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Method for producing sized paper or cardboard |
A method is disclosed for manufacturing a surface sized web of paper or paperboard, the method comprising the step of applying to at least one side of the web an aqueous furnish of size. According to the method, the solids content of the size furnish being applied is at least 15% and the size frunish is applied to one side of the web by an amount not greater than 5 g/m2 as aqueous furnish of size applied to the web. |
1. A method for manufacturing a surface sized web of paper or paperboard, comprising. applying to at least one side of the web an aqueous furnish of size wherein the solids content of the aqueous size furnish is at least 15% and the size furnish is applied to one side of the web by an amount not greater than 5 g/m2 as aqueous furnish of size applied to the web; and pressing the applied size furnish into the surface of the web. 2. The method of claim 1, wherein the solids content of the size furnish is 15% to 40%. 3. The method of claim 1, wherein the size furnish is applied to at least one side of the web by means of a film-transfer press. 4. The method of claim 3, wherein the size furnish is applied simultaneously to both sides of the web by means of a film-transfer press, whereby the overall amount of size furnish applied to the web is 10 g/m2. 5. The method of claim 1, wherein the size furnish is applied to the web by means of a spray or jet applicator. 6. The method of claim 5, wherein the web is pressed in a nip downstream of the application of size in order to ensure size penetration. 7. The method of claim 6, wherein size penetration into the web is controlled by adjusting the machine-direction distance between application of the size and the press nip. 8. The method of claim 1, wherein the weight profile of size furnish adhered to the web is gauged at least in the cross-machine direction of the web and application of size is adjusted based on the gauging result. 9. The method of claim 1, wherein the size furnish being applied contains cooked starch. 10. The method of claim 2, wherein the size furnish is applied to at least one side of the web by means of a film-transfer press. 11. The method of claim 10, wherein the size furnish is applied simultaneously to both sides of the web by means of a film-transfer press, whereby the overall amount of size furnish applied to the web is 10 g/m2. 12. The method of claim 2, wherein the size furnish is applied to the web by means of a spray or jet applicator. 13. The method of claim 12, wherein the web is pressed in a nip downstream of the application of size in order to ensure size penetration. 14. The method of claim 13, wherein size penetration into the web is controlled by adjusting the machine-direction distance between application of the size and the press nip. 15. The method of claim 2, wherein the weight profile of size furnish adhered to the web is gauged at least in the cross-machine direction of the web and application of size is adjusted based on the gauging result. 16. The method of claim 4, wherein the weight profile of size furnish adhered to the web is gauged at least in the cross-machine direction of the web and application of size is adjusted based on the gauging result. 17. The method of claim 2, wherein the size furnish being applied contains cooked starch. 18. The method of claim 4, wherein the size furnish being applied contains cooked starch. |
Pyridazinone |
The invention relates to novel compounds, to a method for the production of said compounds and to the use thereof as medicaments, more particularly as antiviral agents, especially against cytomegalovirus. |
1. A compound of the general formula (I) in which A is attached via position 2, 3, 5 or 6 to the aromatic ring and A represents oxygen or NR6, E represents oxygen, CR9R10 or NR7, Y represents oxygen or NR8, D and X are identical or different and represent in each case oxygen or sulfur, G represents hydrogen, or G represents C6-C10-aryl, where C6-C10-aryl may optionally be substituted by up to three substituents selected from the group consisting of halogen, hydroxyl, nitro, cyano, C1-C6-alkoxy, hydroxy-carbonyl, C1-C6-alkoxycarbonyl, amino, mono- or di-C1-C6-alkylamino, mono- or di-C1-C6-alkylaminocarbonyl and C1-C6-alkyl, where C1-C6-alkoxy, C1-C6-alkoxycarbonyl, mono- or di-C1-C6-alkylamino, mono- or di-C1-C6-alkylaminocarbonyl or C1-C6-alkyl may optionally be substituted by up to three substituents selected from the group consisting of halogen, hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl, mono- or di-C1-C6-alkylaminocarbonyl and C6-C10-aryl, or G represents C6-C10-aryl, where C6-C10-aryl may optionally be substituted by phenyl, where phenyl may optionally be substituted by up to three substituents selected from the group consisting of halogen, hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl, mono- or di-C1-C6-alkylaminocarbonyl and C1-C6-alkyl, where C1-C6-alkyl for its part may optionally be substituted by up to three substituents selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or G represents C6-C10-aryl, where C6-C10-aryl may optionally be substituted by phenyl, where phenyl may optionally be substituted by C5-C6-heteroaryl or C5-C7-heterocyclyl, where C5-C6-heteroaryl or C5-C7-heterocyclyl for their part may optionally be substituted by up to three substituents selected from the group consisting of halogen, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or G represents C6-C10-aryl, where C6-C10-aryl may optionally be substituted by a group of the following formula G represents C5-C10-heteroaryl or C5-C7-heterocyclyl, where C5-C10-heteroaryl or C5-C7-heterocyclyl may optionally be substituted by up to three substituents selected from the group consisting of halogen, nitro, cyano, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono or di-C1-C6-alkylaminocarbonyl, or G represents C3-C10-cycloalkyl, where C3-C10-cycloalkyl may optionally be substituted by up to three substituents selected from the group consisting of halogen, nitro, cyano, hydroxyl, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono or di-C1-C6-alkylaminocarbonyl, R1, R2, R3 and R4 are identical or different and each represent hydrogen, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, C6-C10-aryl or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono or di-C1-C6-alkylaminocarbonyl, and where C6-C10-aryl may optionally be substituted by up to three substituents selected from the group consisting of halogen, hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl, mono or di-C1-C6-alkylaminocarbonyl and C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or R1 and R2 or R3 and R4 together with the carbon atom to which they are attached form a C3-C6-cycloalkyl ring, where the C3-C6-cycloalkyl ring may optionally be substituted by up to three substituents selected from the group consisting of halogen, hydroxyl, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or R1 and R3 together with the carbon atoms to which they are attached form a C3-C6-cycloalkyl ring, where the C3-C6-cycloalkyl ring may optionally be substituted by up to three substituents selected from the group consisting of halogen, hydroxyl, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R5 represents hydrogen, halogen, hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino or C1-C6-alkyl, where C1-C6-alkoxy, mono- or di-C1-C6-alkylamino or C1-C6-alkyl may optionally be substituted by up to three substituents selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R6, R7 and R8 are identical or different and represent in each case hydrogen or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R9 and R10 are identical or different and represent in each case hydrogen, NR11R12, OR13 or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R11, R12 and R13 are identical or different and represent in each case hydrogen or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or a tautomer, a stereioisomer, a stereoisomeric mixture or a pharmacologically acceptable salt thereof. 2. A compound of the general formula (I) as claimed in claim 1, in which A is attached via position 2, 3, 5 or 6 to the aromatic ring, and A represents oxygen or NR6, E represents oxygen, CR9R10 or NR7, Y represents oxygen or NR8, D and X are identical or different and represent in each case oxygen or sulfur, G represents hydrogen, or G represents C6-C10-aryl, where C6-C10-aryl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, hydroxyl, nitro, cyano, C1-C6-alkoxy, hydroxycarbonyl, C1-C6-alkoxycarbonyl, amino, mono- or di-C1-C6-alkylamino, mono- or di-C1-C6-alkylaminocarbonyl and C1-C6-alkyl, where C1-C6-alkoxy, C1-C6-alkoxycarbonyl, mono- or di-C1-C6-alkylamino, mono- or di-C1-C6-alkylaminocarbonyl or C1-C6-alkyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl, mono- or di-C1-C6-alkylaminocarbonyl and C6-C10-aryl, or G represents C6-C10-aryl, where C6-C10-aryl may optionally be substituted by phenyl, where phenyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl, mono- or di-C1-C6-alkylaminocarbonyl and C1-C6-alkyl, where C1-C6-alkyl for its part may optionally be substituted by up to three substituents independently of one another selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or G represents C6-C10-aryl, where C6-C10-aryl may optionally be substituted by phenyl, where phenyl may optionally be substituted by C5-C6-heteroaryl or C5-C7-heterocyclyl, where C5-C6-heteroaryl or C5-C7-heterocyclyl for their part may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or G represents C6-C10-aryl, where C6-C10-aryl may optionally be substituted by a group of the following formula G represents C5-C10-heteroaryl or C5-C7-heterocyclyl, where C5-C10-heteroaryl or C5-C7-heterocyclyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, nitro, cyano, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or G represents C3-C10-cycloalkyl, where C3-C10-cycloalkyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, nitro, cyano, hydroxyl, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R1, R2, R3 and R4 are identical or different and represent in each case hydrogen, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, C6-C10-aryl or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, and where C6-C10-aryl may optionally be substituted by up to three substituents selected from the group consisting of halogen, hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl, mono- or di-C1-C6-alkylaminocarbonyl and C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino,-mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, where R1, R2, R3 and R4 are not simultaneously hydrogen, or R1 and R2 or R3 and R4 together with the carbon atom to which they are attached form a C3-C6-cycloalkyl ring, where the C3-C6-cycloalkyl ring may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, hydroxyl, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or R1 and R3 together with the carbon atoms to which they are attached form a C3-C6-cycloalkyl ring, where the C3-C6-cycloalkyl ring may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, hydroxyl, C1-C6-alkyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R5 represents hydrogen, halogen, hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino or C1-C6-alkyl, where C1-C6-alkoxy, mono- or di-C1-C6-alkylamino or C1-C6-alkyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R6, R7 and R8 are identical or different and represent in each case hydrogen or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R9 and R10 are identical or different and represent in each case hydrogen, NR11R12, OR13 or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R11, R12 and R13 are identical or different and represent in each case hydrogen or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, or a tautomer, a stereoisomer, a stereoisomeric mixture or a pharmacologically acceptable salt thereof. 3. A compound of the general formula (I) as claimed in claim 1 or 2, in which A is attached via position 2, 3, 5 or 6 to the aromatic ring and A represents NR6, E represents NR7, Y represents NR8, D and X represent oxygen, G represents C6-C10-aryl, where C6-C10-aryl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, hydroxyl, cyano and C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents of halogen, or G represents C5-C6-heteroaryl, where C5-C6-heteroaryl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen and C1-C3-alkyl, or G represents C3-C10-cycloalkyl, where C3-C10-cycloalkyl may optionally be substituted by up to three substituents C1-C6-alkyl, R1, R2 and R3 are identical or different and represent in each case hydrogen or represent C1-C3-alkyl, R4 represents hydrogen, C6-C10-aryl or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, C1-C6-alkylcarbonylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, and where C6-C10-aryl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of halogen, hydroxyl, C1-C6-alkoxy and C1-C6-alkyl, where R1, R2, R3 and R4 do not simultaneously represent hydrogen, R5 represents hydrogen, halogen, hydroxyl, amino, mono- or di-C1-C6-alkylamino or C1-C6-alkyl, where C1-C6-alkyl may optionally be substituted by up to three substituents independently of one another selected from the group consisting of hydroxyl, C1-C6-alkoxy, amino, mono- or di-C1-C6-alkylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and mono- or di-C1-C6-alkylaminocarbonyl, R6, R7 and R8 represent hydrogen, or a tautomer, a stereoisomer, a stereoisomeric mixture or a pharmacologically acceptable salt thereof. 4. A compound of the general formula (I) as claimed in claim 1, 2 or 3, where the radical A is attached via position 3 to the aromatic ring. 5. A compound of the general formula (I) as claimed in claim 1, 2 or 3, where D and X and represent oxygen. 6. A compound of the general formula (I) as claimed in claim 1, 2 or 3, where A, E and Y represent NH. 7. A compound of the general formula (I) as claimed in claim 1, 2 or 3, where G represents substituted phenyl. 8. A compound of the general formula (I) as claimed in claim 1, 2 or 3, where R1, R2 and R5 represent hydrogen and R3 and R4 represent methyl. 9. A process for preparing the compounds of the formula (I) as claimed in claim 1, which comprises [A] reacting compounds of the general formula (II) in which A is attached via position 2, 3, 5 or 6 to the aromatic ring and R1, R2, R3, R4, R5, A, X and Y are as defined above, with compounds of the general formula (III) D=C═N-G (III) in which D and G are as defined above to give compounds of the general formula (Ia) in which A is attached via position 2, 3, 5 or 6 to the aromatic ring and R1, R2, R3, R4, R5, A, D, G, X and Y are as defined above, or [B] reacting compounds of the general formula (II) with compounds of the general formula (IV) in which D, E and G are as defined above and L1 represents p-nitrophenyl or halogen, preferably bromine or chlorine, to give compounds of the general formula (I) in which A is attached via position 2, 3, 5 or 6 to the aromatic ring and R1, R2, R3, R4, R5, A, D, E, G, X and Y are as defined above, or [C] reacting compounds of the general formula (V) in which —NCD is attached via position 2, 3, 5 or 6 to the aromatic ring and R1, R2, R3, R4, R5, D, X and Y are as defined above with compounds of the general formula (VI) H-M-G (VI) in which G is as defined above and M represents oxygen or NR7, where R7 is as defined above, to give compounds of the general formula (Ib) in which —NH—C(D)-M-G is attached via position 2, 3, 5 or 6 to the aromatic ring and R1, R2, R3, R4, R5, D, G, M, X and Y are as defined above. 10. A compound of the general formula (I) as claimed in claim 1, 2 or 3 for controlling disorders. 11. A medicament, comprising compounds of the general formula (I) as claimed in claim 1, 2 or 3 in combination with at least one pharmaceutically acceptable, pharmaceutically safe carrier or excipient. 12. The use of compounds of the general formula (I) as claimed in claim 1, 2 or 3 for preparing as medicament for treating viral disorders. 13. A medicament as claimed in claim 12, for treating viral disorders. 14. A method for controlling viral disorders in humans and animals by administration of an antivirally effective amount of at least one compound as claimed in any of claims 1 to 3. |
Methods and pharmaceutical compositions for immune deception, particularly useful in the treatment of cancer |
An immuno-molecule which comprises a soluble human MHC class I effector domain; and an antibody targeting domain which is linked to the soluble human MHC class I effector domain, methods of making same and uses thereof. |
1. An immuno-molecule comprising: a soluble human MHC class I effector domain; and a targeting domain being linked to said soluble human MHC class I effector domain. 2. The immuno-molecule of claim 1, wherein said targeting domain is an antibody targeting domain. 3. The immuno-molecule of claim 1, wherein said targeting domain is a ligand targeting domain. 4. The immuno-molecule of claim 3, wherein said ligand targeting domain is selected from the group consisting of PDGF, EGF, KGF, TGFα, IL-2. IL-3, IL-4, IL-6, VEGF and its derivatives and TNF. 5-29. (Canceled) 30. A nucleic acid construct encoding an immuno-molecule, the construct comprising: a first polynucleotide encoding a soluble human MHC class I effector domain; and a second polynucleotide encoding a targeting domain; said first polynucleotide and said second polynucleotide are selected and being joined such that said soluble human MHC class I effector domain and said antibody targeting domain are translationally fused optionally via a peptide linker in-between. 31. The nucleic acid construct of claim 30, wherein said targeting domain is an antibody targeting domain. 32. The nucleic acid construct of claim 30, wherein said targeting domain is a ligand targeting domain. 33. The nucleic acid construct of claim 32, wherein said ligand targeting domain is selected from the group consisting of PDGF, EGF, KGF, TGFα, IL-2. IL-3, IL-4, IL-6, VEGF and its derivatives and TNF. 34-53. (Canceled) 54. A nucleic acid construct encoding an immuno-molecule, the construct comprising: a first polynucleotide encoding a soluble human MHC class I effector domain; and a second polynucleotide encoding a variable region of one of a light chain or a heavy chain of an antibody targeting domain; said first polynucleotide and said second polynucleotide are selected and being joined such that said soluble human MHC class I effector domain and said variable region of said one of a light chain and heavy chain of said antibody targeting domain are translationally fused optionally via a peptide linker in-between; a third polynucleotide encoding said other of said light chain and heavy chain of said antibody targeting domain. 55. A nucleic acid construct system comprising: a first nucleic acid construct which comprises: a first polynucleotide encoding a soluble human MHC class I effector domain; and a second polynucleotide encoding a variable region of one of a light chain or a heavy chain of an antibody targeting domain; said first polynucleotide and said second polynucleotide are selected and being joined such that said soluble human MHC class I effector domain and said variable region of said one of a light chain and heavy chain of said antibody targeting domain are translationally fused optionally via a peptide linker in-between; a second nucleic acid construct which comprises: a third polynucleotide encoding said other of said light chain and heavy chain of said antibody targeting domain. 56. An isolated preparation of bacterial derived inclusion bodies comprising over 30 percent by weight of an immuno-molecule, the immuno-molecule comprises: a soluble human MHC class I effector domain; and a targeting domain being linked to said soluble human MHC class I effector domain. 57. The isolated preparation of claim 56, wherein said targeting domain is an antibody targeting domain. 58. The isolated preparation of claim 56, wherein said targeting domain is a ligand targeting domain. 59. The isolated preparation of claim 56 claim 58, wherein said ligand targeting domain is selected from the group consisting of PDGF, EGF, KGF, TGFα, IL-2. IL-3, IL-4, IL-6, VEGF and its derivatives and TNF. 60-82. (Canceled) 83. A method of producing an immuno-molecule comprising: expressing, in bacteria, the immuno-molecule which comprises: a soluble human MHC class I effector domain which includes a functional human β-2 microglobulin and a functional human MHC class I heavy chain linked thereto; and a targeting domain being linked to said soluble human MHC class I effector domain; and isolating the immuno-molecule. 84. The method of claim 83, wherein said targeting domain is an antibody targeting domain. 85. The method of claim 83, wherein said targeting domain is a ligand targeting domain. 86-94. (Canceled) 95. A method of selectively killing a cell in a patient, the cell presenting an antigen, the method comprising administering to the patient an immuno-molecule which comprises: a soluble human MHC class I effector domain complexed with an MHC-restricted peptide; and a targeting domain being linked to said soluble human MHC class I effector domain, said targeting domain being selectively binding to said antigen; whereby, said soluble human MHC class I effector domain completed with an MHC-restricted peptide initiates a CTL mediated immune response against said cell, thereby selectively killing the cell in vivo. 96. The method of claim 95, wherein said targeting domain is an antibody targeting domain. 97. The method of claim 95, wherein said targeting domain is a ligand targeting domain. 98-122. (Canceled) |
<SOH> FIELD AND BACKGROUND OF THE INVENTION <EOH>The present invention relates to a novel concept in immunotherapy, by which deception of the immune system results in specific and most efficient destruction of cells of interest, cancer cells in particular. There is strong evidence that tumor progression in cancer patients is controlled by the immune system. This conclusion is based on observations that tumor progression is often associated with secretion of immune suppressive factors and/or downregulation of MHC class I antigen presentation functions (1-5). The inference is that tumors must have elaborated strategies to circumvent an apparently effective immune response. Importantly, a tumor-specific immune response can be detected in individuals (6-8). The apparent inefficiency of anti tumor immune responses that results in failure to combat the disease laid the foundation to current concepts of immunotherapy. It is suggested that boosting the anti-tumor immune response by deliberate vaccination or by other immunotherapeutic approaches may increase the potential benefits of immune-based therapies (6,9-11). The MHC class I-restricted CD8 cytotoxic T cell (CTL) effector arm of the adaptive immune response is best equipped to recognize the tumor as foreign and initiate the cascade of events resulting in tumor destruction (12,13). Therefore, the most attractive approach in cancer immunotherapy is centered on vaccination strategies designed to enhance the CTL arm of the antitumor response and consequently overcome the mechanisms of tumor escape from CTL (9-11). One of the best-studied escape mechanisms by which tumor cells evade immune attack is by downregulation of the MHC class I molecules which are go the antigens recognized by CTLs (1-5,14). Mutations along the class I presentation pathway should be the simplest way for tumors to escape CTL-mediated elimination since it can be achieved by one or two mutational events (two mutations to inactivate both alleles or one mutation to create a dominant negative inhibitor) (1-3). Downregulation of NHC class I expression is frequently observed in human tumors, and is particularly pronounced in metastatic lesions (3, 14-17). This is circumstantial but nevertheless compelling evidence of the role of CTL in controlling tumor progression in cancer patients. MIC class I expression has been mainly analyzed in surgically removed tumor specimens using immunohistochemical methods (14-15). Partial reduction or complete loss of MHC have been reported, encompassing all MHC molecules or limited to particular alleles (14-15). MHC loss can be seen in some but not all lesions of the same patient. Downregulation of MHC class I expression has been attributed to mutations in β2-microglobulin (β2-m), transporter associated with antigen presentation (TAP) proteins, or the proteosomal LMP-2 and LMP-7 proteins (2, 18-21). Additional evidence implicating loss of MHC class I expression as a mechanism for tumor escape from CTL-mediated elimination comes from a longitudinal study of a melanoma patient. Tumor cells removed during initial surgery presented nine different antigens restricted to four separate HLA class I alleles to CTL clones established from the patient (1). The patient remained disease free for 5 years after which a metastasis was detected. Notably, a cell line established from the metastatic lesion had lost all four alleles that had previously been shown to present melanoma antigens. Thus, the downregulation of class I MHC molecule is a severe limiting problem for cancer immunotherapy and the application of anti-cancer vaccines. There is thus a widely recognized need for, and it would be highly advantageous to have, an novel approach of immunotherapy devoid of the above limitations, namely an approach of immunotherapy which is independent of the level of expression of MHC class I molecules by cancer cells. |
