this methodology, nucleic acids each encoding a different peptide plus a signal for display of the peptide on the outer surface of a biological system, are introduced into the system. The peptides are expressed and binding domains in the peptides are displayed on the outer surface of the biological system. The system is exposed to target molecules and those members of the system which bind target molecules are isolated and the nucleic acids amplified. Successful binding domains are then characterized. This general method of exposing a variety of peptides, each displaying a different putative binding region, is termed “panning” herein. To date, phage is the preferred biological system for use in panning methodologies. This is partly due to difficulties in the use of bacterial systems for expression and display of heterologous peptides. |
<SOH> SUMMARY OF THE INVENTION <EOH>It has now been discovered using studies involving full-length hybrid proteins that a protease is synthesized by Caulobacter with an unusual structural feature: the enzyme possesses a domain sharing sequence similarity with the S-layer protein monomer. This native protease is responsible for the cleavage phenomenon seen with Caulobacter expressing heterologous peptides inserted into a full-length S-layer protein. This discovery provides the means for optimizing use of Caulobacter as a system for expression and cell-surface display of heterologous peptides and for use of such a system for expression of unknown or uncharacterized peptides with good fidelity. This facilitates the use of Caulobacter as an expression system for producing random libraries of peptides or gene fragments for display and panning purposes. The methodologies known in the art for expression of heterologous peptides within an S-layer monomer followed by assembly and adherence of hybrid S-layer protein on a Caulobacter cell surface may now be used in a bacterial-mediated library display system. This has significant advantages over conventional phage display, notably because Caulobacter are of a sufficient size to be sorted through use of conventional cell sorting techniques such as fluorescent activated cell sorting (FACS). Such techniques cannot be used for sorting phage. This invention provides an isolated Caulobacter deficient in a protease native to Caulobacter that cleaves hybrid Caulobacter S-layer protein monomers. Also provided is a library of such Caulobacter in which members of the library differ by the amino acid sequence of heterologous peptides in hybrid S-layer protein monomers expressed by the members. This invention also provides a method of producing a Caulobacter optimized for use as a host cell for expression of a hybrid Caulobacter S-layer protein monomer, comprising: i) providing a Caulobacter which exhibits protease activity whereby hybrid S-layer protein monomers expressed by the Caulobacter are cleaved; ii) mutating the Caulobacter ; and iii) selecting and culturing a mutated Caulobacter which does not exhibit said activity. Mutation may be by insertion, deletion, or substitution and may be performed through use of radiation and/or chemical mutagens, transposon insertion, homologous recombination or other methods. Also provided are mutated Caulobacter or descendents thereof produced by this method and libraries of such Caulobacter. The native Caulobacter protease cleaves hybrid S-layer protein monomers as defined herein, in preference to wild-type S-layer protein monomers. The protease appears to be specific for S-layer protein monomers. Within this context, the protease appears to have general specificity for cleavage of hybrid S-layer protein monomers, but tends to cleave C-terminal of Arg and Phe residues. The protease may comprise one or more RTX sequences characteristic of known calcium binding consensus sequences and metalloprotease active site comprising a sequence characteristic of a metalloprotease active site consensus sequence. In this specification, the native Caulobacter protease is termed “Sap”, an acronym for “S-layer associated protease”. In this specification, a gene encoding this protease is termed “sap”. The Sap protease may comprise a C-terminal portion of about 150 to about 250, preferably about 190 to about 230, and more preferably about 200 amino acids having about 30% or more sequence identity to a portion of the amino acid sequence of a Caulobacter S-layer protein monomer, as determined using the Blast™ search algorithm at default settings (see: Altschul, S. F., et al. 1997). The protease may comprise from about 600 to about 700 amino acids, preferably about 630 to about 670 amino acids, and more preferably about 650 to about 660 amino acids, depending upon the source strain. Sap protease sequences described herein have about 658 amino acids. The portion of a Caulobacter S-layer protein monomer to which the protease exhibits sequence identity will be of similar size to the protease C-terminal portion and will be located toward the N-terminus of the S-layer protein monomer. A portion of an S-layer protein monomer exhibiting sequence identity as described above includes amino acids 23-242 of RsaA. A region of the protease exhibiting such sequence identity to RsaA may be a region corresponding to amino acids 451-650 of the specific Sap amino acid sequence described herein. The protease may further comprise an N-terminal region exhibiting about 50% or more sequence identity (determined as described above) with a P. aeruginosa alkaline protease region, as described below. The protease may comprise an amino acid sequence that has about 75%, 80%, 85%, 90%, 95% or more sequence identity to the specific Sap amino acid sequence disclosed herein. This invention provides Caulobacter in which a native protease as described above is not expressed or is inactive (a “protease-negative” strain). These Caulobacter may comprise a mutated gene or coding region for the native protease. Protease-negative strains of this invention may also comprise recombinant DNA operably linked to a promoter recognized by Caulobacter , wherein the recombinant DNA comprises a nucleic acid sequence encoding at least a C-terminal secretion signal of Caulobacter S-layer protein monomer. The recombinant DNA may further comprise a nucleic acid sequence encoding sufficient part of Caulobacter S-layer protein monomer N-terminal region to provide for attachment of the coding product of the recombinant DNA to the cell surface of the Caulobacter . The recombinant DNA may comprise substantially all of a Caulobacter S-layer protein gene. The DNA may further comprise restriction sites to facilitate insertion of DNA encoding one or more peptides (including polypeptides and proteins) heterologous to Caulobacter S-layer protein into the recombinant DNA whereby the recombinant DNA encodes a hybrid S-layer protein monomer comprising all or part of an S-layer protein monomer and the one or more heterologous peptides. The recombinant DNA may further comprise DNA encoding a peptide, polypeptide, or protein heterologous to Caulobacter S-layer protein monomer. The recombinant DNA may be present in the Caulobacter on a plasmid or integrated into the Caulobacter genome. This invention provides a panel or library of Caulobacter wherein individual members of the panel or library express different heterologous peptides, polypeptides, or proteins as part of a hybrid S-layer protein monomer, as described above. Preferably, the Caulobacter in the panel or library will be one or more protease-negative strains of this invention. Also provided are methods of sorting such a library according to whether a target to which the library is exposed binds to hybrid S-layer protein monomers on members of the library. This invention also provides methods for detecting binding of a target compound to the surface of a Caulobacter that is a member of a library or panel as described above. Preferably, the target compound will be labelled, preferably with a fluorescent label. Detection of members of the panel or library to which the compound is bound may be done by FACS and performed by a flow cytometer. These methods may further comprise selection of Caulobacter to which the target binds, amplification of DNA encoding the heterologous peptide, polypeptide, or protein and sequencing of said DNA. This invention provides a method of sorting members of a Caulobacter library according to ability of S-layer on a Caulobacter to bind to a target, wherein members of the library express and secrete different hybrid S-layer protein monomers, comprising: i) exposing the library to a fluorescent labelled target; and ii) segregating members of the library with a flow cytometer according to whether the labelled target is bound to a member or is not bound to a member. |
Diagnosis of vascular disease susceptibility using bacteriophage phi-cpn1 host chlamydia |
Methods are provided for determining susceptibility of a patient to vascular disease according to presence of chlamydia infected with bacteriophage ΦCpn1, and compositions for use in such methods. The compositions include bacteriophage ΦCpn1 proteins and nucleic acids, elementary bodies from chlamydia infected with ΦCpn1, and antibodies thereto. |
1. A method for determining whether a subject is susceptible to vascular disease, comprising testing a biological sample from said subject for the presence or absence of bacteriophage ΦCpn1, a component of a host Chlamydia bacterium or an antibody to said bacteriophage or said Chlamydia component, wherein detection of the presence of said bacteriophage. Chlamydia component or antibody is indicative of the presence of said host Chlamydia bacteria in the subject and susceptibility of the subject to vascular disease. 2. (cancel) 3. The method of claim 1, wherein the sample is a bodily fluid from the subject to be tested, and the method further comprises the step of obtaining the bodily fluid. 4. The method of claim 1, wherein the sample is a fraction of a bodily fluid from said subject, which fraction contains bacteria, virus, antibody, peptide or nucleic acid. 5. The method of claim 1, wherein the host Chlamydia is C. pneumoniae. 6. The method of claim 5, wherein the C. pneumoniae is of the strain AR39. 7. The method of claim 1, wherein detection of the host Chlamydia component being detected comprises a Chlamydia elementary body. 8. The method of claim 7, wherein the presence of said elementary body is detected by detecting in the sample an antibody to said Chlamydia elementary body. 9. The method of claim 7, wherein the presence of said elementary body is detected by measuring antibody binding to the elementary body. 10. The method of claim 1, wherein at the time of testing, it is unknown whether the subject has a risk factor or predisposition to vascular disease. 11. The method of claim 1, wherein the subject does not have a hypercholesterolemic condition. 12. The method of claim 1, wherein detection of the bacteriophage host comprises detecting one or more of: (a) a ΦCpn1 peptide (b) a peptide having an amino acid sequence substantially identical to a ΦCpn1 peptide, or (c) an antibody to the ΦCpn1 peptide. 13. The method of claim 12 that measures antibody binding to the peptide. 14. The method of claim 13 that measures, antibody binding to a ΦCpn1 peptide. 15. The method of claim 14, wherein the ΦCpn1 peptide is a ΦCpn1 capsid peptide. 16. The method of claim 13 that measures binding of an antibody that is capable of binding to a polypeptide or peptide having the sequence of any one of SEQ ID NO:8- SEQ ID NO:13. 17. The method of claim 16 that measures binding of an antibody that is capable of binding to Vp1 (SEQ ID NO:8). 18. The method of claim 12, wherein the antibody being detected is present in the sample. 19. The method of claim 12, wherein the detecting comprises sequencing peptides in the sample and comparing sequences so obtained to the sequences designated as SEQ ID NO:8- EQ ID NO:13, to determine whether peptides in the sample comprise (i) one or more of SEQ ID NO:8- SEQ ID NO: 13, or (ii) a sequence substantially identical to one or more of SEQ ID NO:8- SEQ ID NO:13. 20. The method of claim 1, wherein detection of the bacteriophage comprises detecting a ΦCpn1 nucleic acid, or a nucleic acid having a sequence substantially identical to a ΦCpn1 nucleic acid. 21. The method of claim 20, further comprising the step of amplifying ΦCpn1 nucleic acids in the sample. 22. The method of claim 20, further comprising the step or steps of recombinantly expressing ΦCpn1 nucleic acids in the sample to provide the nucleic acid. 23. The method of claim 20, wherein the detecting comprises measuring hybridization of a complementary oligonucleotide to the nucleic acid. 24. The method of claim 20, wherein the detecting comprises sequencing of nucleic acids in the sample and comparing sequences so obtained to SEQ ID NO: 1. 25. The method of claim 1, further comprising after determining the presence of said bacteriophage, Chlamydia component or antibody, the additional step of selecting a course of treatment for said subject's Chlamydia infection. 26. The method of claim 1, further comprising treating the subject for Chlamydia infection. 27. The method of claim 26, wherein the treatment comprises administering to the subject an antibiotic effective against Chlamydia. 28. An isolated antibody that binds to a bacteriophage ΦCpn1 polypeptide or peptide having a sequence SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12 or SEQ ID NO:13. 29. The antibody of claim 28, produced by a method which comprises: (a) immunizing a mammal with a polypeptide or peptide comprising one of more of: (i) SEQ ID NO:8. (ii) SEQ ID NO:9, (iii) SEQ ID NO:10, (iv) SEQ ID NO: 11, (v) SEQ ID NO:12, (vi) SEQ ID NO:13, (vii) an immunogenic fragment of any of (i)-(vi), or (viii) a polypeptide or peptide having substantial sequence identity to any of (i)-(vi); and, (b) isolating said antibody or a cell producing said antibody from the mammal. 30. The antibody of claim 28, that binds to a ΦCpn1 capsid protein. 31. The antibody of claim 28, that binds to Vp1 (SEQ ID NO:8). 32. An immunogenic composition for eliciting an immune response in a mammal against bacteriophage ΦCpn1 or against Chlamydia bacteria that comprise bacteriophage ΦCpn1, the composition comprising (A) a pharmaceutically acceptable carrier and (B) an immunogen which comprises: (i) one or more polypeptides or peptides having a sequence (a) SEQ ID NO:8, (b) SEQ ID NO:9, (c) SEQ ID NO:10, (d) SEQ ID NO:11, (e) SEQ ID NO:12, or (f) SEQ ID NO:13, (ii) an immunogenic fragment of any of (a)-(f), (iii) a polypeptide or peptide having substantial sequence identity to any of (a)-(f); or (iv) a nucleic acid molecule encoding any one or more of said polypeptides, peptides or fragments of (i), (ii) or (iii). 33. The composition of claim 32 wherein the immunogen is one of more of said polypeptides or peptides of (i). 34. A kit for use in a method for testing a biological sample for the presence of a bacteriophage ΦCpn1, a component of a chlamydia host of said bacteriophage, or an antibody to said bacteriophage or said Chlamydia component, which kit is useful for determining whether a subject is susceptible to vascular disease, the kit comprising, in a commercial package: (i) one or more detecting moieties selected from the group consisting of: (a) a ΦCpn1 polypeptide or a peptide fragment thereof; (b) a peptide having substantial sequence identity to the ΦCpn1 polypeptide or peptide fragment of (a); (c) an antibody that binds to a ΦCpn1 peptide; (d) one or more oligonucleotides complementary to a ΦCpn1 nucleic acid; (e) an antibody that binds to a Chlamydia AR39 elementary body; and (f) a peptide that binds to the elementary body-binding antibody of (e); and (ii) instructions for use of the one or more detecting moieties for detecting the presence of said bacteriophage ΦCpn1 or Chlamydia host component in said sample. 35. The kit of claim 34, wherein the detecting moiety is an antibody that binds to a ΦCpn1 peptide. 36. The kit of claim 35, further comprising a peptide that binds to the ΦCpn1 peptide-binding antibody. 37. The kit of claim 34, wherein the detecting moiety is said ΦCpn1 polypeptide, said peptide fragment, or said peptide that has substantial sequence identity to the ΦCpn1 polypeptide or peptide fragment thereof. 38. The kit of claim 37, further comprising an antibody that binds to an immunoglobulin molecules of the subject from whom the sample is derived. 39. The method of 25 further comprising treating the subject for Chlamydia infection in accordance with the selected course of treatment. 40. The method of claim 39, wherein the treatment comprises administering to the subject an antibiotic effective against Chlamydia. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The etiopathology of abdominal aortic aneurysm (AAA) is still largely unknown. It has been postulated that localized inflammation and destruction of structural connective tissue by a direct infectious or autoimmune process is involved. There is also some evidence of a possible association between Chlamydia infection and vascular disease (including atherosclerosis and AAA). Animal studies have demonstrated that following inoculation in the respiratory tract, C. pneumoniae disseminates throughout the vascular system, and in some animal models (such as models for hypercholesterolemic conditions) may result in the development of atherosclerotic lesions [1-6]. There is some suggestion that antibiotic therapy might lower the risk of acute coronary events [7, 8]. Observational studies have also shown some relationship between the presence of C. pneumoniae and AAA [9, 10]. Most people will be infected by C. pneumoniae by the age at which the clinical manifestations of atherosclerosis usually appear [11] and some argue that C. pneumoniae is simply an organism that more readily infects diseased arteries [12], or that its association with vascular disease is confounded by its association with other atherosclerotic risk factors [13]. Further complicating the situation was the apparent genetically-linked variability of host response to genital Chlamydia trachomatis infection as manifested by differences in risk of development of pelvic inflammatory disease [14]. Variability in host response could be why some persons are more prone to develop vascular disease when infected with C. pneumoniae . There could also be variations in the vasculotrophism and pathogenicity of different C. pneumoniae strains. DNA sequencing has shown genetic variation in C. pneumoniae [15-17]. One C. pneumoniae strain (AR39) exhibits a 4524 nucleotide single-stranded DNA bacteriophage ΦCpn1 [16]. Phage-bearing strains of some bacteria have been found to be more pathogenic than phage-free strains [18] but the implications of phage presence in Chlamydia was unknown. |
