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1. A nucleic acid vector for concurrently imparting herbicide resistance to a plant and cross protecting the plant, the vector comprising; (a) sufficient potyvirus nucleic acid sequence to permit a potyvirus to replicate and spread within the plant infected by the vector; (b) a first mutation in said potyvirus nucleic acid sequence which attenuates symptoms of said potyvirus in the plant infected by the vector; (c) a second mutation in said potyvirus nucleic acid sequence which abolishes transmission of said potyvirus by an insect vector; and (d) an additional nucleic acid sequence encoding a protein which imparts resistance to an herbicide when expressed in the plant infected by the vector. 2. The nucleic acid vector of claim 1, wherein said first mutation includes an amino acid substitution of the conserved FRNK box in the HC-Pro gene (SEQ ID NO.: 4) of said potyvirus nucleic acid sequence. 3. The nucleic acid vector of claim 2, wherein said amino acid substitution in the conserved FRNK box of the potyvirus nucleic acid sequence includes a substitution of a Arg to Ile at position 180 within the HC-pro gene product (SEQ ID NO.: 5) of said potyvirus. 4. The nucleic acid vector of claim 1, wherein said second mutation includes an alteration of a conserved DAG triplet at position 8 in an N terminal region of the coat protein (SEQ ID NO.: 3) of said potyvirus. 5. The nucleic acid vector of claim 4, wherein said alteration of said DAG triplet includes a substitution for an alanine residue in said DAG triplet. 6. The nucleic acid vector of claim 1, wherein said insect vector is an aphid. 7. The nucleic acid vector of claim 1, wherein said sufficient potyvirus nucleic acid sequence is derived from zucchini yellow mosaic virus (ZYMV). 8. The nucleic acid vector of claim 1, wherein said additional nucleic acid sequence is at least a portion of a phosphinothricin acetyltransferase coding sequence. 9. The nucleic acid vector of claim 1, wherein said protein is at least a functional portion of a phosphinothricin acetyltransferase. 10. A method of concurrently imparting herbicide resistance to a plant and cross protecting the plant against at least one potyvirus, the method comprising inoculating at least a portion of the plant with a vector comprising; (a) sufficient potyvirus nucleic acid sequence to permit a potyvirus to replicate and spread within the plant infected by the vector; (b) a first mutation in said potyvirus nucleic acid sequence which attenuates symptoms of said potyvirus in the plant infected by the vector; (c) a second mutation in said potyvirus nucleic acid sequence which eliminates transmission of said potyvirus by an insect vector; and (d) an additional nucleic acid sequence encoding a protein which imparts resistance to an herbicide when expressed in the plant infected by the vector. 11. The method of claim 10, wherein said first mutation includes an amino acid substitution of the conserved FRNK box in the HC-Pro gene (SEQ ID NO.: 4) of the potyvirus nucleic acid sequence. 12. The method of claim 10, wherein said amino acid substitution of the conserved FRNK box in the HC-Pro gene of the potyvirus nucleic acid sequence includes a substitution of a Arg to Ile at position 180 within the HC-pro gene product (SEQ ID NO.: 5) of said potyvirus. 13. The method of claim 10, wherein said second mutation includes an alteration of a conserved DAG triplet at position 8 in an N terminal region of the coat protein (SEQ ID NO.: 3) of said potyvirus. 14. The method of claim 13, wherein said alteration of said DAG triplet includes a substitution for an alanine residue in said DAG triplet. 15. The method of claim 10, wherein said insect vector is an aphid. 16. The method of claim 10, wherein said sufficient potyvirus nucleic acid sequence is derived from zucchini yellow mosaic virus (zymv). 17. The method of claim 10, wherein said additional nucleic acid sequence is at least a portion of a phosphinothricin acetyltransferase coding sequence. 18. The method of claim 10, wherein said protein is at least a functional portion of a phosphinothricin acetyltransferase. 19. The method of claim 10, wherein said at least a portion of a plant includes an item selected from the group consisting of at least one cell in the plant and at least one plant cell in tissue culture. 20. At least a portion of a plant treated according to the method of claim 10. 21. An infectious virion harvested from a plant treated according to the method of claim 10.

<SOH> FIELD AND BACKGROUND OF THE INVENTION <EOH>The present invention relates to nucleic acid vectors capable of imparting herbicide resistance and viral cross protection, methods of use thereof and plants expressing same and, more particularly, to vectors based on sequences derived from attenuated potyvirus sequences and further including sequences which impart resistance to a chosen herbicide. Zucchini yellow mosaic virus is a member of the potyviridae family (Shukla et al. (1989) Adv. Virus Res. 36:273-314). Potyviridae is the largest group of plant viruses and its members infect most commercial or cultivated crops. Worldwide, ZYMV is one of the most devastating diseases of cucurbit species (e.g., squash, melon, watermelon, cucumber etc.; Desbiez and Lecoq, (1997) Plant Pathol. 46:809-829). As in all potyviruses, the ZYMV genome consists of a single messenger-polarity RNA molecule of about 9.6 kb, encapsidated by ˜2000 units of coat protein (CP), forming a helical, flexuous, filamentous particle of about 750 nm long and 11 nm wide (Desbiez, and Lecoq, (1997) Plant Pathol. 46:809-829 and Lisa et al. (1981) Phytopathology 71:667-672). Means for attenuating potyviruses in general, and ZYMV in particular, have been described in WO 99/51749 and in Gal-On (2000, Phytopathology 90:467-473). However, these earlier teachings contain neither a hint nor a suggestion that a single vector might be employed to concurrently cross protect a plant and render the plant resistant to an herbicide. U.S. Pat. No. 5,958,422 as well as WO9602649 and WO9218618 teach modified plant viruses as vectors for heterologous peptides, including peptides useful for vaccination. However, these patents relate to use of plants as bioreactors for vaccine production and do not teach cross protection of the plants themselves. Further, these patents do not teach introduction of herbicide resistance genes into the plants. While use of Glufosinate resistance genes in commercial agriculture is well known (e.g. AgrEvo's Liberty-Link-Oilseed Rape, -Canola, -Maize etc.), this resistance has typically been accomplished by germ line transformation of plants. Such germ line transformation raises concerns about unwanted spread of herbicide resistant plants and or transfer of the herbicide resistance gene to wild plant relatives via pollination (Quist D. and Chapela I. H. (2001) Nature 414(6863): 541-3.) Whitham et al. (Proc. Natl. Acad. Sci. USA (1999) 96: 772-777) teaches use of a potyvirus expressing an herbicide resistance gene in plants in order to identify plant mutants that affect viral replication, cell to cell and long distance movement within a plant. The teachings of Whitham preclude use of attenuated strains of virus to impart cross protection against subsequent wild type viral infection. In addition, use of the teachings of Whitham in commercial agriculture is infeasible because of the pathogenic outcome of viral infection on agriculturally important plants and the high probability of transmission by insect vectors in the field. U.S. Pat. No. 6,303,848 to Kumagai et al. teaches use of nucleic acid vectors to impart herbicide resistance to crops using a tobamovirus vector. The teachings of Kumagai require use of a subgenomic plant viral promoter which precludes application of his teachings to potyvirus. Further, the teachings of Kumagai do not include expression of a phosphinothricin acetyltransferase gene which confers resistance to glufosinate ammonium based herbicides. Further, Kumagai teaches use of a tobamavirus which is devastating to plants so that its use in commercial agriculture is infeasible. Further, Kumagai does not teach mutants which are impaired in their ability to be transmitted from plant to plant by their normal mode of transmission i.e. mechanically via infected tissue and contaminated soil. Thus spread of viral vectors according to the teachings of Kumagai cannot be controlled increasing the likelihood of uncontrolled infection in untreated plants. U.S. Pat. No. 5,766,885 to Carrington et al. teaches expression of foreign genes in a potyvirus vector. However, Carrington fails to teach use of attenuated strains of potyvirus in order to minimize viral symptoms. Further, Carrington fails to teach use of potyvirus vectors to impart herbicide resistance. The scope of the teachings of Carrington is limited to use of plants as bioreactors and added value agricultural traits are not found in his teachings. There is thus a widely recognized need for, and it would be highly advantageous to have, non-insect transmissible nucleic acid vectors capable of concurrently imparting herbicide resistance and cross protection and, methods of use thereof and plants expressing same devoid of the above limitations.

<SOH> SUMMARY OF THE INVENTION <EOH>According to one aspect of the present invention there is provided a nucleic acid vector for concurrently imparting herbicide resistance to a plant and cross protecting the plant. The vector includes: (a) sufficient potyvirus nucleic acid sequence to permit a potyvirus to replicate and spread within the plant infected by the vector; (b) a first mutation in the potyvirus nucleic acid sequence which attenuates symptoms of the potyvirus in the plant infected by the vector; (c) a second mutation in the potyvirus nucleic acid sequence which abolishes transmission of the potyvirus by an insect vector; and (d) an additional nucleic acid sequence encoding a protein which imparts resistance to an herbicide when expressed in the plant infected by the vector. According to another aspect of the present invention there is provided a method of concurrently imparting herbicide resistance to a plant and cross protecting the plant against at least one potyvirus and. The method includes inoculating at least a portion of the plant with a vector including; (a) sufficient potyvirus nucleic acid sequence to permit a potyvirus to replicate and spread within the plant infected by the vector; (b) a first mutation in the potyvirus nucleic acid sequence which attenuates symptoms of the potyvirus in the plant infected by the vector; (c) a second mutation in the potyvirus nucleic acid sequence which eliminates transmission of the potyvirus by an insect vector; and (d) an additional nucleic acid sequence encoding a protein which imparts resistance to an herbicide when expressed in the plant infected by the vector. According to further features in preferred embodiments of the invention described below, the first mutation includes an amino acid substitution in the HC-Pro gene (SEQ ID NO.: 4) of the conserved FRNK box of the potyvirus nucleic acid sequence. According to still further features in the described preferred embodiments the amino acid substitution in the HC-Pro gene of the conserved FRNK box of the potyvirus nucleic acid sequence includes a substitution of an Arg to Ile at position 180 within the HC-pro gene product (SEQ ID NO.: 5) of the potyvirus. According to still further features in the described preferred embodiments the insect vector is an aphid. According to still further features in the described preferred embodiments the sufficient potyvirus nucleic acid sequence is derived from zucchini yellow mosaic virus (ZYMV). According to still further features in the described preferred embodiments the additional nucleic acid sequence is at least a portion of a phosphinothricin acetyltransferase coding sequence. According to still further features in the described preferred embodiments the protein is at least a functional portion of a phosphinothricin acetyltransferase. According to still further features in the described preferred embodiments at least a portion of a plant treated according to the claimed method is an integral part of the invention, as are progeny of at least a portion of a plant treated according to the method. According to still further features in the described preferred embodiments an infectious virion harvested from a plant treated according to the claimed method is an integral part of the invention. The present invention successfully addresses the shortcomings of the presently known configurations by providing a vector for concurrently imparting herbicide resistance and cross protection against wild type virus. Concurrent receipt of these two effects from a single treatment contributes to increased crop yield and significantly reduces production costs. Reduction in production costs stein from both elimination of crop damage and from ease of introducing these traits into a wide variety of commercial plant strains.

Virus causing respiratory tract illness in susceptible mammals

The invention relates to the field of virology. The invention provides an isolated essentially mammalian negative-sense single stranded RNA virus (MPV) within the sub-family Pneumovirinae of the family Paramyxoviridae and identifiable as phylogenetically corresponding to the genus Metapneumovirus and components thereof.

1. An isolated essentially mammalian negative-sense single stranded RNA virus (MPV) belonging to the sub-family Pneumovirinae of the family Paramyxouiridae and identifiable as phylogenetically corresponding to the genus Metapneumovirus 2. An isolated negative-sense single stranded RNA virus (MPV) belonging to the sub-family Pneumovirinae of the family Paramyxovirdae and identifiable as phylogenetically corresponding to the genus Metapneumovirus by determining a nucleic acid sequence of said virus and testing it in phylogenetic tree analyses wherein maximum likelihood trees are generated using 100 bootstraps and 3 jumbles and finding it to be more closely phylogenetically corresponding to a virus isolate deposited as I-2614 with CNCM, Paris than it is corresponding to a virus isolate of avian pneumovirus (APV) also known as turkey rhinotracheitis virus (TRTV), the aetiological agent of avian rhinotracheitis. 3. A virus according to claim 2 wherein said avian pneumovirus comprises APV type C (APV-C). 4. A virus according to claim 1 to 3 wherein said nucleic acid sequence comprises an open reading frame (ORF) encoding a viral protein of said virus. 5. A virus according to claim 4 wherein said open reading frame is selected from the group of ORFs encoding the N, P, M, and F proteins. 6. A virus according to claim 5 wherein said open reading frame is selected from the group of ORFs encoding the SH or G proteins. 7. A virus according to anyone of claims 1 to 6 comprising a nucleic acid or functional fragment phylogenetically corresponding to a sequence shown in FIG. 6. 8. A virus according to anyone of claims 1 to 7 comprising an MPV isolate deposited as I-2614 with CNCM, Institute Pasteur, Paris or a virus isolate phylogenetically corresponding therewith. 9. A virus according to claim 8 isolatable from a human with respiratory tract illness. 10. An isolated or recombinant nucleic acid or MPV-specific functional fragment thereof obtainable from a virus according to anyone of claims 1 to 9. 11. A vector comprising a nucleic acid according to claim 10. 12. A host cell comprising a nucleic acid according to claim 10 or a vector according to claim 11. 13. An isolated or recombinant proteinaceous molecule or MPV-specific functional fragment thereof encoded by a nucleic acid according to claim 10. 14. An antigen comprising a proteinaceous molecule or MPV-specific functional fragment thereof according to claim 13. 15. An antibody specifically directed against an antigen according to claim 14. 16. A method for identifying a viral isolate as an MPV comprising reacting said viral isolate or a component thereof with an antibody according to claim 15. 17. A method for identifying a viral isolate as an MPV comprising reacting said viral isolate or a component thereof with a nucleic acid according to claim 10. 18. A method according to claim 16 or 17 wherein said MPV comprises a human MPV. 19. A viral isolate identifiable with a method according to anyone of claims 16 to 18 as a mammalian negative-sense single stranded RNA virus within the sub-family Pneumovirinae of the family Paramyxoviridae and identifiable as phylogenetically corresponding to the genus Metapneumovirus. 20. A method for virologically diagnosing an MPV infection of a mammal comprising determining in a sample of said mammal the presence of a viral isolate or component thereof by reacting said sample with a nucleic acid according to claim 10 or an antibody according to claim 15. 21. A method for serologically diagnosing an MPV infection of a mammal comprising determining in a sample of said mammal the presence of an antibody specifically directed against an MPV or component thereof by reacting said sample with a proteinaceous molecule or fragment thereof according to claim 13 or an antigen according to claim 14. 22. A diagnostic kit for diagnosing an MPV infection comprising a virus according to anyone of claims 1 to 9, a nucleic acid according to claim 10, a proteinaceous molecule or fragment thereof according to claim 13, an antigen according to claim 14 and/or an antibody according to claim 15. 23. Use of a virus according to any one claims 1 to 9, a nucleic acid according to claim 10, a vector according to claim 11, a host cell according to claim 12, a proteinaceous molecule or fragment thereof according to claim 13, an antigen according to claim 14, or an antibody according to claim 15 for the production of a pharmaceutical composition. 24. Use according to claim 23 for the production of a pharmaceutical composition for the treatment or prevention of an MPV infection. 25. Use according to claim 23 or 24 for the production of a pharmaceutical composition for the treatment or prevention of respiratory tract illnesses. 26. A pharmaceutical composition comprising a virus according to any one claims 1 to 9, a nucleic acid according to claim 10, a vector according to claim 11, a host cell according to claim 12, a proteinaceous molecule or fragment thereof according to claim 13, an antigen according to claim 14, or an antibody according to claim 15. 27. A method for the treatment or prevention of an MPV infection comprising providing an individual with a pharmaceutical composition according to claim 26. 28. A method for the treatment or prevention of a respiratory illness comprising providing an individual with a pharmaceutical composition according to claim 26. 29. A method according to claim 27 or 28 wherein said individual comprises a human. 30. A method to obtain an antiviral agent useful in the treatment of respiratory tract illness comprising establishing a cell culture or experimental animal comprising a virus according to any one of claims 1 to 9, treating said culture or animal with an candidate antiviral agent, determining the effect of said agent on said virus or its infection of said culture or animal, and selecting an anitviral agent with the desired effect. 31. An antiviral agent obtainable according to the method of claim 30. 32. Use of an antiviral agent according to claim 31 for the preparation of a pharmaceutical composition. 33. Use according to claim 33 for the preparation of a pharmaceutical composition for the treatment of respiratory tract illness. 34. Use according to claim 32 or 33 for the preparation of a pharmaceutical composition for the treatment of an MPV infection. 35. A pharmaceutical composition comprising an antiviral agent according to claim 31. 36. A method for the treatment or prevention of an MPV infection comprising providing an individual with a pharmaceutical composition according to claim 35. 37. A method for the treatment or prevention of a respiratory illness comprising providing an individual with a pharmaceutical composition according to claim 35. 38. A method according to claim 36 or 37 wherein said individual comprises a human. 39. A method for virologically diagnosing an MPV infection of an animal comprising determining in a sample of said animal the presence of a viral isolate or component thereof by reacting said sample with a nucleic acid or an antibody specifically reactive with a component of an avian pneumovirus (APV), said nucleic acid or antibody being cross-reactive with a component MPV. 40. A method for serologically diagnosing an MPV infection of an animal comprising determining in a sample of said animal the presence of an antibody directed against an MPV or component thereof by reacting said sample with a proteinaceous molecule or fragment thereof or antigen derived from an APV isolate or component thereof, said molecule, fragment or antigen selected for being essentially homologous with a component of MPV. 41. A method for virologically diagnosing an APV infection of a bird comprising determining in a sample of said bird the presence of a viral isolate or component thereof by reacting said sample with a nucleic acid according to claim 10 or an antibody according to claim 15 said nucleic acid or antibody being cross-reactive with a component of APV. 42. A method for serologically diagnosing an APV infection of a bird comprising determining in a sample of said bird the presence of an antibody specifically directed against an APV or component thereof by reacting said sample with a proteinaceous molecule or fragment thereof according to claim 13 or an antigen according to claim 14, said molecule, fragment or antigen selected for being essentially homologous with a component of APV. 43. A method according to anyone of claims 39 to 42 wherein said APV comprises APV-C. 44. Use of a diagnostic test designed to detect APV specific antibodies for the detection of an antibody directed against MPV. 45 Use according to claim 44 wherein said test comprises an enzyme immune assay (EIA). 46. A method for the detection of an antibody directed against MPV in a sample comprising testing said sample in a diagnostic test designed to detect APV specific antibodies. 47. A method according to claim 46 wherein said test comprises an enzyme immune assay (EIA).

Method and apparatus for solid or solution phase reaction under ambient or inert conditions

The present invention generally provides a novel, automation-compatible solid or solution phase reaction vessel, as well as methods for using such a vessel. Generally, the reaction vessel comprises a microplate assembly with a modular solid phase included within the individual reaction wells. The reaction vessel of the invention allows for the integration of solid phase chemistry with the processing abilities of solution phase chemistry. According to the invention, the microplate assembly and the solid phases are configured so as to integrate together into a single reaction vessel. The combination enables solid phase reactions in a single vessel with full compatibility to liquid handling automation. Further, the combination enables novel methods for performing combination solution phase/solid phase reactions under inert conditions.