<SOH> SUMMARY OF THE INVENTION <EOH>The MHC class I-restricted CD8 cytotoxic T cell (CTL) effector arm of the adaptive immune response is best equipped to recognize tumor cells as foreign and initiate the cascade of events resulting in tumor destruction. However, tumors have developed sophisticated strategies to escape immune effector mechanisms, of which the best-studied is by downregulation of MHC class I molecules which are the antigens recognized by CTLs. To overcome this and develop new approaches for immunotherapy, and while reducing the present invention to practice, a recombinant molecule was constructed in which a single-chain MHC is specifically targeted to tumor cells through its fusion to cancer specific-recombinant antibody fragments or a ligand that binds to receptors expressed by tumor cells. As an exemplary molecule of the present invention, a single-chain HLA-A2 molecule was genetically fused to the variable domains of an anti IL-2 receptor α subunit-specific humanized antibody, anti-Tac (aTac). The construct, termed B2M-aTac(dsFv) was expressed in E. coli and functional molecules were produced by in vitro refolding in the presence of HLA-A2-restricted antigenic peptides. Flow cytometry studies revealed the ability to decorate antigen-positive, HLA-A2-negative human tumor cells with HLA-A2-peptide complexes in a manner that was entirely dependent upon the specificity of the targeting antibody fragment. Most importantly, B2M-aTac(dsFv)-mediated coating of target tumor cells made them susceptible for efficient and specific HLA-A2-restricted, melanoma gp100 peptide-specific CTL-mediated lysis. These results demonstrate the concept that antibody-guided tumor antigen-specific targeting of MHC-peptide complexes on tumor cells can render them susceptible and potentiate CTL killing. This novel approach now opens the way for the development of new imimounotherapeutic strategies based on antibody targeting of natural cognate MHC ligands and CTL-based cytotoxic mechanisms. Hence, while reducing the present invention to practice a novel strategy was developed to re-target class I MHC-peptide complexes on the surface of tumor cells in a way that is independent of the extent of class I MHC expression by the target tumor cells. To this end, in one embodiment of the present invention, two arms of the immune system were employed in fusion. One arm, the targeting moiety, comprises tumor-specific recombinant fragments of antibodies directed to tumor or differentiation antigens which have been used for many years to target radioisotopes, toxins or drugs to cancer cells (22, 23). The second, effector arm, is a single-chain MHC molecule (scMHC) composed of human β2-microglobulin linked to the three extracellular domains of the HLA-A2 heavy chain (24,25, WO 01/72768). By connecting the two molecules into a single recombinant gene and expressing the gene. The new molecule is expressed efficiently in E. coli and produced, for example, by in vitro refolding in the presence of HLA-A2-restricted peptides. This approach, as shown herein, renders the target tumor cells susceptible to lysis by cytotoxic T cells regardless of their MHC expression level and thus may be employed as a new approach to potentiate CTL-mediated anti-tumor immunity. This novel approach will lead to the development of a new class of recombinant therapeutic agents capable of selective killing and elimination of tumor cells utilizing natural cognate MHC ligands and CTL-based cytotoxic mechanisms. According to one aspect of the present invention there is provided an immuno-molecule comprising: a soluble human MHC class I effector domain; and a targeting domain being linked to the soluble human MHC class I effector domain. Thus, according to another aspect of the present invention there is provided a nucleic acid construct encoding an immuno-molecule, the construct comprising: a first polynucleotide encoding a soluble human MHC class I effector domain; and a second polynucleotide encoding a targeting domain; the first polynucleotide and the second polynucleotide are selected and being joined such that the soluble human MHC class I effector domain and the antibody targeting domain are translationally fused optionally via a peptide linker in-between. According to still another aspect of the present invention there is provided a nucleic acid construct encoding an immuno-molecule, the construct comprising: a first polynucleotide encoding a soluble human MHC class I effector domain; and a second polynucleotide encoding a variable region of one of a light chain or a heavy chain of an antibody targeting domain; the first polynucleotide and the second polynucleotide are selected and being joined such that the soluble human MHC class I effector domain and the variable region of the one of the light chain and heavy chain of the antibody targeting domain are translationally fused optionally via a peptide linker in-between; and a third polynucleotide encoding the other of the one of the light chain and heavy chain of the antibody targeting domain. According to an additional aspect of the present invention there is provided a nucleic acid construct system comprising: a first nucleic acid construct which comprises: a first polynucleotide encoding a soluble human MHC class I effector domain; and a second polynucleotide encoding a variable region of one of a light chain or a heavy chain of an antibody targeting domain; the first polynucleotide and the second polynucleotide are selected and being joined such that the soluble human MHC class I effector domain and the variable region of the one of the light chain and heavy chain of the antibody targeting domain are translationally fused optionally via a peptide linker in-between; a second nucleic acid construct which comprises: a third polynucleotide encoding the other of the one of the light chain and heavy chain of the antibody targeting domain. According to a further aspect of the present invention there is provided a method of selectively killing a cell in a patient, the cell presenting an antigen (e.g., a receptor), the method comprising administering to the patient an immuno-molecule which comprises: a soluble human MHC class I effector domain complexed with an MHC-restricted peptide; and a targeting domain being linked to the soluble human MHC class I effector domain, the targeting domain being for selectively binding to the antigen; whereby, the soluble human MHC class I effector domain complexed with the MHC-restricted peptide initiates a CTL mediated immune response against the cell, thereby selectively killing the cell in vivo. According to further features in preferred embodiments of the invention described below, the targeting domain is an antibody targeting domain. According to still further features in the described preferred embodiments the targeting domain is a ligand targeting domain. According to still further features in the described preferred embodiments the ligand targeting domain is selected from the group consisting of PDGF, EGF, KGF, TGFα, IL-2, [IL-3, IL-4, IL-6, VEGF and its derivatives, TNF According to still further features in the described preferred embodiments the soluble human MHC class I effector domain and the antibody targeting domain are translationally fused, optionally with a translationally fused peptide linker in-between. According to still further features in the described preferred embodiments the antibody targeting domain comprises a variable region of a light chain of an antibody linked to the effector domain. According to still further features in the described preferred embodiments the variable region of the light chain of the antibody and the effector domain are translationally fused, optionally with a translationally fused peptide linker in-between. According to still further features in the described preferred embodiments the antibody targeting domain further comprises a variable region of a heavy chain of an antibody linked to the variable region of the light chain of the antibody. According to still further features in the described preferred embodiments the variable region of the heavy chain of the antibody and the variable region of the light chain of the antibody are translationally fused, optionally with a translationally fused peptide linker in-between. According to still further features in the described preferred embodiments the variable region of the heavy chain of the antibody is linked to the variable region of the light chain of the antibody via a peptide linker. According to still further features in the described preferred embodiments the variable region of the heavy chain of the antibody is linked to the variable region of the light chain of the antibody via at least one S—S bond. According to still further features in the described preferred embodiments the antibody targeting domain comprises a variable region of a heavy chain of an antibody linked to the effector domain. According to still further features in the described preferred embodiments the variable region of the heavy chain of the antibody and the effector domain are translationally fused, optionally with a translationally fused peptide linker in-between. According to still further features in the described preferred embodiments the antibody targeting domain further comprises a variable region of a light chain of an antibody linked to the variable region of the heavy chain of the antibody. According to still further features in the described preferred embodiments the variable region of the light chain of the antibody and the variable region of the heavy chain of the antibody are translationally fused, optionally with a translationally fused peptide linker in-between. According to still further features in the described preferred embodiments the variable region of the light chain of the antibody is linked to the variable region of the heavy chain of the antibody via a peptide linker. According to still further features in the described preferred embodiments the variable region of the light chain of the antibody is linked to the variable region of the heavy chain of the antibody via at least one S—S bond. According to still further features in the described preferred embodiments the antibody targeting domain is capable of binding to a tumor associated antigen. According to still further features in the described preferred embodiments the antibody targeting domain is capable of binding to a tumor specific antigen. According to still further features in the described preferred embodiments the soluble-human MHC class I effector domain comprises a functional human β-2 microglobulin and a functional human MHC class I heavy chain linked thereto. According to still further features in the described preferred embodiments the functional human MHC class I heavy chain comprises domains α 1-3. According to still further features in the described preferred embodiments the functional human β-2 microglobulin and the functional human MHC class I heavy chain are translationally fused, optionally with a translationally fused peptide linker in-between. According to still further features in the described preferred embodiments the soluble human MHC class I effector domain further comprises a MHC-restricted peptide. According to still further features in the described preferred embodiments the MHC-restricted peptide is linked to the functional human β-2 microglobulin. According to still further features in the described preferred embodiments the MHC-restricted peptide and the functional human β-2 microglobulin are translationally fused, optionally with a translationally fused peptide linker in-between. According to still further features in the described preferred embodiments the MHC-restricted peptide is complexed with the functional human MHC class I heavy chain. According to still further features in the described preferred embodiments the MHC-restricted peptide is derived from a common pathogen. According to still further features in the described preferred embodiments the MHC-restricted peptide is derived from a pathogen for which there is an active vaccination. According to still further features in the described preferred embodiments the MHC-restricted peptide is derived from a tumor associated or specific antigen. According to further features in preferred embodiments of the invention described below, any of the nucleic acid constructs described herein, further comprising at least one cis acting regulatory sequence operably linked to the coding polynucleotides therein. According to still further features in the described preferred embodiments the cis acting regulatory sequence is functional in bacteria. According to still further features in the described preferred embodiments the cis acting regulatory sequence is functional in yeast. According to still further features in the described preferred embodiments the cis acting regulatory sequence is functional in animal cells. According to still further features in the described preferred embodiments the cis acting regulatory sequence is functional in plant cells. According to still another aspect of the present invention there is provided a transformed cell comprising any of the nucleic acid constructs or the nucleic acid construct system described herein. According to further features in preferred embodiments of the invention described below, the cell is a eukaryotic cell selected from the group consisting of a mammalian cell, an insect cell, a plant cell, a yeast cell and a protozoa cell. According to still further features in the described preferred embodiments the cell is a bacterial cell. According to yet an additional aspect of the present invention there is provided an isolated preparation of bacterial derived inclusion bodies comprising over 30 percent by weight of an immuno-molecule as described herein According to still an additional aspect of the present invention there is provided a method of producing an immuno-molecule comprising: expressing, in bacteria, the immuno-molecule which comprises: a soluble human MHC class I effector domain which includes a functional human β-2 microglobulin and a functional human MHC class I heavy chain linked thereto; and a targeting domain being linked to the soluble human MHC class I effector domain; and isolating the immuno-molecule. According to further features in preferred embodiments of the invention described below, immuno-molecule further comprises an MHC-restricted peptide linked to the functional human β-2 microglobulin, the method further comprising refolding the immuno-molecule to thereby generate an MHC class I-MHC-restricted peptide complex. According to still further features in the described preferred embodiments isolating the immuno-molecule is via size exclusion chromatography. According to still further features in the described preferred embodiments an MHC-restricted peptide is co-expressed along with the immuno-molecule in the bacteria. According to still further features in the described preferred embodiments expressing, in the bacteria, the immuno-molecule is effected such that the immuno-molecule forms inclusion bodies in the bacteria. According to still further features in the described preferred embodiments the MHC-restricted peptide and the immuno-molecule co-form inclusion bodies in the bacteria. According to still further features in the described preferred embodiments isolating the immuno-molecule further comprises: denaturing the inclusion bodies so as to release protein molecules therefrom; and renaturing the protein molecules. According to still further features in the described preferred embodiments renaturing the protein molecules is effected in the presence of an MHC-restricted peptide. According to still further features in the described preferred embodiments the MHC-restricted peptide is co-expressed in the bacteria. The present invention successfully addresses the shortcomings of the presently known configurations by providing a new means with which to combat cancer. |
Method and apparatus for separation of milk, colostrum, and whey |
Apparatus and method for separation of milk and milk products, e.g., involving sequential separation of milk, colostrum, and whey components by cross-flow filtration. The apparatus and method in a preferred aspect employ cross-flow filtration, chromatography and fermentation to separate and fully utilize the components of milk, colostrum, and whey to generate numerous individual components, minimize waste, lower adverse environmental issues and provide enhanced economic benefits to dairy producers. A wide variety of consumer and nutraceutical products can be produced from the fractions and/or sub-fractions of milk products obtained from such separation. The invention further contemplates a methodology for selecting optimum membrane, device, and operating conditions to achieve a desired separation. |
1. A method for sequentially separating components of milk, comprising the steps of: (a) providing a milk source; (b) effectuating a sufficient flow of milk from the milk source through one or more cross-flow filtration modules, using one or more fluid delivery means, wherein each fluid delivery means is connected to at least one cross-flow filtration module; and (c) sequentially capturing one or more filtration fractions generated by the cross-flow filtration modules. 2. A method according to claim 1, wherein each cross-flow filtration module comprises at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels each extending between the inlet and outlet, that are of equal length to one another as measured between the inlet and the outlet. 3. A method according to claim 1, wherein the cross-flow filtration modules comprise filtration membranes selected from the group consisting of cellulose-based membranes, polymer-based membranes, and ceramic-based membranes. 4. A method according to claim 1, wherein the milk from the milk source is flown through a cream separator upstream of the cross-flow filtration modules to remove at least part of a fatty component of the milk. 5. A method according to claim 1, wherein the milk is pasteurized before being flowed to the cross-flow filtration modules. 6. A method according to claim 1, further comprising the step of controlling and monitoring temperature of the fluid within the cross-flow filtration modules. 7. A method according to claim 1, further comprising the step of recycling water generated by the cross-flow filtration modules. 8. A method according to claim 1, wherein the milk is flowed through a cross-flow filtration module to be separated into a casein-rich fraction and a casein-depleted fraction. 9. A method according to claim 8, wherein the casein-rich fraction of the milk is captured as retentate of the cross-flow filtration module, and wherein the casein-depleted fraction of the milk is captured as permeate of the cross-flow filtration module. 10. A method according to claim 8, wherein the cross-flow filtration module comprises a membrane having an average pore size in a range of from about 5 KD to about 3000 KD. 11. A method according to claim 8, wherein the cross-flow filtration module comprises a membrane having an average pore size in a range of from about 50 KD to about 2000 KD. 12. A method according to claim 8, wherein the cross-flow filtration module comprises a membrane having an average pore size in a range of from about 100 KD to about 2000 KD. 13. A method according to claim 8, wherein the cross-flow filtration module comprises a cellulose-based membrane selected from the group consisting of cellulose membranes, cellulose acetate membranes, and regenerated cellulose membranes. 14. A method according to claim 13, wherein the cellulose-based membrane has an average pore size in a range of from about 5 KD to about 100 KD. 15. A method according to claim 8, wherein the cross-flow filtration module comprises a regenerated cellulose membrane having an average pore size of about 100 KD. 16. A method according to claim 8, wherein the cross-flow filtration module comprises a polymer-based membrane selected from the group consisting of polysulfone (PS) membranes, polyethersulfone (PES) membranes, sulfonated-polyethersulfone (S-PES) membranes, sulfonated-polysulfone (S-PS) membranes, polyamide (PA) membranes, polyacrylonitrile (PAN), polytetrafluoroethylene (PTFE), polyolefin (PO), and polyvinylidinedifluoride (PVDF) membranes. 17. A method according to claim 16, wherein the polymer-based membrane has an average pore size in a range of from about 5 KD to about 3000 KD. 18. A method according to claim 8, wherein the cross-flow filtration module comprises a polymer-based membrane having an average pore size of about 2000 KD. 19. A method according to claim 8, wherein the cross-flow filtration module comprises a polysulfone membrane having an average pore size of about 2000 KD. 20. A method according to claim 8, further comprising the step of concentrating and/or diafiltering the casein-rich fraction. 21. A method according to claim 8, further comprising the step of concentrating and/or diafiltering the casein-depleted fraction. 22. A method according to claim 8, wherein the casein-rich fraction is used to manufacture a diary product selected from the group consisting of: casein, micellular casein, caseinates, cheese, process cheese, process cheese food, milk powder, skim milk powder, and substrate for milk protein concentrate. 23. A method according to claim 8, wherein the casein-depleted fraction is used to manufacture a diary product selected from the group consisting of: whey protein isolates (WPI), whey protein subcomponents, and whey protein concentrates (WPC). 24. A method according to claim 8, wherein the casein-depleted fraction is used for manufacturing a product selected from the group consisting of: infant formula, α-lactalbumin enriched WPI, sialyllactose and α-lactalbumin enriched WPC, sialyllactose and α-lactalbumin enriched WPI, β-lactoglobulin enriched WPI, β-lactoglobulin enriched WPC, α-lactalbumin depleted WPI, α-lactalbumin depleted WPC, sialyllactose and α-lactalbumin depleted WPI, sialyllactose and α-lactalbumin depleted WPC, sialyllactose enriched lactose, α-lactalbumin, β-lactoglobulin and sialyllactose. 25. A method according to claim 8, further comprising the steps of: adding fatty component of milk to the casein-rich fraction; and drying said casein-rich fraction to form milk powder having a predetermined concentration of fatty component. 26. A method according to claim 1, comprising the steps of: optionally flowing the milk from the milk source through a first cross-flow filtration module to remove at least a portion of bacteria contained therein; flowing the milk, optionally filtered in the first cross-flow filtration module, through a second cross-flow filtration module to separate the milk into a casein-rich fraction and a casein-depleted fraction; capturing the casein-rich fraction; flowing the casein-depleted fraction of the milk through a third cross-flow filtration module to form a fraction that is enriched with albumin and immunoglobulins and a fraction that is depleted of albumin and immunoglobulins; capturing the fraction that is enriched with albumin and immunoglobulins; flowing the fraction that is depleted of albumin and immunoglobulins of the milk through a fourth cross-flow filtration module to form a β-lactoglobulin-rich fraction and a β-lactoglobulin-depleted fraction; capturing the β-lactoglobulin-rich fraction; flowing the β-lactoglobulin-depleted fraction of the milk through a fifth cross-flow filtration module to form a α-lactalbumin-rich fraction and a α-lactalbumin-depleted fraction; capturing the α-lactalbumin-rich fraction; flowing the α-lactalbumin-depleted fraction of the milk through a sixth cross-flow filtration module to form a complex carbohydrates rich fraction and a complex carbohydrates depleted fraction; capturing the complex carbohydrates rich fraction; flowing the complex carbohydrates depleted fraction through a seventh cross-flow filtration module to form a lactose-rich fraction and a lactose-depleted fraction; capturing the lactose-rich fraction; and discharging and/or recycling the lactose-depleted fraction of milk. 27. A method according to claim 26, further comprising the step of pasteurizing the milk source and/or any fraction of the milk components generated therein. 28. A method according to claim 26, wherein the cross-flow filtration modules comprise filtration membranes selected from the group consisting of cellulose-based membranes, polymer-based membranes, and ceramic-based membranes. 29. A method according to claim 26, wherein the second cross-flow filtration module comprises a cellulose-based membrane selected from the group consisting of cellulose membranes, cellulose acetate membranes, and regenerated cellulose membranes. 