<SOH> SUMMARY OF THE INVENTION <EOH>It has now been discovered that C. pneumoniae containing the phage ΦCpn1 is involved in vascular disease. This invention provides a method for identifying a subject susceptible to vascular disease, comprising: i) providing a sample from a subject to be tested for susceptibility to vascular disease; and ii) detecting the presence or absence of a bacteriophage ΦCpn1 host Chlamydia component in the sample, or an antibody to said component, wherein presence of said component or antibody is indicative of the presence of said host Chlaniydia in the subject and susceptibility of the subject to vascular disease. This invention provides methods to determine whether a patient is susceptible to vascular disease, including abdominal aortic aneurysm (AAA), atherosclerosis, stroke, heart attack, etc. Such a patient could be one with a predisposition or known risk factor to vascular disease (such as high blood lipid levels or smoking) but the methods of this invention determine susceptibility independent of such known risk factors or predispositions and are predictive without prior assessment of such risk factors or predispositions. The methods of this invention may comprise providing a sample of biological material from a patient to be tested. Such biological material may be a tissue sample, fluid sample (e.g. blood or serum sample) or sample of material associated with the body such as sputum. In the methods of this invention, a sample is tested for the presence in the patient of host C. pneumoniae which contains a bacteriophage. The host C. pneumoniae which may be detected by such testing may be strain AR39. This invention may be performed by directly detecting the presence of a host Chlamydia such as AR39. Such an embodiment may be used when it is unknown if a patient has a known risk factor or predisposition. This method is useful even if it is unknown if a subject has elevated lipid levels or does not have a hypercholesterolemic condition. Detection in these embodiments may be by means of antibodies specific to AR39 such as those described herein. Such antibodies may be specific to AR39 elementary bodies. Methods of this invention preferably involve direct detection of the phage ΦCpn1 alone or in combination with direct detection of presence of the host. While the presence of the phage is indicative of the presence of host C. pneumoniae containing the phage, directly testing for the phage has the best correlation to susceptibility to vascular disease. In such embodiments, the detection of phage may be detection of the phage itself, of a phage peptide, or nucleic acid associated with the phage, or of an antibody to a phage peptide that is present in the patient. Known methods may be employed, such as obtaining a nucleic acid from a fraction taken from a sample of bodily fluid from a patient and determining the presence or absence of part or all of a bacteriophage genome. This invention provides an antibody which binds to a phage antigen such as the peptides of SEQ ID NO:8-13. This invention also provides antibodies to phage ΦCpn1 which are suitable for use in the aforementioned methods. Such methods for producing such antibodies may be specific to a peptide associated with the phage. Preferably, such an antibody will be specific to an antigenic epitope present on the surface of the phage, which includes the antibodies to phage capsid proteins and those described herein for protein Vp1. Alternatively, antibodies for use in the aforementioned methods may be anti-idiotypic antibodies in that they are specific for antibodies to the phage or a phage peptide. This invention also provides compositions comprising one or more peptides associated with phage ΦCpn1 for eliciting an immune response against phage ΦCpn1 (or a peptide of said phage). Also provided are nucleic acid compositions and methods of using such compositions for expressing phage peptides and for eliciting an immune response by expression of the nucleic acid in mammalian subjects to which the composition is administered, wherein the composition comprises one or more nucleic acids encoding all or part of a peptide corresponding to a peptide of phage ΦCpn1. These compositions may comprise pharmaceutically acceptable diluents, excipients, and adjuvants to facilitate administration of the composition to a patient or animal and in the case of the nucleic acid composition, may comprise a vector for expression of the nucleic acid in a subject. The invention also provides the use of all or part of a phage ΦCpn1 peptide in an immunoassay to test for the presence of phage or anti-phage antibodies in a biological sample. This invention provides a kit for use in the method of claim 1 , comprising in a commercial package: i) one or more detecting moieties selected from the group consisting of: a ΦCpn1 peptide, or peptide having substantial sequence identity to a peptide or fragment thereof; an antibody that binds to a ΦCpn1 peptide; one or more oligonucleotides complementary to a ΦCpn1 nucleic acid; an antibody that binds to Chlamydia AR39 elementary bodies; and, a peptide that binds to an antibody that binds to such elementary bodies; and, ii) instructions for use of the one or more detecting moieties for detecting the presence of a bacteriophage ΦCpn1 host Chlamydia components in a sample. This invention also provides methods for selecting a course of treatment of (or for treatment of) a patient who is or may become susceptible to vascular disease (or which patient is or may become infected with C. pneumoniae comprising the bacteriophage). The method may comprise administering an agent effective against the C. pneumoniae such as an antibiotic. In this specification, the term “peptide” is meant to encompass a peptide of any length including moieties that may be referred to as polypeptides and proteins. |
Plain bearing provided with a friction coating and method for making same |
A high-performance plain bearing provided with a friction coating including a bronze layer whereof the pores are filled with a PTFE-type fluoroplastic paste filled with molybdenum bisulphide. The production method includes sintering a layer of bronze powder on the metal support of the bearing, a vacuum ionic cleaning, applying a filled fluoroplastic film, passing the component through a die and after sintering the paste, a final pass in the die for final dimensioning. During sintering and passing through the die an anti-adhesive and wear-resistant coating, based on titanium nitride or the like, is applied on the equipment. |
1-4. (Canceled). 5. A method for making a friction coating on a high-performance plain bearing, comprising: (a) sintering a layer of bronze powder onto a metallic support under a reducing atmosphere using tooling comprising a non-stick and wear-resistant coating; (b) ionic cleaning under vacuum to activate a surface of the sintered bronze layer without increasing roughness of the sintered bronze layer; (c) applying a layer of fluoroplastic paste of PTFE type filled with molybdenum disulfide to the sintered bronze surface and installing into a die the support coated with the sintered bronze layer; (d) passing a component of the coated metallic support into a die with addition of a non-stick and wear-resistant coating to compress the paste applied in the applying (c) into pores of the sintered bronze layer; (e) drying the paste: (f) passing the component a second time through the die with addition of the non-stick and wear-resistant coating; (g) sintering the paste under predetermined duration and temperature conditions followed by immersion in water; and (h) passing the component a final time through a die equipped with a non-stick coating to achieve its end dimensions. 6. The method for producing a friction coating on a plain bearing as claimed in claim 5, wherein the non-stick and wear-resistant coating used in the sintering (a), the passing (d), the passing (f) and the passing (h) is based on titanium nitride or on a nitride with equivalent properties such as zirconium nitride, chromium nitride, or aluminum nitride. 7. The method for producing a friction coating on a plain bearing as claimed in claim 5, wherein in the drying (e) the drying of the paste is performed at 90° C. for two hours and in the sintering (g) the sintering of the paste is performed at 375° C. for 40 minutes. 8. The method for producing a friction coating on a plain bearing as claimed in claim 6, wherein in the drying (e) the drying of the paste is performed at 90° C. for two hours and in the sintering (g) the sintering of the paste is performed at 375° C. for 40 minutes. 9. The method for producing a friction coating on a plain bearing as claimed in claim 5, further comprising before applying the bronze powder: (a1) electrolytic deposition of copper onto the metallic support under conditions known per se, followed by tinning. 10. The method for producing a friction coating on a plain bearing as claimed in claim 6, further comprising before applying the bronze powder: (a1) electrolytic deposition of copper onto the metallic support under conditions known per se, followed by tinning. 11. The method for producing a friction coating on a plain bearing as claimed in claim 7, further comprising before applying the bronze powder: (a1) electrolytic deposition of copper onto the metallic support under conditions known per se, followed by tinning. 12. The method for producing a friction coating on a plain bearing as claimed in claim 8, further comprising before applying the bronze powder: (a1) electrolytic deposition of copper onto the metallic support under conditions known per se, followed by tinning. |
Method and system for sustainable digital watermarking |
Digital marks (so-called fingerprints and watermarks) serve two basic purposes: (1) Investigative: the owner reads a fingerprint to determine how the marked entity leaked; and (2) Legal: the owner must prove in court that (a) there is a watermark (a concealed copyright message), and (b) it is the owner's. The main difficulty of item (2) is that the first use of the watermark software reveals the watermarking method to the public so that hostile parties are equipped to remove or damage its watermarks. The invention uses tamper-resistant software encoding techniques to protect the digital mark extractor algorithm, frustrating the attacks of hostile parties in two ways: the resulting code is obscure (that is, its inner workings are incomprehensible; and chaotic (that is, a modification at any point will almost certainly produce a nonsense program. |
1-24. (CANCELLED) 25. A method for performing digital marking comprising the steps of: developing a digital mark injector program for inserting a digital mark into a digital file; developing a complementary digital mark extractor program for identifying said digital mark in said digital file; and encoding said digital mark extractor program using tamper-resistant software encoding techniques. 26. A method for performing digital watermarking comprising the steps of: developing a watermark-injector program for inserting a watermark into a digital file; developing a complementary watermark extractor program for identifying said watermark in said digital file; and encoding said watermark extractor program using tamper-resistant software encoding techniques. 27. The method of claim 25 wherein said step of encoding comprises the step of: encoding said watermark extractor program using data-flow encoding techniques. 28. The method of claim 27 wherein said step of encoding comprises the step of: transforming the data flow in said watermark extractor to dissociate the observable operation of the transformed said watermark extractor from the intent of the original software code. 29. The method of claim 27 wherein said step of encoding comprises the step of: encoding the data flow in said watermark extractor into a domain which does not have a corresponding semantic structure, to increase the tamper-resistance and obscurity of said watermark extractor. 30. The method of claim 25 wherein said step of encoding comprises the step of: encoding said watermark extractor program using control-flow encoding techniques. 31. The method of claim 30 wherein said step of encoding comprises the step of: transforming the control flow in said watermark extractor program to dissociate the observable operation of the transformed watermark extractor program from the intent of the original software code. 32. The method of claim 30 wherein said step of encoding comprises the step of: dispersing subsequences of instructions within said watermark extractor program into a plurality of locations; merging multiple dispersed subsequences into single blocks of code; selecting said subsequences of instructions from merged blocks of code for either functionally effective or decoy execution, as needed, to separate the observable operation of resulting code from the intent of the original software during execution. 33. The method of claim 30 wherein said step of encoding comprises the step of: adding fake-robust control transfers to said watermark extractor program, to increase the tamper-resistance of said watermark extractor program. 34. The method of claim 25 wherein said step of encoding comprises the step of: encoding said watermark extractor program using mass-data encoding techniques. 35. The method of claim 34 wherein said step of encoding comprises the step of: encoding a secret key in said watermark extractor program, using mass-data encoding techniques. 36. The method of claim 34 wherein said step of encoding comprises the step of: responding to a request to store a data value at a virtual address by: mapping said virtual address onto a randomly selected actual address; and storing said data value in a memory location indexed by said actual address. 37. The method of claim 25 wherein said step of encoding comprises the step of: encoding said watermark extractor program using white box encoding techniques. 38. The method of claim 37 wherein said step of encoding comprises the step of: representing one or more algorithmic steps or components as tables, thereby permitting encodings to be completely arbitrary nonlinear bijections. 39. The method of claim 37 wherein said step of encoding comprises the step of: identifying functions and transforms substantive to the targeted software program; generating new functions and transforms which alter the processing activity visible to the attacker; and replacing those identified functions and transforms with the new functions and transforms in the software program. 40. The method of claim 25, in which the level of obscurity is sufficient to make attacks on the watermarks via understanding of the extractor prohibitively expensive for most attackers. 41. The method of claim 40, in which multiple distinct watermarks and corresponding extraction programs are employed to increase security. 42. An apparatus for executing the method of: developing a digital mark injector program for inserting a digital mark into a digital file; developing a complementary digital mark extractor program for identifying said digital mark in said digital file; and encoding said digital mark extractor program using tamper-resistant software encoding techniques. 43. A computer readable memory medium for storing software code executable to perform the method of: developing a digital mark injector program for inserting a digital mark into a digital file; developing a complementary digital mark extractor program for identifying said digital mark in said digital file; and encoding said digital mark extractor program using tamper-resistant software encoding techniques. 44. A carrier signal incorporating software code executable to perform the method of: developing a digital mark injector program for inserting a digital mark into a digital file; developing a complementary digital mark extractor program for identifying said digital mark in said digital file; and encoding said digital mark extractor program using tamper-resistant software encoding techniques. |
<SOH> BACKGROUND OF THE INVENTION <EOH>In recent years, personal computers and digital communication systems have become pervasive in industrialized nations. As well, the speed and power of these systems are now at the point where any subject matter including images, audio, video and software can be stored, transferred and reproduced with ease. Because these systems are digital, the storage, transfer and reproduction can be performed flawlessly; each successive copy of a digital file may be made precisely the same as the original. This is in contrast to analogue systems used in the past, where the quality of each successive copy was poorer than the previous one, and all of the known storage media deteriorated overtime. Obviously, this ability to copy and transfer digital data with virtually no loss in quality is having a great impact on the revenues of many digital rights holders, including music, movie and software producers. Many techniques for protecting the intellectual property rights of these digital content and software producers have been proposed but have had little success. One technique that has shown particular promise is that of “digital marks”. Digital marks such as “digital fingerprints” or “watermarks” are patterns of bits (i.e. 0's and 1's) inserted into a digital file, that can be used to identify the file's owner or originator. Unlike printed watermarks, which are intended to be somewhat visible, digital watermarks are designed to be invisible. The actual bits that make up the watermark are also scattered throughout the digital file so that they cannot be identified and manipulated by an attacker. Digital marks generally serve two basic purposes (see, for example: Protecting ownership rights through digital watermarking , by H. Berghel and L. O'Gorman, 1996, IEEE Computer 29:7, pp. 101-103 and Attacks on copyright marking systems , by Fabien A. P. Petitcolas, Ross J. Anderson, and Markus G. Kuhn, 1998, 2nd Workshop on Information Hiding, LNCS vol. 1525 (isbn 3-540-65386-4), pp. 218-238): 1. Investigative: the owner reads a fingerprint (such as a concealed serial number) to determine how the marked entity (say, a picture in jpeg format) leaked. A trail of marked entities serves the owner in the same way that a trail of fingerprints serves a police detective; so that an owner can identify persons who have purloined the owner's property; and 2. Legal: the owner must prove in court that an infringed data file contains a watermark (a concealed copyright message), and that this watermark is the owner's. The investigative need is the easier of the two needs to meet, since everything may be kept secret: the owner need reveal nothing about how the mark was applied, and legal proof is not required. That is, there is generally no need to justify the manner in which the thief was identified, since the owner is primarily interested in asserting legal rights over a marked entity in the thief's possession. However, addressing the legal purpose is sufficiently difficult that some consider it infeasible against a resourceful, clever attacker (see, for example: Protecting digital media content , by Nasir Memon and Ping Wah Wong, 1998, Communications of the ACM 41:7, pp. 34-43). Greatly exacerbating the difficulty of addressing the legal purpose above is the fact that the first use of a watermark extraction program in a courtroom, based on a particular watermarking method, reveals the watermarking method. After the first legal justification (and hence, revelation) of the method, hostile parties are equipped to remove or damage its watermarks on a permanent basis thereafter. In order to provide legal watermarking the owner of the watermark would proceed roughly along the lines of the following: 1. employing a watermarking method, embodied in an “injector” implementation which applies the watermark to an owned item (possibly using a secret key) and an “extractor” implementation which reveals the watermark on an owned item (requiring the secret key, if one was used by the injector program); 2. keeping secret the keys (if any) used for watermarking owned items; 3. in court, demonstrating the watermark as follows: a. revealing the algorithm, so it can be justified as legitimate; b. loading the extractor and the impugned data/software file to be analysed, and having an agent of the court or a party trusted by the court apply the secret key (if any) to the extractor. Note that the key need not be revealed; and c. demonstrating that the revelation of the watermark by the (now justified) extractor proves the owner's claim to be the rightful owner of the disputed item containing the watermark. This methodology may be implemented in two major variants: 1. either keeping the injector and extractor algorithms a secret until it is necessary to disclose them during a court proceeding; or 2. publishing the injector and extractor algorithms. If the injector and extractor algorithms had been kept secure by the owner, then the public disclosure of these algorithms will reduce the level of security that has been enjoyed before the public disclosure. If the watermarking method had already been published, then the attacker would already have had sufficient information to permit compromise of the watermarks the algorithm applies. There is therefore a need for a watermarking method which allows legal justification of a claim to ownership of a purloined, watermarked entity, without revealing to the world at large the algorithm by which the watermark is applied or revealed. |
<SOH> SUMMARY OF THE INVENTION <EOH>It is therefore an object of the invention to provide a novel method and system of digital watermarking which obviates or mitigates at least one of the disadvantages of the prior art. One aspect of the invention is broadly defined as a method for performing digital marking comprising the steps of: developing a digital mark injector program for inserting a digital mark into a digital file; developing a complementary digital mark extractor program for identifying the digital mark in the digital file; and encoding the digital mark extractor program using tamper-resistant software encoding techniques. Another aspect of the invention is defined as a system for performing digital marking comprising a computer operable to perform the steps of: developing a digital mark injector program for inserting a digital mark into a digital file; developing a complementary digital mark extractor program for identifying the digital mark in the digital file; and encoding the digital mark extractor program using tamper-resistant software encoding techniques. |
Apparatus and method for digital content concealment in a storage medium recorded using a recording device |
A system and process for the concealment of digital content recorded on a storage medium such as a compact disc (CD) and, more specifically, to an apparatus and method for recording a CD or other storage medium using a recording drive, such as a CD-R or CD-RW drive, that is connected to a computer and that conceals the stored digital content on the medium from being read by a compact disc read-only memory (CD-ROM) or other digital-based reader or computer device, for example as done when performing digital audio extraction (also known as ripping) of musical content from an audio CD. The apparatus and method permit improved control of the usage and copying of published content on physical media by an immediate purchaser while limiting copying and distribution of the content to unauthorized recipients. |