1. A reaction vessel assembly comprising: a microplate having a rigid body with a plurality of open reaction wells disposed therein, each of said open reaction wells comprising a fluid vessel with an opening and an interior volume with a sample-holding space located therein; a funnel cap inserted into each of the open reaction wells for at least partially sealing the open well while allowing for venting of the well through a vent passage; a modular solid phase disposed within the interior volume of each of the open wells such that the modular solid phase does not block the passage of the funnel cap; whereby the reaction vessel is accessible through the funnel cap at all times. 2. The reaction vessel of claim 1, wherein the solid phase is an exposed polymer surface object comprising a rigid or polymer-containing, unreactive base, wherein the polymer is attached or contained by the base. 3. The reaction vessel of claim 2, wherein the active polymer attached to the unreactive base is selected from the group consisting of polyethylene, polypropylene, and polytetrafluoroethylene. 4. The reaction vessel of claim 1, wherein each funnel cap comprises a sealing plug and a vent tube; wherein the sealing plug forms a seal at the mouth of the open wells; and the vent tube forms the vent passage, attaches to the sealing plug and terminates in a vent opening. 5. The reaction vessel of claim 4, wherein the solid phase is disposed in the lower portion of the reaction well below the vent tube such that the solid phase does not block-the vent opening. 6. The reaction vessel of claim 4, wherein the solid phase is immobilized in the upper portion of the reaction well above the vent opening such that the solid phase does not block the vent opening. 7. The reaction vessel of claim 1, wherein the funnel cap is configured so as to substantially prevent the escape of a liquid sample contained within the interior volume of the reaction well. 8. The reaction vessel of claim 7, wherein the solid phase is immobilized in the lower portion of the reaction well such that the solid phase does not block the vent passage. 9. The reaction vessel of claim 7, wherein the solid phase is disposed in the upper portion of the reaction well such that the solid phase does not block the vent passage. 10. The reaction vessel of claim 1, wherein the solid phase is directly disposed on at least a portion of the interior walls of the reaction wells, whereby the solid phase adheres to the at least portion of the interior walls of the reaction wells. 11. The reaction vessel of claim 10, wherein the solid phase is disposed on a lower portion of the interior walls of the reaction wells relative to the vent passage. 12. The reaction vessel of claim 10, wherein the solid phase is disposed on an upper portion of the interior walls of the reaction wells relative to the vent passage. 13. The reaction vessel of claim 10, wherein the solid phase covers the entire surface of the interior walls of the reaction wells below the vent passage. 14. The reaction vessel of claim 10, wherein the solid phase covers the entire surface of the interior walls of the reaction wells above the vent passage. 15. The reaction vessel of claim 1, further comprising an upper inert atmosphere cap configured so as to provide a constant positive pressure of inert gas while allowing for access to the reaction wells. 16. The reaction vessel of claim 15, wherein the solid phase is shaped to press fit into the funnel caps inserted into the open reaction wells. 17. The reaction vessel of claim 15, wherein the solid phase is directly disposed on at least a portion of the interior walls of the reaction wells, whereby the solid phase adheres to the at least portion of the interior walls of the reaction wells. 18. The reaction vessel of claim 17, wherein the solid phase is disposed on a lower portion of the interior walls of the reaction wells relative to the vent passage. 19. The reaction vessel of claim 17, wherein the solid phase is disposed on an upper portion of the interior walls of the reaction wells relative to the vent passage. 20. The reaction vessel of claim 17, wherein the solid phase covers the entire surface of the interior walls of the reaction wells below the vent passage. 21. The reaction vessel of claim 17, wherein the solid phase covers the entire surface of the interior walls of the reaction wells above the vent passage. 22. A reaction vessel assembly comprising: a lower microplate assembly having a rigid body comprising a plurality of open reaction wells disposed therein and a funnel vent associated with each open reaction well for at least partially sealing the open well while allowing for venting of the well; and an upper inert atmosphere cap configured so as to provide a constant positive pressure of inert gas while allowing for access to the open reaction wells. 23. The reaction vessel of claim 22, wherein the funnel vent is configured so as to substantially prevent the escape of a liquid sample contained within an interior volume of the reaction well. 24. A reaction vessel assembly comprising: a microplate having a rigid body with a plurality of open reaction wells disposed therein, each of said open reaction wells comprising a fluid vessel with an opening and an interior volume with a sample-holding space located therein and a solid phase disposed within the interior volume of each of the reaction wells, wherein the solid phase is directly disposed on at least a portion of the interior walls of the reaction wells, and whereby the solid phase adheres to the at least portion of the interior walls of the reaction wells. 25. The reaction vessel of claim 24, wherein the solid phase is disposed on a lower portion of the interior walls of the reaction wells. 26. The reaction vessel of claim 24, wherein the solid phase is disposed on an upper portion of the interior walls of the reaction wells. 27. The reaction vessel of claim 24, wherein the solid phase substantially covers the entire surface of the interior walls of the reaction wells. 28. The reaction vessel of claim 24, further comprising an upper inert atmosphere cap configured so as to provide a constant positive pressure of inert gas while allowing for access to the reaction wells. 29. A method for performing a combination solution phase/solid phase reaction using the reaction vessel of claim 8 comprising the steps of: (a) inserting solution phase reagents into the interior volume of the reaction well such that the solution phase reagent mixture is in contact with the solid phase; and (b) allowing a solid phase reaction to proceed to form a product attached to the solid phase. 30. The method of claim 29, further comprising the steps of (c) removing the solution phase mixture from the wells; (d) introducing into the wells a solution capable of cleaving the product of the solid phase reaction from the solid phase and allowing for the cleavage of the product from the solid phase to proceed; and (e) recovering the product cleaved from the solid phase. 31. A method for performing a catalyzed solution reaction using the reaction vessel of claim 8 comprising the steps of: (a) inserting solution phase reagents into the interior volume of the reaction well such that the solution phase reagent mixture is in contact with the solid phase; and (b) allowing a solution phase reaction catalyzed by the solid phase to proceed to form a product in the solution phase. 32. A method for performing a solution phase reaction using the reaction vessel of claim 8 comprising the steps of: (a) inserting solution phase reagents into the interior volume of the reaction well such that the solution phase reagent mixture is in contact with the solid phase; and (b) allowing a solution phase reaction to proceed to form a primary product and one or more secondary products; wherein at least one of the secondary products attaches to the solid phase; and (c) separating the solution phase from the solid phase with one or more secondary products attached to the solid phase. 33. A method for performing a combination solution phase/solid phase reaction using the reaction vessel of claim 11 comprising the steps of: (a) inserting solution phase reagents into the interior volume of the reaction well such that the solution phase reagent mixture is in contact with the solid phase; and (b) allowing a solid phase reaction to proceed to form a product attached to the solid phase. 34. The method of claim 33, further comprising the steps of (c) removing the solution phase mixture from the wells; (d) introducing into the wells a solution capable of cleaving the product of the solid phase reaction from the solid phase and allowing for the cleavage of the product from the solid phase to proceed; and (e) recovering the product cleaved from the solid phase. 35. A method for performing a catalyzed solution reaction using the reaction vessel of claim 11 comprising the steps of: (a) inserting solution phase reagents into the interior volume of the reaction well such that the solution phase reagent mixture is in contact with the solid phase; and (b) allowing a solution phase reaction catalyzed by the solid phase to proceed to form a product in the solution phase. 36. A method for performing a solution phase reaction using the reaction vessel of claim 11 comprising the steps of: (a) inserting solution phase reagents into the interior volume of the reaction well such that the solution phase reagent mixture is in contact with the solid phase; and (b) allowing a solution phase reaction to proceed to form a primary product and one or more secondary products; wherein at least one of the secondary products attaches to the solid phase; and (c) separating the solution phase from the solid phase with one or more secondary products attached to the solid phase. 37. A method for performing a combination solution phase/solid phase reaction using the reaction vessel of claim 18 comprising the steps of: (a) inserting solution phase reagents into the interior volume of the reaction well such that the solution phase reagent mixture is in contact with the solid phase adhered to the interior walls of the reaction wells; and (b) allowing a solid phase reaction to proceed to form a product attached to the solid phase. 38. The method of claim 37, further comprising the steps of (c) removing the solution phase mixture from the wells; (d) introducing into the wells a solution capable of cleaving the product of the solid phase reaction from the solid phase and allowing for the cleavage of the product from the solid phase to proceed; and (e) recovering the product cleaved from the solid phase. 39. A method for performing a catalyzed solution reaction using the reaction vessel of claim 18 comprising the steps of: (a) inserting solution phase reagents into the interior volume of the reaction well such that the solution phase reagent mixture is in contact with the solid phase; and (b) allowing a solution phase reaction catalyzed by the solid phase to proceed to form a product in the solution phase. 40. A method for performing a solution phase reaction using the reaction vessel of claim 18 comprising the steps of: (a) inserting solution phase reagents into the interior volume of the reaction well such that the solution phase reagent mixture is in contact with the solid phase; and (b) allowing a solution phase reaction to proceed to form a primary product and one or more secondary products; wherein at least one of the secondary products attaches to the solid phase; and (c) separating the solution phase from the solid phase with one or more secondary products attached to the solid phase. 41. A method for performing a combination solution phase/solid phase reaction using the reaction vessel of claim 9 comprising the steps of: (a) inserting starting material reagents into the interior volume of the reaction well and inverting the reaction vessel such that the reagents contact the solid phase; (b) allowing a solid phase reaction to proceed to thereby form a solid phase product; (c) turning the reaction vessel to an upright position such that the solution phase is no longer in contact with the solid phase; (d) removing the solution phase from the reaction wells and introducing into the reaction wells a cleaving solution capable of separating the solid phase product from the solid phase; (e) inverting the vessel such that the cleaving solution separates the product from the solid phase. 42. The method of claim 41, further comprising inverting the vessel after the separation of the solid phase product from the solid phase, introducing into the vessel a second reagent solution; and allowing a solution phase reaction to proceed between the solid phase product and the second solution reagent to form a solution phase product. 43. A method for performing a combination solution phase/solid phase reaction using the reaction vessel of claim 19 comprising the steps of: (a) inserting starting material reagents into the interior volume of the reaction well and inverting the reaction vessel such that the reagents contact the solid phase; (b) allowing a solid phase reaction to proceed to thereby form a solid phase product; (c) turning the reaction vessel to an upright position such that the solution phase is no longer in contact with the solid phase; (d) removing the solution phase from the reaction wells and introducing into the reaction wells a cleaving solution capable of separating the solid phase product from the solid phase; (e) inverting the vessel such that the cleaving solution separates the product from the solid phase. 44. The method of claim 43, further comprising inverting the vessel after the separation of the solid phase product from the solid phase, introducing into the vessel a second reagent solution; and allowing a solution phase reaction to proceed between the solid phase product and the second solution reagent to form a solution phase product.

<SOH> BACKGROUND OF THE INVENTION <EOH>Combinatorial chemistry generally relates to a set of techniques for creating a multiplicity of compounds and then testing them for desired activity. More specifically, combinatorial chemistry involves the formation of large libraries of molecules en masse, instead of the synthesis of compounds one by one as had been done traditionally. Once the libraries are obtained, the most promising lead pharmaceutical compounds are identified by high-throughput screening for further evaluation. Generally, combinatorial compounds are created either by solution-phase synthesis or by producing compounds bound covalently to solid phase particles. Solid phase synthesis can make multi-step reactions easier to perform and more reliably allows one to drive reactions to completion because excess reagents can be added and then easily washed away after each reaction step. Further, solid phase synthesis allows for the use of split synthesis, a technique that produces large support-bound libraries in which each solid phase particle holds a single compound. Soluble libraries can then be produced by cleavage of the compounds from the solid support. Nonetheless, a much wider range of organic reactions can be available for solution phase synthesis, and products in solution can often be more easily identified and characterized. As such, solution phase synthesis can still be preferable in some situations. Regardless of the method employed, combinatorial synthesis methods can either be manually performed, or can be automated. Manual synthesis requires repetitions of several relatively simple operations—addition of reagents, incubation and separation of solid and liquid phases, and removal of liquids. This character of the synthetic process renders it optimal for automation. Several designs of automated instruments for combinatorial synthesis have appeared in the patent and non-patent literature. The combinatorial approach to the synthesis of new drug entities has stimulated the development of a wide range of technologies for parallel processing. These have ranged from simple heated agitation systems to fully automated multi-probe synthesizers. Many have been developed to meet the demand for new drug candidates, but the drive towards parallel processing in all areas of laboratory development has also expanded significantly. Historically the discovery and optimization of candidate compounds for development as drugs has been extraordinarily expensive and time-consuming. Although the relatively new approach of “rational drug design” has promise for the future, the pharmaceutical industry has generally relied on mass screening of many-membered “libraries” of chemical compounds for the identification of “lead” compounds worthy of further study and structure-activity relationship (SAR) work. To meet this need high-throughput screening (HTS) technology has been developed that permits pharmaceutical companies to evaluate hundreds of thousands of individual chemical entities per year. Typically, these screens involve measuring some interaction (e.g., binding) between a biological target such as an enzyme or receptor and chemical compounds under test. The screens generally commence with the addition of individual compounds (or mixtures of compounds) to the individual wells in a 96 or higher-well “microtiter” plate that contains the biological target of interest (e.g., a receptor, enzyme or other protein). Ligand/receptor binding or other interaction events are then deduced by, for instance, various spectrophotometric techniques. Those chemical entities that exhibit promise in initial screens (e.g., that bind a biological target with some threshold affinity) are then subjected to chemical optimization, SAR work, other types of testing, and, if warranted, eventual development as drugs. Now that HTS has simplified and made more cost-effective the task of determining whether large chemical libraries contain promising lead compounds or “hits”, many pharmaceutical companies are limited not by their ability to screen candidate compounds but rather by their ability to synthesize them in the first place. At one point, most pharmaceutical companies relied on their historical collections of natural products and individually synthesized chemical entities as compound libraries to be subjected to mass screening. However, expanding these libraries—especially with a view toward increasing the “diversity” of the chemical space that they probe—has proven problematic. For instance the cost of having a synthetic organic or medicinal chemist synthesizes individual molecules in a serial fashion has been estimated to be several thousand dollars, and this is obviously a painstakingly slow process. Thus, the advent of high-throughput screening has created a need for correspondingly high-throughput chemical synthesis (HTCS) to feed this activity. “Combinatorial chemistry” and related techniques for high-throughput parallel syntheses of large chemical libraries were created in response to this need. To simplify the separation of intermediate compounds during multistep organic syntheses, much of this chemistry is generally performed while the compound being synthesized is covalently immobilized on a solid support such as a bead. Once the chemical building blocks have been properly assembled, the desired compounds are usually cleaved from their supports (often highly swellable polymeric resins) before being carried through to HTS. Various definitions of “combinatorial chemistry” and “combinatorial synthesis” have been proposed and are in current use. Some synthesis strategies (e.g., “split-and-mix”) are truly “combinatorial” in nature and have as their hallmark the ability to produce very large libraries; indeed, as many as a million library members can be synthesized in a modest number of reactions (and correspondingly small number of reaction vessels) by virtue of the exponential mathematics involved. One of the several limitations of such approaches, however, is the difficulty of identifying the particular individual chemical species responsible for any activity measured in an assay of what is generally a mixture of compounds. Other approaches such as high-throughput parallel synthesis are typically used to produce somewhat smaller chemical libraries containing, for example, from several hundred to several hundred thousand individual compounds. Here, discrete compounds (and occasionally mixtures) are spatially segregated during chemical synthesis so no ambiguity exists as to the identity of any compound producing a “hit.” However, parallel synthesis requires that chemical reactions be conducted in parallel in a relatively large number of reaction vessels, thus placing a premium on the ability to automate and improve the speed and efficiency of the synthetic process. Most high-throughput chemical syntheses (HTCS) performed in the context of combinatorial chemistry and parallel synthesis are presently conducted in multi-vessel reaction assemblies often referred to as “reaction blocks” by virtue of their monolithic construction. In most solid-phase syntheses, the compound being constructed is covalently attached to resin beads and so many of these multi-vessel reaction blocks include provision for a porous frit to retain the polymer resin beads (and compounds attached thereto) in the reaction vessel during the multiple resin washing steps that are used to remove excess reagents (e.g., building blocks, solvents, catalysts, etc.) after individual reaction steps. Constructions based on specialized reactors connected permanently (or semipermanently) to containers for the storage of reagents are strongly limited in their throughput. The productivity of automated instruments can be dramatically improved by use of disposable reaction vessels, (such as multititer plates or test tube arrays) into which reagents are added by pipetting, or by direct delivery from storage containers. The optimal storage vehicle is a syringe-like apparatus of a material inert to the chemical reactants, etc., e.g., a glass syringe, allowing the storage of the solution without any exposure to the atmosphere, and capable of serving as a delivery mechanism at the same time. See U.S. Pat. No. 6,045,755 issued on Apr. 4, 2000. Liquid removal from the reaction vessel (reactor) is usually accomplished by filtration through a filter-type material. The drawback of this method is the potential clogging of the filter by the solid phase support material, leading to extremely slow liquid removal, or to contamination of adjacent reactor compartments. An alternative technique based on the removal of liquid by suction from the surface above the sedimented solid phase is limited due to incomplete removal of the liquid from the reaction volume. See U.S. Pat. No. 6,045,755 issued on Apr. 4, 2000. U.S. Pat. Nos. 5,202,418; 5,338,831; and 5,342,585 describe methods for liquid removal involving the placement of resin in polypropylene mesh packets, and removal of liquid through the openings of these packets (therefore this process is basically filtration), or removal of the liquid from the pieces of porous textile-like material by centrifugation. Liquid removal by centrifugation was also described in U.S. Pat. No. 6,12,054 issued on Sep. 19, 2000. The method described therein generally involves the use of widely available solid phase organic synthetic protocols and disposable reaction vessel arrays such as microtiter style plates. The reaction vessel array is spun around its axis to create a “pocket” in which the solid material is retained. None of the prior art contemplates the removal of liquid by creation of “pockets” from which material cannot be removed by centrifugal force. Microtiter plates provide convenient handling systems for processing, shipping, and storing small liquid samples. Such devices are especially useful in high throughput screening and combinatorial chemistry applications and are well suited for use with robotic automation systems, which are adapted to selectively deliver various substances into different individual wells of the microtiter plate. As such, microtiter plates have proven especially useful in various biological, pharmacological, and related processes, which analyze and/or synthesize large numbers of small liquid samples. Standard multi-well microtiter plates come in a range of sizes, with shallow well plates having well volumes on the order of 200 to 300 microliters, with deep well plates typically having well volumes of 1.2 ml or 2.0 ml. A common example of a multi-well microtiter plate system is the standard 96-well microplate. Such microplates are typically fabricated from a variety of materials including polystyrene, polycarbonate, polypropylene, PTFE, glass, ceramics, and quartz. Unfortunately, standard microtiter plates suffer from a number of limitations, particularly with regard to chemical synthesis. For example, spillage, leakage, evaporation loss, airborne contamination of well contents, and inter-well cross-contamination of liquid samples are some of the common deficiencies that limit the application of standard microtiter plate assemblies in high throughput synthesis systems. Various techniques such as the inclusion of sealing layers or septums have been used in an attempt to overcome some of these shortcomings. For instance, WO 00/03805 discloses a microtiter reaction system comprising a support rack having an array of reaction wells disposed therein. The microtiter reaction system further includes a porous gas-permeable layer positioned over support rack, wherein the gas-permeable layer has an array of holes therein with each hole being positioned over each of the plurality of reaction wells. Finally, the assembly includes a gasket positioned over the porous gas-permeable layer and a top cover positioned over gasket. While effective to some degree, such microtiter assemblies are complicated, difficult to seal and reseal, and are generally expensive to manufacture. There still remains a need for a simple, efficient means of performing solid phase synthesis, particularly a method and apparatus amenable to use with automated methods for such synthesis. There also remains a need for a simple, efficient means for preventing spillage, leakage, evaporation loss, airborne contamination of well contents, and inter-well cross-contamination of liquid samples in microtiter reaction systems suitable for use in conjunction with automated solid phase/liquid synthesis.