30. A method according to claim 26, wherein the second cross-flow filtration module comprises a membrane having average pore size in the range from about 100 KD to about 3000 KD. 31. A method according to claim 26, wherein the second cross-flow filtration module comprises a membrane having an average pore size in a range of from about 100 KD to about 1000 KD, selected from the group consisting of cellulose-based membranes selected from the group consisting of cellulose membranes, cellulose acetate membranes, and regenerated cellulose membranes. 32. A method according to claim 26, wherein the second cross-flow filtration module comprises a polymeric membrane having an average pore size in a range of between 800 KD and 2500 KD and/or a measured bubble point between 65 and 120 PSIG. 33. A method according to claim 26, wherein the second cross-flow filtration module comprises a regenerated cellulose membrane having an average pore size of about 100 KD. 34. A method according to claim 26, further comprising the step of separating and purifying albumin and immunoglobulins from the fraction that is enriched with albumin and immunoglobulins, using a method selected from the group consisting of chromatography, cross-flow filtration, cross-flow chromatography, and diafiltration. 35. A method according to claim 26, further comprising the step of separating and purifying β-lactoglobulin from the β-lactoglobulin-rich fraction of the milk, using a method selected from the group consisting of chromatography, cross-flow filtration, cross-flow chromatography, and diafiltration. 36. A method according to claim 26, further comprising the step of separating and purifying α-lactalbumin from the α-lactalbumin-rich fraction of the milk, using a method selected from the group consisting of chromatography, cross-flow filtration, cross-flow chromatography, and diafiltration. 37. A method according to claim 36, further comprising the step of adding the separated and purified α-lactalbumin into the casein-depleted fraction of the milk generated by the second cross-flow filtration module to form an α-lactalbumin-enriched soluble milk protein concentrate. 38. A method according to claim 37, further comprising the step of drying the α-lactalbumin-enriched soluble milk protein concentrate to form a powder product. 39. A method according to claim 26, further comprising the step of separating and purifying complex carbohydrates from the complex carbohydrates-rich fraction of the milk, using a method selected from the group consisting of chromatography, cross-flow filtration, cross-flow chromatography, and diafiltration. 40. A method according to 39, further comprising the step of fractioning the complex carbohydrates into one or more subcomponents using a method selected from the group consisting of chromatography, cross-flow filtration, cross-flow chromatography, and diafiltration. 41. A method according to claim 26, further comprising the step of subjecting the lactose-rich fraction of the milk to a bacterial process and/or an enzymatic process. 42. A method according to claim 26, further comprising the step of fermenting the lactose-rich fraction of the milk to produce at least one product selected from the group consisting of lactobacillus, lactic acid, and Vitamin B-12. 43. A method according to claim 26, further comprising the step of crystallizing the lactose-rich fraction of the milk to produce at least one product selected from the group consisting of lactose, food grade lactose, lactobacillus, lactic acid, and Vitamin B-12. 44. A method according to claim 26, further comprising the step of combining the casein-rich fraction from the second cross-flow filtration module with the α-lactalbumin-rich fraction from the fifth cross-flow filtration module to form an α-lactalbumin-enriched substrate. 45. A method according to claim 26, further comprising the step of drying at least one of the captured fractions of milk by a method selected from the group consisting of lyophilization, spray-drying, freeze-drying, crystallization, and evaporation. 46. A method according to claim 26, wherein each cross-flow filtration module is connected to at least one fluid delivery means for flowing the milk or a fraction of the milk therethrough. 47. A method according to claim 26, wherein temperature of each cross-flow filtration module is controlled and monitored by temperature controlling/monitoring means. 48. A method according to claim 1, wherein sialyllactose is isolated from the milk, said method comprising the steps of: optionally flowing the milk from the milk source through a first cross-flow filtration module to filter out at least a portion of bacteria contained therein; flowing the milk, optionally filtered in the first cross-flow filtration module, through a second cross-flow filtration module to separate the milk into a casein-rich fraction and a casein-depleted fraction; capturing the casein-rich fraction; flowing the casein-depleted fraction of the milk through a third cross-flow filtration module to form a fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin, and a fraction that is depleted of said milk proteins; capturing the fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin; flowing the fraction that is depleted of said milk proteins through a fourth cross-flow filtration module to form a sialyllactose-enriched fraction and a sialyllactose-depleted fraction; capturing the sialyllactose-enriched fraction; and discharging the sialyllactose-depleted fraction. 49. A method according to claim 1, wherein the milk source supplies casein-depleted whey, and wherein sialyllactose is separated from said casein-depleted whey, comprising the steps of: optionally flowing the casein-depleted whey from the milk source through a first cross-flow filtration module to filter out at least a portion of bacteria contained therein; flowing the casein-depleted whey, optionally filtered in the first cross-flow filtration module, through a second cross-flow filtration module to form a fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin, and a fraction that is depleted of said milk proteins; capturing the fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin; flowing the fraction that is depleted of said milk proteins through a third cross-flow filtration module to form a sialyllactose-enriched fraction and a sialyllactose-depleted fraction; capturing the sialyllactose-enriched fraction; and discharging the sialyllactose-depleted fraction. 50. A method according to claim 1, wherein immunoglobulins are isolated and purified from the milk, said method comprising the steps of: optionally flowing the milk from the milk source through a first cross-flow filtration module to filter out at least a portion of bacteria contained therein; flowing the milk, optionally filtered in the first cross-flow filtration module, through a second cross-flow filtration module to separate the milk into a casein-rich fraction and a casein-depleted fraction; capturing the casein-rich fraction; flowing the casein-depleted fraction of the milk through a third cross-flow filtration module to form an immunoglobulin-rich fraction and an immunoglobulin-depleted fraction; and capturing both the immunoglobulin-rich fraction and the immunoglobulin-depleted fraction. 51. A method according to claim 50 further comprising the additional step of concentrating and/or diafiltering the immunoglobulin-rich fraction. 52. A method according to any one of claims 50 and 51 further comprising the additional step of purifying immunoglobulins from the immunoglobulin-rich fraction by a method selected from the group consisting of chromatography, cross-flow filtration, cross-flow chromatography, and diafiltration. 53. A method according to any one of claims 50 and 51 further comprising the additional step of concentrating and/or diafiltering the immunoglobulin depleted fraction for further uses. 54. A method according to any one of claims 50 and 51 wherein the immunoglobulins have therapeutic effects. 55. A method according to any one of claims 50 and 51 wherein the immunoglobulins are used to treat gastrointestinal track disorder. 56. A method according to any one of claims 50 and 51 wherein the immunoglobulins are used to treat a mammal of the same species as that of the milk source. 57. A method according to any one of claims 50 and 51 wherein the immunoglobulins are used to treat a mammal of a different species from that of the milk source. 58. A method according to claim 1, wherein the milk source supplies fluid containing mixtures of complex carbohydrates and lactose, and wherein complex carbohydrates are isolated and purified from said mixtures, said method comprising the steps of: flowing the fluid mixtures from the milk source through a first cross-flow filtration module to separate said mixtures into a complex carbohydrates rich fraction and a complex carbohydrate depleted fraction; capturing both the complex carbohydrates rich fraction and the complex carbohydrates depleted fraction; concentrating and/or diafiltering the complex carbohydrates rich fraction to obtain complex carbohydrates; crystallizing and/or drying the complex carbohydrates; and concentrating and/or diafiltering the complex carbohydrates depleted fraction to obtain lactose; and crystallizing and/or drying the lactose. 59. A method according to claim 1, wherein sialyllactose is isolated from the milk, said method comprising the steps of: flowing the milk from the milk source through a first cross-flow filtration module to separate the milk into a casein-rich fraction and a casein-depleted fraction; capturing the casein-rich fraction; flowing the casein-depleted fraction of the milk through a second cross-flow filtration module to form a fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin, and a fraction that is depleted of said milk proteins; capturing the fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin; flowing the fraction that is depleted of said milk proteins through a third cross-flow filtration module to form a sialyllactose-enriched fraction and a sialyllactose-depleted fraction; capturing the sialyllactose-enriched fraction; and discharging the sialyllactose-depleted fraction. 60. A method according to claim 1, wherein the milk source directly supplies casein-depleted whey, and wherein sialyllactose is separated from said casein-depleted whey, comprising the steps of: flowing the casein-depleted whey from the milk source through a first cross-flow filtration module to form a fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin, and a fraction that is depleted of said milk proteins; capturing the fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin; flowing the fraction that is depleted of said milk proteins through a second cross-flow filtration module to form a sialyllactose-enriched fraction and a sialyllactose-depleted fraction; capturing the sialyllactose-enriched fraction; and discharging the sialyllactose-depleted fraction. 61. A method according to claim 1, wherein immunoglobulins are isolated and purified from the milk, said method comprising the steps of: flowing the milk from the milk source through a first cross-flow filtration module to separate the milk into a casein-rich fraction and a casein-depleted fraction; capturing the casein-rich fraction; flowing the casein-depleted fraction of the milk through a second cross-flow filtration module to form an immunoglobulin-rich fraction and an immunoglobulin-depleted fraction; capturing both the immunoglobulin-rich fraction and the immunoglobulin-depleted fraction; 62. A method according to claim 61 further comprising the additional step of concentrating and/or diafiltering the immunoglobulin-rich fraction. 63. A method according to any one of claims 61 and 62 further comprising the additional step of purifying immunoglobulins from the immunoglobulin-rich fraction by a method selected from the group consisting of chromatography, cross-flow filtration, cross-flow chromatography, and diafiltration. 64. A method according to claim 61 further comprising the additional step of concentrating and/or diafiltering the immunoglobulin depleted fraction for further uses. 65. A method according to claim 1, wherein a fraction enriched with α-lactalbumin and sialyllactose are isolated from the milk, said method comprising the steps of: flowing the milk from the milk source through a first cross-flow filtration module to separate the milk into a casein, albumin, immunoglobulins, and β-lactoglobulin enriched fraction and a casein, albumin, immunoglobulins, and β-lactoglobulin depleted fraction; capturing the casein, albumin, immunoglobulins, and β-lactoglobulin enriched fraction; optionally utilizing the captured casein, albumin, immunoglobulins, and β-lactoglobulin enriched fraction to manufacture a diary product selected from the group consisting of: casein, micellular casein, caseinate, cheese, process cheese, process cheese food, milk powder, skim milk powder and substrate for milk protein concentrate; flowing the casein, albumin, immunoglobulins, and β-lactoglobulin depleted fraction of the milk through a second cross-flow filtration module to form a fraction that is enriched with α-lactalbumin and sialyllactose, and a fraction that is depleted of α-lactalbumin and sialyllactose; capturing the fraction that is enriched with α-lactalbumin and sialyllactose; and discharging the fraction that is depleted of α-lactalbumin and sialyllactose. 66. A method according to claim 65, wherein the first cross-flow filtration module comprises at least one filtration membrane selected from the group consisting of cellulose-based membranes, polymer-based membranes, and ceramic-based membranes, and wherein the second cross-flow filtration module comprises at least one filtration membrane selected from the group consisting of polymer-based membranes. 67. A method according to claim 65, wherein the first cross-flow filtration module comprises at least one filtration membrane having an average pore size in a range of from about 30 KD to about 200 KD, and wherein the second cross-flow filtration module comprises at least one polymer-based membrane having an average pore size in a range of from about 1 KD to about 30 KD. 68. A method according to claim 65, wherein the first cross-flow filtration module comprises at least one regenerated cellulose membrane having an average pore size in a range of from about 20 KD to about 100 KD, and wherein the second cross-flow filtration module comprises at least one polymer-based membrane having an average pore size in a range of from about 1 KD to about 30 KD. 69. A method according to claim 65, wherein the first cross-flow filtration module comprises at least one polymer-based membrane having an average pore size in a range of from about 30 KD to about 200 KD, and wherein the second cross-flow filtration module comprises at least one polymer-based membrane having an average pore size in a range of from about 1 KD to about 30 KD. 70. A method according to claim 1, wherein a fraction enriched with α-lactalbumin and sialyllactose are isolated from the milk, said method comprising the steps of: flowing the milk from the milk source through a first cross-flow filtration module to separate the milk into a casein-rich fraction and a casein-depleted fraction; capturing the casein-rich fraction; optionally utilizing the captured casein-rich fraction to manufacture a diary product selected from the group consisting of: casein, micellular casein, caseinate, cheese, process cheese, process cheese food, milk powder, skim milk powder and substrate for milk protein concentrate; flowing the casein-depleted fraction of the milk through a second cross-flow filtration module to form a fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin, and a fraction that is depleted of albumin, immunoglobulins, and β-lactoglobulin; capturing the fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin; optionally utilizing the captured fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin to manufacture a diary product selected from the group consisting of: β-lactoglobulin enriched whey protein isolate (WPI), β-lactoglobulin enriched whey protein concentrate (WPC), α-lactalbumin depleted WPI, α-lactalbumin depleted WPC, sialyllactose and α-lactalbumin depleted WPI, and sialyllactose and α-lactalbumin depleted WPC; flowing the albumin, immunoglobulins, and β-lactoglobulin depleted fraction of the milk through a third cross-flow filtration module to form a fraction that is enriched with α-lactalbumin and sialyllactose, and a fraction that is depleted of α-lactalbumin and sialyllactose; capturing the fraction that is enriched with α-lactalbumin and sialyllactose; discharging the fraction that is depleted of α-lactalbumin and sialyllactose. 71. A method according to claim 70, wherein the first cross-flow filtration module comprises at least one filtration membrane selected from the group consisting of cellulose-based membranes, polymer-based membranes, and ceramic-based membranes, wherein the second cross-flow filtration module comprises at least one filtration membrane selected from the group consisting of cellulose-based membranes, polymer-based membranes, and ceramic-based membranes, and wherein the third cross-flow filtration module comprises at least one filtration membrane selected from the group consisting of polymer-based membranes. 72. A method according to claim 70, wherein the first cross-flow filtration module comprises at least one filtration membrane having an average pore size in a range of from about 500 KD to about 3000 KD, wherein the second cross-flow module comprises at least one filtration membrane having an average pore size in a range of from about 30 KD to about 150 KD, and wherein the third cross-flow filtration module comprises at least one filtration membrane having an average pore size in a range of from about 1 KD to about 30 KD. 73. An apparatus for sequentially separating components of milk, comprising: (a) a milk source; (b) one or more cross-flow filtration modules communicatively connected to said milk source, for generating one or more filtration fractions; (c) one or more fluid delivery means connected to each of said cross-flow filtration modules to effectuate flow of milk through said cross-flow filtration modules for separation of milk components; and (d) one or more means downstream of each of said cross-flow filtration modules for sequentially capturing one or more filtration fractions generated by the cross-flow filtration modules. 74. An apparatus according to claim 73, wherein each cross-flow filtration module comprises at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels that are of equal length between the inlet and the outlet. 75. An apparatus according to claim 73, wherein the one or more cross-flow filtration modules comprise a filtration membrane selected from the group consisting of cellulose-based membranes, polymer-based membranes, and ceramic-based membranes. 76. An apparatus according to claim 73, further comprising a cream separator upstream of said cross-flow filtration modules for removing at least a portion of fatty component from the milk. 77. An apparatus according to claim 73, further comprising a pasteurizer upstream and/or downstream of said one or more cross-flow filtration modules for pasteurizing the milk. 78. An apparatus according to claim 73, further comprising temperature controlling/monitoring means for controlling and monitoring temperature of said milk and/or filtration fractions generated by the one or more cross-flow filtration modules. 79. An apparatus according to claim 73, comprising a cross-flow filtration module for separating the milk from the milk source into a casein-rich fraction and a casein-depleted fraction. 80. An apparatus according to claim 79, wherein the cross-flow filtration module comprises membranes selected from the group consisting of cellulose-based membranes, polymer-based membranes, and ceramic-based membranes. 81. An apparatus according to claim 79, wherein the cross-flow filtration module comprises a membrane having an average pore size in a range of from about 100 KD to about 3000 KD. 82. An apparatus according to claim 79, wherein the cross-flow filtration module comprises a membrane having an average pore size in a range of from about 100 KD to about 1000 KD, selected from the group consisting of cellulose-based membranes selected from the group consisting of cellulose membranes, cellulose acetate membranes, and regenerated cellulose membranes. 83. An apparatus according to claim 79, wherein the cross-flow filtration module comprises a polymeric membrane having an average pore size between 800 KD and 2500 KD and/or a measured bubble point between 65 and 120 PSIG. 84. An apparatus according to claim 79, wherein the cross-flow filtration module comprises a regenerated cellulose membrane having an average pore size of about 100 KD 85. An apparatus according to claim 73, comprising: an optional first cross-flow filtration module downstream of the milk source and communicatively connected thereto for filtering out all or at least a portion of bacteria contained in the milk; a second cross-flow filtration module, downstream of the first cross-flow filtration module if provided and communicatively connected thereto, or if not provided, then communicatively connected directly to the milk source, which separates the milk into a casein-rich fraction and a casein-depleted fraction; means connected to said second cross-flow filtration module for capturing the casein-rich fraction; a third cross-flow filtration module downstream of the second cross-flow filtration module and communicatively connected thereto, which receives the casein-depleted fraction and further separates it into a fraction that is enriched with albumin and immunoglobulins and a fraction that is depleted of albumin and immunoglobulins; means connected to said third cross-flow filtration module for capturing the fraction that is enriched with albumin and immunoglobulins; a fourth cross-flow filtration module downstream of the third cross-flow filtration module and communicatively connected thereto, which receives the fraction that is depleted of albumin and immunoglobulins and further separates it into a β-lactoglobulin-rich fraction and a β-lactoglobulin-depleted fraction; means connected to said fourth cross-flow filtration module for capturing the β-lactoglobulin-rich fraction; a fifth cross-flow filtration module downstream of the fourth cross-flow filtration module and communicatively connected thereto, which receives the β-lactoglobulin-depleted fraction and further separates it into a α-lactalbumin-rich fraction and a α-lactalbumin-depleted fraction; means connected to said fifth cross-flow filtration module for capturing the α-lactalbumin-rich fraction; a sixth cross-flow filtration module downstream of the fifth cross-flow filtration module and communicatively connected thereto, which receives the α-lactalbumin-depleted fraction and further separates it into a complex carbohydrates rich fraction and a complex carbohydrates depleted fraction; means connected to said sixth cross-flow filtration module for capturing the complex carbohydrates rich fraction; a seventh cross-flow filtration module downstream of the sixth cross-flow filtration module and communicatively connected thereto, which receives the complex carbohydrates depleted fraction and further separates it into a lactose-rich fraction and a lactose-depleted fraction; and means connected to said seventh cross-flow filtration module for capturing the lactose-rich fraction; means for discharging and/or recycling the lactose-depleted fraction. 86. An apparatus according to claim 73, comprising: a first cross-flow filtration module downstream of the milk source and communicatively connected thereto, which separates the milk into a casein-rich fraction and a casein-depleted fraction; means connected to said first cross-flow filtration module for capturing the casein-rich fraction; a second cross-flow filtration module downstream of the first cross-flow filtration module and communicatively connected thereto, which receives the casein-depleted fraction and further separates it into a fraction that is enriched with albumin and immunoglobulins and a fraction that is depleted of albumin and immunoglobulins; means connected to said second cross-flow filtration module for capturing the fraction that is enriched with albumin and immunoglobulins; a third cross-flow filtration module downstream of the second cross-flow filtration module and communicatively connected thereto, which receives the fraction that is depleted of albumin and immunoglobulins and further separates it into a β-lactoglobulin-rich fraction and a β-lactoglobulin-depleted fraction; means connected to said third cross-flow filtration module for capturing the β-lactoglobulin-rich fraction; a fourth cross-flow filtration module downstream of the third cross-flow filtration module and communicatively connected thereto, which receives the β-lactoglobulin-depleted fraction and further separates it into a α-lactalbumin-rich fraction and a α-lactalbumin-depleted fraction; means connected to said fourth cross-flow filtration module for capturing the α-lactalbumin-rich fraction; a fifth cross-flow filtration module downstream of the fourth cross-flow filtration module and communicatively connected thereto, which receives the α-lactalbumin-depleted fraction and further separates it into a complex carbohydrates rich fraction and a complex carbohydrates depleted fraction; means connected to said fifth cross-flow filtration module for capturing the complex carbohydrates rich fraction; a sixth cross-flow filtration module downstream of the fifth cross-flow filtration module and communicatively connected thereto, which receives the complex carbohydrates depleted fraction and further separates it into a lactose-rich fraction and a lactose-depleted fraction; and means connected to said sixth cross-flow filtration module for capturing the lactose-rich fraction; means for discharging and/or recycling the lactose-depleted fraction. 