1. A system for concealing digital content on a physical medium, comprising: a personal computer, said personal computer comprising hardware and software that is constructed and arranged to permit a consumer to bum an optical disc; and wherein at least one of said hardware and said software is configured so as to enable the consumer to bum an optical disc that incorporates copy protection. 2. A system according to claim 1, wherein said copy protection is configured so as to permit improved control over the copying and distribution of content that is stored on the optical disc. 3. A system according to claim 1, wherein said copy protection is configured to frustrate further copying of the content from the optical disc. 4. A system according to claim 1, wherein said optical disc comprises a CD, and wherein said copy protection incorporates means for making modifications to control data that is located in a lead-in area of the CD. 5. A system according to claim 1, wherein said optical disc comprises a CD, and wherein said copy protection incorporates means for verifying a lack of errors in control data on the CD. 6. A system according to claim 1, wherein at least one of said hardware and said software is configured so as to enable the consumer to make a multiple session CD having first and second sessions in which the first session actually contains audio data, but is coded to indicate that it contains digital data, and has intentional errors introduced to its control data. 7. A system according to claim 6, wherein said second session contains digital data in a standard format. 8. A system according to claim 1, wherein at least one of said hardware and said software is configured so as to enable the consumer to bum a multisession optical disc. 9. A system according to claim 8, wherein at least one of said hardware and said software is configured so as to enable the consumer to burn a multisession optical disc comprising a first session containing content in a first file format and a second session containing the same content in a second file format. 10. A system according to claim 8, wherein at least one of said hardware and said software is further configured so as to enable the consumer to bum a computer program on to the optical disc. 11. A system according to claim 9, wherein at least one of said hardware and said software is further configured to bum information on said multisession optical disc that relates to the copy protected status of the content. 12. A system according to claim 1 1, wherein at least one of said hardware and said software is further configured to bum information on to said multisession optical disc that will instruct a personal computer as to the specific form of copy protection that has been applied to the optical disc. 13. A personal computer, comprising: accessing means for accessing a digital work; determining means for determining whether said digital work is copy protected; and recording means for recording a copy protected digital work on to a portable digital media in a copy protected format. 14. A personal computer according to claim 13, wherein said accessing means comprises an optical disc reader for accessing said digital work from an optical disc. 15. A personal computer according to claim 14, wherein said determining means comprises a program that runs on said personal computer, and wherein said program is constructed and arranged to receive copy protection information that is encoded on said optical disc. 16. A personal computer according to claim 15, wherein said recording means is constructed and arranged to record said copy protected digital work on to an optical disc together with said encoded copy protection information. 17. A personal computer according to claim 13, wherein said recording means comprises an optical disc recorder that is constructed and arranged to record said digital work in a first format on a first session of an optical disc and to further record said digital work in a second format on a second session of said optical disc. 18. A personal computer according to claim 17, wherein said recording means is further constructed and arranged to record encoded copy protection information on said optical disc. 19. A copy protected optical disc, comprising: a digital work recorded on said optical disc in a copy protected format; and information encoded on said optical disc for a personal computer relating to copying of said digital work by said personal computer on to a recordable optical disc in a copy protected format. 20. A copy protected optical disc according to claim 19, further comprising a computer program recorded on said optical disc that may be executed by the personal computer. 21. A method of distributing digital content, comprising: providing the digital content to a consumer in a copy protected format; and enabling the consumer to record the digital content on to an optical disc in a copy protected format that will limit further dissemination of the digital content. 22. A method of distributing digital content according to claim 21, wherein said step of enabling the consumer to record the digital content comprises providing recording software that is executable on a personal computer for permitting recordation of the digital content on to an optical disc in a copy protected format. 23. A method of distributing digital content according to claim 22, wherein said recording software permits the digital content to be recorded in a first file format on a first session of the optical disc and in a second, different file format on a second session of the optical disc, and wherein said recording software is further constructed and arranged to record information on to the optical disc relating to the nature of the copy protection. |
<SOH> BACKGROUND OF THE INVENTION <EOH>This invention relates in general to the concealment of digital content recorded on a storage medium such as a compact disc (CD) and, more specifically, to an apparatus and method for recording a CD or other storage medium using a recording drive, such as a CD-R or CD-RW drive, that is connected to a computer and that conceals the stored digital content on the medium from being read by a compact disc read-only memory (CD-ROM) or other digital-based reader or computer device, for example as done when performing digital audio extraction (also known as ripping) of musical content from an audio CD. The apparatus and method of the present invention permit improved control of the usage and copying of published content on physical media by an immediate purchaser while limiting copying and distribution of the content to unauthorized recipients. Individuals commonly use a recording drive connected to a personal computer to record audio CDs (this process is also known as burning CDs). So-called burning software, such as sold by Adaptec and other companies, is used to control the computer during the CD burning process. The recorded CDs may be made using standard CD-R and CD-RW recordable storage mediums. Individuals typically use such recorded CDs to create compilations of music that may be obtained from multiple different sources. Such sources include audio CDs purchased by the individual, MP3 music files downloaded from the Internet, and digital music files using other standard formats. These sources may further include music files obtained under a digital rights management (DRM) system. For example, an individual may purchase several audio CDs and then record selected music tracks from each CD to a newly created compilation CD-R. A problem with existing CD burning software is that the audio CDs created can be distributed to other individuals who can use the newly-created CD to create yet other audio CDs or additional digital copies of the music content on the CD. Further, existing CD burning software does not use a copy protection mechanism to control the use and/or subsequent copying of such content. In many cases, the individuals receiving a newly-created CD from an original purchaser of the source CD will not be authorized or otherwise have the right to make or distribute copies of the music content. However, audio CDs made using existing burning software will permit unauthorized individuals to use a newly-created CD to upload the music content to an Internet server or other computer for public distribution or to create additional audio CDs that may be improperly distributed. The music and other entertainment industries have a business interest in promoting sales of CDs and other storage medium containing entertainment content. However, the music and entertainment industries desire to control the burning and handling of CDs by customers in a way that restricts unauthorized distribution to others who have not purchased or acquired a right from the publisher to receive entertainment content. Thus, there is a need for an audio CD recording system that permits audio CD consumers to make compilation CDs for personal use, but that controls or prevents the unauthorized uploading to the Internet or making of subsequent CD copies using the compilation or other audio CD recorded by the consumer without requiring hardware modifications or modification of the actual data content itself There is a further need for compilation CDs made using this audio CD recording system to retain full playability in audio CD players in order to meet current consumer expectations regarding the use of purchased audio CDs and audio CDs burned from these purchased CDs. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG. 1 is a schematic diagram of a compact disc recording system according to the present invention; FIG. 2 illustrates the contents of a multiple session compact disc recorded using the recording system of FIG. 1 ; FIG. 3 is a flow diagram illustrating a recording method for a storage medium using digital content concealment according to the present invention; FIG. 4 illustrates the standard subcode structure for audio CDs; FIG. 5 illustrates the standard channel Q data format for the subcode structure of FIG. 4 ; FIG. 6 illustrates the standard mode I Data-Q lead-in track format for the channel Q data format of FIG. 5 ; FIG. 7 illustrates the standard mode I Data-Q audio and lead-out track format for the channel Q data format of FIG. 5 ; FIG. 8 is a table illustrating an example of a table of contents (TOC) according to the present invention for a first session of a CD; FIG. 9 is a table illustrating an example of a table of contents according to the present invention for a second session of a CD; and FIG. 10 is a table illustrating an example of modified CRC values in the channel Q data of the program area of a CD according to the present invention. FIG. 11 is a table illustrating an example of modified ATime values in the channel Q data of the program area of a CD according to the present invention. detailed-description description="Detailed Description" end="lead"? |
Developer being less susceptible to oxidation and method for preparation thereof |
Provided are an oxidation-suppressed developer which is less apt to be oxidized and a method of manufacturing the oxidation-suppressed developer. The oxidation-suppressed developer contains a divalent-trivalent iron salt in a dissolved state. The oxidation-suppressed developer consists essentially of a compound which is obtained by performing neutralization after magnetite is dissolved in concentrated hydrochloric acid and by adding a crystal obtained by concentrating this solution to a concentrated hydrochloric acid solution of magnetite. The divalent-trivalentiron salt is contained in a concentration of 0.25×10−8 mg/liter to 0.25×10−10 mg/liter. |
1. An oxidation-suppressed developer, characterized in that the oxidation-suppressed developer contains a divalent-trivalent iron salt in a dissolved state. 2. The oxidation-suppressed developer according to claim 1, characterized in that the oxidation-suppressed developer contains pi-water in a volumetric concentration of not less than 0.01 ppm but not more than 100 ppm when converted to a pi-water stock solution with respect to the total volume. 3. The oxidation-suppressed developer according to claim 1, characterized in that said oxidation-suppressed developer contains Fe3O4 ions in a volumetric concentration of not less than 0.01 ppb but not more than 100 ppb when converted to a saturated concentrated hydrochloric acid of Fe3O4 with respect to the total volume. 4. The oxidation-suppressed developer according to claim 1, characterized in that said divalent-trivalent iron salt consists essentially of a compound expressed by the chemical formula Fe+2mFe+3nCl2m+3n (where, m and n are each an arbitrary natural number). 5. The oxidation-suppressed developer according to claim 4, characterized in that said divalent-trivalent iron salt is contained in a concentration of 0.25×10−8 mg/liter to 0.25×10−10 mg/liter. 6. The oxidation-suppressed developer according to claim 5, characterized in that said divalent-trivalent iron salt is a compound which is obtained by dissolving ferric chloride in a strongly alkaline aqueous solution and thereafter performing neutralization. 7. The oxidation-suppressed developer according to claim 1, characterized in that said divalent-trivalent iron salt exhibits magnetic properties. 8. The oxidation-suppressed developer according to claim 7, characterized in that said divalent-trivalent iron salt is a compound which is obtained by dissolving magnetite in concentrated hydrochloric acid, thereafter performing neutralization, and adding a crystal obtained by concentrating this solution to a concentrated hydrochloric acid solution of magnetite. 9. The oxidation-suppressed developer according to claim 1, characterized in that the oxidation-suppressed developer consists essentially of a fast developer. 10. The oxidation-suppressed developer according to claim 1, characterized in that the oxidation-suppressed developer consists essentially of a color developer. 11. A method of manufacturing an oxidation-suppressed developer, characterized in that a solution of a divalent-trivalent iron salt is added to developer components. 12. The method of manufacturing an oxidation-suppressed developer according to claim 11, characterized in that pi-water is added to developer components so as to obtain a volumetric concentration of not less than 0.01 ppm but not more than 100 ppm when converted to a pi-water stock solution with respect to the total volume. 13. The method of manufacturing an oxidation-suppressed developer according to claim 11, characterized in that an aqueous solution of Fe3O4 is added to developer components so as to obtain a volumetric concentration of not less than 0.01 ppb but not more than 100 ppb when converted to a saturated concentrated hydrochloric acid of Fe3O4 with respect to the total volume. 14. The method of manufacturing an oxidation-suppressed developer according to claim 11, characterized in that that said divalent-trivalent iron salt consists essentially of a compound expressed by the chemical formula Fe+2mFe+3nCl2m+3n (where, m and n are each an arbitrary natural number) and is added to developer components so as to obtain an aqueous solution in a concentration of 0.25×10−8 mg/liter to 0.25×10−10 mg/liter. 15. The oxidation-suppressed developer according to claim 6, characterized in that the oxidation-suppressed developer consists essentially of a fast developer. 16. The oxidation-suppressed developer according to claim 7, characterized in that the oxidation-suppressed developer consists essentially of a fast developer. 17. The oxidation-suppressed developer according to claim 8, characterized in that the oxidation-suppressed developer consists essentially of a fast developer. 18. The oxidation-suppressed developer according to claim 6, characterized in that the oxidation-suppressed developer consists essentially of a color developer. 19. The oxidation-suppressed developer according to claim 7, characterized in that the oxidation-suppressed developer consists essentially of a color developer. 20. The oxidation-suppressed developer according to claim 8, characterized in that the oxidation-suppressed developer consists essentially of a color developer. |
<SOH> BACKGROUND ART <EOH>Conventional developers are distributed in a state of a concentrated developer stock solution having high component concentrations and diluted with water when they are used. Incidentally, because the dilution of a developer stock solution requires time and labor and variations occur in quality depending on dilution methods, it is desirable that a developer be circulated in a diluted state. However, a developer diluted with water is rapidly oxidized, posing the problem that the storage period is short. The present invention was made by paying attention to such conventional problems and has as its object the provision of an oxidation-suppressed developer which is less apt to be oxidized and a method of manufacturing the oxidation-suppressed developer. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG. 1 is a graph showing the effect of pi-water on the sulfite ions of a fast developer in Embodiment 1 of the present invention. In FIG. 1 , the numeral 1 denotes pi-water and the numeral 2 denotes an object of comparison; and FIG. 2 is a graph showing the effect of pi-water on the sulfite ions of a color developer in Embodiment 3 of the present invention. In FIG. 2 , the numeral 3 denotes pi-water and the numeral 4 denotes an object of comparison. detailed-description description="Detailed Description" end="lead"? |
Method for transferring a page from a server to a client, a program suitable to carry out the method, and the use thereof |
A method for transferring a page to a client which is coupled to a first server wherein the page comprises at least one request for a page element. The method comprises the step of collecting the page in the first server, in response to a request for transferring the page to the client, and the further steps carried out in the first server of: deriving from the page the at least one request for the page element; collecting the at least one page element, in response to the at least one request for the page element derived from the page; combining the at least one page element with the page; and transferring the page with the collected at least one page element to the client. |
1. A method for transferring a page to a client which is coupled to a first server, the page comprising at least one request for a page element, comprising the step of: collecting the page comprising at least one request for a page element in the first server, in response to a request for transferring the page to the client; deriving in the first server from the page the at least one request for the page element; collecting the at least one page element in the first server, in response to the at least one request for the page element derived from the page; combining in the first server the at least one page element with the page; transferring the page with the collected at least one page element to the client; and charging the client on a basis of the page with the collected at least one page element transferred to the client. 2. The method according to claim 1, wherein the step of collecting the page comprises the step of: transferring a request for the page by the first server to a second server. 3. The method according to claim 1, wherein the step of collecting the page comprises the steps of; checking if the page is stored in a memory, which is accessible for the first server; and transferring a request for the page and/or page element by the first server to a second server, if the page and/or the at least one page element is not stored in the memory. 4. The method according to claim 1, further comprising the step of: prior to transferring the page with the collected at least one page element from the first server to the client, converting the page with the collected at least one page element into a predetermined format. 5. The method according to claim 4, wherein the step of converting the page into the predetermined format comprises the step of removing the at least one request for a page element from the page. 6. The method according to claim 4, wherein the predetermined format is an at least partly compressed format. 7. The method according to claim 4, wherein the predetermined format comprises a predetermined maximum file size. 8. The method according to claim 4, wherein the predetermined format is defined for the client in a table comprised in the first server. 9. The method according to claim 4, wherein the format is determined by the client by transferring a message from the client to the first server. 10. The method according to claim 1, comprising the further steps of: transferring a message to the client comprising a quantity which provides a measure for the costs of transferring the page and the at least one page element to the client, prior to transferring the page with the collected at least one page element to the client. 11. The method according to claim 10, comprising the further step of: transferring a message from the client to the first server which determines whether or not to enable the first server to transfer the page and the collected at least one page element to the client in response to the message comprising the quantity. 12. The method according to claim 10, wherein the quantity comprises the size of the page and with the collected at least one page element. 13. The method according to claim 10, wherein the quantity comprises the estimated time for transferring the page with the collected at least one page element to the client. 14. The method according to claim 10, wherein the quantity comprises the total call charges for transferring the page and with the collected at least one page element. 15-16. (Cancelled) 17. The method according to claim 1, wherein the request for transferring the page to the client is generated by the first server. 18-19. (Cancelled) 20. The method according to claim 1 wherein the page comprises an hypertext mark-up language page. 21-33. (Cancelled) 34. A system for transferring a page to a client within a communication system including a first server, wherein said page further includes at least one request for a page element, comprising: means for collecting the page comprising at least one request for a page element in the first server, in response to a request for transferring the page to the client; means for deriving in the first server the page the at least one request for the page element; means for collecting the at least one page element in the first server, in response to the at least one request for the page element derived from the page; means for combining in the first server the at least one page element with the page; means for transferring the page with the collected at least one page element to the client; and means for charging the client on a basis of the page with the collected at least one page element transferred to the client. 35. The system of claim 34, wherein said means for collecting the page further comprises means for transferring a request for the page by the first server to a second server. 36. The system of claim 34 further comprising, prior to transferring the page with the collected at least one page element from the first server to the client, means for converting the page with the collected at least one page element into a predetermined format. 37. The system of claim 36 wherein said means for converting the page into the predetermined format further comprises means for removing the at least one request for a page element from the page. |