<SOH> SUMMARY OF THE INVENTION <EOH>The present invention allows for the integration of solid phase chemistry with the process of solution phase chemistry. Generally, the present invention provides a novel method and apparatus for solid phase, combination solution/solid phase, and solution phase reactions. In one aspect of the invention, a reaction vessel assembly is provided which comprises a microplate having a rigid body with a plurality of open reaction wells mounted therein, a funnel cap inserted into each of the open reaction wells for at least partially sealing the open wells while allowing for venting of the well through a vent passage, and a modular solid phase immobilized within the interior volume of each of the open wells such that the modular solid phase does not block the passage of the funnel cap. In a preferred embodiment of the invention, the funnel cap is configured so as to substantially prevent the escape of a liquid sample contained within the interior volume of the reaction wells. Further, the discrete solid phase includes a support which is preferably a rigid or self-contained polymer object comprising a rigid or self-contained, unreactive base with an active polymer attached or within containment of the unreactive base, allowing for very high exposed polymer surface area. Examples of this polymer object can be in the form of tea-bags [1], crowns [2] , Irori Kans [3] , lanterns [4] , or sintered resins [5] . As long as the polymer object is rigid (not free-flowing), its support is unreactive, and it can be shaped to fit the funnel insert, it is a candidate for use within this reactor. In another aspect of the invention, a method for performing a combination solution phase/solid phase reaction using the reaction vessel of the invention where the solid support is immobilized in the lower portion of the reaction is provided comprising the steps of: a. Solid phase reaction wherein i. Reactants are added to the vessel and contacted with the solid support ii. A solid phase reaction occurs and the product is attached to the solid phase material iii. The reaction solution is removed from the vessel iv. A Cleaving solution is added v. The solid phase product is released in the solution phase b. Solution phase as a catalyst of a solution phase reaction wherein i. Reactants are added to the vessel and contacted with the solid support ii. A solution phase reaction occurs with the solid support and the product remains in the solution phase iii. The solution phase product is then released in the solution phase c. Solution phase reaction wherein the solid phase is functionalized to serve as a scavenger wherein i. Reactants are added to the vessel and contacted with the solid support ii. A solution phase reaction occurs with the solid support and the product remains in the solution phase iii. By products bond to the solid support iv. The solution phase product is recovered in the solution phase, while the by products are left behind on the solid support In another aspect of the invention, a method for performing a combination solution phase/solid phase reaction using the reaction vessel of the invention where the solid support is immobilized in the upper portion of the reaction is provided comprising the steps of: d. Solid Phase Reaction. i. Reactants are added to the vessel and initially do not contact the solid support ii. Reaction vessel is inverted to contact reactants with solid support iii. A solid phase reaction occurs with the solid support iv. Reaction vessel is turned upright and the reaction mixture is removed v. Cleavage solution is added to the vessel vi. Reaction vessel is inverted to contact the cleavage solution with the solid support vii. Solid phase product is cleaved into the solution viii. Reaction vessel is returned to upright position and product can be removed e. Solution phase reaction (catalyst) i. Reactants are added to vessel and do not contact the solid support ii. Reaction vessel is inverted to contact reactants with solid support iii. A solution phase reaction occurs with the solid support and the product remains in the solution phase iv. Reaction vessel is returned to upright position. v. The solution phase product is then removed in the solution phase f. Solution phase reaction (solid phase functionalized to serve as a scavenger) i. Reactants are added to vessel and do not contact the solid support ii. Reaction vessel is inverted to contact reactants with solid support iii. A solution phase reaction occurs with the solid support and the product remains in the solution phase iv. By products bond to the solid support v. Reaction vessel is returned to upright position vi. The solution phase product can now be removed in the solution phase and while the by products are left behind on the solid support In another aspect, the invention provides a novel microtiter chemical reaction system which allows for reactions to be carried out in an inert atmosphere while minimizing spillage, leakage, evaporation, and cross-contamination. In one embodiment of the invention, a reaction vessel assembly is provided which comprises a lower reaction well microplate assembly having a rigid body with a plurality of open reaction wells disposed therein, and a funnel cap inserted into each of the open reaction wells for at least partially sealing the open well while allowing for venting of the well through a vent passage. The reaction vessel further comprises an upper inert atmosphere cap, which is configured so as to provide a constant positive pressure of inert gas while allowing access to the lower reaction wells. In a preferred embodiment of the invention, the funnel cap is configured so as to substantially prevent the escape of a liquid sample contained within the interior volume of the reaction well. The reaction system can be used as an inert solution phase reactor where reactants are dispensed through the funnel of the upper inert atmosphere cap and through the funnel of the lower reaction vessel. The liquid reaction is contained in the lower reaction vessel while the atmosphere is controlled by the positive pressure of inert gas flowing in and out of the entire vessel through the upper cap. In another embodiment of the invention, the inert reaction system can be used for solid phase reactions using the solid support in the lower reaction vessel. Within that vessel the solid support can be immobilized at the bottom or top of the vessel. A method for performing a combination solution phase/solid phase reaction using the reaction vessel of the invention where the solid support is immobilized in the lower portion of the reaction is provided comprising the steps of: g. Solid phase reaction i. Reactants are added to vessel and contact the solid support ii. A solid phase reaction occurs and the product is attached to the solid phase material iii. The reaction solution is removed from the vessel iv. A Cleaving solution is added v. The solid phase product can now be removed in the solution phase h. Solution phase reaction (catalyst) i. Reactants are added to vessel and contact the solid support ii. A solution phase reaction occurs with the solid support and the product remains in the solution phase iii. The solution phase product can now be removed in the solution phase i. Solution phase reaction (solid phase functionalized to serve as a scavenger) i. Reactants are added to vessel and contact the solid support ii. A solution phase reaction occurs with the solid support and the product remains in the solution phase iii. By products bond to the solid support iv. The solution phase product can now be removed in the solution phase and while the by products are left behind on the solid support In another aspect of the invention, a method for performing a combination solution phase/solid phase reaction using the reaction vessel of the invention where the solid support is immobilized in the upper portion of the reaction is provided comprising the steps of: Inert reaction vessel, solid support is located at the upper end of the reaction vessel. j. Solid Phase Reaction. i. Reactants are added to vessel and do not contact the solid support ii. Reaction vessel is inverted to contact reactants with solid support iii. A solid phase reaction occurs with the solid support iv. Reaction vessel is turned upright and the reaction mixture is removed v. Cleavage solution is added to the vessel vi. Reaction vessel is inverted to expose cleavage solution with solid support vii. Solid phase product is cleaved into the solution viii. Reaction vessel is returned to upright position and product can be removed k. Solution phase reaction (catalyst) i. Reactants are added to vessel and do not contact the solid support ii. Reaction vessel is inverted to contact reactants with solid support iii. A solution phase reaction occurs with the solid support and the product remains in the solution phase iv. Reaction vessel is returned to upright position. v. The solution phase product can now be removed in the solution phase l. Solution phase reaction (solid phase functionalized to serve as a scavenger) i. Reactants are added to vessel and do not contact the solid support ii. Reaction vessel is inverted to contact reactants with solid support iii. A solution phase reaction occurs with the solid support and the product remains in the solution phase iv. By products bond to the solid support v. Reaction vessel is returned to upright position The solution phase product can now be removed in the solution phase while the by products are left behind on the solid support. In another embodiment of the invention, the polymer object can also be shaped to fit into the reaction vessel in such a way that it does not interfere with the action of aspirating or dispensing from this vessel. The polymer object would take the shape of a cylinder or donut and reside on the outer wall of the reaction vessel. The funnel cap can be used to ensure that the accessing device does not contact the polymer object. In another embodiment of the invention, the polymer can be directly attached to the walls of the reaction vessel, perhaps through sintering [5] , whereas the solid support would be the reaction vessel itself. The polymer object would reside on the perimeter of the reaction vessel to avoid interference with the accessing device. Again, the funnel cap could be used to ensure that the accessing device does not contact the polymer object. In yet another embodiment the inert reaction system can utilize the shaped polymer placed into the lower reaction vessels or the polymer can be attached to the walls of the lower reaction vessel.

Setting real time clocks in communications netwroks

A request requesting a real time clock (RTC) value in a communications network is sent from a network element to a management system via a data communications network. The time taken from the sending of the request to the receipt of the RTC value is compared with a predetermined maximum and, if less than or equal to the maximum, the network element RTC is updated. If above the maximum, the RTC value is discarded, and a fresh request is sent. The received acceptable RTC value may be corrected by subtracting either the minimum transmission time or half the actual transmission time.

1. A method of setting a real time clock in a network element of a data communications network having a management system communicating with the network element across the network, the method characterised by: sending a message from the network element to the management system requesting a real time clock (RTC) value; receiving the RTC value at the network element; measuring the time taken between the sending of the RTC request message and receipt of the RTC value; comparing the measured time with a predetermined acceptable time; and if the measured value is acceptable: setting the network element real time clock with the received value. 2. A method according to claim 1, wherein if the comparison of the measured value and the acceptable value shows the measured value to be unacceptable, the received RTC value is rejected and a further RTC value request message is sent by the network to the management system. 3. A method according to claim 2, wherein upon determination that a received RTC value is unacceptable, the network element compares the number of unacceptable values received with a maximum permissible number and, if the number of unacceptable values received equals the maximum permissible number, sends an alarm to the management system. 4. A method according to any preceding claim, wherein the step of setting the network element real time clock with an acceptable received value comprises correcting the received value to compensate for transmission time. 5. A method according to claim 4, wherein the step of correcting the received RTC value comprises subtracting a minimum transmission time from the received value. 6. A method according to claim 4, wherein the step of correcting the received RTC value comprises subtracting half the measured time taken between sending of the RTC request message and receipt of the RTC value. 7. A network element for a data communications network having a management system communicating with the network element across the network, the network element comprising: a real time clock; a message generator for generating and sending real time clock (RTC) value requests to the management system; a message receiver for receiving requested RTC values from the management system; a timer for measuring the time between sending of an RTC request message and receipt of the RTC value; a comparator for comparing the measured time with a predetermined acceptable time; and means for setting the network element real time clock with the received RTC value if that value is acceptable. 8. A data communications system comprising at least one network element and a management system, the network element and the management system communicating across the communications network; the system comprising at the network element: a real time clock set by the management system; a message generator for generating and sending real time clock (RTC) value requests to the management system; a message receiver for receiving requested RTC values from the management system; a timer for measuring the time between sending of an RTC request message and receipt of the RTC value; a comparator for comparing the measured time with a predetermined acceptable time; and means for setting the network element real time clock with the received value if that value is acceptable; and at the management system: means for receiving RTC value request messages from the network element and for sending RTC values to the network element in response thereto. 9. A network element or data communications system according to claim 7 or claim 8, and further comprising: means for rejecting the received RTC value if the measured time is unacceptable. 10. A network element or data communications system according to claim 9, wherein the network element comprises a further comparator for comparing the number of unacceptable received RTC values with a predetermined maximum, and an alarm generator for generating and sending an alarm to the management system if the predetermined maximum number of unacceptable values has been reached. 11. A network element or data communications system according to any of claims 7 to 10, wherein the network element comprises a real time clock value corrector for correcting received acceptable real time clock values to compensate for transmission time from the management system. 12. A network element or data communications system according to claim 11, wherein the real time clock value corrector comprises a subtractor for subtracting from the acceptable RTC value the minimum time between sending the RTC value request message and receiving the RTC value. 13. A network element or data communications system according to claim 11, wherein the real time clock value corrector comprises a subtractor for subtracting half the measured time taken between sending of the RTC request message and receipt of the RTC value. 14. A method of setting a real time clock in a network element of a data communications network comprising: requesting a real time clock value RTC from a remote RTC source; measuring the period between the RTC request and receipt of an RTC value; and updating the real time clock of the network value if the measured period is within a predetermined acceptable range.

Sample carrier

The invention relates to a sample carrier which is used to receive chemical and/or biological samples, comprising a receiving part (30) having recesses (32) for receiving the samples. The recesses (32) are sealed by means of a base part (38). The base part (38) is glued to the receiving part (30). The receiving part has a slit-shaped recess (44) for receiving excess glue in order to prevent glue from spreading to the outside of the base part (38).

1. A sample carrier for receiving chemical and/or biological samples, comprising a receiving part (30) provided with deepened portions (32) for receiving the samples, and a base part (38) closing the deepened portions (32), the base part (38) being connected to the receiving part (30) at connection sites by means of a bonding agent, characterized in that the receiving part (30) and/or the base part (38) are provided, at the connection sites, with at least one recess (44,52,60) for receiving excess bonding agent. 2. The sample carrier according to claim 1, characterized in that the recess (44) is provided in the edge region of the base part (38) and/or of the receiving part (30). 3. The sample carrier according to claim 1 or 2, characterized in that the recess (44) is formed by a stepped portion (42) provided in the receiving part (30) outside the deepened portions (32). 4. The sample carrier according to claim 3, characterized in that the base part (38) extends beyond the stepped portion (42) to form a gap-shaped recess (44). 5. The sample carrier according to claim 4, characterized in that the gap-shaped recess (44) is formed substantially continuously along the edge of the base part (38). 6. The sample carrier according to any one of claims 1 to 5, characterized in that the recess (44) is open in the direction of an outer edge of the receiving part (30). 7. The sample carrier according to any one of claims 1 to 6, characterized in that the base part (38) is permeable to radiation to allow for the examination of the sample. 8. The sample carrier according to any one of claims 1 to 7, characterized in that the base part (38) comprises a plastic film or glass. 9. The sample carrier according to any one of claims 1 to 8, characterized in that the thickness of the base part (38) is smaller than 500 μm, particularly smaller than 300 μm. 10. The sample carrier according to any one of claims 1 to 9, characterized in that the receiving part (30) is provided with centering elements (48) determining the position of the base part.

Method and circuit for forming an atm cell

A pressure pulse generator for use in transmitting pressure signals to surface in a fluid-based drilling system having a housing having an inlet to admit drilling fluid to the interior of housing, and an outlet to discharge fluid from the interior of the housing; a control element slidably mounted in the housing between an open and a closed position, the control element generating a pressure pulse in the supply of pressure fluid when the control element takes-up the closed position; a control passage extending through the control elements and closable by a pilot valve element exposed to the pressure of the fluid in the passage; and an actuator coupled with the valve element to generate a pressure signal, to move the valve element to a position closing said passage and thereby to cause movement of the control element towards the closed position; the coupling between the actuator and the valve element includes a yieldable biassing element which provides control of the amplitude of the pressure signals produces by the generator.

1. A pressure pulse generator for use in transmitting pressure signals to surface in a fluid-based drilling system, said generator being arranged in use in the path of a pressurized fluid to operate a drilling assembly and being capable of being actuated to generate pressure signals in such fluid for transmission to surface pressure monitoring equipment, in which the pulse generator comprises: a housing positionable in the path of the supply of pressurized fluid, said housing having an inlet arrangement for admitting a portion of the fluid to the interior of the housing, and an outlet arrangement for discharging fluid from the interior of the housing for supply of the drilling assembly; a control element slidably mounted in the housing for movement between an open and a closed position with respect to said inlet arrangement, said control element being operative to generate a pressure pulse in the supply of pressure fluid when the control element takes-up the closed position; a control passage extending through the control elements and closable by a pilot valve element arranged to be exposed to the pressure of the fluid in the passage; and an actuator coupled with the valve element and operative, when the pressure generator is activated to generate a pressure signal, to move the valve element to a position closing said passage and thereby to cause movement of the control element towards the closed position; in which the coupling between the actuator and the valve element includes a yieldable biassing element which provides control of the amplitude of the pressure signals produces by the generator. 2. A pressure pulse generator according to claim 1, in which the biasing element comprises a spring arrangement. 3. A pressure pulse generator according to claim 1, in which the biassing element comprises a floating piston assembly incorporated within an actuator link between the actuator and the valve element. 4. A pressure pulse generator according to claim 1 in which the inlet arrangement comprises a ring mounted internally of the housing at an upstream end thereof, and which defines a restricted inlet passage with the valve element. 5. A pressure pulse generator according to claim 4, in which a set of rings is provided, having different internal bores, and selectable for use according to by-pass requirements. 6. A pressure pulse generator according to claim 4, in which the ring includes at least one by-pass port. 7. A pressure pulse generator according to claim 1 in which the actuator comprises an electromagnetic actuator, and the coupling comprises an actuator shaft connected to the actuator a second actuator shaft connected to the valve element, and with said yieldable biassing element (20) located between first actuator shaft and the second actuator shaft.

Human hyperpolarization-activated cyclic nucleotide-gated cation channel hcn1

The present invention is directed to novel human DNA sequences encoding human HCN1 proteins, the protein encoded by the DNA sequences, vectors comprising the DNA sequences, host cells containing the vectors, and methods of identifying inhibitors and activators of cation channels containing the human HCN1 proteins.

1. An isolated DNA comprising a nucleotide sequence encoding human HCN1. 2. The DNA of claim 1 comprising a nucleotide sequence encoding a polypeptide having an amino acid sequence selected from the group consisting of SEQ.ID.NOs.:2, 4, 6, 8, 10, 12, 14, 16, and 18. 3. The DNA of claim 1 comprising a nucleotide sequence selected from the group consisting of: SEQ.ID.NO.:1, SEQ.ID.NO.:3, SEQ.ID.NO.:5, SEQ.ID.NO.:7, SEQ.ID.NO.:9, SEQ.ID.NO.:11, SEQ.ID.NO.:13, SEQ.ID.NO.:15, SEQ.ID.NO.:17, positions 26 to 2695 of SEQ.ID.NO.:1, positions 26 to 2695 of SEQ.ID.NO.:3, positions 26 to 2695 of SEQ.ID.NO.:5, positions 26 to 2695 of SEQ.ID.NO.:7, positions 26 to 2695 of SEQ.ID.NO.:9, positions 26 to 2695 of SEQ.ID.NO.:11, positions 26 to 2695 of SEQ.ID.NO.:13, positions 26 to 2695 of SEQ.ID.NO.:15, and positions 26 to 2695 of SEQ.ID.NO.:17. 4. An isolated DNA that hybridizes under stringent conditions to the DNA of claim 3 and that encodes a protein having substantially the same biological activity as human HCN1. 5. An expression vector comprising the DNA of claim 3. 6. A recombinant host cell comprising the DNA of claim 3. 7. DNA, substantially free of other nucleic acids, comprising a nucleotide sequence encoding a polypeptide having an amino acid sequence selected from the group consisting of SEQ.ID.NOs.:2, 4, 6, 8, 10, 12, 14, 16, and 18. 8. DNA, substantially free of other nucleic acids, comprising a nucleotide sequence selected from the group consisting of: SEQ.ID.NO.:1, SEQ.ID.NO.:3, SEQ.ID.NO.:5, SEQ.ID.NO.:7, SEQ.ID.NO.:9, SEQ.ID.NO.:11, SEQ.ID.NO.:13, SEQ.ID.NO.:15, SEQ.ID.NO.:17, positions 26 to 2695 of SEQ.ID.NO.:1, positions 26 to 2695 of SEQ.ID.NO.:3, positions 26 to 2695 of SEQ.ID.NO.:5, positions 26 to 2695 of SEQ.ID.NO.:7, positions 26 to 2695 of SEQ.ID.NO.:9, positions 26 to 2695 of SEQ.ID.NO.:11, positions 26 to 2695 of SEQ.ID.NO.:13, positions 26 to 2695 of SEQ.ID.NO.:15, and positions 26 to 2695 of SEQ.ID.NO.:17. 9. An isolated human HCN1 protein. 10. The protein of claim 7 comprising an amino acid sequence selected from the group consisting of SEQ.ID.NOs.:2, 4, 6, 8, 10, 12, 14, 16, and 18. 11. The protein of claim 8 containing a single amino acid substitution. 12. The protein of claim 8 containing two or more amino acid substitutions where the amino acid substitutions do not occur in conserved positions. 13. A protein, substantially free of other proteins, comprising an amino acid sequence selected from the group consisting of SEQ.ID.NOs.:2, 4, 6, 8, 10, 12, 14, 16, and 18. 14. An antibody that binds specifically to a human HCN1 protein. 15. A DNA or RNA oligonucleotide probe comprising at least 10 contiguous nucleotides from SEQ.ID.NOs.:1, 3, 5, 7, 9, 11, 13, 15, or 17. 16. A method of identifying substances that bind to cation channels containing human HCN1 protein comprising: (a) providing cells expressing a cation channel containing human HCN1 protein; (b) exposing the cells to a substance that is not known to bind cation channels containing human HCN1 protein; (c) determining the amount of binding of the substance to the cells; (d) comparing the amount of binding in step (c) to the amount of binding of the substance to control cells where the control cells are substantially identical to the cells of step (a) except that the control cells do not express human HCN1 protein; where if the amount of binding in step (c) is greater than the amount of binding of the substance to control cells, then the substance binds to cation channels containing human HCN1 protein. 17. A method of identifying substances that bind cation channels containing human HCN1 protein comprising: (a) providing cells expressing cation channels containing human HCN1 protein; (b) exposing the cells to a compound that is known to bind to the cation channels containing human HCN1 protein in the presence and in the absence of a substance not known to bind to cation channels containing human HCN1 protein; (c) determining the amount of binding of the compound to the cells in the presence and in the absence of the substance; where if the amount of binding of the compound in the presence of the substance differs from that in the absence of the substance, then the substance binds cation channels containing human HCN1 protein. 18. A method of identifying activators or inhibitors of cation channels containing human HCN1 protein comprising: (a) recombinantly expressing human HCN1 protein in a host cell so that the recombinantly expressed human HCN1 protein forms cation channels either by itself or by forming heteromers with other cation channel subunit proteins; (b) measuring the biological activity of the cation channels formed in step (a) in the presence and in the absence of a substance not known to be an activator or an inhibitor of cation channels containing human HCN1 protein; where a change in the biological activity of the cation channels formed in step (a) in the presence as compared to the absence of the substance indicates that the substance is an activator or an inhibitor of cation channels containing human HCN1 protein.