87. An apparatus according to any one of claims 85 and 86, further comprising a pasteurizer upstream and/or downstream of any of the cross-flow filtration modules for pasteurizing the milk source or any one or more filtration fractions generated by the cross-flow filtration modules. 88. An apparatus according to any one claims 85 and 86, comprising multiple fluid delivery means arranged in a manner that each cross-flow filtration module is connected to at least one fluid delivery means, said fluid delivery means function to effectuate a flow of the milk or a fraction of the milk through each cross-flow filtration module. 89. An apparatus according to any one of claims 85 and 86, further comprising temperature controlling/monitoring means for controlling and monitoring temperature of said milk and/or filtration fractions generated by the cross-flow filtration modules. 90. An apparatus according to any one of claims 85 and 86, further comprising a cream separator upstream of said cross-flow filtration modules for removing all or at least a portion of fatty component from the milk. 91. A method of milk separation, comprising separating milk to recover at least one milk product therefrom, by cross-flow membrane filtration, wherein said method does not include any chromatography or precipitation steps. 92. The method of claim 91, wherein the milk product comprises a material selected from the group consisting of fats, lipids, insoluble casein, immunoglobulins, albumin, beta-lactoglobulin, alpha-lactalbumin, complex carbohydrates, siallyllactose, simple carbohydrates, lactose. 93. The method of claim 91, wherein the cross-flow membrane filtration is carried out in a cross-flow filtration module including a filter with geometrically regular subchannels geometrically corresponding to one another in a flow passage for said filtration, wherein operating conditions and/or said subchannels have been optimized with respect to shear rate and/or permeate diffusion. 94. A α-lactalbumin-enriched soluble milk protein concentrate. 95. A β-lactoglobulin and α-lactalbumin-enriched whey protein isolate. 96. A sialyllactose-enriched whey protein isolate. 97. A method for isolating lactose and sialyllactose from a lactose and sialyllactose source liquid, comprising the steps of: (a) providing a lactose and sialyllactose source liquid comprising both lactose and sialyllactose; (b) optionally, removing proteins from said lactose and sialyllactose source liquid, by flowing said source liquid through a first cross-flow filtration module; (c) crystallizing the lactose and sialyllactose source liquid, so as to form lactose crystals and a soluble liquid; (d) decanting the soluble liquid, so as to isolate the lactose crystals therefrom; (e) passing said soluble liquid through a second cross-flow filtration module to form a sialyllactose-rich retentate fraction and a sialyllactose-depleted permeate fraction; (f) capturing the sialyllactose-enriched retentate fraction; and (g) discharging the sialyllactose depleted fraction. 98. A method according to claim 97, comprising step (b) for removing proteins from said lactose and sialyllactose source liquid, by flowing said source liquid through a first cross-flow filtration module, wherein said first cross-flow filtration module comprises at least one cellulose-based membrane, and wherein the second cross-flow filtration module comprises at least one polymer-based membrane. 99. A method according to claim 97, comprising step (b) for removing proteins from said lactose and sialyllactose source liquid, by flowing said source liquid through a first cross-flow filtration module, wherein said first cross-flow filtration module comprises at least one filtration membrane having an average pore size in a range of from about 5 KD to about 30 KD, and wherein the second cross-flow module comprises at least one filtration membrane having an average pore size in a range of from about 1 KD to about 30 KD. 100. A method for isolating lactose and sialyllactose, comprising the steps of: (a) providing a source liquid that contains lactose and sialyllactose therein, said source liquid being selected from the group consisting of milk, skim milk, milk whey, and colostrum; (b) flowing said source liquid through a first cross-flow filtration module, so as to separate said source fluid into a protein-rich fraction and a protein-depleted fraction; (c) capturing the protein-rich fraction; (d) optionally, utilizing the captured protein-rich fraction to manufacture a diary product selected from the group consisting of: milk, skim milk, reconstituted milk, casein, micellular casein, caseinate, whey protein isolate, whey protein concentrate, cheese, process cheese, process cheese food, milk powder, skim milk powder, and substrate for milk protein concentrate; (e) flowing the protein-depleted fraction through a second cross-flow filtration module to form a sialyllactose-enriched fraction and a sialyllactose-depleted fraction; (f) capturing the sialyllactose-rich fraction; and (g) discharging the sialyllactose-depleted fraction. 101. A method according to claim 100, wherein the first cross-flow filtration module comprises at least one cellulose-based membrane, and wherein the second cross-flow filtration module comprises at least one polymer-based membrane. 102. A method according to claim 100, wherein the first cross-flow filtration module comprises at least one filtration membrane having an average pore size in a range of from about 5 KD to about 30 KD, and wherein the second cross-flow module comprises at least one filtration membrane having an average pore size in a range of from about 1 KD to about 30 KD. 103. A method for fractioning multiple economically valuable components from a skim milk source, comprising the steps of: concentrating said skim milk source, so as to reach a concentration level within a range of from about 1.5 to about 6 times of that of said skim milk source before concentration; diafiltering said concentrated skim milk source for about 1 to about 5 times; flowing said skim milk source through a first cross-flow filtration module, so as to separate the skim milk source into a casein-rich fraction and a casein-depleted fraction; capturing the casein-rich fraction; utilizing the captured casein-rich fraction to manufacture a diary product selected from the group consisting of: casein, micellular casein, caseinate, cheese, process cheese, process cheese food, milk powder, skim milk powder and substrate for milk protein concentrate; flowing the casein-depleted fraction through a second cross-flow filtration module to form a fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin, and a fraction that is depleted of albumin, immunoglobulins, and β-lactoglobulin; capturing the fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin; utilizing the captured fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin to manufacture whey protein isolates; flowing the fraction that is depleted of albumin, immunoglobulins, and β-lactoglobulin through a third cross-flow filtration module, to form a fraction that is enriched with α-lactalbumin and sialyllactose, and a fraction that is depleted of α-lactalbumin and sialyllactose; capturing the fraction that is enriched with α-lactalbumin and sialyllactose; and utilizing said fraction that is enriched with α-lactalbumin and sialyllactose for nutraceutical preparations and/or production of infant formula. 104. A method according to claim 103, further comprising the step of flowing the fraction that is depleted of α-lactalbumin and sialyllactose through a fourth cross-flow filtration module, to form a lactose-enriched fraction and a lactose-depleted fraction. 105. A method according to claim 104, further comprising the steps of (1) fermenting the lactose-enriched fraction to form a biomass, and (2) flowing said biomass through a centrifuge and/or a fifth cross-flow filtration module, to form a biomass-enriched fraction and a lactic acid fraction. 106. A method according to claim 1, wherein the milk source supplies casein-depleted whey, and wherein α-lactalbumin and sialyllactose are separated from said casein-depleted whey, comprising the steps of: flowing the casein-depleted whey from the milk source through a first cross-flow filtration module to form a fraction that is enriched with milk proteins and a fraction that is depleted of milk proteins, wherein said milk proteins are selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin; capturing the fraction that is enriched with milk proteins; optionally, utilizing the captured fraction that is enriched with milk proteins for manufacturing milk products selected from the group consisting of whey protein isolates and whey protein concentrates; flowing the fraction that is depleted of milk proteins through a second cross-flow filtration module to form a fraction that is enriched with α-lactalbumin and sialyllactose, and a fraction that is depleted of α-lactalbumin and sialyllactose; capturing the fraction that is enriched with α-lactalbumin and sialyllactose; and discharging the fraction that is depleted of α-lactalbumin and sialyllactose. 107. A method according to claim 1, wherein the milk source supplies casein-depleted whey, and wherein α-lactalbumin and sialyllactose are separated from said casein-depleted whey, comprising the steps of: flowing the casein-depleted whey from the milk source through a first cross-flow filtration module to form a fraction that is enriched with milk proteins and a fraction that is depleted of milk proteins, wherein said milk proteins are selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin; capturing the fraction that is enriched with milk proteins; optionally, utilizing the captured fraction that is enriched with milk proteins for manufacturing milk products selected from the group consisting of whey protein isolates and whey protein concentrates; flowing the fraction that is depleted of milk proteins through a second cross-flow filtration module to form a α-lactalbumin-enriched fraction and a α-lactalbumin-depleted fraction; capturing the α-lactalbumin-enriched fraction; flowing the α-lactalbumin-depleted fraction through a third cross-flow filtration module to form a sialyllactose-enriched lactose fraction and a sialyllactose-depleted lactose fraction; capturing the sialyllactose-enriched lactose fraction; and discharging the sialyllactose-depleted lactose fraction. 108. A method for fractioning multiple economically valuable components from a skim milk source, comprising the steps of: concentrating said skim milk source, so as to reach a concentration level within a range of from about 1.5 to about 15 times of that of said skim milk source before concentration; flowing the concentrated skim milk source through a first cross-flow filtration module, to form a casein, albumin, immunoglobulins, and β-lactoglobulin rich fraction and a casein, albumin, immunoglobulins, and β-lactoglobulin depleted fraction; optionally, diafiltering the casein, albumin, immunoglobulins, and β-lactoglobulin rich fraction for about 1 to about 10 times; capturing the casein, albumin, immunoglobulins, and β-lactoglobulin rich fraction; utilizing the captured casein, albumin, immunoglobulins, and β-lactoglobulin rich fraction for manufacturing a diary product selected from the group consisting of: cheese, process cheese, process cheese food, milk powder, skim milk powder and substrate for milk protein concentrate; flowing the casein, albumin, immunoglobulins, and β-lactoglobulin depleted fraction through a second cross-flow filtration module, to form a fraction that is enriched with α-lactalbumin and sialyllactose, and a fraction that is depleted of α-lactalbumin and sialyllactose; capturing the fraction that is enriched with α-lactalbumin and sialyllactose; and utilizing the captured fraction that is enriched with α-lactalbumin and sialyllactose to manufacture a nutraceutical product and/or infant formula. 109. A method according to claim 108, further comprising the step of flowing the fraction that is depleted of α-lactalbumin and sialyllactose through a third cross-flow filtration module to form a lactose-enriched fraction, and a lactose-depleted fraction. 110. A method according to claim 109, further comprising the steps of (1) fermenting the lactose-enrich fraction to form a biomass, and (2) flowing said biomass through a centrifuge and/or a fourth cross-flow filtration module, to form a biomass-enriched fraction and a lactic acid fraction. 111. A method according to claim 108, where the said second cross-flow filtration membrane has a retention coefficient for α-lactalbumin, so that the retention rate of α-lactalbumin is within a range of from about 20% to about 80%. 112. A method for isolating lactoferrin from a skim milk source, comprising the steps of: concentrating said skim milk source, so as to reach a concentration level within a range of from about 1.5 to about 15 times of that of said skim milk source before concentration; flowing the concentrated skim milk source through a first cross-flow filtration module, to form a casein and lactoferrin rich retentate fraction and a casein and lactoferrin depleted permeate fraction; optionally, diafiltering the casein and lactoferrin rich retentive fraction for about 1 to about 10 times; treating the casein and lactoferrin rich retentate fraction, by adding acid and/or sodium chloride thereto, so as to facilitate the release of lactoferrin from the casein; diafiltering the treated casein and lactoferrin rich retentive fraction for about 3 to about 10 times, so to separate such into a casein rich retentate and a casein depleted permeate, wherein said casein depleted permeate contains released lactoferrin; flowing the casein depleted permeate fraction through a second cross-flow filtration module, to form a lactoferrin-enriched fraction and a lactoferrin-depleted fraction; capturing the lactoferrin-enriched fraction; and optionally, concentrating the lactoferrin-enriched fraction to form a concentrated lactoferrin product. 113. A method of separating a target substance from a mixture solution containing two or more components, comprising the steps of: flowing the mixture solution through a first cross-flow filtration module at a predetermined shear rate, wherein said first cross-flow filtration module comprises at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels of substantially equal length between the inlet and the outlet, wherein the target substance is passed into the permeate, and wherein other components of larger molecular sizes contained by the mixture solution are retained as retentive; holding the first cross-flow filtration module at a predetermined transmembrane pressure; and when the target substance reaches a maximum percent transmission, diafiltering the mixture solution for about 1.5 to about 20 times, wherein said predetermined shear rate and said predetermined transmembrane pressure are determined so as to maximize the percent transmission of the target substance through said first cross-flow filtration module. 114. A method according to claim 113, wherein said predetermined shear rate is adjusted to be within a range of from about 4,000 to about 7,000 inverse seconds, and wherein said predetermined transmembrane pressure is adjusted to be less than about 13 psig. 115. A method of separating a target substance from a mixture solution containing two or more components, comprising the steps of: flowing the mixture solution through a first cross-flow filtration module at a predetermined shear rate, wherein said first cross-flow filtration module comprises at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels of equal length between the inlet and the outlet, wherein the target substance is retained as a retentate, and wherein other components of smaller molecular sizes contained by the mixture solution are passed into the permeate; holding the first cross-flow filtration module at a predetermined transmembrane pressure; and when the other components of smaller molecular sizes reaches a maximum percent transmission, diafiltering the mixture solution for about 1.5 to about 20 times, wherein said predetermined shear rate and said predetermined transmembrane pressure are determined so as to maximize the percent transmission of the other components of smaller molecular sizes through said first cross-flow filtration module. 116. A method according to claim 115, wherein said predetermined shear rate is adjusted to be within a range of from about 4,000 to about 7,000 inverse seconds, and wherein said predetermined transmembrane pressure is adjusted to be less than about 13 psig. 117. A method of separating a target substance from a mixture solution containing two or more components, comprising the steps of: flowing the mixture solution through a first cross-flow filtration module at a predetermined channel velocity, wherein said first cross-flow filtration module comprises at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels of substantially equal length between the inlet and the outlet, wherein the target substance is passed into the permeate, and wherein other components of larger molecular sizes contained by the mixture solution are retained as retentate; holding the first cross-flow filtration module at a predetermined transmembrane pressure; and when the target substance reaches a maximum percent transmission, diafiltering the mixture solution for about 1.5 to about 20 times, wherein said predetermined channel velocity and said predetermined transmembrane pressure are determined so as to maximize the percent transmission of said target substance through said first cross-flow filtration module. 118. A method according to claim 117, wherein said predetermined channel velocity is adjusted to be within a range of from about 0.5 M/s to about 1.5 M/s, and wherein said predetermined transmembrane pressure is adjusted to be less than about 13 psig. 119. A method of separating a target substance from a mixture containing two or more components comprising the steps of: flowing the mixture solution through a first cross-flow filtration module at a predetermined channel velocity, wherein said first cross-flow filtration module comprises at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels of equal length between the inlet and the outlet, wherein the target substance is retained as a retentate, and wherein other components of smaller molecular sizes contained by the mixture solution are passed into the permeate; holding the first cross-flow filtration module at a predetermined transmembrane pressure; and when the other components of smaller molecular sizes reaches a maximum percent transmission, diafiltering the mixture solution for about 1.5 to about 20 times, wherein said predetermined channel velocity and said predetermined transmembrane pressure are determined so as to maximize the percent transmission of the other components of smaller molecular sizes through said first cross-flow filtration module. 120. A method according to claim 119, wherein said predetermined channel velocity is adjusted to be within a range of from about 0.5 M/s to about 1.5 M/s, and wherein said predetermined transmembrane pressure is adjusted to be less than about 13 psig. 121. A method according to any of the claims 113-120, wherein said predetermined transmembrane pressure is adjusted to be less than about 4 psig. 122. A method according to any of the claims 113-120, wherein said predetermined transmembrane pressure is adjusted to be approximately 0 psig. 123. A method for concentrating one or more target substances in a solution, comprising the steps of: providing a first cross-flow filtration module comprising at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels that are of equal length between the inlet and the outlet; and passing said solution through said first cross-flow filtration module at a shear rate within a range of from about 5,000 to about 20,000 inverse seconds, so as to concentrate the target substances as a retentate thereof, wherein transmembrane pressure of said first cross-flow filtration module is adjusted in such manner that permeate flow rate of said first cross-flow filtration module is maximized. 124. A method of concentrating one or more target substances in a solution, comprising the steps of: providing a first cross-flow filtration module comprising at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels that are of equal length between the inlet and the outlet; and passing said solution through said first cross-flow filtration module at a channel velocity within a range of from about 0.8 M/sec to about 3 M/sec, so as to concentrate the target substances as a retentate thereof, wherein transmembrane pressure of said first cross-flow filtration module is adjusted in such manner that permeate flow rate of said first cross-flow filtration module is maximized. 125. A method according to claim 8, further comprising the steps of: adding lactose component of milk to the casein-rich fraction; and drying said casein-rich fraction to form milk powder having a predetermined lactose concentration. 126. A α-lactalbumin and sialyllactose-enriched whey protein isolate. 127. A method for fractioning multiple economically valuable components from a skim milk source, comprising the steps of: flowing said skim milk source through a first cross-flow filtration module, so as to separate the skim milk source into a casein-rich fraction and a casein-depleted fraction and thereby concentrate said skim milk source, so as to reach a concentration level within a range of from about 1.5 to about 6 times of that of said skim milk source before concentration; optionally, diafiltering the casein-rich fraction on said first cross-flow filtration module or on another cross-flow filtration module for about 1 to about 5 times; capturing the casein-rich fraction; utilizing the captured casein-rich fraction to manufacture a diary product selected from the group consisting of: casein, micellular casein, caseinate, cheese, process cheese, process cheese food, milk powder, skim milk powder and substrate for milk protein concentrate; flowing the casein-depleted fraction through a second cross-flow filtration module to form a fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin, and a fraction that is depleted of albumin, immunoglobulins, and β-lactoglobulin; capturing the fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin; utilizing the captured fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin to manufacture whey protein isolates; flowing the fraction that is depleted of albumin, immunoglobulins, and β-lactoglobulin through a third cross-flow filtration module, to form a fraction that is enriched with α-lactalbumin and sialyllactose, and a fraction that is depleted of α-lactalbumin and sialyllactose; capturing the fraction that is enriched with α-lactalbumin and sialyllactose; and utilizing said fraction that is enriched with α-lactalbumin and sialyllactose for nutraceutical preparations and/or production of infant formula. 128. A method according to claim 127, further comprising the step of subjecting the fraction that is depleted of α-lactalbumin and sialyllactose to a bacterial process and/or an enzymatic process. 129. A method according to claim 127, further comprising the step of fermenting the fraction that is depleted of α-lactalbumin and sialyllactose to produce at least one product selected from the group consisting of lactobacillus, lactic acid, and Vitamin B-12. 130. A method for isolating lactoferrin from a skim milk source, comprising the steps of: flowing said skim milk source through a first cross-flow filtration module, so as to separate the skim milk source into a casein and lactoferrin rich retentate fraction and a casein and lactoferrin depleted fraction and thereby concentrate said skim milk source, so as to reach a concentration level within a range of from about 1.5 to about 6 times of that of said skim milk source before concentration; optionally, diafiltering the casein and lactoferrin rich retentive on said first cross-flow filtration module or on another cross-flow filtration module for about 1 to about 5 times; treating the casein and lactoferrin rich retentate fraction, by adjusting the pH and/or salt content thereto, so as to facilitate the release of lactoferrin from the casein; diafiltering the treated casein and lactoferrin rich retentive fraction for about 3 to about 10 times, so to separate such into a casein rich retentate and a casein depleted permeate, wherein said casein depleted permeate contains released lactoferrin; flowing the casein depleted permeate fraction through a second cross-flow filtration module, to form a lactoferrin-enriched fraction and a lactoferrin-depleted fraction; capturing the lactoferrin-enriched fraction; and optionally, concentrating the lactoferrin-enriched fraction to form a concentrated lactoferrin product. 