Hiv-1 vpr interactions with mitochondrial apoptosis inducing factor and methods of using the same |
Assays to identify Vpr/AIF interaction and translocation inhibitors are disclosed. |
1. A method of identifying compounds that inhibit HIV Vpr binding to AIF comprising a test assay that comprises the steps of: i) contacting a) HIV Vpr or a fragment of Vpr known to interact with AIF and b) AIF or a fragment of AIF which interacts with Vpr in the presence of c) a test compound and ii) comparing the level of HIV Vpr binding to AIF to the level of HIV Vpr binding to AIF in the absence of said test compound. 2. The method of claim 1 wherein a) HIV Vpr and b) AIF or a fragment of AIF which interacts with Vpr are contacted in the presence of c) a test compound. 3. The method of claim 1 wherein a) HIV Vpr or a fragment of Vpr known to interact with AIF and b) AIF are contacted in the presence of c) a test compound. 4. The method of claim 1 wherein a) HIV Vpr and b) AIF are contacted in the presence of c) a test compound. 5. The method of claim 1 further comprising a positive control assay that comprises the steps of: i) contacting a) HIV Vpr or a fragment of Vpr known to interact with AIF and b) AIF or a fragment of AIF which interacts with Vpr in the presence of c) anti-Vpr antibodies which competitively bind to Vpr with respect to AIF and/or anti-AIF antibodies which competitively bind to AIF with respect to Vpr. 6. The method of claim 1 wherein the concentration of test compound is between 1 μM and 500 μM. 7. The method of claim 1 wherein the concentration of test compound is between 10 μM to 100 μM. 8. The method of claim 1 wherein a series of test assays are performed using a series of dilutions of test compounds. 9. A kit for performing the method of claim 1 comprising: a) a first container comprising HIV Vpr or a fragment of Vpr known to interact with AIF; and, b) a second container comprising AIF or a fragment of AIF which interacts with Vpr in the presence of c) a test compound; and optionally, c) instructions for performing the test assay. 10. The kit of claim 9 further comprising a fourth container comprising anti-Vpr antibodies which competitively bind to Vpr with respect to AIF and/or anti-AIF antibodies which competitively bind to AIF with respect to Vpr. 11. A method of identifying compounds that inhibit HIV Vpr/AIF nuclear translocation comprising a test assay that comprises the steps of: i) contacting, in the presence of a test compound, cells that comprise HIV Vpr or a fragment of Vpr known to interact with AIF and AIF or a fragment of AIF which interacts with Vpr in the presence and AIF; and ii) comparing the level of Vpr/AIF in the nucleus to the level of Vpr/AIF in the nucleus in the absence of said test compound and/or comparing the level of Vpr/AIF in the cytoplasm to the level of Vpr/AIF in the cytoplasm in the absence of said test compound. 12. The method of claim 10 wherein the cells comprise a) HIV Vpr and b) AIF or a fragment of AIF which interacts with Vpr. 13. The method of claim 10 wherein the cells comprise a) HIV Vpr or a fragment of Vpr known to interact with AIF and b) AIF. 14. The method of claim 10 wherein the cells comprise a) HIV Vpr and b) AIF. 15. The method of claim 1 wherein the concentration of test compound is between 1 μM and 500 μM. 16. The method of claim 1 wherein the concentration of test compound is between 10 μM to 100 μM. 17. The method of claim 1 wherein a series of test assays are performed using a series of dilutions of test compounds. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The HIV accessory protein Vpr has been identified as being capable of cell cycle arrest and the induction of apoptosis. This observation is described in PCT application PCT/US01/10028, which is incorporated herein by reference. The interaction of HIV Vpr with the human cellular protein hVIP has been disclosed in PCT application PCT/US98/21432, which is incorporated herein by reference. Mitochondrial apoptosis inducing factor has been identified. There is a need for drug discovery screens and novel drugs to prevent or induce apoptosis and that are useful in the treatment of inflammatory disease and cancer. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention relates to methods of identifying compounds that inhibit Vpr binding to AIF. The present invention relates to methods of identifying compounds that inhibit the nuclear translocation of Vpr in the presence of AIF. detailed-description description="Detailed Description" end="lead"? |
Wnv core protein/capsid interacting protein and uses of the same |
Substantially pure WIP-1 and its use in assays to identify PAP-1/WIP-1 interaction inhibitors or WIP-1/WNV Cp interaction inhibitors are disclosed. Compositions comprising antisense sequences to prevent expression of WIP-1 and methods of treating cancer using the same are disclosed. |
1. A substantially pure protein having the amino acid sequence of WIP-1. 2. A recombinant expression vector comprising a nucleic acid sequence that encodes a protein of claim 1. 3. A host cell comprising the recombinant expression vector of claim 2. 4. An isolated nucleic acid molecule consisting of the WIP-1 cDNA sequence or a fragment thereof having at least 10 nucleotides. 5. An oligonucleotide molecule comprising a nucleotide sequence complementary to a nucleotide sequence of at least 10 nucleotides of the WIP-1 cDNA sequence. 6. The oligonucleotide molecule of claim 5 consisting of a nucleotide sequence complementary to a nucleotide sequence of 10-150 nucleotides of the WIP-1 cDNA sequence. 7. The oligonucleotide molecule of claim 6 consisting of a nucleotide sequence complementary to a nucleotide sequence of 18-28 nucleotides of the WIP-1 cDNA sequence. 8. A pharmaceutical composition comprising a nucleic acid molecule of claim 5 and a pharmaceutically acceptable carrier. 9. An isolated antibody which binds to an epitope on WIP-1. 10. The antibody of claim 8 wherein said antibody is a monoclonal antibody. 11. A pharmaceutical composition comprising a substantially pure protein having the amino acid sequence of WIP-1 or a fragment thereof and a pharmaceutically acceptable carrier. 12. The pharmaceutical composition of claim 11 wherein said composition is sterile and pyrogen free. 13. The pharmaceutical composition of claim 12 comprising a substantially pure protein having the amino acid sequence of WIP-1. 14. The pharmaceutical composition of claim 12 comprising a substantially pure protein having the amino acid sequence of fragment of WIP-1 wherein said fragment is soluble and capable of binding to WNV Cp. 15. The pharmaceutical composition of claim 11 wherein the substantially pure protein of claim 11 having the amino acid sequence of fragment of WIP-1 wherein said fragment is soluble and capable of binding to WNV Cp. 16. A method of identifying compounds that inhibit WNV Cp binding to WIP-1 comprising contacting WNV Cp and WIP-1 in the presence of a test compound and comparing the level of WNV Cp binding to WIP-1 to the level of WNV Cp binding to WIP-1 in the absence of said test compound. 17. A method of identifying compounds that inhibit PAP-1 binding to WIP-1 comprising contacting PAP-1 and WIP-1 in the presence of a test compound and comparing the level of PAP-1 binding to WIP-1 to the level of PAP-1 binding to WIP-1 in the absence of said test compound. 18. A method of treating an individual who has tumor cells with PAP-1 assocaites with WIP-1 comprising administering to said individual a composition which comprises a compound that prevents WIP-1 expression or WIP-1/PAP-1 interaction. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The core protein of West Nile virus (WNV), also referred to as the capsid or Cp, has recently been identified as being capable of inducing apoptosis in cell in which it is present. This observation is described in PCT/US01/31355 and Ser. No. 60/237,885, which are each incorporated herein by reference. There is a need to identify novel compounds which inhibit WNV replication. Specifically, safe and effective compounds are sought which reduce replication by interfering with particular molecular signals mediated by WNV capsid protein. Likewise, safe and effective compounds are sought which interfere with the cofactor with which WNV Cp interacts, which is an essential component of the cell cycle cascade. Moreover, there is a need to identify the co-factor and target it in methods of modulating the cell cycle. There is a need for compounds and methods for inhibiting the activity of WNV Cp. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention relates to substantially pure WIP-1, to recombinant expression vectors comprising a nucleic acid sequence that encodes and host cells that comprise the recombinant expression vector. The present invention relates to isolated nucleic acid molecules consisting of the WIP-1 cDNA sequence or a fragment thereof having at least 10 nucleotides, or a nucleotide sequence complementary to a nucleotide sequence of at least 10 nucleotides of the WIP-1 cDNA sequence. The present invention relates to isolated antibodies which binds to an epitope on the WIP-1. The present invention relates to methods of identifying compounds that inhibit WNV Cp binding to WIP-1. The present invention relates to methods of identifying compounds that inhibit PAP-1 binding to WIP-1. The present invention relates to methods of treating an individual who has tumor cells with PAP-1 associated with WIP-1 comprising administering to said individual a composition which comprises a compound that prevents WIP-1 expression or WIP-1/PAP-1 interaction. |
Boring device and boring method |
A drilling device is composed so as to drill a drilled object composed of a brittle material with the end of a rotating core bit by using a core bit in which a bit, formed by dispersing and arranging a cemented carbide or super abrasive in a binder phase, is provided on the end of a cylindrical tube, and rotating the core bit around an axis with a direct motor. The drilling device is composed so that the core bit is rotated such that the peripheral velocity at the outer periphery of the bit is 300 m/min or more while the core bit presses against the drilled object at a pressure of 0.6 N/mm2 or more during drilling. |
1. A drilling device having a drilling tool, in which a bit formed by dispersing and arranging a cemented carbide or super abrasive in a binder phase, is provided on the end of a rod-shaped or cylindrical tool body, and a rotary drive device that rotates the drilling tool around an axis, and being composed so as to drill a drilled object composed of a brittle material by pressing the end of the rotating drilling tool against the drilled object; wherein, the rotary drive device is composed so as to maintain the peripheral velocity at the outer periphery of the bit at 300 m/min or more while pressing the drilling tool against the drilled object at a predetermined pressure of 0.6 N/mm2 or more during drilling. 2. A drilling device according to claim 1 wherein, the diameter of the drilling tool is 3 mm to 200 mm. 3. A drilling device according to claim 1 wherein, the diameter of the drilling tool is 3 mm to less than 15 mm. 4. A drilling device according to claim 1 wherein, the diameter of the drilling tool is 15 mm to less than 50 mm. 5. A drilling device according to claim 1 wherein, the diameter of the drilling tool is 50 mm to 200 mm. 6. A drilling device according to claim 1 wherein, the rotary drive device is provided with a tube-shaped rotor in which a rotating shaft, to which the aforementioned drilling tool is attached on its end, is integrally provided passing through it, and a cylindrical stator provided on the outer peripheral surface of the rotor. 7. A drilling method for drilling a drilled object composed of a brittle material by rotating around an axis a drilling tool in which a bit, formed by dispersing and arranging a cemented carbide or super abrasive in a binder phase, is provided on the end of a cylindrical tool body, and pressing the end of the rotating drilling tool against the drilled object; wherein, the drilled object is drilled by pressing the drilling tool against the drilled object at a predetermined pressure of 0.6 N/mm2 or more while maintaining the peripheral velocity at the outer periphery of the bit at 300 m/min or more. 8. A drilling method according to claim 7 wherein, the diameter of the drilling tool of 3 mm to 200 mm is used. 9. A drilling method according to claim 7 wherein, the diameter of the drilling tool of 3 mm to less than 15 mm is used. 10. A drilling method according to claim 7 wherein, the diameter of the drilling tool of 15 mm to less than 50 mm is used. 11. A drilling method according to claim 7 wherein, the diameter of the drilling tool of 50 mm to 200 mm is used. 12. A drilling device according to claim 2 wherein, the rotary drive device is provided with a tube-shaped rotor in which a rotating shaft, to which the aforementioned drilling tool is attached on its end, is integrally provided passing through it, and a cylindrical stator provided on the outer peripheral surface of the rotor. 13. A drilling device according to claim 3 wherein, the rotary drive device is provided with a tube-shaped rotor in which a rotating shaft, to which the aforementioned drilling tool is attached on its end, is integrally provided passing through it, and a cylindrical stator provided on the outer peripheral surface of the rotor. 14. A drilling device according to claim 4 wherein, the rotary drive device is provided with a tube-shaped rotor in which a rotating shaft, to which the aforementioned drilling tool is attached on its end, is integrally provided passing through it, and a cylindrical stator provided on the outer peripheral surface of the rotor. 15. A drilling device according to claim 5 wherein, the rotary drive device is provided with a tube-shaped rotor in which a rotating shaft, to which the aforementioned drilling tool is attached on its end, is integrally provided passing through it, and a cylindrical stator provided on the outer peripheral surface of the rotor. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Technical Field The present invention relates to a drilling device and drilling method for drilling holes in drilled objects composed of stone materials, bedrock or other typically brittle materials such as concrete, asphalt, granite and marble, and more particularly, to a drilling device and drilling method suitable for use when drilling tiles and joints of tiled walls or use when drilling concrete walls laid on the inner surfaces of tunnels, sewer pipes and so forth. 2. Background Art A method for reinforcing existing concrete walls consists of first cutting out a large portion of the wall, providing an iron brace in the cut out opening and then reinforcing the entire wall by solidifying this brace and an anchor arranged on the inner peripheral surface of the opening with concrete. At this time, the anchor is arranged by containing in a hole provided in the inner peripheral surface of the opening. The hole for arranging this anchor is formed as shown in FIG. 11 , for example, by a drilling device provided with a core bit 80 (drilling tool), composed by providing a tip-shaped bit 80 a , which is formed by dispersing and arranging a cemented carbide or super abrasive in a binder phase comprised by sintering a binder, on the end of a cylindrical tool body, and a motor 81 (rotary drive device) for rotating this core bit 80 around an axis. Namely, during drilling, a drilled object in the form of concrete 82 is drilled by pressing bit 80 a provided on the end of core bit 80 against concrete 80 while rotating to form a columnar core 83 a . By then extracting core 83 after braking off base 83 a of core 83 remaining inside concrete 82 , a hole having a diameter of, for example, about 15-50 mm and depth of about 50-500 mm is formed corresponding to the diameter of core bit 80 . In addition, in order to prevent collapse of a concrete wall laid on the inner surface of a tunnel, a hole is drilled through this concrete wall to bedrock on the back side of the concrete wall, and a grouting material and so forth is injected through this hole between the concrete wall and bedrock to reinforce the concrete wall. When drilling into a concrete wall, conventional rock drills, which drill holes in bedrock, are not used because the vibrations generated by the rock drill act to promote collapse, and in their stead, a drilling device as shown in FIG. 11 is similarly used to drill concrete structures. In this case, holes having a diameter of, for example, about 70-100 mm are drilled corresponding to the diameter of core bit 80 . In addition, in order to prevent separation of tiles accompanying dilapidation of structures having tiled outer walls, holes are drilled in the tiles and joints between tiles to form holes that reach to the underlying concrete wall, after which resin is injected behind the almost separated tiles through these holes to adhere the tiles to the concrete wall. A small impact drill for drilling concrete, for example, is used to drill holes in such tiles and tile joints. However, since ordinary impact drills cause the drill to vibrate during drilling and drill while pounding the drilled object in the manner of a hammer, they conversely promote separation of the tiles resulting in the disadvantage of damaging the outer wall. Therefore, a drilling device is used that is provided with a drilling tool, in which a bit is provided on the end of a rod-shaped or cylindrical drill body, and a rotary drive device for rotating this drilling tool around an axis. In the case of a drilling device of the prior art as shown in the drawing, a rotary shaft attached with a core bit is rotated by lowering the rotating speed with a gear and so forth in order to increase the generated torque obtained at a predetermined output power of the motor. The output power referred to here indicates the output power that can be extracted outside the motor but excluding the loss within the motor. Although this output power is decreased due to friction and so forth during the course of rotation being transmitted by a gear or other rotation transmission mechanism, it is ultimately converted to output power of the drilling device that rotates the core bit. This output power of the drilling device is then supplied for drilling holes. Namely, if the sum of the force in the tangential direction applied to the end of the core bit due to resistance received from the drilled object during drilling is taken to be F t , and the radius of the core bit is taken to be r, then the work required for making one revolution of the core bit during drilling can be expressed as 2πrF t . Therefore, when the core bit rotates f N per unit time, the power of the drilling device can be expressed as 2πF t f N . This relationship is more accurate if expressed as 2πF t f N =vF t since rω is the peripheral velocity v at the outer periphery of the core bit. However, since rF t is the generated torque required for rotating the core bit, if this generated torque is taken to be T, then the output power of the drilling device can be represented as P output ∝Tf N proportional to the product of rotating speed and generated torque. In this manner, under conditions in which output power P output of the drilling device is a certain fixed value, in order to increase generated torque T, the rotating speed f N of the drilling tool is reduced by lowering the rotating speed of the motor with gears and so forth, even though transmission loss of the output power attributable to the gears is present. A drilling device of the prior art as previously described had the shortcoming of slow drilling speed. Consequently, it invited the problems of prolonging the construction period and worsening the surrounding environment due to noise and vibrations generated during drilling. For example, in the case of performing tunnel repair, a large number of holes having a depth of 500-1000 mm must be drilled. However, in the case of using a drilling device of the prior art, it takes about 30 minutes to drill a single hole, thereby resulting in the problem of requiring enormous construction costs in terms of labor costs alone to complete drilling of all the holes. In addition, construction work has also recently been performed involving not only the concrete walls of tunnels, but also drilling holes in the concrete wall on the inner surfaces of sewer pipes followed by injecting a corrosion-resistant material behind the sewer pipes. In this manner, there has been a need to develop a technology suitable for drilling a large number of holes in a short period of time in concrete walls over long distances. In addition, since drilling devices of the prior art as mentioned above drill holes while reducing the rotating speed of the drilling tool without using impact vibrations like those used in impact drills, they had the disadvantage of a slow drilling speed as compared with ordinary impact drills. There are cases in which nearly all of the tiles of outer walls are typically separated or beginning to be separated in the case of poorly constructed buildings and so forth. Since the task of completely removing all of the tiles and then reattaching them is actually quite bothersome, resin is ultimately injected behind all of the separated tiles. In this case, an extremely large number of holes must be drilled in the tiles. Consequently, there were the problems of a prolonged construction period and increased costs due to the increase in drilling time. In view of these reasons, there was a desire to develop a drilling device having low levels of vibrations capable of rapidly drilling holes comparable to impact drills and particularly without promoting separation of the tiles due to vibrations generated during drilling. Therefore, the object of the present invention is to provide a drilling device and drilling method capable of drilling a drilled object in a short period of time by reducing the value of the work required to drill holes of a predetermined depth without waste. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG. 1 is a lateral view showing an example of a drilling device as a first embodiment as claimed in the present invention. FIG. 2 is a partially cutaway lateral view showing a drilling device body of a drilling device as a first embodiment as claimed in the present invention. FIG. 3 is a cross-sectional view of a support column section for explaining the structure of the support column section of a drilling device of the present embodiment. FIG. 4 is a cross-sectional view of a movement mechanism for explaining the constitution and structure of the movement mechanism of a drilling device of the present embodiment. FIG. 5 is a block diagram schematically showing the electrical circuit connections of a drilling device of the present embodiment. FIG. 6 is a graph showing the relationship between bit peripheral velocity and drilling speed standardized according to torque value. FIG. 7 is a graph showing the relationship between bit peripheral velocity and the amount of drilling work by a drilling device. FIG. 8 is lateral view showing an example of a drilling device as a second embodiment as claimed in the present invention. FIG. 9 is a partially cutaway lateral view showing the drilling device body of a drilling device as a second embodiment as claimed in the present invention. FIG. 10 is a partially cutaway lateral view showing the drilling device body of a drilling device as a third embodiment as claimed in the present invention. FIG. 11 is a cross-sectional view of a drilling device for explaining the structure of a drilling device of the prior art. detailed-description description="Detailed Description" end="lead"? |