<SOH> BACKGROUND OF THE INVENTION <EOH>The HCN genes encode a family of cation channels that are believed to carry a current known as I h or I q in neural tissue and I f in cardiac tissue. This current is activated by hyperpolarization beyond about −50 to −70 mV, does not inactivate, is carried by both Na + and K + , exhibits a small single channel conductance (about 1 pS), and has the effect of slowly depolarizing a cell toward the I h reversal potential of about −30 mV. The voltage dependence of I h can be modulated by cyclic nucleotides such as cAMP or cGMP. The I h current can contribute significantly to the total current at subthreshold membrane potentials, and thus can be an important factor in the regulation of neuronal firing and cardiac contraction. Three major roles for the I h current have been postulated in neurons: (a) I h contributes to the cell's resting membrane potential; (b) I h can modulate the summation of synaptic inputs into the neuron, e.g., by counteracting hyperpolarizing signals from inhibitory postsynaptic potentials; and (c) I h contributes to the generation of “pacemaker” or oscillatory activity (i.e., rhythmic, spontaneous firing of action potentials). In the heart, the I f current arises following repolarization of an action potential, which returns the cell to its hyperpolarized resting membrane potential. In pacemaker regions of the heart, such as the sinoatrial node, this hyperpolarization activates I f , which leads to a slow depolarization of the myocyte, eventually returning the membrane potential to the action potential threshold, and triggering another action potential. The larger the I f current, the more rapid the return to the action potential threshold and the faster the heart will beat. Agents that stimulate the heart by stimulating the β-adrenergic receptor act, in part, through the I f current. Such agents lead to an increase in intracellular cAMP which shifts the voltage dependence of the If current towards more positive (i.e., depolarized) levels, resulting in faster entry of this current into its role in moving the cell back toward the action potential threshold. For reviews of the I h /I f current, see Clapham, 1998, Neuron 21:5-7; Luthi & McCormick, 1998, Neuron 21:9-12; Pape, 1996, Ann. Rev. Physiol. 58:299-327; DiFrancesco, 1993, Ann. Rev. Physiol. 55:455-472. Certain HCN genes and their encoded protein products have been identified. The DNA and deduced amino acid sequences, as well as some electrophysiological properties, of human HCN2 and human HCN4 have been disclosed (Vaccari et al., 1999, Biochim. Biophys. Acta 1446:419425; Seifert et al., 1999, Proc. Natl. Acad. Sci. USA 96:9391-9396; Ludwig et al., 1999, EMBO J. 18:2323-2329; GenBank accession nos. AF065164 and AJ012582 (HCN2); GenBank accession nos. AJ132429 and AJ238850 (HCN4)). GenBank accession no. AF064876 represents a partial, internal fragment of human HCN1, lacking 5′ and 3′ ends. GenBank accession no. AW054787 represents an EST containing only the carboxy terminal sequences of human HCN1. GenBank accession no. AC013384 represents human chromosome 2 genomic DNA sequences that encompass HCN1 but there is no indication of which portion of the disclosed sequence represents HCN1 coding sequence. Certain fragments of human HCN3 have appeared in certain databases (GenBank accession no. AI571225 is an amino terminal EST; AQ625620 is a partial genomic sequence). Full length mouse HCN1, HCN2, and HCN3 have been cloned as has a partial mouse cDNA encoding HCN4 (Santoro et al., 1998, Cell 93:717-729; Ludwig et al., 1998, Nature 393:587-591). Mouse (GenBank accession no. AJ225123), rat (GenBank accession no. AJ247450), and rabbit (GenBank accession no. AF168122) HCN1 sequences have been deposited in databases. Examination of the cDNAs encoding HCN channels revealed that the HCN proteins represent a family of ion channels having six putative transmembrane domains (S1-S6) and a cAMP binding domain. Functional expression of human HCN2 in a kidney cell line produced currents with properties similar to those of the heart I f current (Vaccari et al., 1999, Biochim. Biophys. Acta 1446:419-425). It is desirable to discover as wide a variety as possible of novel cation channels, especially those from humans and those exhibiting restricted tissue expression. Such novel cation channels would be attractive targets for drug discovery, useful in counterscreens for a variety of other drug targets, and would be valuable research tools for understanding more about ion channel biology.

<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is directed to a novel human DNA sequence encoding human HCN1, a hyperpolarization-activated cyclic nucleotide-gated cation channel. The present invention also includes certain polymorphic variants of human HCN1. The present invention includes DNA comprising the nucleotide sequences shown as SEQ.ID.NOs.:1, 3, 5, 7, 9, 11, 13, 15, and 17 as well as DNA comprising the coding regions of SEQ.ID.NOs.:1, 3, 5, 7, 9, 11, 13, 15, and 17. Also provided are proteins encoded by the novel DNA sequences. The human HCN1 proteins of the present invention comprise the amino acid sequences shown as SEQ.ID.NOs.:2, 4, 6, 8, 10, 12, 14, 16, and 18 as well as fragments thereof. Methods of expressing the novel human HCN1 proteins in recombinant systems are provided. Also provided are methods of using human HCN1 as a drug target by identifying activators and inhibitors of cation channels comprising human HCN1 proteins. Also provided are methods of using the novel human HCN1 proteins and DNA encoding these HCN1 proteins in counterscreens for assays designed to identify activators and inhibitors of other drug targets.

Transient eutectic phase process for ceramic-metal bonding metallization and compositing

A method for directly joining ceramics (10) and metals (12). The method involves forming a structure having a ceramic component (10), a more refractory metallic component and a less refractory metallic-material-based interlayer (14) disposed between the ceramic component (10) and the metallic component (12); adding a eutectic forming reactant to the metallic interlayer (14); and heating the structure to approximately a eutectic melting temperature of the reactant and the interlayer to form a metallic-material-based eutectic liquid that interacts with the metallic component to form a bond that directly joins the ceramic and metallic components to one another.

1. A method for directly joining ceramics and metals, the method comprising: forming a structure having a ceramic component, a metallic component and a metallic interlayer disposed between the ceramic component and the metal metallic component, the metallic interlayer being less refractory than the metallic component; adding a eutectic liquid forming reactant to the metallic interlayer; and heating the structure to approximately a eutectic melting temperature of the reactant and the interlayer to form metallic-material-based eutectic liquid that interacts with the ceramic component and the metallic component to form a bond that directly joins the ceramic and metallic components to one another. 2. The method according to claim 1, wherein the structure further includes a barrier layer that controls the interaction between the metallic interlayer and the metallic component. 3. The method according to claim 1, wherein the adding step is performed prior to the heating step. 4. The method according to claim 1, wherein the adding step is performed substantially concurrent with the heating step. 5. The method according to claim 1, wherein the reactant comprises a gas. 6. The method according to claim 5, wherein the gas comprises oxygen. 7. The method according to claim 1, wherein the metallic interlayer comprises copper. 8. The method according to claim 1, wherein the ceramic component comprises alumina. 9. The method according to claim 1, wherein the metallic component comprises nickel. 10. The method according to claim 1, wherein the reactant comprises oxygen, the metallic interlayer comprises copper, the ceramic component comprises alumina, and the metallic component comprises nickel. 11. The method according to claim 1, wherein the ceramic component is selected from the group consisting of a ceramic layer, ceramic particles, ceramic fibers, ceramic fibrous structures, and combinations thereof; the metallic component is selected from the group consisting of a metal layer, a metal alloy layer, an intermetallic layer, metal particles, metal alloy particles, intermetallic particles, metal fibers, metal alloy fibers, intermetallic fibers, metal fibrous structures, metal alloy fibrous structures, intermetallic fibrous structures and combinations thereof; and the metallic interlayer is selected from the group consisting of a metal, a metal alloy, an intermetallic, and combinations thereof. 12. A method for directly joining ceramics and metals, the method comprising: forming a structure having a ceramic component and a metallic component; and reacting a metallic-material-based eutectic liquid with the metallic component, which is more active than the eutectic liquid, such that active metal specie diffuse to the ceramic component thereby enhancing bonding between the ceramic component and the metallic component. 13. The method according to claim 12, wherein the ceramic component is selected from the group consisting of a ceramic layer, ceramic particles, ceramic fibers, ceramic fibrous structures, and combinations thereof; the metallic component is selected from the group consisting of a metal layer, a metal alloy layer, an intermetallic layer, metal particles, metal alloy particles, intermetallic particles, metal fibers, metal alloy fibers, intermetallic fibers, metal fibrous structures, metal alloy fibrous structures, intermetallic fibrous structures and combinations thereof; and the metallic-material-based eutectic liquid is selected from the group consisting of a metal, a metal alloy, an intermetallic, and combinations thereof. 14. A method for directly joining ceramics and metals, the method comprising: forming a structure having a ceramic component and a metallic component; and reacting a metallic-material-based eutectic liquid with the metallic component, which is more refractory than the eutectic liquid, to form a liquid composition that solidifies isothermally as a transient liquid phase joining the ceramic component and the metal component to one another. 15. The method according to claim 14, wherein the ceramic component is selected from the group consisting of a ceramic layer, ceramic particles, ceramic fibers, ceramic fibrous structures, and combinations thereof; the metallic component is selected from the group consisting of a metal layer, a metal alloy layer, an intermetallic layer, metal particles, metal alloy particles, intermetallic particles, metal fibers, metal alloy fibers, intermetallic fibers, metal fibrous structures, metal alloy fibrous structures, intermetallic fibrous structures and combinations thereof; and the metallic-material-based eutectic liquid is selected from the group consisting of a metal, a metal alloy, an intermetallic, and combinations thereof. 16. A method for directly joining ceramics and metals, the method comprising: forming a structure having a ceramic component and a metallic component; reacting the metallic component with a metallic-material-based eutectic liquid that transitions into a transient liquid phase that solidifies; and further reacting the solidified transient liquid phase with the metallic component, which is more refractory than the metallic component, at elevated temperature to form a solid metallic composition with a melting point that is greater than the solidified transient liquid phase. 17. The method according to claim 16, wherein the ceramic component is selected from the group consisting of a ceramic layer, ceramic particles, ceramic fibers, ceramic fibrous structures, and combinations thereof; the metallic component is selected from the group consisting of a metal layer, a metal alloy layer, an intermetallic layer, metal particles, metal alloy particles, intermetallic particles, metal fibers, metal alloy fibers, intermetallic fibers, metal fibrous structures, metal alloy fibrous structures, intermetallic fibrous structures and combinations thereof; and the metallic-material-based eutectic liquid is selected from the group consisting of a metal, a metal alloy, an intermetallic, and combinations thereof. 18. A method for directly joining ceramics and metals, the method comprising: forming a structure having a ceramic component and a metallic component; providing a metallic-material-based eutectic liquid that transitions into a transient liquid phase that solidifies; and reacting the solidified transient liquid phase with the metallic component, which is more refractory than the solidified transient liquid phase, at an elevated temperature to form a homogeneous metallically bonded material. 19. The method according to claim 16, wherein the ceramic component is selected from the group consisting of a ceramic layer, ceramic particles, ceramic fibers, ceramic fibrous structures, and combinations thereof; the metallic component is selected from the group consisting of a metal layer, a metal alloy layer, an intermetallic layer, metal particles, metal alloy particles, intermetallic particles, metal fibers, metal alloy fibers, intermetallic fibers, metal fibrous structures, metal alloy fibrous structures, intermetallic fibrous structures and combinations thereof; and the metallic-material-based eutectic liquid is selected from the group consisting of a metal, a metal alloy, an intermetallic, and combinations thereof. 20. A method of fabricating a composite structure or material, the method comprising: providing a ceramic component selected from the group consisting of ceramic particles, ceramic fibers, and ceramic fibrous structures and combinations thereof; providing a metallic component selected from the group consisting of metal particles, metal alloy particles, intermetallic particles, metal fibers, metal alloy fibers, intermetallic fibers, metal fibrous structures, metal alloy fibrous structures, intermetallic fibrous structures, and combinations thereof, the metallic component coated with a less refractory metallic interlayer selected from the group consisting of a metal, a metal alloy, an intermetallic, and combinations thereof; mixing the ceramic component with the metallic component, the metallic interlayer being disposed between the ceramic component and the metallic component; adding a eutectic liquid forming reactant to the metallic interlayer; and heating the structure to approximately a eutectic melting temperature of the reactant and the metallic interlayer to form a metallic-material-based eutectic liquid that interacts with the ceramic component and the metallic component to form a bond that directly joins the ceramic component and metallic component to one another. 21. The method according to claim 20, wherein the eutectic liquid transitions into a transient liquid phase that solidifies; and reacting the solidified transient liquid phase with the metallic component, which is more refractory than the solidified transient liquid phase, at an elevated temperature to form a homogeneous metallically bonded material better.

<SOH> BACKGROUND OF THE INVENTION <EOH>No single material is available today that possesses all of the material properties to meet the stringent demands of many traditional and advanced applications. Metals, although ductile with high thermal and electrical conductivity, often cannot withstand high temperatures or corrosion, and expand significantly with increasing temperature. An alternative to metals are ceramics, which are brittle insulators. Ceramics are refractory, hard, and wear-resistant, with excellent hot properties and relatively low thermal expansion. By joining ceramics and metals, composite components that may employ the desired properties of each material, can be manufactured to meet these increasing requirements. The technology required to bond these dissimilar materials effectively, reliably, and economically is in high demand. Several joining technologies, which utilize interfacial methods, have proven effective for bonding ceramics and metals, but their high processing costs limit their penetration of some potential markets. Direct joining or bonding requires few processing steps and therefore significantly reduces cost and eliminates interfacial joining material that may compromise properties. It can also provide property advantages such as hermeticity, stress transfer, stress reduction, continuity of strain, electrical response, interfacial properties, and mechanical interlocking. Therefore, an effective method of directly joining or bonding ceramics and metals is needed.

<SOH> SUMMARY OF THE INVENTION <EOH>A method is described herein for directly joining ceramics and metals. The method comprises forming a structure having a ceramic component, a metallic component and a metallic interlayer disposed between the ceramic component and the metallic component, the metallic interlayer being less refractory than the metallic component by means of a eutectic melt formed by adding a eutectic-forming reactant, such as a gas, an oxide of the metallic material of the interlayer or other compound, to the metallic interlayer (this is commonly termed gas-metal eutectic); and heating the structure to approximately a eutectic melting temperature of the eutectic-based interlayer system to form a metallic-material-based eutectic liquid that interacts with the metallic component to form a bond that directly joins the ceramic and metallic components to one another. The components may be bulk parts, metallization, ceramic coating layers, or compositing materials

Network selection in a mobile telecommunications system

A mobile telephone (1) in a cellular telecommunications system uses data stored in a SIM cards (10) select a network (4) for registration and to perform optimise call routing to a call destination. If data relevant to networks which are currently available is not stored, a request message is sent to a control centre (14) and updating information received in a response message. The subscriber identity data stored in a memory file (200) of the SIM can also be replaced to enable a preferred subscriber identifier to be used with the currently registered network, selection being made using a look-up table (201) updated when required using request and response messages. Greater control of the network selection and cost saving by avoiding roaming agreements are thereby achievable.