131. A method according to claim 130 further comprising the steps of: flowing the casein and lactoferrin depleted fraction and the optional diafiltrate of the casein and lactoferrin rich fractions of the first cross-flow filtration module through a second cross-flow filtration module to form a fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin, and a fraction that is depleted of albumin, immunoglobulins, and β-lactoglobulin; capturing the fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin; utilizing the captured fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin to manufacture whey protein isolates; flowing the fraction that is depleted of albumin, immunoglobulins, and β-lactoglobulin through a third cross-flow filtration module, to form a fraction that is enriched with α-lactalbumin and sialyllactose, and a fraction that is depleted of α-lactalbumin and sialyllactose; capturing the fraction that is enriched with α-lactalbumin and sialyllactose; and utilizing said fraction that is enriched with α-lactalbumin and sialyllactose for nutraceutical preparations and/or production of infant formula. 132. A method according to claim 131, further comprising the step of subjecting the fraction that is depleted of α-lactalbumin and sialyllactose to a bacterial process and/or an enzymatic process. 133. A method according to claim 131, further comprising the step of fermenting the fraction that is depleted of α-lactalbumin and sialyllactose to produce at least one product selected from the group consisting of lactobacillus, lactic acid, and Vitamin B-12. 134. A method according to claim 1, wherein the milk source directly supplies casein-depleted whey, and wherein α-lactalbumin and sialyllactose is separated from said casein-depleted whey, comprising the steps of: flowing the casein-depleted whey from the milk source through a first cross-flow filtration module to form a fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin and a fraction that is depleted of said milk proteins; capturing the fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin; flowing the fraction that is depleted of said milk proteins through a second cross-flow filtration module to form a α-lactalbumin and sialyllactose enriched fraction and a α-lactalbumin and sialyllactose depleted fraction; capturing the α-lactalbumin and sialyllactose enriched fraction; and discharging the sialyllactose-depleted fraction. 135. A method according to claim 134, further comprising the step of subjecting the fraction that is depleted of α-lactalbumin and sialyllactose to a bacterial process and/or an enzymatic process. 136. A method according to claim 134, further comprising the step of fermenting the fraction that is depleted of α-lactalbumin and sialyllactose to produce at least one product selected from the group consisting of lactobacillus, lactic acid, and Vitamin B-12. 137. A method according to claim 1, wherein the milk source directly supplies casein-depleted whey, and wherein α-lactalbumin and sialyllactose is separated from said casein-depleted whey, comprising the steps of: flowing the casein-depleted whey from the milk source through a first cross-flow filtration module to form a fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin and partially depleted of α-lactalbumin and a fraction that is depleted of said milk proteins and containing a portion of α-lactalbumin; capturing the fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin and partially depleted of α-lactalbumin; flowing the fraction that is depleted of said milk proteins and containing a portion of α-lactalbumin through a second cross-flow filtration module to form a α-lactalbumin and sialyllactose enriched fraction and a α-lactalbumin and sialyllactose depleted fraction; capturing the α-lactalbumin and sialyllactose enriched fraction; and discharging the sialyllactose-depleted fraction. 138. A method according to claim 137, further comprising the step of subjecting the fraction that is depleted of α-lactalbumin and sialyllactose to a bacterial process and/or an enzymatic process. 139. A method according to claim 137, further comprising the step of fermenting the fraction that is depleted of α-lactalbumin and sialyllactose to produce at least one product selected from the group consisting of lactobacillus, lactic acid, and Vitamin B-12. 140. A method according to claim 1, wherein the milk source directly supplies casein-depleted whey, and wherein whey protein isolate (WPI), α-lactalbumin and sialyllactose are separated from said casein-depleted whey, comprising the steps of: flowing the casein-depleted whey from the milk source through a first cross-flow filtration module to form a fraction that is enriched with the fatty components of the casein-depleted whey and a fraction that is depleted of said fatty components of the casein-depleted whey; flowing the fraction that is depleted of said fatty components of the casein-depleted whey through a second cross-flow filtration module to form a fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin and a fraction that is depleted of said milk proteins; capturing the fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin; utilizing the captured fraction that is enriched with albumin, immunoglobulins, and β-lactoglobulin to manufacture whey protein isolates; flowing the fraction that is depleted of said milk proteins through a third cross-flow filtration module to form a α-lactalbumin and sialyllactose enriched fraction and a α-lactalbumin and sialyllactose depleted fraction; capturing the α-lactalbumin and sialyllactose enriched fraction; and discharging the sialyllactose-depleted fraction. 141. A method according to claim 140, further comprising the step of subjecting the fraction that is depleted of α-lactalbumin and sialyllactose to a bacterial process and/or an enzymatic process. 142. A method according to claim 140, further comprising the step of fermenting the fraction that is depleted of α-lactalbumin and sialyllactose to produce at least one product selected from the group consisting of lactobacillus, lactic acid, and Vitamin B-12. 143. A method according to claim 1, wherein the milk source directly supplies casein-depleted whey, and wherein whey protein isolate (WPI), α-lactalbumin and sialyllactose are separated from said casein-depleted whey, comprising the steps of: flowing the casein-depleted whey from the milk source through a first cross-flow filtration module to form a fraction that is enriched with the fatty components of the casein-depleted whey and a fraction that is depleted of said fatty components of the casein-depleted whey; flowing the fraction that is depleted of said fatty components of the casein-depleted whey through a second cross-flow filtration module to form a fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin and partially depleted of α-lactalbumin and a fraction that is depleted of said milk proteins and containing a portion of α-lactalbumin; capturing the fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin and partially depleted of α-lactalbumin; utilizing the captured fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, and β-lactoglobulin and partially depleted of α-lactalbumin to manufacture whey protein isolates; flowing the fraction that is depleted of said milk proteins and containing a portion of α-lactalbumin through a third cross-flow filtration module to form a α-lactalbumin and sialyllactose enriched fraction and a α-lactalbumin and sialyllactose depleted fraction; capturing the α-lactalbumin and sialyllactose enriched fraction; and discharging the sialyllactose-depleted fraction. 144. A method according to claim 143, further comprising the step of subjecting the fraction that is depleted of α-lactalbumin and sialyllactose to a bacterial process and/or an enzymatic process. 145. A method according to claim 143, further comprising the step of fermenting the fraction that is depleted of α-lactalbumin and sialyllactose to produce at least one product selected from the group consisting of lactobacillus, lactic acid, and Vitamin B-12. 146. A method according to claim 1, wherein milk protein concentrate (MPC) and α-lactalbumin and sialyllactose are isolated from the milk, said method comprising the steps of: flowing the milk from the milk source through a first cross-flow filtration module to separate the milk into a fraction enriched with milk proteins selected from the group consisting of casein, albumin, immunoglobulins, and β-lactoglobulin and a fraction that is depleted of said milk proteins casein, albumin, immunoglobulins, and β-lactoglobulin; capturing the casein, albumin, immunoglobulins, and β-lactoglobulin rich fraction; utilizing the captured fraction that is enriched with milk proteins selected from the group consisting of casein, albumin, immunoglobulins, and β-lactoglobulin to manufacture milk protein concentrate; optionally, utilizing the captured milk protein concentrate to manufacture a diary product selected from the group consisting of: milk, skim milk, reconstituted milk, casein, micellular casein, caseinate, whey protein isolate, whey protein concentrate, cheese, process cheese, process cheese food, milk powder, and skim milk powder; flowing the casein, albumin, immunoglobulins, and β-lactoglobulin depleted fraction of the milk through a second cross-flow filtration module to form a α-lactalbumin and sialyllactose-enriched fraction and a α-lactalbumin and sialyllactose-enriched depleted fraction; capturing the α-lactalbumin and sialyllactose-enriched enriched fraction; and discharging the α-lactalbumin and sialyllactose-enriched depleted fraction. 147. A method according to claim 146, further comprising the step of subjecting the fraction that is depleted of α-lactalbumin and sialyllactose to a bacterial process and/or an enzymatic process. 148. A method according to claim 146, further comprising the step of fermenting the fraction that is depleted of α-lactalbumin and sialyllactose to produce at least one product selected from the group consisting of lactobacillus, lactic acid, and Vitamin B-12. 149. A method according to claim 1, wherein milk protein concentrate (MPC) and α-lactalbumin and sialyllactose are isolated from the milk, said method comprising the steps of: flowing the milk from the milk source through a first cross-flow filtration module to separate the milk into a fraction enriched with milk proteins selected from the group consisting of casein, albumin, immunoglobulins, and α-lactoglobulin and partially depleted of α-lactalbumin and a fraction that is depleted of said milk proteins and containing a portion of α-lactalbumin capturing the casein, albumin, immunoglobulins, and β-lactoglobulin and partially depleted of α-lactalbumin fraction; utilizing the captured fraction that is enriched with milk proteins selected from the group consisting of casein, albumin, immunoglobulins, and β-lactoglobulin and partially depleted of α-lactalbumin to manufacture milk protein concentrate; optionally, utilizing the captured milk protein concentrate to manufacture a diary product selected from the group consisting of: milk, skim milk, reconstituted milk, casein, micellular casein, caseinate, whey protein isolate, whey protein concentrate, cheese, process cheese, process cheese food, milk powder, and skim milk powder; flowing the fraction that is depleted of said milk proteins and containing a portion of α-lactalbumin through a second cross-flow filtration module to form a α-lactalbumin and sialyllactose-enriched fraction and a α-lactalbumin and sialyllactose-enriched depleted fraction; capturing the α-lactalbumin and sialyllactose-enriched enriched fraction; and discharging the α-lactalbumin and sialyllactose-enriched depleted fraction. 150. A method according to claim 149, further comprising the step of subjecting the fraction that is depleted of α-lactalbumin and sialyllactose to a bacterial process and/or an enzymatic process. 151. A method according to claim 149, further comprising the step of fermenting the fraction that is depleted of α-lactalbumin and sialyllactose to produce at least one product selected from the group consisting of lactobacillus, lactic acid, and Vitamin B-12. 152. A α-lactalbumin enriched product selected from the group consisting of nutraceutical preparation, juvenile formula, infant formula and senior formula. 153. A α-lactalbumin and sialyllactose enriched product selected from the group consisting of nutraceutical preparation, juvenile formula, infant formula and senior formula. 154. A sialyllactose enriched product selected from the group consisting of nutraceutical preparation, juvenile formula, infant formula and senior formula. 155. A β-lactoglobulin enriched product selected from the group consisting of nutraceutical preparation, juvenile formula, infant formula and senior formula. 156. A lactoferrin enriched product selected from the group consisting of nutraceutical preparation, juvenile formula, infant formula and senior formula. 157. A α-lactalbumin, sialyllactose and lactoferrin enriched product selected from the group consisting of nutraceutical preparation, juvenile formula, infant formula and senior formula. 158. A α-lactalbumin and lactoferrin enriched product selected from the group consisting of nutraceutical preparation, juvenile formula, infant formula and senior formula. 159. A sialyllactose and lactoferrin enriched product selected from the group consisting of nutraceutical preparation, juvenile formula, infant formula and senior formula. 160. A β-lactoglobulin and lactoferrin enriched product selected from the group consisting of nutraceutical preparation, juvenile formula, infant formula and senior formula. 161. A α-lactalbumin, sialyllactose and lactoferrin enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 162. A α-lactalbumin and sialyllactose enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 163. A α-lactalbumin and lactoferrin enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 164. A sialyllactose and lactoferrin enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 165. A sialyllactose enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 166. A lactoferrin enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 167. A α-lactalbumin enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 168. A β-lactoglobulin and lactoferrin enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 169. A β-lactoglobulin enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 170. A lactoferrin enriched product selected from the group consisting of milk protein concentrate, whey protein isolate and whey protein concentrate. 171. A method according to any of the claims 1-50, 58-62, 64-72, 73, 97-120, 123-125 and 127-151, wherein at least one of the cross-flow filtration module(s) is of uniform geometry. 172. A method according to any of the claims 1-50, 58-62, 64-72, 73, 97-120, 123-125 and 127-151, wherein each of the cross-flow filtration module(s) comprises at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels that are of substantially equal length between the inlet and the outlet. 173. An apparatus according to any of the claims 73-86, wherein at least one of the cross-flow filtration module(s) is of uniform geometry. 174. An apparatus according to any of the claims 73-86, wherein each of the cross-flow filtration module(s) comprises at least one permeate, at least one inlet, at least one outlet, and multiple fluid-flow sub-channels that are of substantially equal length between the inlet and the outlet. |
<SOH> FIELD OF THE INVENTION <EOH>The present invention relates to method and apparatus for sequential separation of various nutritional components of milk, particularly sequential separation of various milk proteins, carbohydrates, enzymes, and minerals contained in milk, colostrum, whey, or other diary products, using cross-flow filtration modules. |
<SOH> SUMMARY OF THE INVENTION <EOH>The invention relates in one broad aspect to a method and apparatus for separating raw milk, milk-based diary product, or dairy waste into multiple components in a sequential fashion, using cross-flow filtration modules, as described more fully hereinafter. In one specific aspect, the present invention relates to a method for separating milk by cross-flow filtration, comprising the steps of: a) providing a milk source; b) effectuating flow of milk from the milk source through one or more cross-flow filtration modules, using a fluid delivery means, wherein each fluid delivery means is connected to at least one cross-flow filtration module; and c) sequentially capturing one or more filtration fractions generated by the cross-flow filtration modules. The term “milk” in the present application is intended to be broadly construed to encompass any type of natural or modified dairy products, including, but not limited to: milk per se (i.e., whole milk), skim milk, milk fat, colostrum, whey, milk fractions, milk concentrates, milk dilutions, milk subcomponents, milk isolates, and other lactic materials, unless the context otherwise requires. Such natural or modified dairy products may originally be obtained from bovine, ovine, human, goat, rabbit, deer, or other mammal sources, and they may include mixtures of lactic materials from two or more of such mammals. In another specific aspect, the present invention relates to an apparatus for isolating and purifying one or more milk components, comprising: a) a milk source; b) one or more cross-flow filtration modules communicatively connected to the milk source, for generating one or more filtration fractions; c) one or more fluid delivery means connected to each of the cross-flow filtration modules for creating sufficient flow of milk through the cross-flow filtration modules to effect separation of milk components; and d) one or more means downstream of each of the cross-flow filtration modules for sequentially capturing one or more fractions generated by the cross-flow filtration modules. “Cross-flow filtration module” refers herein to a type of filter module or filter cassette that comprises a porous filter element across a surface of which the liquid medium to be filtered is flowed in a tangential flow fashion, for permeation through the filter element of selected component(s) of the liquid medium. In a cross-flow filtration module employed in accordance with the present invention, the shear force exerted on the filter element (separation membrane surface) by the flow of the liquid medium serves to oppose accumulation of solids on the surface of the filter element. Useful cross-flow filters include microfiltration, ultrafiltration, nanofiltration and reverse osmosis filter systems. The cross-flow filter may comprise a multiplicity of filter sheets (filtration membranes) in an operative stacked arrangement, e.g., wherein filter sheets alternate with permeate and retentate sheets, and as a liquid to be filtered flows across the filter sheets, impermeate (non-permeating) species, e.g., solids or high-molecular-weight species of diameter larger than the filter sheet's pore size(s), are retained in the retentate flow, and the liquid along with any permeate species diffuse through the filter sheet and enter the permeate flow. In a preferred embodiment of the present invention, such cross-flow filtration module comprises a permeate collection and discharge arrangement, a feed inlet, a retentate outlet, and multiple fluid-flow sub-channels that may for example be equidistant to the inlet and the outlet, i.e., the length of each such sub-channel as measured between its inlet and outlet is equal to other such sub-channels. Cross-flow filtration modules and cross-flow filter cassettes useful in practice of the present invention are commercially available from North Carolina SRT, Inc. (Cary, N.C.), and are variously described in the following U.S. patents of Henry B. Kopf: U.S. Pat. No. 4,867,876, “Filter Plate, Filter Plate Element, and Filter Comprising Same, issued Sep. 19, 1989; U.S. Pat. No. 4,882,050, same title, issued Nov. 21, 1989; U.S. Pat. No. 5,034,124, same title, issued Sep. 11, 1990; U.S. Pat. No. 5,049,268, same title, issued Sep. 17, 1991; U.S. Pat. No. 5,232,589, “Filter Element and Support, issued Aug. 3, 1993; U.S. Pat. No. 5,342,517, “Filter Cassette Article,” issued Aug. 30, 1994; U.S. Pat. No. 5,593,580, same title, issued Jan. 14, 1997; and U.S. Pat. No. 5,868,930, same title, issued Feb. 9, 1999; the disclosures of all of which are hereby incorporated herein by reference in their respective entireties. One specific aspect of the present invention relates to separation of a casein-rich fraction and a casein-depleted fraction of milk, comprising the steps of: a) providing a source of milk; b) optionally flowing the milk through a cream separator to remove all or at least a portion of the fatty component of the milk; c) optionally pasteurizing the milk, using a pasteurizer; d) flowing the milk through a cross-flow filtration module to separate the milk into a casein-rich retentate fraction and a casein-depleted permeate fraction; and e) recovering both the casein rich fraction and the casein depleted fraction generated by the cross-flow filtration module. The casein-rich fraction generated by such process can be used for manufacturing various dairy products, such as cheese, milk powder, and substrate for cheese production or milk protein concentrate. The casein-depleted fraction generated by such process contains various soluble whey proteins, such as IgG, albumin, alpha-lactalbumin and beta-lactoglobulin, and it can be used for manufacturing of whey protein isolates, subcomponents, and concentrates. During prior art cheese-making processes, whey proteins are usually harvested from the supernatant waste of cheese manufacturing and therefore contain casein-precipitants such as rennet or acid, which deleteriously reduce the quality and nutritional value of the whey proteins thus obtained. By contrast, the method of the present invention separates casein from the milk without introducing any chemical precipitants that will undermine the nutritional integrity of natural milk. Thus, the casein-separation process according to the present invention creates two liquid fractions, one being enriched in casein and the other being depleted of casein, in which both are free of chemical precipitants. The casein-depleted fraction is a clear yellow-green liquid containing unaltered immunoglobulins, α-lactalbumin, β-lactoglobulin, bovine serum albumin, lactoferrin, lactoperoxidase, immunoglobulins, carbohydrates, peptides, sialyllactose and lactose, which can be subject to further uses. Moreover, in the mass production of milk proteins and powder milk, it is desirable to utilize all of the beneficial components of the milk feedstock. A preferred aspect of the present invention therefore relates to an integral process for sequentially isolating each of multiple useful components of milk, thereby separating milk into multiple fractions to facilitate efficient uses of each fraction, with minimal waste of beneficial components. Such integral process comprises the steps of: 1) providing a milk source; 2) optionally removing all or at least a portion of fatty component of the milk supplied by the milk source, using a cream separator; 3) optionally pasteurizing the milk, using a pasteurizer; 4) optionally flowing the milk through a first cross-flow filtration module, which filters out matter that is not natural component(s) of milk, such as bacteria; 5) flowing the (optionally filtered) milk through a second cross-flow filtration module to separate it into a retentate casein-rich fraction and a permeate casein-depleted fraction; 6) capturing the retentate casein-rich fraction; 7) flowing the permeate casein-depleted fraction of the milk through a third cross-flow filtration module suitable to form a retentate fraction that is enriched with macromolecules such as albumin and immunoglobulins and a permeate fraction depleted in such macromolecules; 8) capturing the retentate fraction that is enriched with macromolecules such as albumin and immunoglobulins; 9) flowing the permeate fraction depleted of the macromolecules through a fourth cross-flow filtration module to form a β-lactoglobulin-rich retentate fraction and a β-lactoglobulin-depleted permeate fraction; 10) capturing the β-lactoglobulin-rich retentate fraction; 11) flowing the β-lactoglobulin-depleted permeate fraction through a fifth cross-flow filtration module to form an x-lactalbumin-rich retentate fraction and an α-lactalbumin-depleted permeate fraction; 12) capturing the α-lactalbumin-rich retentate fraction; 13) flowing the α-lactalbumin-depleted permeate fraction through a sixth cross-flow filtration module to form a complex carbohydrates-rich retentate fraction and a complex carbohydrates-depleted permeate fraction; 14) capturing the complex carbohydrates-rich retentate fraction; 15) flowing the complex carbohydrates-depleted permeate fraction through a seventh cross-flow filtration module to form a lactose-rich retentate fraction and a lactose-depleted