Method and apparatus for managing energy in plural energy storage units |
A system for managing energy stored in a plurality of series connected energy storage units has a plurality of energy storage unit controllers, each controller being associated with one of the plurality of energy storage units, a balancing circuit between two of the energy storage units, the balancing circuit being controlled by at least one of the energy storage unit controllers, a serial electrical interface between the energy storage unit controllers for providing voltage isolated bi-directional communication, and a central controller in electrical communication with the energy storage unit controllers. A method for managing charge in a plurality of series connected energy storage units forming an energy storage device includes monitoring a current supplied to the energy storage device, determining at least one of a charging rate and a capacity of a first energy storage unit and a second energy storage unit in the energy storage device, and diverting current from the first energy storage unit to the second energy storage unit in response to the steps of monitoring and determining. |
1. A system for managing energy stored in a plurality of series connected energy storage units, the system comprising: a plurality of energy storage unit controllers, each controller being associated with one of the plurality of energy storage units; a balancing circuit between two of the energy storage units, the balancing circuit being controlled by at least one of the energy storage unit controllers; a serial electrical interface between the energy storage unit controllers for providing voltage isolated bi-directional communication; and a central controller in electrical communication with the energy storage unit controllers. 2. The system of claim 1, wherein the energy storage unit comprises one or more storage cells. 3. The system of claim 1, wherein the energy storage unit controller monitors a parameter of the associated energy storage unit selected from the group consisting of voltage, temperature, and current. 4. The system of claim 1, wherein the balancing circuit includes a reactive element and at least one switch, the switch being controlled by at least one of the energy storage unit controllers. 5. The system of claim 1, wherein a voltage monitored by any one energy storage unit controller is no more than 5 V. 6. The system of claim 1, wherein the communications are digital communications. 7. The system of claim 1, comprising a current sense resistor in series with the energy storage units. 8. The system of claim 7, wherein the central controller monitors a current in the current sense resistor. 9. The system of claim 1, comprising a disconnect switch in series with the energy storage units. 10. The system of claim 9, wherein the disconnect switch is operatively interfaced to the control chip. 11. The system of claim 1, wherein the storage unit controllers can be a separate integrated circuit, a discrete component, or a hybrid of an integrated circuit and a discrete component. 12. The system of claim 1, wherein the integrated circuit can be in a multi-chip module. 13. The system of claim 1, wherein the serial electrical interface provides bi-directional, two way alternate, using bit asynchronous transfer, byte synchronous logical ring with contra flow. 14. A system for managing energy stored in plural series connected energy storage units comprising: first means for monitoring voltage across a first energy storage unit; second means for separately monitoring voltage across a second energy storage unit; and means for balancing charge among the first and second energy storage units by exchanging energy among the first and second energy storage units in response to the first and second monitoring means during at least one of charging and discharging of the first and second energy storage units, the charge balancing means including a reactive equalization circuit. 15. The system of claim 14, wherein the first monitoring means monitors a temperature of the first energy storage unit. 16. The system of claim 14, wherein the second monitoring means monitors a temperature of the second energy storage unit. 17. The system of claim 14, wherein the charge balancing means includes an inductor, at least one switch and a diode connected across each switch. 18. The system of claim 14, comprising means for monitoring current in the plural series connected energy storage units. 19. The system of claim 14, comprising means for voltage isolated bi-directional communications between each means for voltage monitoring. 20. The system of claim 19, wherein the communications are digital communications. 21. The system of claim 14, comprising a disconnect switch in series with the energy storage units. 22. The system of claim 14, wherein each of the first and second monitoring means is a dedicated controller. 23. A method for managing charge in a plurality of series connected energy storage units forming an energy storage device, the method comprising: monitoring a current supplied to the energy storage device; determining at least one of a charging rate and a capacity of a first energy storage unit and a second energy storage unit in the energy storage device; and diverting current from the first energy storage unit to the second energy storage unit in response to the steps of monitoring and determining. 24. The method of claim 23, wherein the storage unit balancing current compensates for an impedance of the storage unit. |
<SOH> BACKGROUND <EOH>1. Field of Invention The present invention relates generally to managing energy in plural energy storage units. More particularly, the present invention relates to a method and system for managing energy in plural energy storage units in which individual energy storage units are each monitored and the charge in each unit equalized during charging and/or discharging. 2. Background Information Battery packs composed of multiple cells can use internal electronics to assist in managing performance and safety issues. Individual cells can be connected in series to provide higher battery pack voltages for high voltage applications (for example, for use in notebook computers and vehicles). Metal hydride cells, such as nickel metal hydride (NiMH), and lithium-based chemistries, such as Li-ion cells, can be damaged or even explode if undercharged or overcharged. Internal electronics can be used for safety reasons to maintain safe voltages, currents and temperatures within the battery pack. Additional examples of uses for internal electronics in battery packs include gauging charge levels to determine remaining run time or charging time, managing the charge and discharge process, and communicating with a host device or appliance. Individual cell voltages may not remain balanced (equal) across a series connected grouping of individual cells or group of cells. Rather, some cells can be over potential and other cells can be under potential as referenced to the voltage measured across all the cells. Therefore, batteries and battery packs can include circuitry to monitor individual cells and, for example, stop the current to the batteries or battery packs before damage occurs. U.S. Pat. No. 5,631,534, the contents of which are herein incorporated by reference, discloses circuitry associated with a battery pack for controlling the voltage balance between cells. A balancing circuit can be used during the charging process to transfer energy from those cells which reach full charge first to those cells that are at a voltage less than full charge. |
<SOH> SUMMARY <EOH>The present invention is directed to a system for managing energy in a plurality of energy storage units. In an exemplary embodiment, a system for managing energy stored in a plurality of series connected energy storage units comprises a plurality of energy storage unit controllers, each controller being associated with one of the plurality of energy storage units, a balancing circuit between two of the energy storage units, the balancing circuit being controlled by at least one of the energy storage unit controllers, a serial electrical interface between the energy storage unit controllers for providing voltage isolated bidirectional communication, and a central controller in electrical communication with the energy storage unit controllers. In another exemplary embodiment, a system for managing energy stored in plural series connected energy storage units comprises first means for monitoring voltage across a first energy storage unit, second means for separately monitoring voltage across a second energy storage unit, and means for balancing charge among the first and second energy storage units by exchanging energy among the first and second energy storage units in response to the first and second monitoring means during at least one of charging and discharging of the first and second energy storage units, the charge balancing means including a reactive equalization circuit. An exemplary method for managing charge in a plurality of series connected energy storage units forming an energy storage device, the method comprises monitoring a current supplied to the energy storage device, determining at least one of a charging rate and a capacity of a first energy storage unit and a second energy storage unit in the energy storage device, and diverting current from the first energy storage unit to the second energy storage unit in response to the steps of monitoring and determining. |
Multiple quantum well broad spectrum gain medium and method for forming same |
A broadband medium (100) for a laser (300) having multiple quantum wells (130). |
1. A broadband gain medium, comprising: a substrate; and a multiple quantum well region on the substrate comprising at least two quantum wells, wherein at least one of the quantum wells exhibits a non-constant thickness profile and a non-constant material composition, and at least one of the quantum wells exhibits a thickness profile that is different than a thickness profile of the other quantum wells. 2. The broadband gain medium of claim 1, wherein the multiple quantum well region comprises an InGaAs/InGaAsP quantum well region. 3. The broadband gain medium of claim 1, wherein the multiple quantum well region comprises a plurality of InGaAs quantum wells, wherein respective InGaAsP layers are positioned between adjacent InGaAs quantum wells. 4. The broadband gain medium of claim 1, wherein the thickness profile of each quantum well is adjusted to broaden a gain spectrum of the broadband gain medium. 5. The broadband gain medium of claim 1, wherein each of the quantum wells has a non-constant thickness profile and a non-constant material composition. 6. The broadband gain medium of claim 1, wherein each quantum well has a different thickness profile. 7. The broadband gain medium of claim 1, wherein each quantum well has a different and non-constant thickness profile and a non-constant material composition. 8. The broadband gain medium of claim 5, wherein each quantum well has a thickness that increases along a resonant cavity direction. 9. The broadband gain medium of claim 7, wherein each quantum well has a thickness that increases along a resonant cavity direction. 10. A tunable semiconductor laser comprising the broadband gain medium of claim 1. 11. A broadband gain medium, comprising: a substrate; a buffer layer on the substrate; a multiple quantum well region on the buffer layer comprising at least two quantum wells, wherein at least one of the quantum wells exhibits a non-constant thickness profile and a non-constant material composition, and at least one of the quantum wells exhibits a thickness profile that is different than a thickness profile of the other quantum wells; a cladding layer on the multiple quantum well region; and a contact layer on the cladding layer. 12. The broadband gain medium of claim 11, wherein the substrate comprises an n-doped InP substrate, the buffer layer comprises an n-doped InP buffer layer, the multiple quantum well region comprises an InGaAs/InGaAsP multiple quantum well region, the cladding layer comprises a p-doped InP layer, and the contact layer comprises a p-doped InGaAs layer. 13. The broadband gain medium of claim 11, wherein the multiple quantum well region comprises a plurality of InGaAs quantum wells, and respective InGaAsP layers positioned between adjacent InGaAs quantum wells. 14. The broadband gain medium of claim 11, wherein the multiple quantum well region comprises: a first InGaAsP layer on the buffet layer; a first InGaAs quantum well on the first InGaAsP layer; a second InGaAsP layer on the first InGaAs quantum well; a second InGaAs quantum well on the second InGaAsP layer; a third InGaAsP layer on the second InGaAs quantum well; a third InGaAs quantum well on the third InGaAsP layer; a fourth InGaAsP layer on the third InGaAs quantum well; a fourth InGaAs quantum well on the fourth InGaAsP layer; and a fifth InGaAsP layer on the fourth InGaAs quantum well. 15. The broadband gain medium of claim 11, wherein each of the quantum wells has a non-constant thickness profile and a non-constant material composition. 16. The broadband gain medium of claim 11, wherein each of the quantum wells has a different thickness profile. 17. The broadband gain medium of claim 11, wherein each of the quantum wells has a different and non-constant thickness profile, and a non-constant material composition. 18. The broadband gain medium of claim 14, wherein each of the quantum wells has a thickness that increases along a resonant cavity direction. 19. The broadband gain medium of claim 18, wherein, for any point along the resonant cavity direction, the thickness of the second InGaAs quantum well is larger than the thickness of the first InGaAs quantum well, the thickness of the third InGaAs quantum well is larger than the thickness of the second InGaAs quantum well, and the thickness of the fourth InGaAs quantum well is larger than the thickness of the third InGaAs quantum well. 20. A tunable semiconductor laser comprising the broadband gain medium of claim 19. 21. A tunable semiconductor laser, comprising: a broadband gain medium, comprising: a substrate, a buffer layer on the substrate, a multiple quantum well region on the buffer layer comprising at least two quantum wells, wherein at least one of the quantum wells exhibits a non-constant thickness profile and a non-constant material composition, and at least one of the quantum wells exhibits a thickness profile that is different than a thickness profile of the other quantum wells, a cladding layer on the multiple quantum well region, and a contact layer on the cladding layer; and a wavelength tuning device optically coupled to the broadband gain medium. 22. The tunable laser of claim 21, wherein the wavelength tuning device comprises a grating. 23. A method of fabricating a broadband gain medium, comprising the steps of: growing at least two quantum wells by metalorganic chemical vapor deposition (MOCVD) selective area growth such that at least one of the quantum wells has a non-constant thickness profile and a non-constant material composition; wherein a growth time for each of the quantum wells is adjusted such that a thickness profile of at least one of the quantum wells is different than a thickness profile of the other quantum wells. 24. The method of claim 23, wherein the step of growing at least two quantum wells by MOCVD selective area growth comprises the steps of: forming an oxide mask on a substrate, wherein the oxide mask comprises first and second tapered oxide mask regions spaced apart on the substrate; and subjecting the substrate to MOCVD, wherein a quantum well growth rate between the first and second tapered oxide regions varies as a function of the width of the first and second tapered oxide regions. 25. The method of claim 23, wherein at least one of the quantum wells is formed so that its thickness increases along a resonant cavity direction. 26. The method of claim 23, wherein four quantum wells are formed with respective thicknesses that increase along a resonant cavity direction. 27. A method of fabricating a semiconductor optical amplifier, comprising the steps of: providing a substrate; forming a buffer layer on the substrate; growing a multiple quantum well region, comprising a plurality of quantum wells, on the buffet layer using metalorganic chemical vapor deposition (MOCVD) selective area growth, such that at least one of the quantum wells has a non-constant thickness profile and a non-constant material composition, wherein a growth time for each of the quantum wells is adjusted such that a thickness profile of at least one of the quantum wells is different than a thickness profile of the other quantum wells; and forming a cladding layer on the multiple quantum well region. 28. The method of claim 27, wherein the step of forming a multiple quantum well region by MOCVD selective area growth comprises the steps of: forming an oxide mask on the substrate, wherein the oxide mask comprises first and second tapered oxide mask regions spaced apart on the substrate; and subjecting the substrate to MOCVD, wherein a quantum well growth rate between the first and second tapered oxide regions varies as a function of the width of the first and second tapered oxide regions. 29. The method of claim 27, wherein at least one of the quantum wells is selectively grown so that its thickness increases along a resonant cavity direction. 30. The method of claim 27, wherein four quantum wells are selectively grown with respective thicknesses that increase along a resonant cavity direction. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention This invention relates to semiconductor optical gain media and, more particularly, to a multiple quantum well semiconductor optical gain medium that exhibits a broad gain spectrum. 2. Background of the Related Art Broadly tunable optical devices, such as broadly tunable semiconductor lasers and broadband wavelength converters, are desired for various optical communication applications, such as optical networking, wavelength-division-multiplexing and other telecommunications applications. Multiple quantum well (MQW) gain materials have been used because of their relatively broad gain spectra. However, there is a continuing need for broader tuning ranges than prior art MQW materials offer. The above references are incorporated by reference herein where appropriate for appropriate teachings of additional or alternative details, features and/or technical background. |