1. A method of operating a mobile telecommunications apparatus in a telecommunications system wherein the apparatus comprises means for making telephone calls via the system, the method comprising: detecting a number of available networks; receiving network identification information from the available networks; selecting one of the available networks by comparing the network identification information with stored network information comprising at least one of a preferred network table and a barred network table; registering with the selected network; determining whether the stared information requires updating, and outputting a request message for receiving updating information for updating the stored network information if the stored information is determined to require updating. 2. A method as claimed in claim 1, wherein the determining step comprises determining whether the network identification information indicates that the mobile telecommunications apparatus has moved to a country which is different from a previous country in which the apparatus was previously registered. 3. A method as claimed in claim 1, wherein the determining step determines that the stored network information needs updating if the mobile telecommunications apparatus has registered with a network which is different from a network which the apparatus was last registered. 4. A method as claimed in claim 1 including the step of receiving a response message comprising the requested updating information for updating the stored routing information. 5. A method as claimed in claim 4 including the step of decrypting the received response message. 6. A method as claimed in claim 4 including the step of updating the stored network information with the updating information. 7. A method as claimed in claim 6, wherein the updating information is stored in a first portion of memory to constitute the updated stored network information, and including the further step of storing a duplicate of the updating information in a second portion of the memory to constitute a backup memory. 8. A method as claimed in claim 7, wherein, when accessing the stored network information, the contents of the first and second portions of memory are compared and, if different, the contents of the backup memory in the second portion of the memory are copied to the first portion of memory to constitute the updated stored network information. 9. A method as claimed in claim 7, wherein the stored network information is updated by an updating application which causes both first and second portions of memory to be updated and wherein other applications are prohibited from updating the second portion of memory. 10. A method as claimed in claim 1, wherein updating the stored information comprises storing in a cache memory of the apparatus the information which is superseded by updating information. 11. A mobile telecommunications apparatus for use in a telecommunications system, comprising: detecting means for detecting a number of available networks; receiving means for receiving network identification information from the available networks; selecting means for selecting one of the available networks by comparing the network identification information with stored network information comprising at least one of a preferred network table and a barred network table; and registering means for registering with the selected network; wherein the apparatus further comprises: determining means for determining whether the stored information requires updating, and output means for outputting a request message for receiving updating information for updating the stored information if the stored information is determined to require updating. 12. Apparatus as claimed in claim 11, wherein the determining means is operable to determine whether the network identification information indicates that the mobile telecommunications apparatus has moved to a country which is different from a previous country in which the apparatus was previously registered. 13. Apparatus as claimed in claim 11, wherein the determining means is operable to determine that the stored network information needs updating if the apparatus has registered with a network which is different from a network with which the apparatus was last registered. 14. Apparatus as claimed in claim 11 including message receiving means for receiving a response message comprising the requested updating information for updating the stored information. 15. Apparatus as claimed in claim 14 including decrypting means for decrypting the received response message. 16. Apparatus as claimed in claim 14 including updating means for updating the stored routing information with the updating information. 17. Apparatus as claimed in claims 16, wherein the updating means is operable to store the updating information in a first portion of the memory, and is further operable to store a duplicate of the updating information in a second portion of the memory to constitute a backup memory. 18. Apparatus as claimed in claim 17, wherein the selecting means comprises accessing means for accessing the stored network information in the first portion of memory and means for comparing the contents of the first and second portions of memory and means for copying the contents of the backup memory in the second portion of the memory to the first portion of memory if the contents are different. 19. Apparatus as claimed in claim 17, wherein the stored information is updated by an application which causes both first and second portions of memory to be updated and wherein an operating system prohibits other applications from updating the second portion of memory. 20. Apparatus as claimed in claim 11 comprising a cache memory (146) for storing the stored network information which is superseded by updating information. 21. A method of operating a mobile telecommunications apparatus in a telecommunications system wherein the apparatus comprises means for making outgoing telephone calls via the system, means for receiving a user generated input defining a call destination and routing means for selecting a preferred route to the call destination via the system for an outgoing telephone call by referring to routing information stored in the apparatus, the method comprising: registering with a network of the system which is local to the current location of the apparatus; receiving network identification information from the local network; determining from the network identification information whether the stored routing information requires updating, and outputting a request message for receiving updating information for updating the stored routing information if the stored routing information is determined to require updating. 22. A method as claimed in claim 21, wherein the determining step comprises determining whether the stored routing information can be utilised in selecting a preferred route via the local network identified by the network identification information. 23. A method as claimed in claim 21, wherein the determining step comprises determining whether the network identification information indicates that the mobile telecommunications apparatus has moved to a country which is different from a previous country in which the apparatus was previously registered. 24. A method as claimed in claim 21, wherein the determining step comprises determining whether stored routing information relating to routing outgoing calls via the local network is valid by referring to expiry time information indicating a time at which the validity of the routing information expires. 25. A method as claimed in claim 24, wherein the expiry information is stored in the apparatus in the form of a time at which the validity of the routing information expires and wherein the method comprises determining the current time with reference to a clock and comparing the time with the expiry information. 26. A method as claimed in claim 24, wherein the determining step comprises determining whether a predetermined number of calls has been made by the apparatus since the last update of the routing information. 27. A method as claimed in claim 24, wherein the determining step comprises counting the number of intervals of predetermined length for which the apparatus remains in operation and determining the validity of the routing information to have expired when a predetermined number of the intervals has been counted. 28. A method as claimed in claim 24, wherein the determining step comprises outputting a request message to obtain the expiry information, receiving a response message including the expiry information and determining from the expiry information whether the validity of the routing information has expired. 29. A method as claimed in claim 21 wherein the determining step comprises determining whether the network identification information indicates that the mobile telecommunications apparatus has registered with a network which is different from a network with which the apparatus was last registered. 30. A method as claimed in claim 21 including the step of receiving a response message comprising the requested updating information for updating the stored routing information. 31. A method as claimed in claim 30 including the step of decrypting the received response message. 32. A method as claimed in claim 30 including the step of updating the stored routing information with the updating information. 33. A method as claimed in claim 32, wherein the stored routing information comprises a routing table and comprising the step of updating the routing table. 34. A method as claimed in claim 33, wherein the routing table is updated so as to include a field determining the time of expiry of the validity of the updated routing table. 35. A method as claimed in claim 30 including the step of updating a preferred network table which defines a list of preferred networks in order of preference for use in the registering step when selecting a network from a plurality of available networks for registration. 36. A method as claimed in claim 30 comprising updating a barred network table comprising a list of networks in respect of which the mobile telecommunications apparatus is barred from registration for use in the registering step when selecting a network from a plurality of available networks for registration. 37. A method as claimed in claim 30, wherein the updating step comprises updating data stored in a SIM card of the mobile telecommunications apparatus. 38. A method as claimed in claim 35 comprising the step of obtaining network identification information for a plurality of networks including the local network which are available for registration, comparing the network identification information for the available networks with the updated preferred network table, selecting a preferred network from the available networks and, if the selected network is not the local network, re-registering with the selected network. 39. A method as claimed in claim 36 including the step of comparing the network identification information for the local network with the updated barred network table, and, if the local network is one of the barred networks, de-registering from the local network. 40. A method as claimed in claim 39 including the step of registering with a further one of the available networks. 41. A method as claimed in claim 1, wherein the telecommunications system is a GSM system. 42. A method as claimed in claim 41, wherein the request message is output using the SMS protocol. 43. A method as claimed in claim 41, wherein the request message is output using a USSD protocol. 44. A method as claimed in claim 41, wherein the request message is output using a UMTS protocol. 45. A method as claimed in claim 41, wherein a request message is output using a TCP/IP protocol. 46. A method as claimed in claim 41, wherein the request message is output using a WAP protocol. 47. A method as claimed in claim 1, wherein the apparatus comprises a mobile telephone. 48. A method as claimed in claim 1, wherein the apparatus comprises a SIM card having a processor and wherein the determining step is performed by the processor of the SIM card. 49. A method as claimed in claim 1 including the step of generating the request message to include call log information comprising accumulated data relating to use of the apparatus. 50. A method as claimed in claim 21, wherein the updating information is stored in a first portion of memory, and including the further step of storing a duplicate of the information in a second portion of the memory to constitute a backup memory. 51. A method as claimed in claim 50, wherein, when accessing the stored information, the contents of the first and second portions of memory are compared and, if different, the contents of the backup memory in the second portion of the memory are copied to the first portion of memory. 52. A method as claimed in claim 50, wherein the stored information is updated by an application which causes both first and second portions of memory to be updated and wherein other applications are prohibited from updating the second portion of memory. 53. A method as claimed in claim 22, wherein, if it is determined that the stored routing information cannot be utilised in selecting a preferred route via the local network, the apparatus is disabled from making outgoing telephone calls. 54. A method as claimed in claim 21 for use with a prepaid subscriber service in which the subscriber makes prepayment to acquire credit for making calls, wherein the stored routing information provides routing of calls via a prepayment platform operable to allow connection if credit remains and provides for call disconnection if credit expires. 55. A method as claimed in claim 54 including the step of outputting a validation request message to the prepaid platform and receiving a verification response therefrom before initiating the making of an outgoing call. 56. A method as claimed in claim 21, wherein updating the stored information comprises storing in a cache memory the information which is superseded by updating information. 57. A mobile telecommunications apparatus for use in a telecommunications system, comprising: registering means for registering with a network of the system which is a local network with respect to the current location of the apparatus; receiving means for receiving network identification information from the local network; storing means for storing routing information; routing means for determining a preferred route via the system for an outgoing call made by the apparatus based on a user input call number, the network identification information and the stored routing information; wherein the apparatus further comprises: determining means for determining whether the stored routing information requires updating, and output means for outputting a request message for receiving updating information for updating the stored routing information if the stored routing information is determined to require updating. 58. Apparatus as claimed in claim 57, wherein the determining means is operable to determine whether the stored routing information can be utilised in selecting a preferred route via the local network identified by the network identification information. 59. Apparatus as claimed in claim 57, wherein the determining means is operable to determine whether the network identification information indicates that the mobile telecommunications apparatus has moved to a country which is different from a previous country in which the apparatus was previously registered. 60. Apparatus as claimed in claim 57, wherein the determining means is operable to determine whether stored routing information relating to routing outgoing calls via the local network is valid by referring to expiry time information indicating a time at which the validity of the routing information expires. 61. Apparatus as claimed in claim 60, wherein the expiry information is stored in the apparatus in the form of a time at which the validity of the routing information expires and wherein the determining means is operable to determine the current time with reference to a clock and comparing the time with the expiry information. 62. Apparatus as claimed in claim 60, wherein the determining means is operable to determine whether a predetermined number of calls has been made by the apparatus since the last update of the routing information. 63. Apparatus as claimed in claim 60, wherein the determining means is operable to count the number of intervals of predetermined length for which the apparatus remains in operation and to determine the validity of the routing information to have expired when a predetermined number of the intervals has been counted. 64. Apparatus as claimed in claim 60, wherein the determining means comprises means for outputting a request message to obtain the expiry information, means for receiving a response message including the expiry information, and wherein the determining means is operable to determine from the expiry information whether the validity of the routing information has expired. 65. Apparatus as claimed in claim 57, wherein the determining means is operable to determine whether the network identification information indicates that the mobile telecommunications apparatus has registered with a network which is different from a network with which the apparatus was last registered. 66. Apparatus as claimed in claim 57 including receiving means for receiving a response message comprising the requested updating information for updating the stored routing information. 67. Apparatus as claimed in claim 66 including decrypting means for decrypting the received response message. 68. Apparatus as claimed in claim 66 including updating means for updating the stored routing information with the updating information. 69. Apparatus as claimed in claim 68, wherein the stored routing information comprises a routing table and wherein the updating means is operable to update the routing table. 70. Apparatus as claimed in claim 69, wherein the updating means is operable to update the routing table so as to include a field determining the time of expiry of the validity of the updated routing table. 71. Apparatus as claimed in claim 66 comprising a preferred network table which defines a list of preferred networks in order of preference for use in the registering step when selecting a network from a plurality of available networks for registration, and wherein the updating means is operable to update the preferred network table. 72. Apparatus as claimed in claim 66 comprising a barred network table comprising a list of networks in respect of which the mobile telecommunications apparatus is barred from registration for use in the registering step when selecting a network from a plurality of available networks for registration, and wherein the updating means is operable to update the barred network table. 73. Apparatus as claimed in claim 66 comprising a SIM card storing data which is updated by the updating means. 74. Apparatus as claimed in claim 71, wherein the receiving means is operable to obtain network identification information for a plurality of networks including the local network which are available for registration, the apparatus further comprising means for comparing the network identification information for the available networks with the updated preferred network table, selecting a preferred network from the available networks and, if the selected network is not the local network, re-registering with the selected network. 75. Apparatus as claimed in claim 72 including means for comparing the network identification information for the local network with the updated barred network table, and, if the local network is one of the barred networks, de-registering from the local network. 76. Apparatus as claimed in claim 75 including means for registering with a further one of the available networks after de-registering from the local network. 77. Apparatus as claimed in claim 11, wherein the telecommunications system is a GSM system. 78. Apparatus as claimed in claim 77, wherein the output means is operable to output the request message using the SMS protocol. 79. Apparatus as claimed in claim 77, wherein the output means is operable to output the request message using a USSR protocol. 80. Apparatus as claimed in claim 77, wherein the output means is operable to output the request message using a UMTS protocol. 81. Apparatus as claimed in claim 77, wherein the output means is operable to output the request message using a TCP/IP protocol. 82. Apparatus as claimed in claim 41, wherein the output means is operable to output the request message using a WAP protocol. 83. Apparatus as claimed in claim 11, wherein the apparatus comprises a mobile telephone. 84. Apparatus as claimed in claim 11, wherein the apparatus comprises a SIM card having a processor and wherein the determining means is constituted by the processor of the SIM card. 85. Apparatus as claimed in claim 11, wherein the output means generates the request message to include call log information comprising accumulated data relating to use of the apparatus. 86. Apparatus as claimed in claim 76, wherein the updating means is operable to store the updating information in a first portion of memory, and to store a duplicate of the information in a second portion of the memory to constitute a backup memory. 87. Apparatus as claimed in claim 86 comprising means for comparing the contents of the first and second portions of memory when and, if different, copying the contents of the backup memory to the first portion of memory. 88. Apparatus as claimed in claim 86, wherein the updating means is operable to update the stored information using an application which causes both first and second portions of memory to be updated and to prohibit other applications from updating the second portion of memory. 89. Apparatus as claimed in claim 58 comprising disabling means operable if it is determined that the stored routing information cannot be utilised in selecting a preferred route via the local network, to disable the apparatus from making outgoing telephone calls. 90. Apparatus as claimed in claim 57 for use with a prepaid subscriber service in which the subscriber makes prepayment to acquire credit for making calls, wherein the stored routing information provides routing of calls via a prepayment platform operable to allow connection if credit remains and provides for call disconnection if credit expires. 91. Apparatus as claimed in claim 90 comprising means for outputting a validation request message to the prepaid platform and means for initiating the making of an outgoing call which is conditional upon receiving a verification response therefrom. 92. Apparatus as claimed in claim 57, wherein the updating means comprises a cache memory for storing the information which is superseded by updating information. 93. A method of operating a mobile telecommunications apparatus in a telecommunications system wherein the apparatus comprises means for making telephone calls via the system and subscriber identity data stored in a first memory of the apparatus is used to associate telephone calls from the apparatus with a respective subscriber account, a set of subscriber identifiers being associated with the same account for use as the subscriber identity data, the method comprising: obtaining a network identifier from a network with which the apparatus is registered; determining which one of the set of subscriber identifiers is preferred for use as the subscriber identity data when the apparatus is registered to the network identified by the network identifier; and, if different from the subscriber identity data; replacing the subscriber identity data stored in the first memory with the preferred subscriber identifier. 94. A method as claimed in claim 931 wherein the step of replacing the subscriber identity data comprises obtaining the preferred subscriber identifier by sending a request message via the network and receiving a response message containing the preferred subscriber identifier. 95. A method as claimed in claim 93 further comprising storing in a second memory a look-up table of subscriber identifiers corresponding to respective network identifiers and wherein the determining step comprises accessing the look-up table. 96. A method as claimed in claim 95 comprising the step of determining whether an entry exists in the look-up table for the network identifier, determining whether the entry remains valid and, if not, sending a request message via the network and receiving a response message containing updating information for populating the table and including an entry for the network identifier. 97. A method as claimed in claim 93 comprising de-registering from the network when it is determined that the subscriber identity data does not correspond to the preferred subscriber identifier for the network identifier and subsequently re-registering after the subscriber identity data stored in the first memory has been replaced by the preferred subscriber identifier. 98. A method as claimed in claim 93, wherein the system comprises a GSM system and wherein the subscriber identity data is an IMSI. 99. A method as claimed in claim 93, wherein the first memory comprises a data file in a SIM card of the apparatus. 100. A mobile telecommunications apparatus for use in a telecommunications system wherein the apparatus comprises means for making telephone calls via the system and subscriber identity data is stored in a first memory of the apparatus for associating telephone calls from the apparatus with a respective subscriber account, a set of subscriber identifiers being associated with the same account for use as the subscriber identity data, the apparatus comprising: registering means for obtaining a network identifier from a network with which the apparatus is registered; means for determining which one of the set of subscriber identifiers is preferred for use as the subscriber identity data when the apparatus is registered to the network identified by the network identifier; and replacing means operable, if different from the subscriber identity data, to replace the subscriber identity data stored in the first memory with the preferred subscriber identifier. 101. Apparatus as claimed in claim 100, wherein the replacing means comprises sending means for sending a request message via the network and receiving means for receiving a response message containing the preferred subscriber identifier. 102. Apparatus as claimed in claim 100 further comprising a second memory storing a look-up table of subscriber identifiers corresponding to respective network identifiers and wherein the determining means is operable to access the look-up table. 103. Apparatus as claimed in claim 102 comprising means for determining whether an entry exists in the look-up table for the network identifier, means for determining whether the entry remains valid. 104. Apparatus as claimed in claim 100 comprising registering means operable to de-register from the network when it is determined that the subscriber identity data does not correspond to the preferred subscriber identifier and further operable to subsequently re-register after the subscriber identity data stored in the first memory has been replaced by the preferred subscriber identifier. 105. Apparatus as claimed in claim 100, wherein the system comprises a GSM system and wherein the subscriber identifier is an IMSI. 106. Apparatus as claimed in claim 100, wherein the first memory comprises a data file in a SIM card of the apparatus. 107. A telecommunications system for use in a method as claimed in claim 1, the system comprising a plurality of networks for providing communication with mobile telecommunications apparatus; and a control center for receiving request messages via one of said networks from mobile telecommunication apparatus registered with said network and for generating response messages containing updated data for storage in said apparatus. 108. A control center for use in a method as claimed in claim 1 comprising: message receiving means for receiving request messages from mobile telecommunications apparatus; a response generator for generating a response message; a transmitter for transmitting the response message; and a database containing data for inclusion in the response message and comprising updating data for updating at least one of: preferred network data; forbidden network data; routing data; and preferred subscriber identifiers. 109. A storage medium storing processor implementable instructions for instructing a processor to control a mobile telecommunications apparatus to perform all of the steps of claim 1. 110. A signal comprising processor implementable instructions for instructing a processor to control a mobile telecommunications apparatus to perform all of the steps of claim 1. 111. A SIM card comprising a processor and a memory 20 storing processor implementable instructions for operating the processor to control an apparatus to perform the steps of a method as claimed in claim 1.

Network conduit for providing access to data services

A web service conduit (3) receives data access requests from browsers (11) from hyperlinks on pages generated by web sites (4), converts the data access requests to web service access requests, and invokes the corresponding web services (40) with the web service access requests.

1. A method of providing access to a remote data service (40) over a network (14), comprising: a. receiving (S2; S11) from a remote terminal (11) a data access request, in a first format, identifying one or more parameters; b. converting the data access request into a second format; and c. forwarding (S5; S14) the data access request in the second format to the remote data service so as to perform a data access using the parameters; wherein the data access request is forwarded by the terminal (11) from a network service (4) remote from the terminal (11) and from the data service (40). 2. The method of claim 1, wherein the data access request is a read access request, the method further comprising: d. receiving (S5) from the remote data service (40), in said second format, a data message containing parameter values corresponding to the parameters identified in said data access request; e. converting the data message from said second format to said first format; and f. forwarding (S6) the data message in the first format to the remote terminal (11). 3. The method of claim 2, wherein the data access request is a write access request and contains parameter values corresponding to the parameters, and step c causes the parameter values to be written to the data service (40). 4. A method of providing access to a web service (40) over the Internet (14), comprising: a. receiving (S2; S11) from a browser (11) a data access request, in a first format, identifying one or more parameters; b. converting the data access request into a second format; and c. forwarding (S5; S14) the data access request in the second format to the web service (40) so as to perform a data access using the parameters; wherein the data access request is forwarded by the browser (I 1) from a web site (4) remote from the web service (40). 5. A method of providing access to a remote data service (40) over a network (14) using a link protocol, comprising sending to a remote terminal (11) a link to a network conduit (3), the link identifying one or more parameters and being formatted so as to cause the network conduit (3) to perform a data access at the remote data service (40) using the parameters when forwarded to the network conduit (3) by the remote terminal (11). 6. The method of claim 5, wherein the link is formatted to provide a data read access at the remote data service (40) such that parameter values corresponding to the parameters are read (S5) from the data service (40) by the network conduit (3) and are forwarded to the remote terminal (11). 7. The method of claim 6, further comprising sending to the remote terminal (11) a link to a program to cause the program to be loaded and executed by the remote terminal (11) so as to control the handling of the data read access by the terminal (11). 8. The method of claim 7, further including the step of receiving the parameter values from the remote terminal (11). 9. The method of claim 5, wherein the link is formatted to provide a data write access at the remote data service (40) and includes parameter values corresponding to the parameters, such that the parameter values are written to the data service (40) by the network conduit (3). 10. A method of providing access to a web service (40) over the Internet (14), comprising sending to a browser (11) an HTML link to a conduit (3), the link identifying one or more parameters and being formatted so as to cause the network conduit (3) to perform a data access at the web service (40) using the parameters when forwarded to the conduit (3) by the browser (11). 11. A method of performing a data access operation at a remote data service (40) over a network (14) from a terminal (11) using a link protocol, including: a. receiving (S1; S10) at the terminal (11) from a network service (4) a link directed to a network conduit (3) and identifying one or more parameters; and b. in response to user activation of the link, connecting (S2) the terminal (11) to the network conduit (3) over the network (14) and identifying the parameters to the network conduit (3), such that the parameters are forwarded by the network conduit (3) to the remote data service (40) so as to perform the data access operation. 12. The method of claim 11, wherein the data access operation is a read access operation, the method further including receiving (S6) parameter values corresponding to the parameters from the network conduit (3). 13. The method of claim 12, further including forwarding (S7) the parameter values to the network service (4). 14. The method of any one of claims 11 to 13, including receiving a link to a program, the link causing the program to be loaded and executed by the terminal (11) so as to control the handling of the read access operation by the terminal (11). 15. The method of claim 11, wherein the data access operation is a write access operation, and the link includes parameter values corresponding to the identified parameters, whereby the parameter values are written to the remote data service (40) by the network conduit (3). 16. A method of performing a web service access operation over the Internet (14) from a browser (11), including: a. receiving (S1; S10) at the browser (11) from a web site (4) a hyperlink to a conduit (3), the hyperlink identifying one or more parameters; and b. in response to user activation of the hyperlink, connecting (S2) the browser (11) to the conduit (3) over the Internet (14) and identifying the parameters to the conduit (3), such that the parameters are forwarded by the conduit (3) to the web service (40) so as to perform the data access operation. 17. A computer program for performing the method of any preceding claim. 18. A carrier bearing a computer program according to claim 17. 19. A system for providing access to a data service (40) over a network (14) using a link protocol, comprising: a. a network service (4) which provides a link to a conduit (3) including one or more parameters; b. a terminal (11) which receives said link and, in response to a user selection, sends the one or more parameters to the conduit (3); and c. said conduit (3) which receives said one or more parameters and sends a data access request including said one or more parameters to the data service (40) over the network (14).

<SOH> BACKGROUND OF THE INVENTION <EOH>Web services are a class of computer program that runs on a server computer connected to the Internet. Instead of using protocols such as HTTP and FTP to communicate with a user, web services are invoked by other programs which may be running on clients or other servers connected to the Internet. Web services may use an XML-based protocol such as SOAP, with a transport protocol such as HTTP. In a conventional architecture shown in FIG. 1 , a client browser 11 accesses web pages over the Internet 14 on a web site 4 , which invokes one or more web services 40 over the Internet 14 as part of the page generation process. Taking for example a web-based timetable lookup service, the user of the client browser 11 downloads a form page from the server 4 and fills in the lookup details of a timetable request. When the form is submitted, the client browser 11 sends a page request including the lookup details to the web site 4 using HTTP. The web site 4 invokes an underlying timetable lookup web service 40 using HTTP to get the data requested by the user, and formats the XML result into a form to be returned in a web page to the browser 11 . This architecture gives great flexibility, and allows the functionality of complex web sites to be distributed as underlying web services across geographic and commercial boundaries. However, there are certain technical requirements for the web site 4 to be able to invoke web services 40 : it must be able to make outgoing HTTP requests and to handle XML, SOAP and other protocols. These requirements can be a significant barrier to the use of web services. Furthermore, there is a great deal of freedom of data formats and protocols within web service standards such as HTTP, SOAP and XML, which makes the migration from one web service to another very difficult for a web site operator.

<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>Specific embodiments of the present invention will now be described with reference to the accompanying drawings, in which: FIG. 1 is a diagram of a conventional web service architecture; FIG. 2 is a diagram of a web service architecture in a general embodiment of the present invention; FIG. 3 is a diagram of the steps in a read operation in an embodiment of the present invention; FIG. 4 shows a web form as displayed on a browser for initiating the read operation; FIG. 5 shows a log-in form as displayed by the browser during the read operation; FIG. 6 shows a trusted web service check form as displayed by the browser during the read operation; FIG. 7 is a diagram of the steps in a write operation in an embodiment of the present invention; and FIG. 8 shows a web page displaying details of an event and a hyperlink to add the event to a web-based calendar. detailed-description description="Detailed Description" end="lead"?

Chemical vapor deposition devices and methods

Apparatus is described for rapidly coating a large area, or for rapidly producing a powder. In one embodiment, a liquid having a coating chemical is pumped from a liquid reservoir to a distribution manifold. From the distribution manifold, the liquid is carried under pressure to a geometric array, e.g., linear, of atomization nozzles. Flow equalization means are provided for equalizing the flow of the liquid delivered to each nozzle, and, preferably, means are provided for equalizing the temperature of the liquid delivered to each nozzle. The liquid, upon exiting the nozzles with the attendant pressure drop atomizes. The atomized liquid coats a substrate either in non-reacted or reacted form, or forms a powder. In a preferred embodiment, a solution of precursor chemical is reacted in a geometric array of flames produced at the nozzles, and a coating material produced in the flame coats the substrate, or a powder is formed. In another embodiment, vaporized precursor and vaporized are fed to a burner chamber having a linear exit slit. The vapor exiting the slit is burned, and material produced in a flame reaction are deposited on a substrate, or the powder formed is collected.