permeate fraction; 16) capturing the lactose-enriched retentate fraction; and 17) discharging the lactose-depleted permeate fraction out of the system. Such integral process enables a maximal utilization of beneficial components contained in milk. It also achieves the purpose of minimizing waste, prolonging the shelf life of the milk product, and maintaining the natural nutritional integrity of milk. In one preferred embodiment of the present application, each of the cross-flow filtration modules comprises a permeate collection structure, an inlet, an outlet, and multiple fluid-flow sub-channels that may for example be equidistant (equally long) between the inlet and outlet. The cross-flow filtration modules are preferably connected to one or more fluid delivery (feed) means, which facilitates the flow of milk or fraction of the milk through the cross-flow filtration module at a sufficient shear rate. It is also preferred to provide temperature controlling/monitoring means to control and monitor the temperature of the fluids processed by the cross-flow filtration modules. Since the flow rates of milk or fraction of milk through each cross-flow filtration module correlate with temperatures, such temperature controlling/monitoring means function so as to specifically enhance the speed of the separation process. Moreover, the temperature controlling/monitoring means can be used to control microbial growth and to increase membrane performance and separation characteristics. One specific embodiment of the present invention provides means for (1) cleaning the milk-processing equipment, such as the cross-flow filtration modules and the fluid delivery means, and (2) recycling water generated by both the milk-separation process as well as the equipment-cleaning process. In another embodiment of the present application, one or more fractions generated by the integral separation process of the invention can be further fractionated, isolated, purified, or otherwise modified. For example, the retentate fraction enriched with albumin and immunoglobulins from the third cross-flow filtration module can be further separated and purified to form albumin and immunoglobulins, using a method such as chromatography, cross-flow chromatography, cross-flow filtration, etc. It is also preferable in respective aspects of the invention to separate and purify β-lactoglobulin and α-lactalbumin from the β-lactoglobulin and α-lactalbumin-rich fractions generated by the separation process, or to separate and purify complex carbohydrates from the complex carbohydrates-rich fraction, using the methods described hereinabove. The lactose-rich retentate fraction from the seventh cross-flow filtration module can also be crystallized or fermented to form additional useful products, such as for example lactobacillus , lactic acid, and Vitamin B-12. It is also preferable in various embodiments of the invention to subject such lactose-rich fraction to a bacterial or enzymatic process to further improve its nutritional value. Another aspect of the present invention relates to production of novel dairy products, by combining two or more milk fractions obtained from the integral separation process of the present invention. For example, one can add the fatty component of milk isolated by the cream separator to the casein-rich fraction generated by the second cross-flow filtration module, and then dry it to form milk powder enriched with milk fat. As another example, it is also desirable in various embodiments of the invention to add α-lactalbumin to the casein-depleted fraction of the milk generated by the second cross-flow filtration module, to form a α-lactalbumin-enriched soluble milk protein concentrate. Various other combinations of one or more milk fractions produced by the method of the present invention, are readily determinable by a person ordinarily skilled in the art. In various specific embodiments of the invention, it is desirable to dry or otherwise condense the milk components that have been separated and purified by the methods described hereinabove, for ease of preservation, storage, and transportation. Various techniques can be employed, including, but not limited to, lyophilization, spray-drying, freeze-drying, crystallization, and evaporation. In further embodiments of the invention, therapeutic components from milk (for example, blood clotting Factor VIII, proteins, hormones, monoclonal antibodies) of transgenic and/or hyper-immunized mammals are produced. Either column and/or cross-flow chromatography steps can be utilized in order to yield products of necessary purities, e.g., as ethical human therapeutic compounds for direct intravenous and/or intramuscular injection. The process of generating such an ethical human therapeutic compound of appropriate purity in one embodiment of the invention comprises the steps of: a) providing a source of milk from either a transgenic and/or hyper-immunized mammal; b) optionally flowing the milk from the milk source through a cream separator to remove all or at least a portion of the fatty component of such milk; c) optionally pasteurizing the milk, using a pasteurizer; d) optionally flowing the milk through a first cross-flow filtration module to filter out foreign matter that is not natural component(s) of milk, such as bacteria; e) flowing the filtered milk through a second cross-flow filtration module to form a casein-rich retentate fraction and a casein-depleted permeate fraction; f) capturing the casein-rich retentate fraction; g) flowing the casein-depleted permeate fraction through a chromatographic resin that is capable of binding at least one target component of the milk; and h) concentrating and/or diafiltering the eluting target component using a cross-flow chromatographic process. The term “target component” as used herein is defined as a human therapeutic agent, e.g., a compound such as a monoclonal antibody, immunoglobulin, etc. Such target compound can be used to treat or prevent various diseases, such as gastrointestinal tract disorder, hemophillia, leukemia, liver disease, diabetes, PKU, viral diseases, bacterial diseases, osteoarthritis, enzymatic deficiencies, protein deficiencies, Alzheimers, infection and cancer. The target compound may be used to treat a mammal of the same species as that of the milk source, or a mammal of a different species from that from which the milk source is derived. Another aspect of the present invention relates to a process for isolating siallylactose from milk, comprising: a) optionally flowing the milk from the milk source through a first cross-flow filtration module to filter out all or at least a portion of bacteria contained therein; b) flowing the filtered milk through a second cross-flow filtration module to separate the milk into a casein-rich fraction and a casein-depleted fraction; c) capturing the casein-rich fraction; d) flowing the casein-depleted fraction of the milk through a third cross-flow filtration module to form a fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin, and a fraction that is depleted of said milk proteins; e) capturing the fraction that is enriched with milk proteins selected from the group consisting of albumin, immunoglobulins, β-lactoglobulin, and α-lactalbumin; f) flowing the fraction that is depleted of said milk proteins through a fourth cross-flow filtration module to form a sialyllactose-enriched fraction and a sialyllactose-depleted fraction; g) capturing the sialyllactose-enriched fraction; and f) discharging the sialyllactose-depleted fraction. The milk separation process of the present invention enables production of many improved or entirely new dairy products which may not have been economically feasible or technically possible prior to the advent of the present invention, such as: 1) fresh or powdered milk of controlled and regulated protein content, particularly fresh or powdered milk enriched with one or more specific proteins such as α-lactalbumin, immunoglobulin, and/or lactoferrin, 2) milk protein concentrate, 3) carbohydrate-enriched milk, 4) lactose-depleted milk, 5) bovine immunoglobulin isolates; 6) drinks, shakes, milk, powders, baby food, or infant formula enriched with α-lactalbumin, carbohydrate, and/or sialyllactose, 7) purified natural sialyllactose, 8) milk enriched with various antibodies, such as Escherichia coli antibody, antibody to gastrointestinal tract disorders, 9) reformulated milk of one mammal which has a similar composition to another mammal's milk, particularly reformulated non-human mammalian milk having a similar composition to human breast milk, etc. Other aspects, features and embodiments of the present invention will be more fully apparent from the ensuing disclosure and appended claims. |
Gene expression in plastids based on replicating vectors |
A process of generating transgenic plants or plant cells transformed on their plastome and being devoid of a selection marker, comprising the following steps: (a) transforming plastids of a plant or plant cell with a DNA comprising: (i) a nucleotide sequence conferring replication of said DNA in a plant cell, (ii) at least one sequence of interest, (iii) sequences flanking said at least one sequence of interest necessary for stable integration of said at least one sequence of interest into the plastid genome, and (iv) a selection marker outside of said sequences flanking said sequence(s) of interest; (b) allowing integration of said at least one sequence of interest into the plastome in the presence of selective pressure; (c) allowing loss of said selection marker sequence by releasing selective pressure; and (d) recovering cells and/or plants being genetically transformed on their plastomes and being devoid of said selection marker. |
1. A process of generating transgenic plants or plant cells transformed on their plastome and being devoid of a selection marker, comprising the following steps: (a) transforming plastids of a plant or plant cell with a DNA comprising: (i) a nucleotide sequence conferring replication of said DNA in a plant cell, (ii) at least one sequence of interest, (iii) sequences flanking said at least one sequence of interest necessary for stable integration of said at least one sequence of interest into the plastid genome, and (iv) a selection marker outside of said sequences flanking said sequence(s) of interest; (b) allowing integration of said at least one sequence of interest into the plastome in the presence of selective pressure; (c) allowing loss of said selection marker sequence by releasing selective pressure; and (d) recovering cells and/or plants being genetically transformed on their plastomes and being devoid of said selection marker. 2. The process of claim 1, wherein said nucleotide sequence conferring replication of said DNA confers replication of said DNA in plastids. 3. The process of claim 2, wherein said replication is provided non-autonomously by reversible integration of said DNA into the plastome. 4. The process of claim 3, wherein the reversible integration is provided by an integration sequence of said DNA which is sufficiently homologous to a sequence of the plastome for homologous recombination. 5. The process of claim 4, wherein the integration sequence is contained in a sequence flanking said at least one sequence of interest (iii) and/or in one or more further sequence(s) other than said flanking sequence. 6. The process of claim 1, wherein said nucleotide sequence conferring replication of said DNA contains a sequence functional as an origin of replication in said plastids, thus conferring autonomous replication to said DNA. 7. The process of claim 6, wherein said DNA contains a further nucleotide sequence conferring replication containing a further sequence functional as an origin of replication in said plastids. 8. The process of claim 1, wherein said nucleotide sequence conferring replication of said DNA causes autonomous replication of said DNA outside of a plant plastid, thus generating multiple copies of said DNA for subsequent transfer to plastids. 9. The process of claim 1, wherein said sequence of interest comprises a sequence which allows visual identification of cells containing transformed plastomes. 10. The process of claim 1 wherein the plant or plant cells to be transformed has a mutation in the plastome which causes an easily distinguishable phenotype, and whereby said process restores said easily distinguishable mutant phenotype, thereby allowing identification of transformed cells. 11. The process of claim 1, wherein, in a previous step, a photosynthesis-related gene is rendered dysfunctional or is eliminated. 12. The process of claim 11, wherein said photosynthesis related gene is selected from the group consisting of rpoA, petA, ycf3, ycf9, and rpoB. 13. The process of claim 11, wherein said DNA contains said photosynthesis-related gene functional for allowing positive selection of said transformation. 14. The process of claim 1, wherein the selection marker is bacterial aminoglycoside phosphotransferase A-6. 15. The process of claim 1, wherein a nucleotide sequence of the plastome is deleted. 16. The process of claim 15, wherein said DNA does not contain a sequence of interest and said flanking sequences (iii) are homologous to plastome sequences that flank said nucleotide sequence of the plastome to be deleted. 17. The process of claim 1, wherein a point mutation is created in the plastome. 18. The process of claim 1 wherein steps (c) and/or (d) comprise allowing segregation of leaf sectors each containing a particular type of plastome. 19. A process of genetic transformation of plant plastids, comprising the following steps: (a) providing a plant plastid with DNA, which: (i) comprises a nucleotide sequence conferring autonomous replication of said DNA in a plant cell, (ii) comprises at least one sequence of interest; and (iii) said DNA is for transcription (a) devoid of transcription initiation and/or termination control elements operably linked to said at least one sequence of interest; or (b) devoid of transcription termination control elements operably linked to said at least one sequence of interest, while it comprises a transcription initiation control element operably linked to said at least one sequence of interest; (b) allowing for replication of said DNA; and (c) recovering cells and/or plants carrying genetically transformed plastids. 20. The process according to claim 19, wherein said nucleotide sequence conferring replication of said DNA causes autonomous replication of said DNA in a plant plastid. 21. The process according to claim 20, wherein transcription of said sequence(s) of interest is not terminated. 22. The process according to claim 21, wherein said transcription generates mRNA comprising multiple units of the transcribed sequence(s) of interest. 23. The process according to claim 19, wherein said nucleotide sequence conferring replication of said DNA causes autonomous replication of said DNA outside of a plant plastid, thus generating multiple copies of said DNA for subsequent transfer to plastids. 24. The process according to claim 19, wherein said DNA further contains one or more sequences providing for selection of transformed plants, cells, or plastids. 25. The process of claim 19, wherein, in a previous step, a photosynthesis-related gene is rendered dysfunctional or is eliminated. 26. The process of claim 25, wherein said photosynthesis-related gene is selected from the group consisting of rpoA, petA, ycf3, ycf9, and rpoB. 27. The process according to claim 25, wherein said DNA contains said photosynthesis-related gene functional for allowing positive selection of said transformation. 28. The process according to claim 19, wherein said DNA further contains sequences allowing stable integration of at least a portion of said DNA into a plastid genome. 29. The process according to claim 28, wherein, after introduction of at least a portion of said DNA into the plastid genome, it becomes free of undesired sequence(s) by recombination. 30. The process according to claim 28, wherein said at least a portion of said DNA comprises said at least one sequence of interest. 31. The process according to claim 30, wherein integration of said at least a portion of said DNA into the plastid genome leaves a sequence providing for selection on a second portion of said DNA which is not integrated into the plastid genome. 32. The process according to claim 31, wherein, after introduction of said at least a portion of said DNA into the plastid genome, selection pressure is released allowing loss of said sequence providing for selection. 33. The process according to claim 19, wherein said DNA is designed so that its maintenance in said plant plastid requires a selective pressure. 34. The process according to claim 19, wherein interaction of said at least one sequence of interest or its expression product with the plastid genome, or expression product(s) of the plastid genome generates a desired function. 35. A transformation vector having a DNA as defined in item (a) of claim 1. 36. Transplastomic plant cells or plants and progeny thereof generated by the process of claim 1. |
<SOH> BACKGROUND OF THE INVENTION <EOH>According to generally accepted knowledge, two classes of cell organelles, i.e. plastids and mitochondria, are derived from initially independent prokaryotes that were taken up into a predecessor of present day eukaryotic cells by separate endosymbiotic events (Gray, 1991). As a consequence, these organelles contain their own DNA, DNA transcripts in the form of messenger RNA, ribosomes, and at least some of the necessary tRNAs that are required for decoding of genetic information (Marechal-Drouard et al., 1991). While, shortly after endosymbiotic uptake, these organelles were genetically autonomous since they contained all the elements necessary to drive prokaryotic life, this autonomy was reduced during evolution by transfer of genetic information to the cell's nucleus. Nevertheless, their genetic information is of sufficient complexity to make recent cell organelles an attractive target for gene technology. This is particularly the case with plastids, because these organelles still encode about 50% of the proteins required for their main function inside the plant cell, photosynthesis. Plastids also encode their ribosomal RNAs, the majority of their tRNAs and ribosomal proteins. In total, the number of genes in the plastome is in the range of 120 (Palmer, 1991). The vast majority of proteins that are found in plastids are, however, imported from the nuclear/cytosolic genetic compartment. Plastids Can Be Genetically Transformed With the development of general molecular cloning technologies, it became soon possible to genetically modify higher plants by transformation. The main emphasis in plant transformation was and still is on nuclear transformation, since the majority of genes, ca. 26.000 in the case of Arabidopsis thaliana , the complete sequence of which was recently published (The Arabidopsis Genome Initiative, 2000), is found in the cell's nucleus. Nuclear transformation was easier to achieve, since biological vectors such as Agrobacterium tumefaciens were available, which could be modified to efficiently enable nuclear transformation (Galvin, 1998). In addition, the nucleus is more directly accessible to foreign nucleic acids, while the organelles are surrounded by two envelope membranes that are, generally speaking, not permeable to macromolecules such as DNA. A capability of transforming plastids is highly desirable since it could make use of the high gene dosage in these organelles that bears the potential of extremely high expression levels of transgenes. In addition, plastid transformation is attractive because plastid-encoded traits are not pollen transmissible; hence, potential risks of inadvertent transgene escape to wild relatives of transgenic plants are largely reduced. Other potential advantages of plastid transformation include the feasibility of simultaneous expression of multiple genes as a polycistronic unit and the elimination of positional effects and gene silencing that may result following nuclear transformation. Methods that allow stable transformation of plastids could indeed be developed for higher plants. To date, two different methods are available, i.e. particle bombardment of tissues, in particular leaf tissues (Svab et al., 1990), and treatment of protoplasts with polyethylene glycol (PEG) in the presence of suitable transformation vectors (Koop et al., 1996). Both methods mediate the transfer of plasmid DNA across the two envelope membranes into the organelle's stroma, Conventional plastid transformation technology is described in Heifetz, 2000 and Koop et al., 1996. Conventional plastid transformation vectors usually need to serve at least two purposes: (1) introduction of one or more desired foreign genes to be expressed by the genetic machinery of the plastids, and (2) selection of cells containing transformed plastomes by inhibitor selection and/or by screening for a detectable phenotype. Plastid transformation vectors usually contain complete gene cassettes consisting of four operable linked elements: a promoter sequence, a 5′ untranslated region, a coding region, and a 3′ untranslated region. These cassettes, however, do not make use of the potential to co-express several genes in an operon under the control of a single promotor. Selection is achieved either by replacing a complete resident plastid gene by a mutant gene, which confers resistance to selection inhibitors (U.S. Pat. No. 5,451,513) or by introducing a complete expression cassette, which leads to enzymatic inactivation of an inhibitor (U.S. _Pat. No. 5,877,402). These marker genes that are needed for the selection of transgenic plant cells from a vast background of untransformed cells code for antibiotic or herbicide resistance genes. Examples for plastid resistance genes are aadA conferring resistance to spectinomycin and streptomycin (Svab & Maliga, 1993), or nptII conferring resistance to kanamycin (Carrer et al., 1993). As these marker genes are stably integrated into the genome together with the genes of interest (GOI), they will stay in the homoplastomic transgenic plants although they are not required for GOI function. These remaining marker genes are a main issue of criticism of plant biotechnology as they could theoretically increase antibiotic resistance of pathogens or herbicide resistance of weeds. Construction of a selection system which does not result in a resistance gene in the transgenic plant is, therefore, highly desirable (Iamtham and Day, 2000). In addition to the two or more gene cassettes, transformation vectors usually contain flanking regions of the insertion site, which are necessary for the stable introduction of engineered sequences into the plastome by two reciprocal homologous recombination events. To this end, chloroplast transformation vectors contain chloroplast genome sequences to serve as homologous flanks. Since the chloroplast genomes of different species differ in their sequences, species-specific transformation vectors have to be used. This requires substantial effort, when cloning transformation vectors, and is in contrast to the situation in nuclear transformation. In all conventional transformation vectors, the selection marker is flanked by sequences homologous to plastid DNA; therefore, it is stably integrated into the plastome, although it is not needed for the desired function of the sequence(s) of interest. These remaining marker genes could theoretically spread to other organisms by giving a selective advantage. Increased antibiotic resistance in pathogens might cause problems in clinical treatment. Thus, the development of a system which results in transplastomic plants not containing any selection marker is highly desirable. A further advantage of such a system would be the possibility of re-using the same marker gene for subsequent transformations, which is at present difficult due to the limited number of efficient selection markers. Furthermore, the copy number of any transgene stably integrated into plastome molecules can obviously never exceed the plastome copy number, thus limiting the potential transgene expression level to a certain degree. Consequently, the copy number of the transgene(s) can be further increased when located on an extra-chromosomal element. U.S. Pat. No. 5,693,507 discloses a process for introducing a heterologous DNA into a chloroplast whereby the heterologous DNA contains operably linked control elements allowing for expression in the chloroplast. The process according to U.S. Pat. No. 5,693,507 has not provided long-term maintenance of the heterologous DNA in a plastid. Moreover, the expression of the heterologous DNA in the plastid is insufficient for practical application. Therefore, it is an object of the invention to provide an efficient and highly versatile process of genetic transformation of plant plastids whereby genetically stable transgenic plants may be produced. It is another object of the invention to provide a process of genetic transformation of plant plastids, which gives stably transformed plants and allows very high transgene expression levels. It is another object of the invention to provide a process of genetic transformation of plant plastids, which allows expression of multiple genes of interest (polycistronic expression). It is a further object of the invention to provide a novel process of genetic transformation of plant plastids, which gives transgenic plants not containing a marker gene such as an antibiotic resistance gene. It is a further object to provide vectors capable of replicating in plant cells, preferably in plastids, whereby the replication frequency of the vector is tuneable. |