<SOH> SUMMARY OF THE INVENTION <EOH>An object of the invention is to solve at least the above problems and/or disadvantages and to provide at least the advantages described hereinafter. The present invention provides a broadband gain medium, and a method for forming the same, that exhibits a broader gain spectrum than prior art semiconductor materials. The broadband gain medium of the present invention includes a multiple quantum well region made up of at least two quantum wells, with at least one of the quantum wells having a thickness and composition that vary as a function of position along the resonant cavity direction, and at least one quantum well having a thickness profile that is different than the other quantum wells. Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention. The objects and advantages of the invention may be realized and attained as particularly pointed out in the appended claims. |
Customization of error handling based on type of user agent |
A World Wide Web (web) server built-in service access function (SAF) is overridden with a service having intelligence about the type of user-agent to which an error page is being served. In this way, error messages are served to a client in a platform appropriate manner. |
1. A method for a World Wide Web (Web) server to process an error encountered by a client, said method comprising: determining an appropriate error file to send to said client, said error file associated with said error; using a replaced SAF, said replaced SAF: determining type of client; adjusting said error file by incorporating said client type; and sending said adjusted error file to said client. 2. The method of claim 1, said determining type of client, further comprising: requesting and reading corresponding headers. 3. The method of claim 1, said determining type of client, further comprising: determining device and running software of said client; and determining content type of said client. 4. The method of claim 3, further comprising: said adjusting error file replacing a default error file extension with an appropriate error file extension, said appropriate file extension corresponding to said running software of said client; and said sending said adjusted error file with said determined content type. 5. The method of claim 1, wherein said client is any of, but not limited to: a personal computer, a cell phone, a personal digital assistant (PDA), and the like. 6. The method of claim 3, wherein said content type is any of, but not limited to: Html, wml, and hdml. 7. An apparatus for a World Wide Web (Web) server to process an error encountered by a client, said apparatus comprising: means for determining an appropriate error file to send to said client, said error file associated with said error; means for using a replaced SAF, said replaced SAF providing: means for determining type of client; means for adjusting said error file by incorporating said client type; and means for sending said adjusted error file to said client. 8. The apparatus of claim 7, said means for determining type of client, further comprising: means for requesting and reading corresponding headers. 9. The apparatus of claim 7, said means for determining type of client, further comprising: means for determining device and running software of said client; and means for determining content type of said client. 10. The apparatus of claim 9, further comprising: said means for adjusting error file providing means for replacing a default error file extension with an appropriate error file extension, said appropriate file extension corresponding to said running software of said client; and said means for sending said adjusted error file with said determined content type. 11. The apparatus of claim 7, wherein said client is any of, but not limited to: a personal computer, a cell phone, a personal digital assistant (PDA), and the like. 12. The apparatus of claim 7, wherein said content type is any of, but not limited to: HTML, wml, and hdml. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Technical Field The invention relates to error handling. More particularly, the invention relates to a World Wide Web (Web) site customizing error handling for a variety of end user devices. 2. Description of the Prior Art Currently, Web servers can customize error messages for different error codes resulting on resident Web sites. That is, Web servers can serve customized error pages on specific errors. However, such Web servers serve only HTML MIME type error messages because only HTML MIME type messages are supported. Thus, WAP phones, PDA's, and other such devices that do not understand HTML MIME type are not able to display their corresponding custom error pages having appropriately customized content, and may, instead, display unpredictable behavior. Such unpredictable behavior can be detrimental to an organization or individual with an interest in a Web site accessed by such devices. W. Doviak, D. L. Whitmore, and F. Houvig, Apparatus and Method for Intelligent Routing of Data Between a Remote Device and a Host System, U.S. Pat. No. 6,198,920 (Mar. 6, 2001) discloses an apparatus and method for transparent communication between a remote or mobile device and a fixed communication host network. The apparatus and method may include a remote network controller that logically resides between the host network and the existing infrastructure(s) that are used to provide communications network contact with one or more remote devices. The remote network controller is connected to the host communication network as a protocol-appropriate communications controller so that remote devices are indistinguishable to the host network from the locally-attached devices. Each remote device may be provided with an asynchronous serial data interface to communicate with a mobile data controller. The mobile data controller, in combination with the remote network controller, provides end-to-end data communication such that incompatible protocols are transparent to the remote device and host communication network. A router may be provided which selects a communications network in accordance with user configured parameters. The router communicates over a plurality of incompatible networks and is capable of using a variety of different protocols. Switching between the plurality of incompatible networks is transparent to the remote device and host communication network. Doviak et al teach a complicated communications system that includes at least the use of special routers and interfaces. T. F. LaPorta. K. K. Sabnani, and T. Y. C. Woo, Two-way Wireless Messaging System Having User Agent, U.S. Pat. No. 5,970,122 (Oct. 19,1999) discloses a two-way wireless messaging system that includes a messaging network having at least one user agent corresponding to a subscriber of a two-way wireless messaging service. The subscriber receives messages from the messaging network along a first communication channel. The user agent includes a plurality of messages stored therein wherein a predetermined message is stored in the user agent and forwarded to a desired destination in response to an originating message code that is received from a two-way messaging device of the subscriber along a second communication return channel. The originating message code can be expanded by the user agent. The messages stored by the user agent can be modified so that different messages can be forwarded to the predetermined destination. The user agent also maintains location information of the two-way messaging device of the subscriber. LaPorta et al teaches communication on a two-way wireless messaging system, and does not teach a simple and elegant way for a Web server to handle errors experienced by a variety of linked devices. T. F. LaPorta. K. K. Sabnani, and T. Y. C. Woo, Two-way Wireless Messaging System, U.S. Pat. No. 5,918,158 (Jun. 29,1999) discloses a way to improve upon a two-way messaging system allowing messages to be originated from a wireless messaging device and replies to be dynamically customizable by the recipients. LaPorta et al teach a hybrid system between traditional paging and electronic mail, improving on traditional paging and messaging services as well as any proposed two-way messaging services by adding direct message origination and reply capability with various capabilities for some message modification and response. However, LaPorta et al does not teach customizing error handling by Web servers for a variety of end user devices. It would be advantageous to provide a method and apparatus that allows sending a correct error page to a linked device and that is both simple and elegant in design and implementation. It would be advantageous to provide a method and apparatus that allow Web serves to manage error handling for a variety of different types of linked end users, such as personal computers, hand-held devices, and the like. |
<SOH> SUMMARY OF THE INVENTION <EOH>A method and apparatus is provided that enables a Web server to customize error messages for a variety of linked communicating devices, such as, for example, cell phones and personal digital assistants (PDAs). A Web server built-in service access function (SAF) is overridden with a service having intelligence about the type of user-agent to which an error page is being served. In this way, error messages are served to a client in a platform appropriate manner. |
Microstructures and use thereof for the directed evolution of biomolecules |
The invention relates to microstructures and the use thereof for the directed evolution of biomolecules. |
1. A method for the cell-free selection of genotype variants from genotype libraries in a microstructure, comprising the following sequence of reaction steps: (a) combining a test fluid comprising a genotype library in an expressible form and expression aids suitable for cell-free expression with a separation fluid in the microstructure to form individual compartments of the test fluid; (b) transporting the compartments through the microstructure, the expression of the genotype into the phenotype being effected in the compartments; (c) detecting the phenotype in the compartments; and (d) selecting the compartments in accordance with their phenotypes. 2-20. (canceled) 21. The method according to claim 1, wherein an assay fluid with reagents suitable for detecting the phenotype is added to the compartments after step (b). 22. The method according to claim 1, wherein said test fluid and expression aids suitable for cell-free expression are selected from aqueous solutions and suspensions of complex compositions. 23. The method according to claim 22, wherein said test fluid contains a cell extract suitable for in-vitro protein expression. 24. The method according to claim 1 wherein said separation fluid is a water-immiscible fluid. 25. The method according to claim 24, wherein the water-immiscible fluid is selected from aliphatic and aromatic hydrocarbons, higher alcohols, higher alkanones, esters and ethers of higher hydrocarbons, halogenated hydrocarbons and silicones and mixtures of these substances. 26. The method according to claim 1 wherein the transport speed of the compartments within the microstructure is from 1×10−7 to 1×10−2 m/s. 27. The method according to claim 26 wherein the transport speed of the compartments with the microstructure is from 1×10−6 to 1×10−4 m/s. 28. The method according to claim 1, wherein the concentration of said genotype library and said combining of the test fluid and the separation fluid are selected in such a way that a statistic average of only one genotype variant is contained per compartment. 29. The method according to claim 1, wherein the compartment volume is from 0.01 fl to 10 pl. 30. The method according to claim 29, wherein the compartment volume is from 0.1 fl to 1 pl. 31. The method according to claim 29, wherein the compartment volume is from 1 to 100 fl. 32. The method according to claim 21, wherein said assay fluid is miscible with said test fluid and immiscible with said separation fluid, and the time of the addition is selected in such a way that an amount of gene product sufficient for detection will already have formed by then. 33. The method according to claim 32 wherein said assay fluid is selected from aqueous solutions, suspensions and emulsions. 34. The method according to claim 1, wherein the assay reagents are specific for the function to be selected. 35. The method according to claim 34, wherein the assay reagents are suitable for analyzing the function to be selected with optical measuring methods. 36. The method according to claim 34, wherein the assay reagent are suitable for analyzing the function to be selected with fluorimetric measuring methods. 37. The method according to claim 1, wherein the detection of the phenotype in the compartments comprises the qualitative determination of the phenotypical properties. 38. The method according to claim 37, wherein the determination of the phenotype is effected by optical methods. 39. The method according to claim 37, wherein the determination of the phenotype properties is effected by fluorimetric methods. 40. The method according to claim 1, wherein the detection of the phenotype in the compartments comprises the quantitative determination of the phenotypical properties. 41. The method according to claim 40, wherein the determination of the phenotype is effected by optical methods. 42. The method according to claim 40, wherein the determination of the phenotype properties is effected by fluorimetric methods. 43. The method according of claim 1, wherein said phenotype is manifested by endonucleolytic activity. 44. The method according of claim 1, wherein said selecting of the compartments is effected by sorting. 45. The method according to claim 44, which further comprises the reaction step of (e) isolating the genotype of the selected compartments to form a new genotype library. 46. The method according to claim 45, wherein the genotype library obtained is subjected to one or more further reaction cycles (a) to (d). 47. The method according to claim 1 which further comprises the reaction step of (e) isolating the genotype of the selected compartments to form a new genotype library. 48. The method according to claim 47, wherein the genotype library obtained is subjected to one or more further reaction cycles (a) to (d). 49. A microstructure for performing the method according to claim 1, comprising: a first supply channel for supplying a test fluid (102) to a reaction channel; a second supply channel for supplying at least one separation fluid (101) to the reaction channel; a detection means (205) provided at the end of the reaction channel for detecting a reaction proceeded in the test fluid; and a selection means for selecting the test fluid compartments (109). 50. The microstructure according to claim 49, wherein the first supply channel is for supplying a fluid containing a genotype. 51. The microstructure according to claim 49, characterized by a first metering means (221, 222) connected with the first or second supply channel for the volume-limited supply of test fluid (102) or separation fluid (101), so that compartments (109, 111) are formed in the reaction channel. 52. The microstructure according to claim 51, characterized by a second metering means (222) which is arranged in such a way that one metering means (221, 222) is provided in each of the first and second supply channels. 53. The microstructure according to claim 52, characterized by a control means connected with said metering means (221, 222) by which said metering means (221, 222) can be controlled in such a way that only test fluid (102) and separation fluid (101) are supplied to the reaction channel alternately. 54. The microstructure according to claim 49, characterized by a third supply channel for supplying assay fluid (103) to the reaction channel. 55. The microstructure according to claim 54, characterized in that a third metering means (223) for the volume-limited supply of assay fluid (103) is connected with the third supply channel and further connected with a recognition means for recognizing a test fluid compartment (109) and can be controlled through a signal transmitted from the recognition means so that the assay fluid (103) is supplied to the test fluid compartment (109). 56. The microstructure according to claim 49, characterized in that said selection means has at least two selection channels (112) connected with the reaction channel, and a selection means (224, 225) for selecting one of the two selection channels (112) depending on the detection result. 57. The microstructure according to claim 56, characterized in that a metering means (224, 225) is provided as a selection means in at least one of said selection channels (112). 58. The microstructure according to claim 49, characterized in that said reaction channels (108, 110) have several individual channels (408, 410) which can be switched in parallel. 59. The microstructure according to claim 58, characterized in that each individual channel has at least one inlet metering means and/or one outlet metering means. 60. The microstructure according to claim 49, characterized in that said metering means (222, 223, 224) are microstructured valve elements. |
<SOH> BACKGROUND OF THE INVENTION <EOH>In methods of directed molecular evolution, libraries containing a wide variety of variants of a biomolecule are used for the selection of variants which correspond to a predetermined goal of evolution. The cyclic repetition of variation, amplification and selection of variants generates optimized biomolecules. Methods of directed molecular evolution are primarily based on the generation of a large number of DNA variants (genotype library). Starting from such a library of genotypes, the corresponding gene products are prepared, screened for their properties (phenotype) and accordingly selected. For examination, screening methods are found to be particularly advantageous due to their flexibility and general applicability as compared to other selection methods, for example, growth-coupled ones. Screening methods are based on the spatial isolation of the genotype variants. This isolation of the genotypes ensures both the possibility to separately measure properties of the different phenotypes and the assignment of the genotype to the phenotype, which is indispensable for the selection and amplification of optimum genotypes. Since the number of variants to be examined can be very high, the segregation of the genotypes is usually effected in sample supports which include a large number of sample compartments in methods performed to date. For example, commercially available sample supports comprise 96, 384 or 1536 sample compartments. The number of sample compartments is chosen as high as possible in order to limit the number of sample supports required and the quantity of necessary assay reagents. A limiting factor in the development of sample supports having an even higher number of sample compartments is the handling of low quantities of liquids in such compartments. Viscosity and surface tension factors as well as the evaporation of fluid samples, which are of minor importance in larger volumes, put strong limits on this procedure. It is the object of the invention to provide a screening method for directed molecular evolution which avoids the mentioned drawbacks of screening methods based on sample supports. In principle, a method for achieving the same object has been known from WO 95/35492. The method described is suitable for separating by their properties sample components of a fluid mixture of samples conveyed in a capillary. A disadvantage of this method is the fact that a possibility for reducing the diffuse mass transport within the fluid stream is not provided. This drawback prevents the use of very small sample compartments because the diffusive loss of sample components prevents the practicability of the sought reactions in the sample compartment especially for small dimensions. Further, the technical realization of the method requires high demands on the mechanical positioning of the components. Such solutions are frequently found to be unstable and error-prone. Further, from DE-A-19950385, an in-vivo screening method is known which enables the identification of per se unselectable activities in a target cell. The nucleic acid sequence to be examined is introduced in these target cells by transfection together with a reporter vector. The activity in the target cells or in their culture supernatant which results from the reporter is employed as a measure of the unselectable activity of the nucleic acid sequence examined and their identification. However, it is known that the use of methods for in-vivo expression is limited by some factors. Thus, intracellular nucleases or proteases can destroy the introduced genotype or the gene product. The gene products expressed can have a toxic or inhibiting effect on the host cells and thus adversely affect their effectiveness. In addition, the gene products can be expressed as an “inclusion body” in an insoluble form or biologically inactive form. In contrast, the cell-free in-vitro expression is not bound to any cellular control mechanisms and enables a direct access to the expressed gene products without isolation operations. In addition, the preparation of artificial gene products is possible by incorporating modified non-proteinogenic amino acids. The present invention describes a screening method in the microstructure according to the invention for the selective identification of genotypes based on cell-free in-vitro expression. |