1. Apparatus for producing a uniform flow aerosol of a liquid comprising: liquid reservoir means containing the liquid at a first temperature; means for pressurizing said liquid; liquid distribution manifold means, said pressurizing means delivering pressurized liquid to said liquid distribution manifold means, an array of thermally controlled nozzles, said nozzles each being configured such that the liquid reaches a second temperature in each of said nozzles and at the exit of each of the nozzles undergoes a pressure drop to a pressure below the liquid's boiling point relevant to the second temperature, and is thereby atomized, means to deliver the liquid from said manifold means to each of said nozzles, said delivery means having flow equalization means which use back pressure, whereby liquid flow is more uniformly delivered to and from each of said nozzles. 2. Apparatus according to claim 1 having means to preheat the liquid delivered to each of said nozzles at an equalized temperature. 3. Apparatus according to claim 1 having energy means associated with each of said nozzels to promote a chemical reaction of at least one component of said liquid so as to produce a powder or coating material. 4. Apparatus according to claim 1 wherein each of said nozzles has associated means for delivering an oxidizing agent in the region of said nozzle, whereby at least one component of said atomized liquid is oxidized to produce a powder or coating material. 5. Apparatus according to claim 1 wherein each of said nozzles has associated means for delivering a combustible agent in the region of said nozzle, whereby at least one component of said atomized liquid is combusted to produce a powder or coating material. 6. Apparatus according to claim 1 wherein said flow equalization means comprises a plurality of conduits, a conduit from said manifold leading to each of said nozzles, the dimensions of each of said conduits being such that liquid of substantially equal flow is delivered to each of said nozzles. 7. Apparatus according to claim 1 wherein said array comprises a linear array of said nozzles. 8. Apparatus according to claim 1 wherein said array comprises a circular array of said nozzles. 9. Apparatus according to claim 1 wherein said array comprises a curved array of said nozzles. 10. Apparatus according to claim 1 wherein the pressure of the liquid in said manifold means is less than 5,000 psi. 11. Apparatus according to claim 1 wherein the pressure of the liquid in said manifold means is less than 10,000 psi. 12. Apparatus according to claim 1 wherein the pressure of the liquid in said manifold means is less than 20,000 psi. 13. Apparatus according to claim 1 wherein the variation of liquid flow rate between any two of said nozzles is less than 10%. 14. Apparatus according to claim 1 wherein the variation of liquid flow rate between any two of said nozzles is less than 5%. 15. Apparatus according to claim 1 wherein the variation of liquid flow rate between any two of said nozzles is less than 2%. 16. Apparatus for depositing a coating material comprising a burner unit comprising a liquid chamber and a liquid opening means exiting said liquid chamber wherein said opening means is elongated in at least one direction, means to provide to said liquid chamber at least one precursor chemical in liquid form and, means to ignite said liquid to produce said coating material from said precursor chemical. 17. The apparatus of claim 16 further having means to provide an oxidizing agent to said chamber in gaseous form. 18. The apparatus of claim 16 having pre-heating means to pre-heat at least one precursor chemical in liquid form fed to said gas chamber. 19. The apparatus of claim 16 wherein said elongated opening means is linear. 20. The apparatus of claim 16 wherein said opening means is an elongated slit 21. The apparatus of claim 16 wherein said opening means comprises a plurality of orifices. 22. The apparatus of claim 21 having means to provide said precursor chemical to said chamber in pressurized liquid form and said orifices are sized sufficiently small such that pressurized liquid exiting said chamber atomizes via some boiling as it leaves said orifices. 23. Apparatus for depositing a coating material comprising a burner unit comprising a fluid chamber and a fluid opening means exiting said fluid chamber wherein said opening is elongated in at least one direction, means to provide to said fluid chamber at least one precursor chemical in fluid form, and means to ignite said fluid to produce said coating material from said precursor chemical.

<SOH> BACKGROUND OF THE INVENTION <EOH>Vapor deposition is a well known method of producing coatings on substrates by exposing at least one surface of the substrate to a vapor phase of the deposition precursor. In CVD a chemical reaction of the precursor occurs on the surface of the substrate, or prior to deposition on the substrate to thereby form the coating on the substrate. The conventional methods of CVD require a chamber in which the substrate is held while the vaporized coating constituents are fed into the chamber. The portion of the vapor that does not deposit on the substrate to form the coating is exhausted out of the chamber where it must be collected for reuse or released into the atmosphere. In the more recently developed CCVD techniques, a combustion source (flame, plasma etc.) is used to promote the chemical reaction in the vicinity of the substrate. In this manner the coating species is formed in close proximity to the substrate such that a larger portion of the coating precursor is deposited on the substrate. This is due to the increased control of the deposition reactions and temperatures. Many coatings will only form at a specific deposition temperature, and below this temperature, the coating will not form on the substrate and is exhausted away. With CCVD, changes of this deposition temperature can be made much quicker, as the surface of the substrate is (in some cases) directly heated by the combustion source such that as the combustion source forms the coating species, and the majority of the precursor forms the coating with very little of the precursor material needing to be exhausted. This can allow open atmosphere depositions without the need for reclamation of the exhausted materials. These processes are detailed in U.S. Pat. Nos. 5,652,021; 5,858,465; 5,863,604; 5,997,956; 6,013,318 and 6,132,653, and issued to Hunt et al. These patents, which are hereby incorporated by reference, disclose methods and apparatus for CCVD of films and coatings wherein a reagent and a carrier medium are mixed together to form a reagent mixture. The mixture, along with an oxidizing agent, is then ignited to create a flame or the mixture is fed to a plasma torch. The energy of the flame or torch vaporizes the reagent mixture and heats the substrate as well. These CCVD techniques have enabled a broad range of new applications and provided new types of coatings, with novel compositions and improved properties. In addition, these technologies are also useful in the formation of powders, as described in the above-referenced patents. A limitation of these previous CCVD processes is that the area coated by the combustion source is somewhat limited to the size of the combustion source, at least within reasonable time frames. The present invention is directed to overcoming this limitation by increasing the effective area coated by the combustion source, thereby increasing the overall rate of the deposition or powder production to a level appropriate for manufacturing processes. The apparatus of the invention is useful in any deposition process in which a liquid is atomized and the atomized liquid is used to form a coating. While a flame is one energy source that may be used to promote chemical reaction of a precursor chemical(s) in liquid form, other energy sources, such as heated gases, induction heaters, etc. may be used, particularly if a non-oxidizing reaction is to be promoted.

<SOH> SUMMARY OF THE INVENTION <EOH>To achieve the above objectives, the present invention provides for methods and apparatus that include at least one increased dimension of the combustion source, particularly for CCVD processes. It should be understood that the various embodiments of the present invention are useful for other methods of deposition such as pyrolytic spray or CVD, and the following detailed description is most specific to the CCVD method for simplicity only. Furthermore, the described devices are also useful in the production of powders when used in conjunction with well known powder collection apparatus. When used to deposit coatings, the present invention allows a greater area to be coated by a single pass of the CCVD apparatus. A first embodiment is an integrated discrete linear flame that is comprised of a plurality of CCVD nozzles aligned in a linear array. Each of the discrete nozzles must be precisely controlled in terms of pressure and temperature to insure a uniform deposition rate and composition across the width of the CCVD apparatus. A second embodiment of the apparatus is in the form of a continuous linear flame wherein vaporized coating material is fed into an extended tube with a flame slit extending along the length of the tube. The coating material ignites as it exits the slit, thereby forming a continuous linear flame that provides a uniform deposition rate and composition along the length of the flame. Both of the embodiments provide an efficient method of producing a large area uniform coating on large substrates from a single chemistry solution, thus increasing deposition rates to a level suitable for manufacturing purposes. In the first embodiment, several CCVD nozzles are arranged side-by-side. The typical deposition area or “footprint” of a single CCVD nozzle is dependent on several factors including but not limited to the material being deposited and distance from the substrate. In its simplest form, the multiple nozzle array consists of two deposition nozzles. The second nozzle may only increase the deposition area by a factor of 1.25, due to interaction effects between the two nozzles. The actual material throw rate, however, will typically increase by a factor close to 2.0. In order to maximize the coating uniformity, deposition area, and overall deposition material throw rate, the spacing between nozzles must be determined through experimentation for each particular application. Should the distance between nozzles for one of these factors interfere with another requirement, (such as maximizing deposition area at the expense of uniformity), two banks of nozzles may be arranged in succession, with the centerlines of the nozzles being offset to provide uniform coverage. To provide uniform deposition between the multiple nozzles, the temperature and flow of each nozzle must be held constant with reference to the other nozzles. To achieve this requirement, back-pressure regulators and a block heater are employed. Each of the nozzles is fed from a common, chemistry solution, distribution manifold. Between the manifold and each nozzle, a back-pressure regulator is provided. The back-pressure regulator may be a standard pressure regulator, a needle valve, or a coiled tubing. Each of the nozzles has an inherent pressure drop in the fluid as it flows from the common manifold and out the exit of the nozzle. If this pressure drop is for example 100 psi in one nozzle, and 200 psi in a second nozzle, then the flow rate differential between these nozzles would be 50% (assuming an equal cross sectional flow area). By providing back-pressure regulators that increase the pressure drop to around 1000 psi, this same pressure drop difference of 100 psi only causes a pressure drop variation of 900 to 100 psi or a flow rate differential of 10%. In the preferred embodiment, the back-pressure regulators are in the form of coiled, small inner diameter tubes, the lengths of which determine the pressure drop for each nozzle. Thus to adjust for equal pressure drops and resulting uniform deposition material flow, the relative length of each tube is adjusted accordingly. More specifically, since the effective flow rate of each orifice from a central manifold is inversely related to its back pressure, it is desired to maintain the back pressure to each line as closely as possible. Of particular importance to the present invention is the capability to maintain a uniform flow through multiple orifices from a single delivery system when the pressure drop across the orifices is not equal or as the pressure variation of an orifice varies over time (accumulation of material). Such a system exists, as per the present invention, when atomizing by releasing a thermally controlled liquid into a volume that is at a pressure below its boiling point relative to the controlled temperature of the liquid. Products to form the orifice are made with a large variation which results in different back pressures at the desired flow rate. Used alone, these orifices do not provide the desired control of the liquid through each nozzle. This is further complicated by the issue of heated liquids that can have variable amounts of material forming on the inside surfaces of the nozzles which causes a time variable back pressure at a constant flow rate. It is not desired to have a pump or liquid mass flow controllers (if one were found that works at the required pressures and flow rates) for each nozzle as the cost and maintenance becomes prohibitive. The current system works by have a precise pressure drop up stream from the nozzle to act in providing uniform flow to each nozzle. Downstream of this flow controlling pressure drop the liquid state would still be maintained so that material will not form over time on the walls of the flow controlling section. The pressure drop across this flow control region needs to be substantially higher than that of the orifice, so that any orifice pressure changes are minor in comparison to the constant pressure of the flow control section. Thus if orifice back pressure can vary from 100 psi to 300 psi, then the flow control pressure drop must be at least 2000 psi (10×) to maintain at least a 10% or better flow control through each nozzle. For even more uniform flow control, a 4000 psi (20×) pressure drop can be used (5% flow variation). For yet even more uniform flow control, a 10000 psi (50×) pressure drop can be used (2% flow variation). In some cases it will be desired to go to even a 100× factor (this would be a manifold pressure of 20,000 psi with a resultant pressure of 100 to 300 psi after the flow control section for the above example). Of course as higher pressures are needed the cost of the system increases, so added tolerance comes with a cost, complexity and size impact. This above example, without the flow control system of the present invention, would have a flow variation of about +/−50%. Available components (tubing, fittings, pumps, valves, etc.) for the present systems have significant price jumps at 5000, 10000, and 20000 psi. Thus there is desire to design a system with the exact level of flow variation required to produce the desired coating or powder, to avoid excessive costs. The flow rate of the deposition material is also dependent on other factors, such as density and viscosity, that are in turn dependent on the temperature of the fluid. In order to maintain a similar temperature between the nozzles, a block heater is used. Each of the nozzles and a thick walled tube leading to the nozzle from the coiled tube is encased in a block of material that is thermally conductive (such as a dense metal). Alternatively, precision-machined orifices may be drilled into the material to form the nozzles and passageway between the nozzles and the coiled tubes. Resistive element heaters are used to heat the block of material such that the fluid flowing through the nozzles is brought to a thermodynamically metastable state. A liquid is in a metastable state if its temperature at the exit of the nozzle is higher than its saturation temperature for a given pressure. By heating the fluid to this temperature, rapid expansion of the liquid is achieved which results in quick and uniform atomization of the precursor material. It should also be understood from the following description, that the apparatus and methods of the present invention can be used to form coatings using deposition techniques other than CCVD, as the use of a combustion source is not necessary for forming some materials. The linear deposition apparatus provides a uniform material deposition rate along its length, thereby forming a more uniform coating than could be previously achieved using prior art devices and techniques. This is due to the pressure and temperature regulation provided between the array of nozzles that form the integrated linear deposition apparatus. A second embodiment of the present invention provides a continuous linear deposition apparatus for applying coatings using precursor chemical-containing fluids.

Gel emulsions in the form of o/w emulsions having a hydrocolloid content

The invention relates to cosmetic preparations in the form of gel emulsions on preparations of the O/W emulsion type having a hydrocolloid content. Said hydrocolloids are selected from the group of preparations based on (i) hydroxyethylcellulose, (ii) xanthan gum, (iii) carbomer. Said preparations further contain (iv) one or more lipids, (v) one or more non-ionic and/or anionic emulsifiers with HLB values between 8 and 16, the overall content of the emulsifiers not exceeding 1.5% by weight.

1-9. (canceled) 10. A composition which is present as a gel emulsion, wherein the composition is based on an O/W emulsion and comprises the following components: (i) hydroxyethylcellulose; (ii) xanthan gum; (iii) carbomer; (iv) one or more lipids; and (v) one or more emulsifiers having HLB values of from 8 to 16, said emulsifiers comprising at least one of a nonionic and an anionic emulsifier; wherein a ratio of (i):(ii):(iii) is a:b:c and a, b and c independently are numbers of from 1 to 5 and b may additionally be equal to 0; components (i), (ii) and (iii) are present in a total concentration of from 0.25% to 1.5% by weight, component (iv) is present in a concentration of from 3.0% to 20.0% by weight, and component (v) is present in a concentration not exceeding 1.5% by weight, all percentages being based on a total weight of the composition. 11. The composition of claim 10, wherein a, b and c independently are numbers of from 1 to 3. 12. The composition of claim 11, wherein components (i), (ii) and (iii) are present in a total concentration of from 0.5% to 1.0% by weight. 13. The composition of claim 10, wherein component (iv) is present in a concentration of from 7.5% to 15% by weight. 14. The composition of claim 10, wherein the one or more emulsifiers (v) comprise one or more emulsifiers having HLB values of from 10 to 12. 15. The composition of claim 10, wherein component (v) is present in a concentration of from 0.5% to 1.0% by weight. 16. The composition of claim 15, wherein component (v) comprises glyceryl stearate citrate. 17. The composition of claim 10, wherein the composition further comprises (vi) one or more fatty alcohols. 18. The composition of claim 17, wherein component (vi) is present in a concentration of up to 10% by weight, based on the total weight of the composition. 19. The composition of claim 17, wherein component (vi) is present in a concentration of up to 5.0% by weight. 20. The composition of claim 17, wherein component (vi) is present in a concentration of up to 3.0% by weight. 21. The composition of claim 13, wherein component (iv) comprises less than about 30% by weight of polar lipids. 22. The composition of claim 13, wherein component (iv) comprises at least one lipid which has an interfacial tension with water of from about 20 to about 30 mN/m. 23. The composition of claim 10, wherein the composition comprises an emulsion having an oil phase which comprises at least 50% by weight of at least one of petrolatum, paraffin oil and polyolefins. 24. The composition of claim 23, wherein the oil phase comprises at least 75% by weight of at least one of petrolatum, paraffin oil and polyolefins. 25. The composition of claim 10, wherein the composition comprises an emulsion having an oil phase which comprises a silicone oil. 26. The composition of claim 10, wherein the composition further comprises at least one antioxidant. 27. The composition of claim 26, wherein the at least one antioxidant is present in a concentration of from 0.001% to 30% by weight, based on the total weight of the composition. 28. The composition of claim 12, wherein the composition further comprises at least one antioxidant in a concentration of from 0.05% to 20% by weight, based on the total weight of the composition. 29. The composition of claim 10, wherein the composition further comprises at least one antioxidant in a concentration of from 1% to 10% by weight, based on the total weight of the composition. 30. The composition of claim 27, wherein the at least one antioxidant comprises an oil-soluble antioxidant. 31. The composition of claim 26, wherein the at least one antioxidant comprises at least one of vitamin A and derivatives thereof. 32. The composition of claim 31, wherein the at least one of vitamin A and derivatives thereof is present in a concentration of from 0.001% to 10% by weight, based on the total weight of the composition. 33. The composition of claim 26, wherein the at least one antioxidant comprises at least one of vitamin E and derivatives thereof. 34. The composition of claim 33, wherein the at least one of vitamin E and derivatives thereof is present in a concentration of from 0.001% to 10% by weight, based on the total weight of the composition. 35. The composition of claim 10, wherein the composition further comprises at least one UV filter substance which is at least one of a UV-A filter substance, a UV-B filter substance and an inorganic pigment. 36. The composition of claim 35, wherein the at least one UV filter substance is present in a total concentration of from 0.1% to 30% by weight, based on the total weight of the composition. 37. A composition which is present as a gel emulsion, wherein the composition is based on an O/W emulsion and comprises the following components: (i) hydroxyethylcellulose; (ii) xanthan gum; (iii) carbomer; (iv) one or more lipids; (v) one or more emulsifiers having HLB values of from 10 to 12, said emulsifiers comprising at least one of a nonionic and an anionic emulsifier; and (vi) one or more fatty alcohols; wherein a ratio of (i):(ii):(iii) is a:b:c and a, b and c independently are numbers of from 1 to 3 and b may additionally be equal to 0; components (i), (ii) and (iii) are present in a total concentration of from 0.5% to 1.0% by weight, component (iv) is present in a concentration of from 7.5% to 15% by weight, component (v) is present in a concentration of from 0.5% to 1.0% by weight, and component (vi) is present in a concentration of up to 3.0% by weight, all percentages being based on a total weight of the composition. 38. The composition of claim 37, wherein component (v) comprises glyceryl stearate citrate. 39. The composition of claim 37, wherein component (iv) comprises less than about 30% by weight of polar lipids. 40. The composition of claim 37, wherein the composition further comprises at least one antioxidant in a concentration of from 1% to 10% by weight, based on the total weight of the composition. 41. The composition of claim 40, wherein the at least one antioxidant comprises at least one of vitamin A, vitamin E and derivatives thereof. 42. The composition of claim 37, wherein the composition further comprises at least at least one of a UV-A filter substance, a UV-B filter substance and an inorganic pigment in a total concentration of 1.0% to 6.0% by weight, based on the total weight of the composition. 43. A cosmetic preparation which comprises the composition of claim 10. 44. The cosmetic preparation of claim 43, which comprises one of a skin protection cream, a cleansing cream, a cleansing milk, a sunscreen lotion, a nourishing cream, a day cream and a night cream. 45. A dermatological preparation which comprises the composition of claim 10. 46. A sunscreen preparation which comprises the composition of claim 35. 47. A lip care product which comprises the composition of claim 10. 48. A hair care product which comprises the composition of claim 10. 49. An aerosol container which comprises the composition of claim 10 and a propellant. 50. The aerosol container of claim 49, wherein the propellant comprises a liquefied hydrocarbon. 51. A roll-on device which comprises the composition of claim 10. 52. A method of treating and protecting skin, wherein the method comprises an application onto the skin of a composition which is present as a gel emulsion, is based on an O/W emulsion and comprises the following components: (i) hydroxyethylcellulose; (ii) xanthan gum; (iii) carbomer; (iv) one or more lipids; and (v) one or more emulsifiers having HLB values of from 10 to 12, said emulsifiers comprising at least one of a nonionic and an anionic emulsifier; wherein a ratio of (i):(ii):(iii) is a:b:c and a, b and c independently are numbers of from 1 to 3 and b may additionally be equal to 0; components (i), (ii) and (iii) are present in a total concentration of from 0.25% to 1.5% by weight, component (iv) is present in a concentration of from 3.0% to 20.0% by weight, and component (v) is present in a concentration not exceeding 1.5% by weight, all percentages being based on a total weight of the composition.

Antisense modulation of wrn espression

Antisense compounds, compositions and methods are provided for modulating the expression of WRN. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding WRN. Methods of using these compounds for modulation of WRN expression and for treatment of diseases associated with expression of WRN are provided.