Controlled agglomeration |
A process for the preparation of a particulate material by a controlled agglomeration method, i.e. a method that enables a controlled growth in particle size. The method is especially suitable for use in the preparation of pharmaceutical compositions containing a therapeutically and/or prophylactically active substance which has a relatively low aqueous solubility and/or which is subject to chemical decomposition. The process comprising i) spraying a first composition comprising a carrier, which has a melting point of about 5° C. or more which is present in the first composition in liquid form, on a second composition comprising a material in solid form, the second composition having a temperature of at the most a temperature corresponding to the melting point of the carrier and/or the carrier composition and ii) mixing or others means of mechanical working the second composition onto which the first composition is sprayed to obtain the particulate material. |
1. A process for the preparation of a particulate material, the method comprising i) spraying a first composition comprising a carrier, which has a melting point of about 5° C. or more such as, e.g., about 10° C. or more, about 20° C. or more or about 25° C. or more and which is present in the first composition In liquid form, on a second composition comprising a material In solid form, the second composition having a temperature of at the most a temperature corresponding to the melting point of the carrier and/or of the carrier composition such as, e.g., a temperature of at least about 2° C., at least about 5° C. or at least about 10° C. lower than the melting point of the carrier and/or of the carrier composition, and ii) mixing or other means of mechanical working the second composition onto which the first composition is sprayed to obtain the particulate material, whereby the second composition is agglomerated into particles in a controlled manner. 2. A process according to claim 1, wherein the process is carried out in a high or low shear mixer or in a fluid bed. 3. A process according to claim 1 or 2, wherein the process is carried out In a fluid bed and the spraying of the carrier composition is performed on the second composition in a fluidised state. 4. A process according to claims 1 or 2, wherein the spraying is performed through a spraying device equipped with temperature controlling means. 5. A process according to any of the preceding claims, wherein the particulate material obtained has a geometric weight mean diameter dgw of ≧10 μm such as, e.g. ≧20 μm, from about 20 to about 2000, from about 30 to about 2000, from about 50 to about 2000, from about 10 to about 2000, from about 75 to about 2000 such as, e.g. from about 100 to about 1500 μm, from about 100 to about 1000μm or from about 100 to about 700 μm, or at the most about 400 μm or at the most 300 μm such as, e.g., from about 50 to about 400 μm such as, e.g., from about 50 to about 350 μm, from about 50 to about 300 μm, from about 50 to about 250 μm or from about 100 to about 300 μm. 6. A process according to any of the preceding claims, wherein the concentration of the carrier in the particulate material is from about 5 to about 95% v/v such as, e.g. from about 5 to 90% v/v, from about 5 to about 85% v/v, from about 5 to about 80% v/v, from about 10 to about 75% v/v, from about 15 to about 75% v/v, from about 20 to abut 75% v/v, from about 25% to about 75% v/v, from about 30% to about 75% v/v, from about 35% to about 75% v/v, from about 25% to about 70% v/v, from about 30% to about 70% v/v, from about 35% to abut 70% v/v, from about 40% to about 70% v/v, from about 45% to about 65% v/v or from about 45% to about 60% v/v. 7. A process according to any of the preceding claims, wherein the carrier composition is brought on liquid form by heating the carrier and/or the carrier composition to a temperature, which causes the carrier and/or the carrier composition to melt. 8. A process according to claim 7, wherein the carrier and/or the carrier composition in liquid form has a viscosity (Brookfield DV-III) of at the most about 800 mPas at a temperature of at the most 100° C. such as, e.g., at the most 700, at the most 600, at the most 500 mPas. 9. A process according to any of the preceding claims, wherein the carrier composition is essentially non-aqueous and it contains at the most about 20% w/w water such as at the most about 15% w/w, at the most abut 10% w/w, at the most about 5% w/w or at the most about 2.5% w/w. 10. A process according to any of the preceding claims, wherein the carrier has a melting point of at least about 30° C. such as, e.g., at least about 35° C. or at least about 40° C. 11. A process according to any of the preceding claims, wherein the carrier has a melting point of at the most about 300° C. such as, e.g., at the most about 250° C., at the most about 200° C., at the most about 150° C. or at the most about 100° C. 12. A process according to any of the preceding claims, wherein the carrier is a hydrophilic carrier, a hydrophobic carrier, a surfactant or mixtures thereof. 13. A process according to claim 12, wherein the carrier is selected from the group consisting of: polyether glycols such as, e.g., polyethylene glycols, polypropylene glycols; polyoxyethylenes, polyoxypropylenes; poloxamers and mixtures thereof. 14. A process according to claim 12, wherein the carrier is selected from the group consisting of: xylitol, sorbitol, potassium sodium tartrate, sucrose tribehenate, glucose, rhamnose, lactitol, behenic acid, hydroquinon monomethyl ether, sodium acetate, ethyl fumarate, myristic acid, citric acid, Gelucire 50/13, Gelucire 44/14, Gelucire 50/10, Gelucire 62/05, Sucro-ester 7, Sucro-ester 11, Sucro-ester 15, maltose, mannitol and mixtures thereof. 15. A process according to claim 12, wherein the carrier is selected from the group consisting of: straight chain saturated hydrocarbons, sorbitan esters paraffins; fats and oils such as e.g., cacao butter, beef tallow, lard, polyether glycol esters; higher fatty acid such as, e.g. stearic acid, myrisic acid, palmitic acid, higher alcohols such as, e.g., cetanol, stearyl alcohol, low melting piont waxes such as, e.g., glyceryl monostearate, hydrogenated tallow, myristyl alcohol, stearyl alcohol, substituted and/or unsubstituted monoglycerides, substituted and/or unsubstituted diglycerides, substituted and/or unsubstituted triglycerides, yellow beeswax, white beeswax, carnauba wax, castor wax, japan wax, acetylate monoglycerides; NVP polymers, PVP polymers, acrylic polymers, or a mixture thereof. 16. A process according to any of claims 1-12, wherein the carrier is polyethylene glycol having an average molecular weight in a range of from about 400 to about 35,000 such as, e.g., from about 800 to about 35,000, from about 1,000 to about 35,000 such as, e.g., polyethylene glycol 1,000, polyethylene glycol 2,000, polyethylene glycol 3,000, polyethylene glycol 4,000, polyethylene glycol 5,000, polyethylene glycol 6000, polyethylene glycol 7,000, polyethylene glycol 8,000, polyethylene glycol 9,000 polyethylene glycol 10,000, polyethylene glycol 15,000, polyethylene glycol 20,000, or polyethylene glycol 35,000. 17. A process according to any of claims 1-12, wherein the carrier is polyethylene oxide having a molecular weight of from about 2,000 to about 7,000,000 such as, e.g. from about 2,000 to about 100,000, from about 5,000 to about 75,000, from about 10,000 to about 60,000, from about 15,000 to about 50,000, from about 20,000 to about 40,000, from about 100,000 to about 7,000,000 such as, e.g., from about 100,000 to about 1,000,000, from about 100,000 to about 600,000, from about 100,000 to about 400,000 or from about 100,000 to about 300,000. 18. A process according to any of claims 1-12, wherein the carrier is a poloxamer such as, e.g. Poloxamer 188, Poloxamer 237, Poloxamer 338 or Poloxamer 407. 19. A process according to any of claims 1-12, wherein the carrier is a sorbitan ester such as, e.g., sorbitan di-isostearate, sorbitan dioleate, sorbitan monolaurate, sorbitan monoisostearate, sorbitan monooleate, sorbitan monopalmitate, sorbitan monostearate, sorbitan sesqui-isostearate, sorbitan sesquioleate, sorbitan sesquistearate, sorbitan tri-isostearate, sorbitan trioleate, sorbitan tristearate or mixtures thereof. 20. A process according to any of the preceding claims, wherein the carrier composition comprises a mixture of hydrophilic and/or hydrophobic carriers. 21. A process according to any of the preceding claims, wherein the carrier composition comprises one or more therapeutically and/or prophylactically active substances. 22. A process according to any of the preceding claims, wherein the carrier composition comprises one or more pharmaceutically acceptable excipients. 23. A process according to any of the preceding claims, wherein the second composition comprises one or more pharmaceutically acceptable excipients. 24. A process according to any of the preceding claims, wherein the second composition comprises one or more therapeutically and/or prophylactically active substances. 25. A process according to claim 22 or 23, wherein the pharmaceutically acceptable excipient is selected from the group consisting of fillers, binders, disintegrants, glidants, colouring agents, taste-masking agents, pH-adjusting agents, solubilizing agents, stabilising agents, wetting agents, surface active agents, antioxidants etc. 26. A process according to claim 21 or 24, wherein the therapeutically and/or prophylactically active substance has an aqueous solubility at 25° C. and pH of 7.4 of at the most about 3 mg/ml such as, e.g., at the most about 2 mg/ml, at the most about 1 mg/ml, at the most about 750 μg/ml, at the most about 500 μg/ml, at the most about 250 μg/ml, at the most about 100 μg/ml, or at the most about 50 μg/ml, or at the most about 25 μg/ml, or at the most about 20 μg/ml or or at the most about 10 μg/ml. 27. A process according to any of the preceding claims, wherein the active substance is a cosmetically active or beneficial substance or a food or nutrient substance. 28. A process according to any of the preceding claims, wherein the second composition comprises magnesium aluminosilicate and/or magnesium aluminometasilicate such as, e.g. Neusilin S1, Neusilin FH2, Neusilin US2, Neusilin UFL2 or the like. 29. A process according to any of the preceding claims, wherein the second composition comprises magnesium aluminosilicate and/or magnesium aluminometasilicate such as, e.g., Neusilin, and the particulate material obtained has an content of carrier of at least about 30% v/v such as, e.g, at least about 40% v/v, at least about 50% v/v, at least about 60% v/v, at least about 70% v/v, at least about 75% v/v, at least about 80% v/v, at least about 85% v/v or at least about 90% v/v. 30. A process according to any of the preceding claims for the preparation of a particulate material for the manufacture of a phannaceutical, cosmetic or food composition. 31. A process according to claim 30, wherein the pharmaceutical, cosmetic or food composition is in a liquid, semi-solid or solid form. 32. A process according to any of the preceding claims for the preparation of a particulate material suitable for use in the preparation of tablets. 33. A method for controlled agglomeration of a finely dispersed solid material, the method comprises subjecting the finely dispersed solid material in the form of a second composition to a process claimed in any of claims 1-32 employing a predetermined amount of carrier, whereby a particulate material having a predetermined geometric weight mean diameter is obtained. 34. A method for improving the bioavailability of a therapeutically and/or prophylactically active substance having an aqueous solubility at 25° C. and pH of 7.4 of at the most about 3 mg/ml, the method comprising subjecting the therapeutically and/or prophylactically active substance in a first and/or a second composition to a process claimed in any of claims 1-32. 35. A method for improving the shelf-life of a pharmaceutical composition which comprises an oxidation sensitive therapeutically and/or prophylactically active substance, the method comprising subjecting the therapeutically and/or prophylactically active substance in a first and/or a second composition to a process claimed in any of claims 1-32 before or during manufacture of the pharmaceutical composition. 36. A particulate material obtainable according to a process claimed in any of claims 1-32. 37. A particulate material according to claim 36 for pharmaceutical or cosmetic use. 38. A particulate material according to claims 36 or 37 for use in the preparation of a solid dosage form. 39. A particulate material according to claim 36 for use in the preparation of tablets. 40. A particulate material according to any of claims 36-39 coated with a coating selected from the group consisting of film coatings, modified release coatings, enteric coatings, protective coatings and anti-adhesive coatings. 41. A particulate material according to any of claims 36-40 for use in the manufacture of tablets obtained by direct compression. 42. A pharmaceutical composition comprising a particulate material obtainable by a process as claimed in any of claims 1-32. 43. A pharmaceutical composition according to claim 42 in the form of a fluid, semi-solid or solid composition. 44. A pharmaceutical composition according to claim 42 in the form of powders, tablets, capsules or sachets. 45. A pharmaceutical composition according to claim 41 in the form of a liquid such as, e.g., a solution or a dispersion including an emulsion and a suspension. 46. Use of a carrier claimed in any of claims 12-21 for the preparation of a solid particulate material, which comprises at least about 25% v/v of the carrier, and which is suitable for further processing into tablets. 47. A pharmaceutical particulate material obtained by mixing a first and a second composition as defined in any of claims 1-32 and heating to a temperature that is below the melting point of a carrier contained in the first composition, the particulate material having a geometric weight mean diameter dgw of ≧10 μm such as, e.g. ≧20 μm, from about 20 to about 2000, from about 30 to about 2000, from about 50 to about 2000, from about 60 to about 2000, from about 75 to about 2000 such as, e.g. from about 100 to about 1500 μm, from about 100 to about 1000μm or from about 100 to about 700 μm, or at the most about 400 μm or at the most 300 μm such as, e.g., from about 50 to about 400 μm such as, e.g., from about 50 to about 350 μm, from about 50 to about 300 μm, from about 50 to about 250,μm or from about 100 to about 300 μm; and the concentration of the carrier in the particulate material being at least about 40% v/v. 48. A particulate material according to claim 47 suitable for use in the preparation of solid dosage form. 49. A particulate material according to claim 47 suitable for use in the preparation of tablets. 50. A particulate material according to claims 48 or 49, wherein the particulate material has sufficient properties with respect to flowability and/or anti-adhesion so that addition of e.g. a lubricant can be omitted when preparing a solid dosage form. 51. A particulate material according to any of claims 47-50 comprising magnesium aluminosilicate and/or magnesium aluminometasilicate. 52. Use of magnesium aluminosilicate and/or magnesium aluminometasilicate as a lubricant 53. Use according to claim 52 for the preparation of pharmaceutical products. 54. Use of magnesium aluminosilicate and/or magnesium aluminometasilicate as an anti-adhesive agent. 55. Use according to claim 54 for the preparation of pharmaceutical products. 56. Use according to any of claims 52-55, wherein magnesium aluminometasilicate is Neusilin such as Neusilin UFL2 or the like. |
<SOH> BACKGROUND OF THE INVENTION <EOH>There is a need for developing new and improved methods which enable preparation of pharmaceutical compositions for oral use that release the active substance from the composition in a suitable manner to enable an absorption of the active substance into the circulatory system. |
Security system and method for computers |
A computer security device (35) comprising a processor (37) that is independent of the host CPU (13) for controlling access between the host CPU (13) and the storage device (21). A program memory (41) that is independent of the computer memory and the storage device (21) unalterably stores and provides computer programs for operating the processor (37) in a manner so as to control access to the storage device (21). The security device (35) is connected only in line with the data access channel (33) between the host CPU (13) and the storage device (21), and off the main data and control bus (15) of the host CPU (13). All data access by the host CPU (13) to the data storage device (21) is blocked before initialization of the security device (35) and is intercepted immediately after the initialization under the control of the processor (37). The processor (37) effects independent control of the host CPU (13) and configuration of the computer (11) to prevent unauthorized access to the storage device (21) during the interception phase. All users of the computer (11) are authenticated with a prescribed profile of access to the storage device (21) and data access to the storage device remains blocked until a user of the computer (11) is correctly authenticated. |
1. A security device for a computer having a host central processing unit (CPU), memory used by the host CPU to load programs in order to operate the computer and a storage device for storing data to be handled by the computer, the security device comprising: processing means independent of the host CPU for controlling access between the host CPU and the storage device; and program memory means independent of the memory of the computer and the storage device to unalterably store and provide computer programs for operating the processing means in a prescribed manner to control said access; wherein the security device is to be connected only in line with the data access channel between the host CPU and the storage device, and off the main data and control bus of the host CPU. 2. A security device as claimed in claim 1, including memory store means independent of the memory means and the storage device of the computer to store critical data and control elements associated with the basic operation of the computer and access to the storage device. 3. A security device as claimed in claim 2, wherein said critical data and control elements are supplied to and used by the host CPU for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer. 4. A security device as claimed in any one of the preceding claims, including authentication means to authenticate a user of the computer having a prescribed profile of access to the storage device. 5. A security device as claimed in claim 4, wherein the authenitication means includes a login verifying means to enable a user of the computer to enter a login identification and password and have that login identification and password verified to authenticate said user being an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. 6. A security device as claimed in claim 5, wherein said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and said login verifying means accesses said critical data and control elements to effect authentication of a user. 7. A security device as claimed in claim 5 or 6, wherein the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user of the computer to prescribed partitions or zones of the storage device. 8. A security device as claimed in any one of the preceding claims, the security device includes intercepting means to block all data access by the host CPU to the data storage device before initialization of the security device and intercept all said data access immediately after said initialization under the control of said processing means. 9. A security device as claimed in claim 3, or any one of claims 4 to 8 as dependent on claim 3, wherein said critical data and control elements include identification data in respect of the storage device for enabling the computer to complete its peripheral check during said start up sequence. 10. A security device as claimed in claim 4, or any one of claims 5 to 9 as dependent on claim 4, wherein said critical data and control elements are supplied to and used by the host CPU for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer, and include a custom boot sector that includes invoking said authentication means for assuming operation of the computer during said start up sequence. 11. A security device as claimed in any one of claims 4 to 7, or any one of claims 8 to 10 as dependent on claim 4, wherein the authentication means includes an authentication application program stored in the program memory means, the memory store means or the storage device. 12. A security device as claimed in claim 11, wherein the authentication application program includes user editing means to enable an authorized user having a particular prescribed level of access to create and edit authorized users for accessing the storage device. 13. A security device as claimed in claim 12 or 13, wherein the authentication application program includes access profile editing means to enable said authorized user having a particular prescribed level of access to allocate and edit particular predetermined levels of access to said prescribed partitions or zones for all authorized users having access to the storage device. 14. A method for securing and protecting a storage device for storing data to be handled by a computer from unauthorized access, the computer having a host central processing unit (CPU) and memory used by the host CPU to load programs in order to operate the computer and storage device, the method comprising: controlling access between the host CPU and the storage device independently of the host CPU; and unalterably storing computer programs for effecting said controlling access in a location separate from the memory and not addressable by the host CPU. 15. A method as claimed in claim 14, including storing critical data and control elements associated with the basic operation of the computer and access to the storage device in a location separate from the memory and the storage device and not addressable by the host CPU. 16. A method as claimed in claim 15, including independently supplying the host CPU with said critical data and control elements for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer. 17. A method as claimed in any one of claims 14 to 16, including authenticating a user of the computer having a prescribed profile of access to the storage device. 18. A method as claimed in claim 17, wherein said authenticating includes enabling a user of the computer to enter a login identification and password and verifying the same to establish whether the user is an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. 19. A method as claimed in claim 18, wherein said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and the verifying includes comparing the entered login identification and password with the login identification and passwords within said critical data and control elements and authenticating a user if there is match. 20. A method as claimed in claim 18 or 19, wherein the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user to prescribed partitions or zones of the storage device. 21. A method as claimed in any one of claims 14 to 20, including blocking all data access by the host CPU to the data storage device during initialization of the computer and intercepting all said data access during the start up sequence after said initialization. 22. A method as claimed in claim 15 or any one of claims 16 to 21 as dependent on claim 15, wherein said critical data and control elements include identification data in respect of the storage device for enabling the computer to complete its peripheral check during said start up sequence. 23. A method as claimed in any one of claims 17 to 20, or claim 21 or 22 as dependent on claim 17, wherein said critical data and control elements include a custom boot sector for the computer that includes invoking the authenticating step; and the method includes assuming operation of the computer during said start up sequence with the custom boot sector and authenticating the user of the computer at such time. 24. A method as claimed in any one of claims 17 to 20, or any of claims 21 to 23 as dependent on claim 17, wherein said authenticating includes enabling a particular prescribed level of authorized user to create and edit login identifications and passwords within the critical data and control elements for specifying authorized users having access to the storage device. 