<SOH> SUMMARY OF THE INVENTION <EOH>It has been found that the screening of a set of samples and the selection or sorting of individual samples from this set can be effected in an in-vitro method in channel structures produced by microstructuring techniques. In these microstructures, the division into individual samples is effected by segregation using various fluid phases. Thus, the present invention relates to: (1) a method for the cell-free (i.e., in-vitro) selection of genotype variants from genotype libraries in a microstructure, comprising the following sequence of reaction steps: (a) combining a test fluid comprising a genotype library in an expressible form and expression aids suitable for cell-free expression with a separation fluid in the microstructure to form individual compartments of the test fluid; (b) transporting the compartments through the microstructure, the expression of the genotype into the phenotype being effected in the compartments; (c) detecting the phenotype in the compartments; and (d) selecting the compartments in accordance with their phenotypes; and (2) a microstructure for performing the method as defined above under (1), comprising: a first supply channel for supplying a test fluid ( 102 ), especially a fluid containing a genotype, to a reaction channel; a second supply channel for supplying at least one separation fluid ( 101 ) to the reaction channel; a detection means ( 205 ) provided at the end of the reaction channel for detecting a reaction proceeded in the test fluid; and a selection means for selecting the test fluid compartments ( 109 ). |
Hybrid optical component for x ray applications and method associated therewith |
One aspect of the invention relates to a multi-layered reflective optical system for the reflection of X rays at a low angle of incidence, producing a two-dimensional optical effect. The inventive optical system comprises: a component having a surface which is reflective in such a way that a first optical effect is produced according to a first direction in space; and means for producing a second optical effect according to a second direction in space which is different from the first direction, characterized in that said means for producing a second optical effect are borne by the reflective surface. A second aspect of the invention relates to a method for the production of said optical system. |
1. A multilayer reflective optical assembly for reflecting X-rays at a low angle of incidence and producing a two-dimensional optical effect, said optical assembly comprising: a component having a reflecting surface shaped so as to produce a first optical mono-dimensional effect in a first direction in space; and means for producing a second mono-dimensional optical effect in a second direction in space, different from said first direction, said means for producing said second optical effect being carried by said reflecting surface, said optical assembly enabling production of a two-dimensional optical effect by making said X-rays undergo a single reflection. 2. The optical assembly of claim 1, wherein said reflecting surface is shaped: as a cylinder with a circular directrix, a cylinder with a parabolic directrix, a cylinder with an elliptical directrix, or a sphere. 3. The optical assembly of either claim 1 or 2, wherein said optical assembly comprises a laterally-graded multilayer. 4. The optical assembly of claim 1, wherein said means for producing said second optical effect comprise a diffractive pattern. 5. The optical assembly of claim 4, wherein said reflecting surface is a cylinder with an axis and said diffractive pattern comprises diffraction gratings made perpendicular to said axis of said cylinder. 6. The optical assembly of claim 4, wherein said diffractive pattern is produced in relief on a multilayer, or etched into a structure of said multilayer. 7. The optical assembly of claim 1, wherein said means for producing said second optical effect comprise a refractive pattern. 8. The optical assembly of claim 7, wherein said refractive pattern is of the Kino lens type. 9. The optical assembly of claim 7, wherein said refractive pattern is produced in relief on a multilayer or etched into a structure of said multilayer. 10. The optical assembly of claim 1, wherein said first mono-dimensional optical effect and said second mono-dimensional optical effect are each a collimation in two respective directions or a focusing in two respective directions. 11. (canceled) 12. (canceled) 13. (canceled) 14. (canceled) 15. (canceled) 16. A process for manufacturing an optical assembly as claimed in claim 1, comprising, in either order on a substrate, the steps of: generating a shaped surface with a desired geometry, depositing a multilayer and generating a pattern. 17. The process of claim 16 further comprising the steps of: depositing a multilayer on a planar substrate then shaping this multilayer so as to give it the desired geometry for the reflective surface and, finally, forming a pattern on the curved multilayer mirror thus created. 18. The process of claim 16 further comprising the steps of: depositing a multilayer on a planar substrate, creating the pattern on top and then shaping the combination to the desired geometry. 19. The process of claim 16 further comprising the steps of: generating a shaped surface with the desired geometry, then depositing a multilayer on top and forming a pattern on the combination. 20. The process of claim 16 further comprising the steps of: generating a shaped surface with the desired geometry, then forming a pattern on the top and depositing a multilayer on the combination. 21. The process of claim 16 further comprising the steps of: generating the pattern first, directly on an initial planar substrate, and then carrying out, in either order, the shaping of the surface and the deposition of a multilayer. 22. The process of claim 16, wherein the pattern comprises a diffractive pattern and the process comprises the creation of a diffractive structure by changing a multilayer structure following the exposure of the structure to an energy beam, the process comprising the control of the exposure of the desired regions of the multilayer structure to said beam, so as to shift the value of the reflective peak of each region of the structure in the desired manner within the wavelength spectrum. 23. The process of claim 22, wherein said exposure control is performed by adapting the duration of exposure of each individual region of the structure to the beam. 24. The process of claim 22, wherein said exposure control involves a temporary and controlled modification of the energy of the beam. 25. The process of claim 16, wherein the pattern comprises a refractive pattern and the process comprising the etching of a resist or of a light element in order to create the refractive pattern. |
Fungal isolates and biological control compositions for the control of weeds |
The present invention discloses fungal isolates of Phoma spp. or extracts obtained therefrom, useful for the control of broad leaf weeds, including Canada thistle, perennial sowthistle, dandelion, scentless chamomile, false cleavers, chickweed, wild buckwheat, and field bindweed. Te present invention also discloses biological control compositions comprising fungal isolates formulated in a growth medium for maintaining the viability of the fungal isolates when the biological control composition is applies to soil. |
1. A method of controlling one or more broad leaf weeds comprising administering Phoma, an extract therefrom, an inoculated broth therefrom, or a combination thereof, to said weeds, or to soil where said weeds grow. 2. The method of claim 1, wherein said Phoma is Phoma cf. macrostoma. 3. The method of claim 2, wherein said one or more broad leaf weeds is a species of a family selected from the group consisting of Compositae, Caryophyllaceae, Convolvulaceae, Plantaginaceae and Rubiaceae. 4. The method of claim 2, wherein said one or more broad leaf weeds is selected from the group consisting of Canada thistle, perennial sowthistle, dandelion, scentless chamomile, false cleavers, chickweed, wild buckwheat, field bindweed, plantain and prairie sunflower. 5. A biocontrol agent comprising one or more than one Phoma cf macrostoma isolate, an extract therefrom, an inoculated broth therefrom, or a combination thereof, said biocontrol agent exhibiting weed control activity, growth enhancement activity, or a combination thereof 6. The biocontrol agent of claim 5, wherein said one or more than one Phoma isolate is selected from the group consisting of a) 85-24B (IDAC 230201-1, deposited Feb. 23, 2001), b) 89-25A (IDAC 110401-1, deposited Apr. 11, 2001), c) 94-26 (IDAC 230201-2, deposited Feb. 23, 2001) d) 94-44B (IDAC 230201-3, deposited Feb. 23, 2001) e) 94-134 (IDAC 230201-4, deposited Feb. 23, 2001), f) 94-359A (IDAC 110401-2, deposited Apr. 11, 2001), g) 95-54A1 (IDAC 230201-5, deposited Feb. 23, 2001), h) 95-268B (IDAC 110401-3, deposited Apr. 11, 2001), i) 97-12B (IDAC 230201-6, deposited Feb. 23, 2001), j) 97-15B2 (IDAC 110401-4, deposited Apr. 11, 2001), and a combination thereof. 7. The biocontrol agent of claim 5, wherein said extract, said inoculated broth, or a combination thereof is obtained from one or more than one Phoma cf. macrostoma isolate. 8. The biocontrol agent of claim 7, wherein said extract, inoculated broth, or a combination thereof is obtained from one or more Phoma isolates selected from the group consisting of: a) 85-24B (IDAC 230201-1, deposited Feb. 23, 2001), b) 89-25A (IDAC 110401-1, deposited Apr. 11, 2001), c) 94-26 (IDAC 230201-2, deposited Feb. 23, 2001), d) 94-44B (IDAC 230201-3, deposited Feb. 23, 2001) e) 94-134 (IDAC 230201-4, deposited Feb. 23, 2001), f) 94-359A (IDAC 110401-2, deposited Apr. 11, 2001), g) 95-54A1 (IDAC 230201-5, deposited Feb. 23, 2001), h) 95-268B (IDAC 110401-3, deposited Apr. 11, 2001), i) 97-12B (IDAC 230201-6, deposited Feb. 23, 2001), and j) 97-15B2 (IDAC 110401-4, deposited Apr. 11, 2001). 9. A biocontrol composition, comprising the biocontrol agent of claim 5, and a medium for supporting viability of said one or more than one Phoma isolate. 10. The biocontrol composition of claim 9, wherein said medium is selected from the group consisting of Agar, pesta, peat prill, vermiculite, clay, starches, potato dextrose broth, V8® juice broth, cereal grain and legume grain. 11. The biocontrol composition of claim 9, wherein said one or more than one Phoma isolate is selected from the group consisting of a) 85-24B (IDAC 230201-1, deposited Feb. 23, 2001), b) 89-25A (IDAC 110401-1, deposited Apr. 11, 2001), c) 94-26 (IDAC 230201-2, deposited Feb. 23, 2001), d) 94-44B (IDAC 230201-3, deposited Feb. 23, 2001) e) 94-134 (IDAC 230201-4, deposited Feb. 23, 2001), f) 94-359A (IDAC 110401-2, deposited Apr. 11, 2001), g) 95-54A1 (IDAC 230201-5, deposited February. 23, 2001), h) 95-268B (IDAC 110401-3, deposited Apr. 11, 2001), i) 97-12B (IDAC 230201-6, deposited Feb. 23, 2001), j) 97-15B2 (IDAC 110401-4, deposited Apr. 11, 2001), and a combination thereof. 12. The biocontrol composition of claim 9, wherein said extract, said inoculated broth, or a combination thereof is obtained from one or more than one Phoma cf. macrostoma isolate. 13. The biocontrol composition of claim 12, wherein said extract, inoculated broth, or a combination thereof obtained from one or more Phoma isolates selected from the group consisting of: a) 85-24B (IDAC 230201-1, deposited Feb. 23, 2001), b) 89-25A (IDAC 110401-1, deposited Apr. 11, 2001), c) 94-26 (IDAC 230201-2, deposited Feb. 23, 2001), d) 94-44B (IDAC 230201-3, deposited Feb. 23, 2001) e) 94-134 (IDAC 230201-4, deposited Feb. 23, 2001), f) 94-359A (IDAC 110401-2, deposited Apr. 11, 2001), g) 95-54A1 (IDAC 230201-5, deposited Feb. 23, 2001), h) 95-268B (IDAC 110401-3, deposited Apr. 11, 2001), i) 97-12B (IDAC 230201-6, deposited February. 23, 2001), and j) 97-15B2 (IDAC 110401-4, deposited Apr. 11, 2001). 14. A method of controlling weed development during crop growth comprising: a) adding an effective amount of said biocontrol agent of claim 5 to produce a treated soil, b) planting said crops in said treated soil, c) growing said crop. 15. A method of controlling weed development during crop growth comprising: a) planting said crop, b) adding an effective amount of said biocontrol agent of claim 5 to soil where said crop is planted, c) growing said crop. 16. A method of controlling weed development during crop growth comprising: a) adding an effective amount of said biocontrol agent of claim 5 to crop seeds, to produce treated crop seed; b) planting said treated crop seed; and c) growing said crop. 17. A method of controlling weed development during established crop growth comprising: a) adding an effective amount of said biocontrol agent of claim 5 to the established crop; and b) growing the crop. 18. A method of controlling weed development during crop growth comprising: a) adding an effective amount of said biocontrol composition of claim 9 to produce a treated soil, b) planting said crops in said treated soil, c) growing said crop. 19. A method of controlling weed development during crop growth comprising: a) planting said crop, b) adding an effective amount of said biocontrol composition of claim 9 to soil where said crop is planted, c) growing said crop. 20. A method of controlling weed development during crop growth comprising: a) adding an effective amount of said biocontrol composition of claim 9 to crop seeds to produce treated crop seed; b) planting said treated crop seed; and c) growing said crop. 21. A method of controlling weed development during established crop growth comprising: a) adding an effective amount of said biocontrol composition of claim 9 to the established crop; and b) growing the crop. 22. A method of controlling weed development comprising applying the biocontrol agent of claim 5 to soil where said weed grows. 23. The method of claim 22, wherein said biocontrol agent is an extract from said one or more than one Phoma macrostoma isolate. 24. A method of controlling weed development comprising applying the biocontrol composition of claim 9 to soil where said weed grows. 25. The method of claim 24, wherein said biocontrol agent is said one or more than one Phoma macrostoma isolate. 26. The method of claim 22, wherein said step of applying comprises dusting, rubbing, spreading, drilling, banding, broadcasting, spraying, liquid injection, pouring or soil drenching. 27. The method of claim 24, wherein said step of applying comprises dusting, rubbing, spreading, drilling, banding, broadcasting, spraying, liquid injection, pouring or soil drenching. 28. A method of controlling weed development comprising applying the biocontrol agent of claim 5 to said weed. 29. A method of controlling weed development comprising applying the biocontrol composition of claim 9 to said weed. 30. The method of claim 28, wherein said step of applying comprises dusting, rubbing, spreading, broadcasting, spraying, or pouring. 31. The method of claim 29, wherein said step of applying comprises dusting, rubbing, spreading, broadcasting, spraying, or pouring. 32. The agent of claim 7, wherein said extract is selected from the group consisting of heat killed barley inoculum, a chloroform extract of said Phoma isolate, a methanol extract of said Phoma isolate, and an ethyl-acetate extract of said Phoma isolate. 33. The agent of claim 7, wherein said inoculated broth is selected from the group consisting of a crude inoculated broth, a filtered inoculated broth, a concentrated inoculated broth or a centrifuged inoculated broth. 34. The agent of claim 8, wherein said extract is selected from the group consisting of heat killed barley inoculum, a chloroform extract of said Phoma isolate, a methanol extract of said Phoma isolate, and an ethyl-acetate extract of said Phoma isolate. 35. The agent of claim 8, wherein said inoculated broth is selected from the group consisting of a crude inoculated broth, a filtered inoculated broth, a concentrated inoculated broth or a centrifuged inoculated broth. 36. The method of claim 23, wherein said extract is selected from the group consisting of heat killed barley inoculum, a chloroform extract of said Phoma isolate, a methanol extract of said Phoma isolate, and a ethyl-acetate extract of said Phoma isolate. 37. The method of claim 22, wherein said inoculated broth is selected from the group consisting of a crude inoculated broth, a filtered inoculated broth, a concentrated inoculated broth or a centrifuged inoculated broth. 38. A coated crop seed, comprising one or more Phoma isolates and a binder. 39. A coated crop seed, comprising an inoculated broth, or an extract obtained from one or more Phoma, isolates and a binder. 40. The coated crop seed of claim 38, wherein one or more than one of said one or more Phoma isolates is a Phoma cf macrostoma isolate. 41. The coated crop seed of claim 40, wherein said Phoma cf macrostoma isolate is selected from the group consisting of: a) 85-24B (IDAC 230201-1, deposited Feb. 23, 2001), b) 89-25A (IDAC 110401-1, deposited Apr. 11, 2001), c) 94-26 (IDAC 230201-2, deposited Feb. 23, 2001), d) 94-44B (IDAC 230201-3, deposited Feb. 23, 2001) e) 94-134 (IDAC 230201-4, deposited Feb. 23, 2001), f) 94-359A (IDAC 110401-2, deposited Apr. 11, 2001), g) 95-54A1 (IDAC 230201-5, deposited Feb. 23, 2001), h) 95-268B (IDAC 110401-3, deposited Apr. 11, 2001), i) 97-12B (IDAC 230201-6, deposited Feb. 23, 2001), and j) 97-15B2 (IDAC 110401-4, deposited Apr. 11, 2001). 42. The coated crop seed of claim 39, wherein one or more than one of said one or more Phoma isolates is a Phoma cf macrostoma isolate. 43. The coated crop seed of claim 42, wherein said Phoma cf macrostoma isolate is selected from the group consisting of: a) 85-24B (IDAC 230201-1, deposited Feb. 23, 2001), b) 89-25A (IDAC 110401-1, deposited Apr. 11, 2001), c) 94-26 (IDAC 230201-2, deposited Feb. 23, 2001), d) 94-44B (IDAC 230201-3, deposited Feb. 23, 2001) e) 94-134 (IDAC 230201-4, deposited Feb. 23, 2001), f) 94-359A (IDAC 110401-2, deposited Apr. 11, 2001), g) 95-54A1 (IDAC 230201-5, deposited Feb. 23, 2001), h) 95-268B (IDAC 110401-3, deposited Apr. 11, 2001), i) 97-12B (IDAC 230201-6, deposited Feb. 23, 2001), and j) 97-15B2 (IDAC 110401-4, deposited Apr. 11, 2001). 44. The method of claim 14, wherein said crop is a perennial crop. 45. The method of claim 44, wherein said perennial crop is selected from the group consisting of a turf, a perennial grass, and a winter cereal. 46. The method of claim 15, wherein said crop is a perennial crop. 47. The method of claim 46, wherein said perennial crop is selected from the group consisting of a turf, a perennial grass, and a winter cereal. 48. The method of claim 16, wherein said crop is a perennial crop. 49. The method of claim 48, wherein said perennial crop is selected from the group consisting of a turf, a perennial grass, and a winter cereal. 50. The method of claim 17, wherein said crop is a perennial crop. 51. The method of claim 50, wherein said perennial crop is selected from the group consisting of a turf, a perennial grass, and a winter cereal. 52. The method of claim 18, wherein said crop is a perennial crop. 53. The method of claim 52, wherein said perennial crop is selected from the group consisting of a turf, a perennial grass, and a winter cereal. 54. The method of claim 19, wherein said crop is a perennial crop. 55. The method of claim 54, wherein said perennial crop is selected from the group consisting of a turf, a perennial grass, and a winter cereal. 56. The method of claim 20, wherein said crop is a perennial crop. 57. The method of claim 56, wherein said perennial crop is selected from the group consisting of a turf, a perennial grass, and a winter cereal. 58. The method of claim 21, wherein said crop is a perennial crop. 59. The method of claim 58, wherein said perennial crop is selected from the group consisting of a turf, a perennial grass, and a winter cereal. 