1. A compound 8 to 50 nucleobases in length targeted to a nucleic acid molecule encoding WRN, wherein said compound specifically hybridizes with and inhibits the expression of WRN. 2. The compound of claim 1 which is an antisense oligonucleotide. 3. The compound of claim 2 wherein the antisense oligonucleotide has a sequence comprising SEQ ID NO: 13, 14, 15, 17, 18, 19, 20, 21, 22, 23, 25, 26, 27, 28, 29, 30, 31, 33, 34, 35, 36, 38, 39, 40, 42, 44, 45, 46, 47, 48, 49, 51, 52, 53, 54, 55, 56, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 85, 86, 87, 89 or 90. 4. The compound of claim 2 wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage. 5. The compound of claim 4 wherein the modified internucleoside linkage is a phosphorothioate linkage. 6. The compound of claim 2 wherein the antisense oligonucleotide comprises at least one modified sugar moiety. 7. The compound of claim 6 wherein the modified sugar moiety is a 2′-O-methoxyethyl sugar moiety. 8. The compound of claim 2 wherein the antisense oligonucleotide comprises at least one modified nucleobase. 9. The compound of claim 8 wherein the modified nucleobase is a 5-methylcytosine. 10. The compound of claim 2 wherein the antisense oligonucleotide is a chimeric oligonucleotide. 11. A compound 8 to 50 nucleobases in length which specifically hybridizes with at least an 8-nucleobase portion of an active site on a nucleic acid molecule encoding WRN. 12. A composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier or diluent. 13. The composition of claim 12 further comprising a colloidal dispersion system. 14. The composition of claim 12 wherein the compound is an antisense oligonucleotide. 15. A method of inhibiting the expression of WRN in cells or tissues comprising contacting said cells or tissues with the compound of claim 1 so that expression of WRN is inhibited. 16. A method of treating an animal having a disease or condition associated with WRN comprising administering to said animal a therapeutically or prophylactically effective amount of the compound of claim 1 so that expression of WRN is inhibited. 17. The method of claim 16 wherein the disease or condition is a hyperproliferative disorder. 18. The method of claim 17 wherein the hyperproliferative disorder is cancer. 19. The method of claim 16 wherein the disease or condition involves premature aging. 20. The method of claim 16 wherein the disease or condition is viral infection.

<SOH> BACKGROUND OF THE INVENTION <EOH>Genomic integrity is critical to the health and survival of any organisms and cells have evolved multiple pathways for the repair of DNA damage. One class of enzymes involved in the maintenance of genomic integrity and stability are DNA helicases. These proteins play important roles in DNA replication, repair, recombination and transcription by unwinding duplex genomic strands allowing the repair machinery access to damaged or mispaired DNA. For example, the RecQ family of helicases has been shown to be important players in linking cell cycle checkpoint responses to recombination repair (Chakraverty and Hickson, BioEssays, 1999, 21, 286-294; Frei and Gasser, J. Cell Sci., 2000, 113, 2641-2646; Wu et al., Curr. Biol., 1999, 9, R518-520). More recently, these helicases have been implicated in the process of posttranscriptional gene silencing (PTGS) (Cogoni and Macino, Science, 1999, 286, 2342-2344). In this process, the helicase is required to separate the double-stranded DNA (dsDNA) before any hybridization and silencing mechanism could be initiated. The RecQ family consists of five members and can be divided into two distinct groups according to whether they contain an additional carboxy- or amino-terminus group. One class containing the longest members of the family include genes known to be defective in several syndromes including the BLM gene in Bloom's syndrome, the WRN gene in Werner's syndrome and the RECQ4 gene in Rothmund-Thompson syndrome. Mutations in these genes lead to an increase in the incidence of cancer as well as other physiologic abnormalities (Karow et al., Curr. Opin. Genet. Dev., 2000, 10, 32-38; Kawabe et al., Oncogene, 2000, 19, 4764-4772). The second class contains the RECQL gene and the RECQ5 gene which encode little more than the central helicase domain and have not been associated with any human disease. WRN (also known as RECQL3) was originally identifed by positional cloning as the gene responsible for Werner's syndrome and localized to chromosome 8p12 (Yu et al., Science, 1996, 272, 258-262.). Werner's syndrome is a rare autosomal recessive disorder characterized by symptoms similar to premature aging including atherosclerosis, osteoporosis, type II diabetes, cataracts and cancers (Goto, Clin. Exp. Rheumatol., 2000, 18, 760-766; Oshima, BioEssays, 2000, 22, 894-901; Shen and Loeb, Trends Genet., 2000, 16, 213-220). In an effort to better define the role of the WRN gene, Marciniak et al. determined the subcellular localization of the protein using indirect immunofluorescence and a polyclonal antibody. These studies revealed a predominant nuclear localization and no difference in localization was detected in normal compared to transformed human cell lines (Marciniak et al., Proc. Natl. Acad. Sci. U.S.A., 1998, 95, 6887-6892). In addition to having helicase activity, WRN also contains intrinsic exonuclease activity and has been shown to bind single-stranded DNA with higher affinity than double-stranded DNA (Orren et al., Nucleic Acids Res., 1999, 27, 3557-3566; Shen et al., J. Biol. Chem., 1998, 273, 34139-34144). Disclosed in U.S. Pat. No. 6,090,620 are the nucleic acid molecules encoding the WRN gene as well as WRN gene products, expression vectors, viral vectors and host cells suitable for expressing the WRN gene products (Fu et al., 2000). Disclosed and claimed in European Patent EP 1135502 is a nucleotide sequence encoding a protein characterized in having a silencing activity comprising a recQ helicase domain, or a functional portion of, wherein the domain is at least 30% homologous with the amino acid sequence of the recQ helicase domain of the product of the qde-3 gene from Neurospora crassa, or its complementary sequence, an expression vector comprising said nucleotide sequence in the sense or antisense orientation for expression of said sequence in bacteria, fungi, plants, and animals, organisms transformed said vector, and use of the nucleotide sequence to modulate the gene silencing in plants, animals and fungi. The human WRN protein is also generally disclosed. Disclosed and claimed in U.S. Pat. No. 6,228,583 are methods of identifying an agent which inhibits the replication or accumulation of ribosomal DNA circles in a yeast cell or in a mammalian cell, wherein the agent inhibits the formation of ribosomal DNA circles in a yeast cell with a mutation in the SGS1 gene, and wherein the agent extends the lifespan of the yeast or mammalian cell. The yeast SGS1 gene is the homolog of human WRN. Further disclosed is the homologous mouse gene, mWRN, and in one embodiment, the invention relates to a method of screening a compound for the ability to alter life span comprising administering the compound to a mouse with a suppressed level of mWRN expression and assaying the mouse for altered life span, wherein antisense nucleic acids are generally disclosed as a means of reducing gene expression. Currently, there are no known therapeutic agents which effectively inhibit the synthesis of WRN. There are reports of DNA minor groove-binding drugs which inhibit the helicase activity of WRN (Brosh et al., Nucleic Acids Res., 2000, 28, 2420-2430). WRN-deficient mice display reduced embryonic survival while live-born mice otherwise appear normal during the first year of life (Lebel and Leder, Proc. Natl. Acad. Sci. U.S.A., 1998, 95, 13097-13102). While mutations and targeted disruptions resulting in altered protein expression in the WRN gene are responsible for Werner's syndrome, the normal function of the WRN gene product and its regulation are still unclear. It is, however, believed to be involved in DNA metabolism and is therefore a potential therapeutic target in conditions involving the production of aberrant DNA products, including the recognition of foreign DNA products as is the case upon viral infection. Consequently, there remains a long felt need for agents capable of effectively inhibiting and/or modulating WRN function. Antisense technology is emerging as an effective means for reducing the expression of specific gene products and may therefore prove to be uniquely useful in a number of therapeutic, diagnostic, and research applications for the modulation of WRN expression. The present invention provides compositions and methods for modulating WRN expression.

<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is directed to compounds, particularly antisense oligonucleotides, which are targeted to a nucleic acid encoding WRN, and which modulate the expression of WRN. Pharmaceutical and other compositions comprising the compounds of the invention are also provided. Further provided are methods of modulating the expression of WRN in cells or tissues comprising contacting said cells or tissues with one or more of the antisense compounds or compositions of the invention. Further provided are methods of treating an animal, particularly a human, suspected of having or being prone to a disease or condition associated with expression of WRN by administering a therapeutically or prophylactically effective amount of one or more of the antisense compounds or compositions of the invention. detailed-description description="Detailed Description" end="lead"?

Method and Device for Carrying Out the Functional Check and Functional Checking of a Technical Unit

The invention relates to a method for carrying out the functional check and a functional checking of a technical unit, comprising the following steps, creation of a check program, by selection and storage of control commands from a number of control commands held in a data bank, depending upon the unit for checking, input of the check program which defines a number of control commands and the sequence thereof into a controller, output of the control commands to the unit in the defined sequence and receiving and displaying of returned information from the unit. The invention further relates to a corresponding device.

1-47. (canceled) 48. A method for performing a functionality test of a technical unit including: providing a plurality of control commands stored in a data base; selecting control commands from said plurality of control commands as a function of a technical unit to be tested; storing said selected control commands; preparing a testing program for the technical unit using said stored, selected control commands; providing a control unit; transferring said testing program to said control unit; connecting said control unit with the technical unit to be tested; outputting said selected control commands to the technical unit in a defined sequence; and receiving and displaying acknowledgment information from the technical unit. 49. A method of performing a functionality test of a technical unit including: providing a plurality of control commands stored in a data base; selecting control commands from said plurality of control commands as a function of a technical unit to be tested; storing said selected control commands; preparing a testing program for the technical unit using said stored, selected control commands; providing a control unit; transferring said testing program to said control unit; providing a bus identical to a bus usable with said technical unit for communicating with other components of a finished printing press; outputting said selected control commands to the technical unit using said bus in a defined sequence; and receiving and displaying acknowledgment information from the technical unit. 50. The method of claim 48 further including preparing a recording of said acknowledgment information received from the technical unit. 51. The method of claim 49 further including preparing a recording of said acknowledgment information received from the technical unit. 52. The method of claim 50 further including all of said selected control commands in said recording. 53. The method of claim 51 further including all of said selected control commands in said recording. 54. The method of claim 48 further including represented said control commands in one of text form and graphic symbols. 55. The method of claim 49 further including represented said control commands in one of text form and graphic symbols. 56. The method of claim 48 further including displaying said control commands during performance of said testing program. 57. The method of claim 49 further including displaying said control commands during performance of said testing program. 58. The method of claim 48 further including providing a bus and using said bus for transmitting said control commands of said testing program to said technical unit. 59. The method of claim 48 further including providing said testing program with only control commands intended for said technical unit. 60. The method of claim 49 further including providing said testing program with only control commands intended for said technical unit. 61. The method of claim 48 including testing at least one of the functions of the technical unit selected from the functions including: engaging and disengaging inking rollers, engaging and disengaging inking ductors, engaging and disengaging ink aspirators, engaging and disengaging dampening ductors, operating drive units of inking rollers, operating drive units of inking ductors, operating drive units of ink aspirators, operating drive units of dampening ductors, operating drive units of cylinders, operating drive units for circumferential registrations of cylinders, operating drive units for diagonal registrations of cylinders, operating drive units for axial registrations of cylinders, operating locking devices of dressings on cylinders, operating dampening water control, and the addition of printing plates. 63. A device for functionality testing of a technical unit comprising: at least one displaying screen for displaying input masks; at least one memory unit; and a control unit with a testing program, said input masks representing a plurality of control commands in one of text form and graphic symbols, said memory unit being adapted to store said control commands selected by said input masks, as a function of a unit to be tested, as a testing program, said testing program being prepared using one of a program different from said testing program and a development computer. 64. The device of claim 63 further including a first interface adapted to transmit control commands to said technical unit and to secure acknowledgment information from said technical unit, said first interface containing a second interface, said second interface being adapted for receiving said testing program, said testing program defining a plurality of control commands and their sequence, said second interface being adapted to output said control commands defined in said testing sequence via said first interface in a defined sequence. 65. The device of claim 64 further including a third interface adapted for outputting acknowledgment information received from said technical unit. 66. The device of claim 65 further wherein said third interface is adapted to output all of said control commands issued by said first interface. 67. The device of claim 66 further including a writing device for a data carrier, not able to be overwritten, connected to said third interface. 68. The device of claim 63 including a second display screen adapted to display control commands and parameters of control commands. 69. The device of claim 63 wherein said memory unit stores said input masks. 70. The device of claim 63 further including a buffer memory adapted for the buffer memory adapted for the intermediate storage of not yet executed memory commands. 71. The device of claim 63 wherein said technical unit is a part of a printing press. 72. The device of claim 63 wherein said technical unit is a printing unit. 73. The device of claim 63 wherein said technical unit is a folding unit.

<SOH> BACKGROUND OF THE INVENTION <EOH>At present, so-called test control consoles are employed for the functionality testing of various units, such as, for example, printing presses or parts thereof, which test control consoles make it possible for an operator, identified as consumer, to control the individual functions of the unit to be tested and to check whether the unit reacts in the intended manner to the settings input at the test control console. Customarily the settings required for a complete functionality test are listed in a test protocol, in which test protocol the consumer can enter, if desired, the reactions of the unit to the settings performed and reported by the test control console in order to document, in this way, the correct or incorrect functioning of the unit. This procedure is lengthy, time-consuming and contains uncertainties which never completely eliminate the possibility that the consumer does not make erroneous settings, does not correctly enter results in the protocol, or omits test steps. DE 197 25 916 A1 describes a diagnostic device for electrically controlled systems, in which additional libraries are added to a diagnostic program. The automatic running of several functions and the input of plain language are not disclosed. DE 195 22 937 C2 discloses a diagnostic system for a motor vehicle with a “fixed” diagnostic program. No plain language input is provided. A testing device for an electro-medical apparatus is known from DE 36 02 171 A1. Access to a data bank for preparing a test program is not possible. U.S. Pat. No. 5,588,109 discloses a method and a device for the remote diagnosis of units already placed in an installation by the customer. A unit to be diagnosed can be selected from a predetermined number. Parameters are assigned to the unit, whose measured values are then detected by the device by use of a remote diagnosis and are shown on a display. A user can either make notes in a “memo field”, or can select further parameters to be measured from a predetermiend number of parameters and display them. By use of a further tool, the user can select a testing program from a predetermined number, which has a fixed sequence of steps.

<SOH> SUMMARY OF THE INVENTION <EOH>The object of the present invention is directed to providing a method and a device for performing a functionality test, as well as to a functionality test of a technical unit. In accordance with the present invention, this object is attained by the provision of a method for performing a functionality test of a technical unit. A testing program is prepared using control commands selected from a larger group of control commands which are stored in a data bank. The testing program is prepared as a function of the unit to be tested. The testing program can be transferred to a control unit. The technical unit can be connected to the control unit and can be tested. A bus may be used for transmitting the control commands, with the bus being the same as the type of bus which the technical unit uses to communicate with other components of a finished printing press. A display screen is used to display input masks that represent control commands in text form, or as graphic symbols. A memory element stores the control commands selected by the input masks, as a function of the unit to be tested, as a testing program. The advantages to be gained by the present invention lie, in particular, in that it assures a complete and correct performance of the functionality test with only little outlay for time and work. Since the control unit includes a complete testing program, the accidental omission of individual test steps is not possible. A time savings results, inter alia, from the fact that the control unit can immediately issue a command, which is already known to the control unit from the testing program, to the unit to be tested as soon as the previous test step is completed. It is thus not necessary to wait until an operator has made the settings required for performing the next steps. In order to be able to generate a complete and a dependable protocol regarding the results of the individual test steps, it is useful for the control unit to have a third interface for issuing acknowledgement information which it has received from the unit, in response to an issued control command. The totality of the acknowledgement information can be considered to be a testing protocol, by the use of which, the correct performance of the functionality test can be verified at any desired later time. To make this later test easier, it is useful not only if the acknowledgement information can be issued via this third interface, but moreover also the issuance of all control commands issued to the unit via the first interface, which have resulted in the respective acknowledgements. In this way, it is possible to provide proof, beyond all doubt, regarding the performance of every individual step of the test program. For creating the protocol, a writing device for a data carrier, which cannot be overwritten, is preferably connected to the third interface. In the simplest embodiment, this writing device can be a printer. For storing extensive protocols in a space-saving manner, the employment of a CD burner can also be considered. To make it possible for a user to follow the course of the testing program, the control unit can be equipped with a display screen for use in displaying control commands issued to the unit, if desired also inclusive of the parameters of such control commands. In the simplest configuration this control unit can be realized as a computer, and in particular as an inexpensive workplace computer, which computer communicates with the technical unit to be tested via a bus. In order to prevent any tampering with the progress of the testing program by the user, the control unit may be configured so that it does not have any interface for use for entering control commands by a user. This can be realized, in the case of a control unit in the form of a workplace computer, if the conversion of the testing program into commands, which commands can be performed by the unit, and the issuance of such commands, is performed under the control of a control program, which control program does not accept any commands from a user during the running of the testing program. It is also possible to provide the user the possibility, within reason, of affecting the control commands sent to the unit, in particular if these control commands are also entered into the protocol to be prepared. To make the fixing of such control commands, or their parameters, easier for the user, it is possible to provide a memory element for storing input masks, each of which input masks contains input fields for specifying a control command, or the parameters of a control command specified by the user.

Androgenic 7-substituted 11-halogen steroids

The invention relates to 11β-halogen steroids with general formula (I), whereby R11 is halogen, X—Y-Z represents a group with one of the two structures CH═C—C or CH2—C═C and the other radicals have the meaning that is indicated in the claims, also the production and use of these compounds for the production of pharmaceutical agents as well as pharmaceutical preparations that contain 11β-halogen steroids.

1. 11β-Halogen steroids with the general formula I: in which X—Y-Z represents a group with one of the two structures CH═C—C or CH2—C═C R1 can be in α- or β-position, and stands for hydrogen, R or P-Q-R that is bonded via P to the basic ring structure, whereby P and Q represent straight-chain or branched-chain, optionally partially or completely fluorinated C1 to C8 alkylene, -alkenylene, -alkinylene groups and can be the same or different, and R represents a CH3 radical or CF3 radical, provided that no substituent R1 is present at Z if X—Y-Z represents the group CH2—C═C, R6 is a hydrogen atom or can have the meanings that are indicated under R7, R7 stands for R or P-Q-R bonded via P to the basic ring structure, whereby these groups have the above-mentioned meanings, R11 represents a halogen, R13 is methyl or ethyl, and R17 is hydrogen or stands for C(O)—R18, whereby R18 is a straight-chain or branched-chain C1 to C18 alkyl, -alkenyl, -alkinyl radical or an aryl radical, or stands for T-U—V bonded via T to the C(O) group, whereby T and U represent straight-chain or branched-chain C, to C18 alkylene, -alkenylene, -alkinylene groups, alicyclic C3 to C12 groups or aryl groups and are the same or different, and V is a straight-chain or branched-chain C, to C18 alkyl, -alkenyl or -alkinyl radical or an aryl radical, or R18 has one of the above-mentioned meanings and in addition is substituted with one or more groups NR19R20 or one or more groups SOxR21, whereby x=0, 1 or 2, and R19, R20 and R21 in each case are hydrogen or T-U—V bonded via T to N, S with the above-mentioned meaning, provided that in addition, the physiologically compatible addition salts with in organic and organic acids are included. 2. 11β-Halogen steroids according to claim 2, characterized in that R11 is fluorine. 3. 11β-Haologen steroids according to claim 2, wherein R6 is hydrogen, methyl, ethyl or fluoromethyl. 4. 11β-Halogen steroids according to claim 2, wherein R7 is methyl, ethyl or fluoromethyl. 5. 11β-Halogen steroids according to claim 2, wherein R1 is hydrogen. 6. 11β-Halogen steroids according to claim 2, wherein R13 is methyl. 7. 11β-Halogen steroids according to claim 2, wherein R17 is hydrogen or C(O)—R18, whereby R18 is a straight-chain or branched-chain C1 to C18 alkyl radical. 8. 11β-Halogen steroids of general formula I, namely 11β-Fluoro 17β-hydroxy-7α-methyl-estr-4-en-3-one, 11β-Chloro-17β-hydroxy-7α-methyl-ester-4-en-3-one, 11β-Bromo-17β-hydroxy-7α-methyl-estr-4-en-3-one, 17β-Hydroxy-11β-iodo-7α-methyl-estr-4-en-3-one, 7α-Ethyl-11β-fluoro-17β-hydroxy-estr-4-en-3-one, 11β-Fluoro-7α-(fluoromethyl)-17β-hydroxy-estr-4-en-3-one, 11β-Fluoro-17β-heptanoyloxy-7α-methyl-estr-4-en-3-one, 11β-Fluoro-7α-methyl-17β-undecanoyloxy-estr-4-ene-3-one, 11β-Fluoro-17β-hydroxy-7α-methyl-estr-5(10)-en-3-one. 9. Use of the structural portion of general formula as a component of compounds with androgenic action, in which X—Y-Z represents a group with one of the two structures CH═C—C or CH2—C═C, the other bonds are saturated in ring A, R7 and R13 are not hydrogen, and R11 is a halogen. 10. Process for the production of 11β-halogen steroids of the general formula I according to claim 1, in which, starting from a compound with general formula B, D whereby R11 means either a hydroxyl group, a halogen or a nucleophilic leaving group, and R13 has the above-mentioned meaning, a compound with general formula B′, E is first formed by 1,6-addition of a metallated alkyl with general formula R7-MX or R7-M, whereby M is an alkali metal, M′ is an alkaline-earth metal, X is a halogen atom, and R7 has the above-mentioned meaning: and this compound then is reduced either selectively to a compound with general formula I or, if R11 is not halogen, is nucleophilically substituted in 11-position with a halogenating agent and then is reduced selectively to the compound with general formula I: 11. Use of the 11β-halogen steroids according to claim 1 for the production of pharmaceutical agents. 12. Pharmaceutical preparations that contain at least one 11β-halogen steroid according to claim 1 as well as at least one pharmaceutically compatible vehicle.