60. A method of controlling weed development during growth of a lawn of grass, the method comprising: a) adding an effective amount of said biocontrol agent of claim 5 to produce a treated soil, b) planting grass seed in said treated soil, c) growing said lawn from said grass seed. 61. A method of controlling weed development during growth of a lawn of grass, the method comprising: a) planting grass seed, b) adding an effective amount of said biocontrol agent of claim 5 to soil where said grass seed is planted, c) growing said lawn from said grass seed. 62. A method of controlling weed development during growth of a lawn of grass comprising: a) adding an effective amount of a biocontrol agent of claim 5 to grass seed to produce treated grass seed; b) planting said treated grass seed; and c) growing said lawn from said treated grass seed. 63. A method of controlling weed development during growth of an established lawn of grass, the method comprising: a) adding an effective amount of the biocontrol agent of claim 5 to the established lawn; and b) growing the lawn. 64. A method of controlling weed development during growth of a lawn of grass, the method comprising: a) adding an effective amount of said biocontrol composition of claim 9 to produce a treated soil, b) planting grass seed in said treated soil, c) growing said lawn from said grass seed. 65. A method of controlling weed development during growth of a lawn of grass, the method comprising: a) planting grass seed, b) adding an effective amount of said biocontrol composition of claim 9 to soil where said grass seed is planted, c) growing said lawn from said grass seed. 66. A method of controlling weed development during crop growth comprising: a) adding an effective amount of a biocontrol composition of claim 9 to grass seed to produce treated grass seed; b) planting said treated grass seed; and c) growing a lawn from said treated grass seed. 67. A method of controlling weed development during growth of an established lawn, the method comprising: a) adding an effective amount of the biocontrol composition of claim 9 to the established lawn; and b) growing the lawn. 68. The method of claim 23, wherein said biocontrol agent is applied to the soil before emergence of the weed. 69. The method of claim 23, wherein said biocontrol agent is applied to the soil after emergence of the weed. 70. The method of claim 25, wherein said biocontrol composition is applied to the soil before emergence of the weed. 71. The method of claim 25, wherein said biocontrol composition is applied to the soil after emergence of the weed. 72. A method of enhancing the growth of a crop, the method comprising: a) adding an effective amount of the biocontrol agent of claim 5 to soil to produce a treated soil; b) planting crop seed in said treated soil, and c) growing said crop from the crop seed. 73. A method of enhancing the growth of a crop, the method comprising: a) planting crop seed in soil; b) adding an effective amount of the biocontrol agent of claim 5 to the soil where said crop seed is planted, and c) growing said crop from the crop seed. 74. A method of enhancing the growth of an established crop, the method comprising: a) adding an effective amount of the biocontrol agent of claim 5 to the established crop; and b) growing the crop. 75. A method of enhancing the growth of a crop, the method comprising: a) adding an effective amount of said biocontrol agent of claim 5 to a crop seed to produce a treated crop seed; b) planting said treated crop seed; and c) growing said crop. 76. The method of claim 72, wherein said crop is a grass. 77. The method of claim 73, wherein said crop is a grass. 78. The method of claim 74, wherein said crop is a grass. 79. The method of claim 75, wherein said crop is a grass, and said crop seed is grass seed. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The use of pesticides to kill insects, weeds and other disease pests is common in agriculture. It has been estimated that Canadian farmers spend more than $750 million on pesticides, and U.S. and European estimates are likely to be several fold higher. On the Canadian prairies, 95% of the land seeded to wheat, barley, canola and flax is treated with one or more pesticides. However, despite extensive pesticide use, weeds continue to cause an estimated one billion dollars in crop losses in Canada alone every year. Weeds are detrimental to agricultural crops because they are capable of outcompeting crop plants for space, sun and nutrients. Particularly troublesome weeds include Canada thistle ( Cirsium arvense ) and other members of the Aster family such as perennial sowthistle ( Sonchus arvense ), and dandelion ( Taraxacum officinale ). Canada thistle ( Cirsium arvense [L.] Scop.) is an aggressive perennial weed in field crops, pastures and roadsides, and is particularly prevalent in Western Canada where it occurs in about 50% of all fields. Canada thistle causes crop yield losses of about 15 to 60% in cereal, oilseed and pulse crops, depending on weed density. In cereal crops, densities of 6 to 20 Canada thistle plants per square metre result in an 18 to 30% loss in grain yield. In 1937, Canada thistle was designated. as a noxious weed by the Canadian Federal Seeds Act. Although weeds of the Aster family, for example Canada thistle and dandelion, can reproduce by flowering, they are difficult to eradicate because their extensive root system. The roots are quite brittle and fragment easily during tillage. This results in greater shoot emergence from stimulated buds. Further adding to the difficulties of control, the root fragments carry sufficient food reserves to survive long periods under adverse conditions. Control of Canada thistle in field crops is currently achieved by pre-seeding, in-crop, and post-harvest chemical control with herbicides, applied at sufficient rates to suppress top growth, or kill the roots. For example, Glyphosate is used as a pre-seeding treatment to kill Canada thistle, or used in-crop on glyphosate tolerant crops. Clopyralid is used for in-crop control to achieve the same effect but has problems with residual activity for some crops in the following year. Other product combinations only provide top growth suppression such as thifensulfuron and tribenuron-methyl or fenoxy-prop and MCPA. Other control options include growing competitive crops and seeding early to get vigorous crop growth before Canada thistle emergence and shallow tilling of soil to reduce root fragmentation and new shoot growth. Also, mowing may be used to control weeds on roadsides, ditches, headlands and fence lines. Controlling patches instead of entire fields is often recommended to reduce costs. There are a number of drawbacks associated with non-chemical control of Canada thistle in addition to those discussed above. First, there are very few crops which are able to outcompete weeds such as Canada thistle and many crops cannot be seeded early enough to provide the crop with a competitive advantage to Canada thistle. Further, seeding crops earlier than usual may be an inconvenience to farmers. Also, shallow tillage of soil and mowing weeds to kill weeds or prevent weed flowering are only temporary solutions and are at best marginally effective in controlling weeds such as Canada thistle. There are also several drawbacks associated with the use of chemical herbicides to control weeds such as Canada thistle. Herbicides are expensive and may be too expensive to be used by some farmers. Further, if a farmer uses less than the required dosage of herbicide to kill the weeds, there is an increased risk that some weeds may develop herbicide resistance. There is also an increased risk of herbicide resistance due to overuse of a herbicide. In addition, herbicides are not available for all crops and all situations. For example, there are no effective herbicides available for crops such as peas and lentil whereas some in-crop chemical herbicides only suppress top growth of weeds without controlling root growth, which is a short-term strategy often used for crops such as wheat, barley and canola. Residual herbicidal activity may also limit crop rotation for some crops and some agronomic herbicide practices may increase weed densities. There are also concerns about the short and long term safety of herbicides, both to consumers and the environment. Environmental issues in the agri-food industry have become a priority with federal and provincial governments, including the development of alternatives for chemical pest control products, with the ultimate goal of reducing chemical pesticide use. Rising economic, environmental and social costs associated with agricultural inputs, spray drift, pesticide residues, government legislation for reduced pesticide use, along with the development of herbicide resistance in weeds make biological control agents attractive strategies for weed control for both agricultural and domestic use. Broad-leaved weeds in turf situations, such as lawns, parks, and golf courses, disrupt the desired visual uniformity (i.e. are unsightly), create problems in the maintenance of the turf due to clumping and growth habits of the weeds, compete with the turf for light, nutrients, and water. Weeds are also are irritants to humans when allergic reactions to their pollen or the chemicals applied for weed control occur. Important weeds in turfgrass belong to the Compositae (such as dandelion, sowthistle), Caryophyllaceae (such as chickweed), and Rubiaceae, and Convolvulaceae. Typically, control of weeds in turf has been with selective, nonselective, systemic, and contact herbicides applied at various times (pre-plant, pre-emergence, and post-emergence). Public pressure is mounting to prevent the use of chemical herbicides in public places such as parks and homeowners lawns, for example, By-laws have recently passed in Calgary, Alberta, and Halifax, Nova Scotia, both in Canada, against their use. Chemical herbicides used in these areas leads to increased chemical exposure to susceptible groups in the population like children, pets, and the elderly. A number of bacteria and fungi are natural pathogens of weeds and it has been suggested that bioherbicides, or weed killers made from biological agents rather than chemical agents, may provide an alternative to chemical pesticides. For example, U.S. Pat. No. 6,008,159 discloses controlling annual weeds using the fungus Pyrenophora . U.S. Pat. Nos. 5,993,802 and 5,472,690 teach suppressing the growth of Calmagrostis canadensis using an isolate of a low temperature basidiomycete fungus, or a mycoherbicide (including at least one or both of Fusarium nivalis and Colletotrichum calamagrostidis ), respectively. U.S. Pat. Nos. 5,952,264 and 5,635,444 teach controlling crabgrass using the fungus Cochliobolus intermedius, or a fungus selected from the genus Culvularia, respectively. U.S. Pat. No. 5,747,029 teaches controlling sicklepod weeds with the fungus Myrothecium verrucaria. U.S. Pat. No. 5,698,491 and WO 98/08389 discloses controlling nutsedge weeds with the fungus Dactylaria higginsii (WO 98/08389 and U.S. Pat. No. 5,698,491). U.S. Pat. No. 4,606,751 teaches controlling Johnson grass and similar weeds with Bipolaris sorghicola spores. The spores are suspended in a solution of water and surfactant and sprayed onto a field onto which the weed is growing. U.S. Pat. No. 5,795,845 discloses a bioherbicidal composition comprising an invert emulsion carrier and a microorganism which is a weakly or non-pathogenic bacterium or fungus. The composition may be used to control pigweed, plumeless thistle, velvet leaf and ground cherry. U.S. Pat. No. 4,636,386 discloses an isolate of Alternaria for the control of Italian thistle. U.S. Pat. No. 5,994,27 discloses a composition comprising a bioherbicide which is an isolate of Sclerotinia minor which produces foliar wilt and rot in broadleaf weed species so as to inhibit their growth. The bioherbicide may be used to control the growth of broadleaf weeds such as dandelion, broadleaf plantain, ragweed, ivy, knotweed sow thistle and white clover. Brebaum and Boland (1999, Plant Disease 83:2000) disclose Phoma exigua and Phoma herbarum as pathogens of dandelion ( Taraxacum officinale ), however, no weed controlling activity was reported using these species. None of the identified references disclose fungal isolates derived from Phoma macrostoma as biocontrol compositions suitable for use to control Canada thistle, dandelion, or other weed species. There is a need in the art for novel bioherbicides and biocontrol compositions for controlling weeds. Further there is a need in the art for novel bioherbicides and biocontrol compositions for controlling weed plants for example Canada thistle, perennial sowthistle, dandelion, prairie sunflower, field bindweed, wild buckwheat, and scentless chamomile, cleavers, and chickweed. Further, there is a need in the art for biocontrol compositions comprising a biological control agent and a growth medium for supporting the viability of the biological control agent when the biocontrol composition is employed to control weeds. It is an object of the present invention to overcome disadvantages of the prior art. The above object is met by a combination of the features of the main claims. The sub claims disclose further advantageous embodiments of the invention. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention relates to bioherbicides. More specifically, the present invention relates to fungal bioherbicides and compositions comprising fungal bioherbicides. According to the present invention there is provided a method of controlling one or more broad leaf weeds comprising administering an isolate of Phoma cf. macrostoma , an extract therefrom, an inoculated broth therefrom, or a combination thereof, to the one or more broad leaf weeds, or to soil where said weeds grow. The present invention is also directed to the method defined above wherein the one or more broad leaf weeds is a species of a family selected from the group consisting of Compositae, Caryophyllaceae, Convolvulaceae, Plantaginaceae and Rubiaceae. Preferably, the one or more broad leaf weeds is selected from the group consisting of Canada thistle, perennial sowthistle, dandelion, scentless chamomile, false cleavers, chickweed, wild buckwheat, plantain, prairie sunflower and field bindweed. The present invention also provides a biocontrol agent comprising one or more than one Phoma isolate, an extract therefrom, an inoculated broth therefrom, or a combination thereof, wherein the one or more than one Phoma isolate, or an extract therefrom, or an inoculated broth therefrom, exhibit weed control activity, growth enhancement activity, or both. The present invention also embraces a biocontrol composition, comprising the biocontrol agent just defined, and a medium for supporting viability of said one or more than one Phoma isolate. Preferably, the one or more than one Phoma isolate, is an isolate of Phoma cf. macrostoma. More preferably, the one or more than one Phoma isolate is selected from the group consisting of a) 85-24B (IDAC 230201-1, deposited Feb. 23, 2001), b) 89-25A (IDAC 110401-1, deposited Apr. 11, 2001), c) 94-26 (IDAC 230201-2, deposited Feb. 23, 2001), d) 94-44B (IDAC 230201-3, deposited Feb. 23, 2001) e) 94-134 (IDAC 230201-4, deposited Feb. 23, 2001), f) 94-359A (IDAC 110401-2, deposited Apr. 11, 2001), g) 95-54A1 (IDAC 230201-5, deposited Feb. 23, 2001), h) 95-268B (IDAC 110401-3, deposited Apr. 11, 2001), i) 97-12B (IDAC 230201-6, deposited Feb. 23, 2001), j) 97-15B2 (IDAC 110401-4, deposited Apr. 11, 2001), and a combination thereof. The present invention also provides a biocontrol agent comprising an extract, or an inoculated broth, from one or more than one Phoma cf. macrostoma isolate. Preferably, the extract or inoculated broth is obtained from the Phoma isolate selected from the group consisting of a) 85-24B (IDAC 230201-1, deposited Feb. 23, 2001), b) 89-25A (IDAC 110401-1, deposited Apr. 11, 2001), c) 94-26 (IDAC 230201-2, deposited Feb. 23, 2001), d) 94-44B (IDAC 230201-3, deposited Feb. 23, 2001) e) 94-134 (IDAC 230201-4, deposited Feb. 23, 2001), f) 94-359A (IDAC 110401-2, deposited Apr. 11, 2001), g) 95-54A1 (IDAC 230201-5, deposited Feb. 23, 2001), h) 95-268B (IDAC 110401-3, deposited Apr. 11, 2001), i) 97-12B (IDAC 230201-6, deposited Feb. 23, 2001), j) 97-15B2 (IDAC 110401-4, deposited Apr. 11, 2001), and a combination thereof. This invention pertains to the above method wherein the extract is selected from the group consisting of heat killed barley inoculum, a chloroform extract of the Phoma isolate, a methanol extract of the Phoma isolate, and a ethyl-acetate extract of the Phoma isolate, and the inoculated broth is selected from the group consisting of a crude inoculated broth, a filtered inoculated broth, or a centrifuged inoculated broth. The present invention also pertains to a method of controlling weed development during crop growth comprising: a) adding an effective amount of a biocontrol agent comprising one or more than one Phoma cf macrostoma isolate, an extract therefrom, an inoculated broth therefrom, or combination thereof to soil to produce treated soil, the one or more than one Phoma cf macrostoma isolate, an extract therefrom, or an inoculated broth therefrom, exhibiting weed control activity; b) planting the crops in the treated soil; and c) growing the crop. According to the present invention there is also provided a method of controlling weed development during crop growth comprising: a) planting the crop, b) adding an effective amount of said biocontrol agent comprising one or more than one Phoma cf macrostoma isolate, an extract therefrom, an inoculated broth therefrom, or a combination therefrom, to soil where the crop is planted, the one or more than one Phoma cf macrostoma isolate, extract therefrom, or inoculated broth therefrom, exhibiting weed control activity; c) growing the crop. The present invention is also directed to a method of controlling weed development during crop growth comprising: a) adding an effective amount of a biocontrol agent comprising one or more than one Phoma cf macrostoma isolate, an extract therefrom, an inoculated broth therefrom, or a combination thereof, to a crop seed to produce treated crop seed, the one or more than one Phoma cf macrostoma isolate, extract therefrom, or inoculated broth therefrom, exhibiting weed control activity; b) planting the treated crop seed; and c) growing the crop. Also included in this invention is the method as just defined wherein the treated crop seed is grass seed, including domestic and specialty turf grass seed, animal pasture or hay seed mixes comprising one or more of timothy, fescue, blue grass, perennial rye grass, bromegrass, canary grass, red top and orchard grass seed. The methods of the present invention are preferably used to control weed development during growth of a perennial crop. Preferably, the perennial crop is selected from the group consisting of a turf, a perennial grass, and a winter cereal. The present invention also provides a method of controlling weed development during established crop growth comprising: a) adding an effective amount of a biocontrol agent comprising one or more than one Phoma cf macrostma isolate, an extract therefrom, an inoculated broth therefrom, or a combination thereof, to the established crop, the one or more than one Phoma cf macrostoma isolate, an extract therefrom, or an inoculated broth therefrom, exhibiting weed control activity, and b) growing the crop. Also included in this invention is the method as just defined wherein the established crop is grass, including domestic and specialty turf grasses, animal pasture or hay mixes comprising one or more of timothy, fescue, blue grass, perennial rye grass, bromegrass, canarygrass, red top and orchard grass. The present invention also provides a method of enhancing the growth of a crop, the method comprising: a) adding an effective amount of a biocontrol agent comprising one or more than one Phoma cf macrostma isolate, an extract therefrom, an inoculated broth therefrom, or a combination thereof to soil to produce a treated soil; b) planting the crop in said treated soil, and c) growing said crop. The present invention also provides a method of enhancing the growth of a crop, the method comprising: a) planting said crop in soil; b) adding an effective amount of a biocontrol agent comprising one or more than one Phoma cf macrostma isolate, an extract therefrom, an inoculated broth therefrom, or a combination thereof to the soil where said crop is planted; and c) growing said crop. The present invention further provides a method of enhancing the growth of an established crop, the method comprising: a) adding an effective amount of a biocontrol agent comprising one or more than one Phoma cf macrostma isolate, an extract therefrom, an inoculated broth therefrom, or a combination thereof to the established crop; and b) growing the crop. Also included in this invention is the methods as just defined wherein the crop is grass, including domestic and specialty turf grasses, animal pasture or hay mixes comprising one or more of timothy, fescue, blue grass, perennial rye grass, bromegrass, canarygrass, red top and orchard grass. The present invention is also directed to a method of enhancing the growth of a crop, the method comprising: a) adding an effective amount of a biocontrol agent comprising one or more than one Phoma cf macrostoma isolate, an extract therefrom, an inoculated broth therefrom, or a combination thereof, to a crop seed to produce treated crop seed; b) planting the treated crop seed; and c) growing the crop. Also included in this invention is the method as just defined wherein the treated crop seed is grass seed, including domestic and specialty turf grass seed, animal pasture or hay seed mixes comprising one or more of timothy, fescue, blue grass, perennial rye grass, bromegrass, canary grass, red top and orchard grass seed. The methods of the present invention also relate to the use of a biocontrol composition comprising one or more than one Phoma cf macrostma isolate, an extract therefrom, an inoculated broth therefrom, or a combination thereof; and a medium for supporting viability of the one or more than one Phoma cf macrostma isolate. The present invention also provides for any of the above methods wherein the biocontrol agent or composition is applied to the soil before or after emergence of the weed, preferably before emergence. The present invention also provides for any of the above methods wherein the biocontrol agent or composition is applied by dusting, rubbing, spreading, drilling, banding, broadcasting, spraying, liquid injection, pouring or soil drenching. The present invention embraces a coated crop seed comprising one or more Phoma isolates and a binder. The invention also includes a coated crop seed comprising an extract obtained from one or more Phoma isolates and a binder. The crop seed that is coated is preferably grass seed, including domestic and specialty turf grass seed, animal pasture or hay seed mixes comprising one or more of timothy, fescue, blue grass, perennial rye grass, bromegrass, canary grass, red top and orchard grass seed. This summary does not necessarily describe all necessary features of the invention but that the invention may also reside in a sub-combination of the described features. |
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