Normalizing circuit with reduced error voltage

A circuit for calibrating a voltage signal. The circuit includes a first voltage-to-current converter receiving the signal to be calibrated and a reference voltage, and providing a current proportional to the voltage difference between the voltage of the signal to be calibrated and the reference voltage, a first current amplifier driven by the first voltage-to-current converter, a second voltage-to-current converter receiving an adjustment voltage and the reference voltage and providing a current proportional to the voltage difference between the adjustment voltage and the reference voltage, a second current amplifier driven by the second voltage-to-current converter, and a means for providing a calibrated voltage signal based on the difference between the currents provided by the first and second current amplifiers.

1. A circuit for calibrating a voltage signal, including: a first voltage-to-current converter receiving a voltage signal to be calibrated and a reference voltage, and providing a current proportional to the voltage difference between the voltage of the signal to be calibrated and the reference voltage, a first current amplifier driven by the first voltage-to-current converter, a second voltage-to-current converter receiving an adjustment voltage and the reference voltage and providing a current proportional to the voltage difference between the adjustment voltage and the reference voltage, a second current amplifier driven by the second voltage-to-current converter, and means for providing a calibrated voltage signal based on the difference between the currents provided by the first and second current amplifiers. 2. The circuit of claim 1, wherein said means for providing a calibrated voltage signal includes: a current conversion means driven by the second current amplifier and providing a current of same amplitude and of sign opposite to that of the current that it receives, and a current-to-voltage converter driven by the first current amplifier and by the current conversion means and providing the calibrated voltage signal. 3. The circuit of claim 1, wherein the first and second current amplifiers, as well as the first and second voltage-to-current converters, have an identical structure. 4. The circuit of claim 3, wherein the first and second current amplifiers have a variable gain and their gain is identically controlled. 5. The circuit of claim 1, wherein each of the first and second current amplifiers includes several amplifier units. 6. The circuit of claim 5, wherein each of the first and second current amplifiers includes a first unit and a second unit, the second unit ensuring a current gain equal to 1 or to 9. 7. The circuit of claim 6, wherein the first unit includes: an input node coupled to ground via a diode-connected transistor, 16 identical branches, each formed of a transistor, and 31 identical branches, each including a transistor in series with a switch, each of the transistors coupled to the input node being flown, when on, by a same current, and an output node coupled to ground via 16 identical branches, each formed of a transistor, and 31 identical branches, each including a transistor in series with a switch, each of the transistors coupled with the output node being flown, when on, by a current of same value as any each transistor coupled with the input node, in the on state. 8. The circuit of claim 1, wherein the adjustment voltage is adjustable and is used to set the minimum value of the calibrated signal. 9. The circuit of claim 1, including a first current source arranged between the output of the first voltage-to-current converter and a fixed voltage, and a second current source arranged between the output of the second voltage-to-current converter and said fixed voltage. 10. (canceled) 11. A circuit for calibrating a voltage signal, including: a first voltage-to-current converter receiving a voltage signal to be calibrated and a reference voltage, and providing a current proportional to the voltage difference between the voltage of the signal to be calibrated and the reference voltage, a first current amplifier driven by the first voltage-to-current converter, a second voltage-to-current converter receiving an adjustment voltage and the reference voltage and providing a current proportional to the voltage difference between the adjustment voltage and the reference voltage, a second current amplifier driven by the second voltage-to-current converter, and means for providing a calibrated voltage signal based on the difference between the currents provided by the first and second current amplifiers, each of the first and second current amplifiers includes a first unit and a second unit, the second unit configured to ensure a current gain equal to a range of 1-9, the first unit including: an input node coupled to ground via a diode-connected transistor, 16 identical branches, each formed of a transistor, and 31 identical branches, each including a transistor in series with a switch, each of the transistors coupled to the input node being flown, when on, by a same current, and an output node coupled to ground via 16 identical branches, each formed of a transistor, and 31 identical branches, each including a transistor in series with a switch, each of the transistors coupled with the output node being flown, when on, by a current of same value as any each transistor coupled with the input node, in the on state. 12. The circuit of claim 11, further comprising a first current source arranged between the output of the first voltage-to-current converter and a fixed 13. A circuit for calibrating a voltage signal, including: a first voltage-to-current converter receiving a voltage signal to be calibrated and a reference voltage, and providing a current proportional to the voltage difference between the voltage of the signal to be calibrated and the reference voltage, a first current amplifier driven by the first voltage-to-current converter, a second voltage-to-current converter receiving an adjustment voltage and the reference voltage and providing a current proportional to the voltage difference between the adjustment voltage and the reference voltage, a second current amplifier driven by the second voltage-to-current converter, and means for providing a calibrated voltage signal based on the difference between the currents provided by the first and second current amplifiers; and a first current source arranged between the output of the first voltage-to-current converter and a fixed voltage, and a second current source arranged between the output of the second voltage-to-current converter and the fixed voltage. 14. The circuit of claim 13, wherein each of the first and second current amplifiers includes a first unit and second unit, the first unit including: an input node coupled to ground via a diode-connected transistor, 16 identical branches, each formed of a transistor, and 31 identical branches, each including a transistor in series with a switch, each of the transistors coupled to the input node being flown, when on, by a same current, and an output node coupled to ground via 16 identical branches, each formed of a transistor, and 31 identical branches, each including a transistor in series with a switch, each of the transistors coupled with the output node being flown, when on, by a current of same value as any, each transistor coupled with the input node, in the on state. 15. A calibration circuit, comprising: first and second current amplifiers receiving as input first and second proportional currents from first and second voltage-to-current converters, respectively; a current converter driven by the second current amplifier and configured to provide a current of the same amplitude and of a sign that is opposite to that of the current that it receives; and a current-to-voltage converter driven by the first current amplifier and by the current conversion circuit and configured to provide on an output thereof a calibrated voltage signal. 16. A calibration circuit, comprising: first and second current amplifiers receiving as input first and second proportional currents from first and second voltage-to-current converters, respectively; a current converter driven by the second current amplifier and configured to provide a current of the same amplitude and of a sign that is opposite to that of the current that it receives; and a current-to-voltage converter driven by the first current amplifier and by the current conversion circuit and configured to provide on an output thereof a calibrated voltage signal; and a first current source arranged between the output of the first voltage-to-current converter and a fixed voltage, and a second current source arranged between the output of the second voltage-to-current converter and a fixed voltage. 17. The circuit of claim 16, wherein each of the first and second current amplifiers includes a first unit and a second unit, the first unit comprising: an input node coupled to ground via a diode-connected transistor, 16 identical branches that are each formed of a transistor, and 31 identical branches that are each comprising a transistor in series with a switch, each of the transistors coupled to an input node supplied by a same current, and an output node coupled to ground via 16 identical branches that are each formed of a transistor, and 31 identical branches that are each comprising a transistor in series with a switch, each of the transistors coupled with the output node being supplied by a current of same value as any of each of the transistors coupled with the input node when in an on state.

<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention The present invention relates to a circuit for calibrating a signal, that is, for providing, based on an input signal, a signal included in a predetermined range. 2. Description of the Related Art There are various applications for calibration circuits. For example, calibration circuits are used in optical disk read systems. In such systems, a calibration circuit is arranged between a photodetector and an analog-to-digital converter to adapt the signal from the photodetector to the analog-to-digital converter input range. The integrated circuit “L6300” of STMicroelectronics is an example of a calibration circuit used in such a system. FIG. 1 illustrates the shape, along time, of a voltage V at the input of a calibration circuit. Voltage V, shown as a sinusoid, oscillates between a minimum value Vmin and a maximum value Vmax. In the mentioned application, voltage V coming from a photodetector oscillates above a reference level Vmin approximately equal to 2.5 volts. Peak-to-peak amplitude Vmax-Vmin of voltage V can vary within a large range from 25 to 500 millivolts, according to whether the photodetector receives all or part of the light emitted by the laser. FIG. 2 illustrates the shape, along time, of voltage VOUT at the output of the calibration circuit. Voltage VOUT has the same frequency as voltage V, but its amplitude is constant and the signal oscillates between a minimum value Vbot and a maximum value Vtop corresponding to desired limiting values. Typical values of Vbot and Vtop, in the application mentioned hereabove, are respectively around 125 and 875 millivolts. Generally, the forming of a calibration circuit must take into account error or offset voltages likely to affect the limiting values of output voltage VOUT. For example, in the mentioned application, an error voltage which can reach ±70 millivolts systematically affects the reference level Vmin. Further, each element of the calibration circuit introduces an error voltage specific to it. This effect is particularly substantial if the circuit includes MOS transistors, since these transistors generate greater error voltages than bipolar transistors. Because of this, many calibration circuits, like circuit “L6300” mentioned hereabove, are formed by means of bipolar transistors. Further, since the signal processing circuits that follow the calibration circuit are generally formed by means of CMOS transistors, the calibration circuit can hardly be formed together with the circuits that follow it on a same silicon wafer to form an integrated circuit. This results in high manufacturing, testing, and interconnection costs. Further, known calibration circuits are powered by relatively high supply voltages, greater than those supplying CMOS circuits.

<SOH> BRIEF SUMMARY OF THE INVENTION <EOH>The disclosed embodiments of the present invention provide a calibration circuit in which the effect of error voltages is decreased. The calibration circuit of the present invention can be easily be made on a same integrated circuit as a digital signal processing circuit, and it can be powered under a reduced voltage. The present invention provides a circuit for calibrating a voltage signal that includes: a first voltage-to-current converter receiving a voltage signal to be calibrated and a reference voltage, and providing a current proportional to the voltage difference between the voltage of the signal to be calibrated and the reference voltage, a first current amplifier driven by the first voltage-to-current converter, a second voltage-to-current converter receiving an adjustment voltage and the reference voltage and providing a current proportional to the voltage difference between the adjustment voltage and the reference voltage, a second current amplifier driven by the second voltage-to-current converter, and a means for providing a calibrated voltage signal based on the difference between the currents provided by the first and second current amplifiers. According to an embodiment of the present invention, the means for providing the calibrated voltage signal includes: a current conversion means driven by the second current amplifier and providing a current of same amplitude and of a direction opposite to that of the current that it receives, and a current-to-voltage converter driven by the first current amplifier and by the current conversion means and providing the calibrated voltage signal. According to an embodiment of the present invention, the first and second current amplifiers, as well as the first and second voltage-to-current converters, have an identical structure. According to an embodiment of the present invention, the first and second current amplifiers have a variable gain and their gain is identically controlled. According to an embodiment of the present invention, each of the first and second current amplifiers includes several amplifier units. According to an embodiment of the present invention, each of the first and second current amplifiers includes a first unit and a second unit, the second unit ensuring a current gain equal to 1 or to 9. According to an embodiment of the present invention, the first unit includes: an input node coupled to ground via a diode-connected transistor, 16 identical branches, each formed of a transistor, and 31 identical branches, each including a transistor in series with a switch, each of the transistors coupled to the input node being run through, when on, by a same current, and an output node coupled to ground via 16 identical branches, each formed of a transistor, and 31 identical branches, each including a transistor in series with a switch, each of the transistors coupled with the output node being run through, when on, by a current of same value as any one, in the on state, of the transistors coupled with the input node. According to an embodiment of the present invention, the adjustment voltage is adjustable and is used to set the minimum value of the calibrated signal. According to an embodiment of the present invention, the circuit includes a first current source arranged between the output of the first voltage-to-current converter and a fixed voltage, and a second current source arranged between the output of the second voltage-to-current converter and said fixed voltage. The present invention also provides a method for calibrating a signal by means of a circuit including a first branch receiving the signal to be calibrated and a second branch receiving an adjustment voltage, the first branch including a first variable-gain current amplifier and the second branch including a second variable-gain current amplifier, the first and second current amplifiers being of identical structure and their gain being adjusted in the same way. The method includes the adjustment steps of: a) the gain of the first and second current amplifiers being set to its maximum value, injecting into the circuit a voltage equal to the minimum value of the voltage to be calibrated and adjusting the adjustment voltage so that the minimum level of the signal at the circuit output correspond to the desired minimum level, then b) injecting the signal to be calibrated and adjusting the gain of the first and second current amplifiers so that the maximum level of the signal at the circuit output corresponds to the desired maximum level.

Process for the polymerization of olefins

The present invention relates to a process for homopolymerization of ethylene or copolymerization of ethylene with alpha-olefins by contacting ethylene or ethylene and alpha-olefin with a catalyst composition comprising: (a) a solid catalyst precursor comprising at least one vanadium compound, at least one magnesium compound and a polymeric material or a solid catalyst precursor comprising at least one vanadium compound, at least one further transition metal compound and/or at least one alcohol, at least one magnesium compound and a polymeric material; and (b) a cocatalyst comprising an aluminum compound.

1. A process for homopolymerization of ethylene or copolymerization of ethylene with alpha-olefins comprising contacting ethylene or ethylene and alpha-olefin with a catalyst composition comprising: (i) a solid catalyst precursor comprising at least one vanadium compound represented by the general formulas V(OR1)nX4-n, V(R2)nX4-n, VX3 and VOX3 wherein R1 and R2 represent an alkyl group, aryl group or cycloalkyl group having 1 to 20 carbon atoms, and X represents a halogen and n represents a number satisfying 0≦n≦4, at least one Grignard compound represented by the general formula R6MgX wherein R6 is a hydrocarbon group having 1 to 20 carbon atoms and X is a halogen atom or a dialkyl magnesium compound represented by the general formula R7R8Mg, wherein R7 and R8 are each a hydrocarbon group having 1 to 20 carbon atoms, and a polymeric support; and (ii) a cocatalyst comprising an aluminum compound represented by the general formula R9nAlX3-n wherein R9 represents a hydrocarbon group having 1 to 10 carbon atoms; X represents a halogen and n represents a number satisfying 0≦n≦3, or by the general formula R10R11Al—O—AlR12R13, wherein R10, R11, R12 and R13 are either the same or different linear, branched or cyclic alkyl group having 1 to 12 carbons, or is an aluminoxane. 2. The process according to claim 1 wherein said solid catalyst precursor further comprises at least transition metal compound represented by the general formulas Tm(OR3)nX4-n, TmOX3 and Tm(R4)nX4-n, wherein Tm is titanium or vanadium, R3 and R4 represent an alkyl group, aryl group or cycloalkyl group having 1 to 20 carbon atoms, X represents a halogen atom and n represents a number satisfying 0≦n≦4 or at least one alcohol represented by the general formula R5OH, wherein R5 is an alkyl group, aryl group or cycloalkyl group having 1 to 20 carbon atoms. 3. (canceled) 4. (canceled) 5. The process according to claim 1, wherein said vanadium compound is selected from the group comprising vanadium tetraethoxide, vanadium tetrapropoxide, vanadium tetrabutoxide, vanadium trichloride, vanadium tetrachloride, vanadium oxytrichloride, and vanadium dichlorodiethoxide. 6. The process according to claim 5, wherein said vanadium compound comprises vanadium tetrachloride or vanadium oxytrichloride. 7. (canceled) 8. The process according to claim 2, wherein the transition metal compound is selected from the group comprising titanium trichloromethoxide, titanium dichlorodimethoxide, titanium tetramethoxide, titanium trichloroethoxide, titanium dichlorodiethoxide, titanium tetraethoxide, titanium trichloropropoxide, titanium dichlorodipropoxide, titanium chlorotripropoxide, titanium tetrapropoxide, titanium trichlorobutoxide, titanium dichlorodibutoxide, titanium tetrabutoxide, vanadium tetrachloride, vanadium tetraethoxide, vanadium tetrapropoxide, vanadium tetrabutoxide, vanadium oxytrichloride, and vanadium dichlorodiethoxide. 9. The process according to claim 8, wherein the transition metal compound comprises titanium tetraethoxide, titanium tetrapropoxide or titanium tetrabutoxide. 10. (canceled) 11. The process according to claim 2, wherein the alcohol is selected from the group comprising methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, cyclohexanol, phenol, methylphenol, and ethylphenol. 12. (canceled) 13. The process according to claim 5, wherein the magnesium compound comprises diethylmagnesium, di-n-propylmagnesium, di-isopropylmagnesium, di-n-butylmagnesium, di-isobutylmagnesium butylethylmagnesium, dihexylmagnesium, dioctylmagnesium, butyloctylmagnesium, ethylmagnesium chloride, butylmagnesium chloride, or hexylmagnesium chloride. 14. The process according to claim 5, wherein the polymeric support is in the form of particles having a mean particle diameter of about 5 to 1000 microns, a pore volume of at least about 0.05 cm3/g, a pore diameter of about 20 to 10000 angstroms and a surface area of about 0.1 to 100 m2/g. 15. The process according to claim 14, wherein said polymeric support has a pore diameter from about 500 to 10000 angstroms and a surface area from about 0.2 to 30 m2/g. 16. The process according to claim 15, wherein the polymeric support is comprised of polyvinylchloride, polyvinylalcohol, polyketone, hydrolyzed polyketone, ethylene-vinylalcohol copolymer, or polycarbonate. 17. (canceled). 18. The process according to claim 15, wherein the polymeric support is comprised of polyvinylchloride having a molecular weight in the range of about 5000 to 500000 g/mole. 19. The process according to claim 18, wherein the Grignard compound is used in the range of about 0.05 to 20 mmol per gram of polymeric support. 20. The process according to claim 1, wherein the molar ratio of vanadium to magnesium in the catalyst precursor is in the range of about 0.01 to 50. 21. The process according to claim 2, wherein the molar ratio of vanadium to transition metal in said transition compound in the catalyst precursor is of about 0.01 to 50. 22. The process according to claim 2, wherein said catalyst precursor comprises an alcohol and the molar ratio of vanadium to OH groups in said alcohol in the catalyst precursor is in the range of about 0.01 to 50. 23. (canceled) 24. The process according to claim 6, wherein the aluminum compound is trimethylaluminum, triethylaluminum, tri-isobutylaluminum or tri-n-hexylaluminum. 25. (canceled) 26. The process according to claim 5, wherein the aluminum compound comprises methyl aluminoxane (MAO) or modified methyl aluminoxane (MMAO). 27. The process according to claim 5, wherein the aluminum compound comprises a mixture of an alkylaluminum and an aluminoxane. 28. The process according to claim 2, wherein the ratio of moles of aluminum in said cocatalyst to the moles of transition metal in said catalyst precursor is about 10 to 5000. 29. (canceled) 30. (canceled) 31. (canceled)

<SOH> FIELD OF THE INVENTION <EOH>The present invention relates to a process for the polymerization of olefins.

<SOH> SUMMARY OF THE INVENTION <EOH>The present invention provides a process for homopolymerization of ethylene or copolymerization of ethylene with alpha-olefins by contacting ethylene or ethylene and alpha-olefin with a catalyst composition comprising: (a) a solid catalyst precursor comprising at least one vanadium compound, at least one magnesium compound and a polymeric material; and (b) a cocatalyst comprising at least one aluminum compound. Most preferably, the component (a) used in the catalyst composition for the process of the present invention further comprises at least one further transition metal compound and/or at least one alcohol. As a result of the present invention polyolefin and especially polyethylenes and copolymers of ethylene and alpha-olefins are provided which have a density of about 0.88 to 0.98 g/cm 3 and weight average molecular weight of about 500 to 900 000 g/mole and molecular weight distribution range of about 2 to about 30. The products have a uniform spherical particle morphology, very low level of fines and catalyst residues, improved thermal stability, excellent optical and improved environmental stress cracking resistance (ESCR). detailed-description description="Detailed Description" end="lead"?