25. A method as claimed in claim 24, wherein said authenticating includes enabling said particular prescribed level of authorized user to allocate and edit particular predetermined levels of access to said prescribed partitions or zones for all authorized users having access to the storage device within the critical data and storage elements. 26. A security device for a computer having a host central processing unit (CPU), memory used by the host CPU to load programs in order to operate the computer and a storage device for storing data to be handled by the computer, the security device comprising: processing means independent of the host CPU for controlling access between the host CPU and the storage device; and; intercepting means to block all data access by the host CPU to the data storage device before initialization of the security device and intercept all said data access immediately after said initialization under the control of said processing means; wherein said processing means effects independent control of the host CPU and configuration of the computer in a manner so as to prevent unauthorized access to the storage device on said intercepting means intercepting said data access immediately after said initialization. 27. A security device as claimed in claim 26, including program memory means independent of the memory of the computer and the storage device to unalterably store and provide computer programs for operating the processing means in a prescribed manner to control said access. 28. A security device as claimed in claim 26 or 27, including memory store means independent of the memory means and the storage device of the computer to store critical data and control elements associated with the basic operation of the computer and access to the storage device. 29. A security device as claimed in claim 28, wherein said critical data and control elements are supplied to and used by the host CPU for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer. 30. A security device as claimed in claim 29, wherein said critical data and control elements include identification data in respect of the storage device for enabling the computer to complete its peripheral check during said start up sequence. 31. A security device as claimed in any one of claims 26 to 30, including authentication means to authenticate a user of the computer having a prescribed profile of access to the storage device. 32. A security device as claimed in claim 31 as dependent on claim 29, said critical data and control elements include a custom boot sector that includes invoking said authentication means for assuming operation of the computer during said start up sequence. 33. A security device as claimed in claim 31 as dependent on claim 29, or claim 32, wherein the authenitication means includes a login verifying means to enable a user of the computer to enter a login identification and password and have that login identification and password verified to authenticate said user being an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. 34. A security device as claimed in claim 33, wherein said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and said login verifying means accesses said critical data and control elements to effect authentication of a user. 35. A security device as claimed in claim 33 or 34, the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user of the computer to prescribed partitions or zones of the storage device. 36. A security device as claimed in any one of claims 31 to 35, wherein the authentication means includes an authentication application program stored in the program memory means, the memory store means or the storage device. 37. A security device as claimed in claim 36, wherein the authentication application program includes user editing means to enable an authorized user having a particular prescribed level of access to create and edit authorized users for accessing the storage device. 38. A security device as claimed in claim 36 or 37, wherein the authentication application program includes access profile editing means to enable said authorized user having a particular prescribed level of access to allocate and edit particular predetermined levels of access to said prescribed partitions or zones for all authorized users having access to the storage device. 39. A security device as claimed in any one of claims 26 to 38, wherein the security device is adapted to be connected only in line with the data access channel between the host CPU and the storage device, and off the main data and control bus of the host CPU. 40. A method for securing and protecting a storage device for storing data to be handled by a computer from unauthorized access, the computer having a host central processing unit (CPU) and memory used by the host CPU to load programs in order to operate the computer and storage device, the method comprising: controlling access between the host CPU and the storage device independently of the host CPU; blocking all data access by the host CPU to the storage device during initialization of the computer; and intercepting all said data access during the start up sequence after said initialization to effect independent control of the host CPU and configuration of the computer in a manner so as to prevent unauthorized access to the storage device thereafter. 41. A method as claimed in claim 40, including unalterably storing computer programs for effecting said controlling access in a location separate from the memory and not addressable by the host CPU. 42. A method as claimed in claim 40 or 41, including storing critical data and control elements associated with the basic operation of the computer and access to the storage device in a location separate from the memory and the storage device and not addressable by the host CPU. 43. A method as claimed in claim 42, including independently supplying the host CPU with said critical data and control elements for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer. 44. A method as claimed in claim 43, wherein said critical data and control elements include identification data in respect of the storage device for enabling the computer to complete its peripheral check during said start up sequence. 45. A method as claimed in any one of claims 40 to 44, including authenticating a user of the computer having a prescribed profile of access to the storage device. 46. A method as claimed in claim 45, as dependent on claim 43, wherein said critical data and control elements include a custom boot sector for the computer that includes invoking the authenticating step; and the method includes assuming operation of the computer during said start up sequence with the custom boot sector and authenticating the user of the computer at such time. 47. A method as claimed in claim 45 or 46, as dependent on claim 43, wherein said authenticating includes enabling a user of the computer to enter a login identification and password and verifying the same to establish whether the user is an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. 48. A method as claimed in claim 47, wherein said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and the verifying includes comparing the entered login identification and password with the login identification and passwords within said critical data and control elements and authenticating a user if there is match. 49. A method as claimed in claim 48, wherein the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user to prescribed partitions or zones of the storage device. 50. A method as claimed in claim 49, wherein said authenticating includes enabling a particular prescribed level of authorized user to create and edit login identifications and passwords within the critical data and control elements for specifying authorized users having access to the storage device. 51. A method as claimed in claim 50, wherein said authenticating includes enabling said particular prescribed level of authorized user to allocate and edit particular predetermined levels of access to said prescribed partitions or zones for all authorized users having access to the storage device within the critical data and storage elements. 52. A security device for a computer having a host central processing unit (CPU), memory used by the host CPU to load programs in order to operate the computer and a storage device for storing data to be handled by the computer, the security device comprising: blocking means for selectively blocking data access between the host CPU and the storage device; and authentication means to authenticate a user of the computer having a prescribed profile of access to the storage device; wherein said blocking means maintains said blocking data access until said authentication means completes correct authentication of the user of the computer. 53. A security device as claimed in claim 52, including processing means independent of the host CPU for controlling the operation of said blocking means for blocking access between the host CPU and the storage device in response to said authentication means. 54. A security device as claimed in claim 53, wherein the blocking means blocks all data access by the host CPU to the data storage device before initialization of the security device and includes intercepting means to intercept all said data access immediately after said initialization under the control of said processing means. 55. A security device as claimed in claim 54, wherein said processing means effects independent control of the host CPU and configuration of the computer in a manner so as to prevent unauthorized access to the storage device, upon said intercepting means intercepting said data access immediately after said initialization and before loading of the operating system of the computer. 56. A security device as claimed in any one of claims 53 to 55, wherein said authentication means enables a software boot of the computer to be effected after correct authentication of the user, and said processing means permits normal loading of the operating system during the start up sequence of the computer following said software boot. 57. A security device as claimed in any one of claims 53 to 56, said processing means controls said blocking means to effect blocking access to the storage device after correct authentication of the user in accordance with the prescribed profile of access of the user. 58. A security device as claimed in any one of claims 53 to 57, including program memory means independent of the memory of the computer and the storage device to unalterably store and provide computer programs for operating the processing means in a prescribed manner to control said access. 59. A security device as claimed in any one of claims 52 to 58 including memory store means independent of the memory means and the storage device of the computer to store critical data and control elements associated with the basic operation of the computer and access to the storage device. 60. A security device as claimed in claim 59, wherein said critical data and control elements are supplied to and used by the host CPU for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer. 61. A security device as claimed in claim 60, wherein said critical data and control elements include identification data in respect of the storage device for enabling the computer to complete its peripheral check during said start up sequence. 62. A security device as claimed in claim 60 or 61, wherein said critical data and control elements include a custom boot sector that includes invoking said authentication means for assuming operation of the computer during said start up sequence. 63. A security device as claimed in any one of claims 60 to 62, wherein the authentication means includes a login verifying means to enable a user of the computer to enter a login identification and password and have that login identification and password verified to authenticate said user being an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. 64. A security device as claimed in claim 63, wherein said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and said login verifying means accesses said critical data and control elements to effect authentication of a user. 65. A security device as claimed in any one of claims 52 to 64, wherein the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user of the computer to prescribed partitions or zones of the storage device. 66. A security device as claimed in any one of claims 52 to 65, wherein the authentication means includes an authentication application program stored in the program memory means, the memory store means or the storage device. 67. A security device as claimed in claim 66, wherein the authentication application program includes user editing means to enable an authorized user having a particular prescribed level of access to create and edit authorized users for accessing the storage device. 68. A security device as claimed in claim 67 as dependent on claim 65, wherein the authentication application program includes access profile editing means to enable said authorized user having a particular prescribed level of access to allocate and edit particular predetermined levels of access to said prescribed partitions or zones for all authorized users having access to the storage device. 69. A security device as claimed in any one of claims 52 to 68, wherein the security device is adapted to be connected only in line with the data access channel between the host CPU and the storage device, and off the main data and control bus of the host CPU. 70. A method for securing and protecting a storage device for storing data to be handled by a computer from unauthorized access, the computer having a host central processing unit (CPU) and memory used by the host CPU to load programs in order to operate the computer and storage device, the method comprising: selectively blocking all data access between the host CPU and the storage device; and authenticating a user of the computer having a prescribed profile of access to the storage device; wherein said blocking of data access is maintained until the user of the computer is correctly authenticated. 71. A method as claimed in claim 70, wherein said selective blocking comprises controlling access between the host CPU and the storage device independently of the host CPU. 72. A method as claimed in claim 71, wherein said selective blocking occurs during initialization of the computer and includes intercepting all said data access during the start up sequence immediately after said initialization and before loading of the operating system of the computer to enable independent control of the host CPU and configuration of the computer in a manner so as to prevent unauthorized access to the storage device. 73. A method as claimed in any one of claims 70 to 72, including performing a software boot of the computer after correct authentication of the user, and allowing normal loading of the operating system during the start up sequence of the computer thereafter. 74. A method as claimed in any one of claims 70 to 73, including controlling blocking access to the storage device after correct authentication of the user in accordance with the prescribed profile of access of the user. 75. A method as claimed in claim 71 or 72, or claim 73 or 74 as dependent on claim 71, including unalterably storing computer programs for effecting said controlling access in a location separate from the memory and not addressable by the host CPU. 76. A method as claimed in any one of claims 70 to 75, including storing critical data and control elements associated with the basic operation of the computer and access to the storage device in a location separate from the memory and the storage device and not addressable by the host CPU. 77. A method as claimed in claim 76, including independently supplying the host CPU with said critical data and control elements for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer. 78. A method as claimed in claim 77, wherein said critical data and control elements include identification data in respect of the storage device for enabling the computer to complete its peripheral check during said start up sequence. 79. A method as claimed in claim 77 or 78, wherein said critical data and control elements include a custom boot sector for the computer that includes invoking the authenticating step; and the method includes assuming operation of the computer during said start up sequence with the custom boot sector and authenticating the user of the computer at such time. 80. A method as claimed in any one of claims 70 to 79, wherein said authenticating includes enabling a user of the computer to enter a login identification and password and verifying the same to establish whether the user is an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. 81. A method as claimed in claim 80, wherein said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and the verifying includes comparing the entered login identification and password with the login identification and passwords within said critical data and control elements and authenticating a user if there is match. 82. A method as claimed in any one of claims 70 to 81, wherein the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user to prescribed partitions or zones of the storage device. 83. A method as claimed in claim 82 as dependent on claim 76, wherein said authenticating includes enabling a particular prescribed level of authorized user to create and edit login identifications and passwords within the critical data and control elements for specifying authorized users having access to the storage device. 84. A method as claimed in claim 83, wherein said authenticating includes enabling said particular prescribed level of authorized user to allocate and edit particular predetermined levels of access to said prescribed partitions or zones for all authorized users having access to the storage device within the critical data and storage elements. 85. A security device for computers substantially as described herein with reference to the accompanying drawings as appropriate. 86. A method for securing and protecting a storage device for storing data to be handled by a computer from unauthorized access substantially as described herein with reference to the accompanying drawings as appropriate. |
<SOH> BACKGROUND <EOH>In these days of widespread computer usage, data stored on a computer system is becoming increasingly accessible to a variety of users. This may occur directly in real time via local and/or remote use of a computer system by different users or indirectly via the loading and running of computer programs at predetermined times automatically or manually by a user of the computer system. With the advent of computer networks allowing remote access to computer systems via local area networks and wide area networks such as the Internet, and the ready transfer of computer programs and data between computer systems, either manually via floppy disks and CD ROMs or automatically via computer networks, the security and integrity of data and information stored on the read/write stores of computers is becoming increasingly of paramount importance. It is now common place for computer systems to incorporate “anti-virus” software in order to protect the data and information stored on the storage device thereof from hostile computer programs, and user authentication procedures allowing predetermined levels of access to data and information stored on the storage device of the computer system, dependent upon the status of the user. A problem with most types of anti-virus software and user authentication protocols used today is the very fact that they are embodied in software, which is required to be executed under the control of the operating system of the computer. Hence, a pre-requisite for such anti-virus or user authentication software to function correctly is that the computer system must be able to power-on, boot-up and invoke the operating system “cleanly”, without any virus or security defeating processes affecting the computer during this time. In the case of anti-virus software, most of this software depends upon having some knowledge of the virus or type of virus that it is attempting to secure the system from. Hence, the anti-virus software needs to be constantly updated and entered onto the computer system before a particular virus finds its way to the computer system. As certain viruses can be extremely hostile and destructive to computer systems, the lag time between the first occurrence of a virus and the production of software to combat the virus still creates a window within which oftentimes irreparable damage can occur to certain computer systems infected with such a virus. Indeed, the production of viruses and anti-virus software does have a tendency to be self-perpetuating. Thus whilst better solutions may have been proposed in the past to combat viruses and ensuring the security of data and information, the state of the art has remained around adopting a software approach to deal with the problem. Notwithstanding this, various hardware-based solutions, which are intrinsically more reliable and resilient in preventing virus or unauthorized access to data stored on a computer system, have been proposed in the past. However, these have been awkward to apply, restricted in their adaptablility to different and changing formatting standards or have required user interaction of a technical nature well beyond the mere loading of executable programs, in order to make them effective or even operational. |
<SOH> SUMMARY OF THE INVENTION <EOH>It is an object of the present invention to provide more robust protection for data and information stored on a computer system from unauthorized access and/or misuse. In accordance with one aspect of the present invention, there is provided a security device for a computer having a host central processing unit (CPU), memory used by the host CPU to load programs in order to operate the computer and a storage device for storing data to be handled by the computer, the security device comprising: processing means independent of the host CPU for controlling access between the host CPU and the storage device; and program memory means independent of the memory of the computer and the storage device to unalterably store and provide computer programs for operating the processing means in a prescribed manner to control said access; wherein the security device is to be connected only in line with the data access channel between the host CPU and the storage device, and off the main data and control bus of the host CPU. Preferably, the security device includes memory store means independent of the memory means and the storage device of the computer to store critical data and control elements associated with the basic operation of the computer and access to the storage device. Preferably, said critical data and control elements are supplied to and used by the host CPU for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer. Preferably, the security device comprises authentication means to authenticate a user of the computer having a prescribed profile of access to the storage device. Preferably, the authenitication means includes a login verifying means to enable a user of the computer to enter a login identification and password and have that login identification and password verified to authenticate said user being an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. Preferably, said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and said login verifying means accesses said critical data and control elements to effect authentication of a user. Preferably, the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user of the computer to prescribed partitions or zones of the storage device. Preferably, the security device includes intercepting means to block all data access by the host CPU to the data storage device before initialization of the security device and intercept all said data access immediately after said initialization under the control of said processing means. Preferably, said critical data and control elements include identification data in respect of the storage device for enabling the computer to complete its peripheral check during said start up sequence. Preferably, said critical data and control elements include a custom boot sector that includes invoking said authentication means for assuming operation of the computer during said start up sequence. Preferably, the authentication means includes an authentication application program stored in the program memory means, the memory store means or the storage device. Preferably, the authentication application program includes user editing means to enable an authorized user having a particular prescribed level of access to create and edit authorized users for accessing the storage device. Preferably, the authentication application program includes access profile editing means to enable said authorized user having a particular prescribed level of access to allocate and edit particular predetermined levels of access to said prescribed partitions or zones for all authorized users having access to the storage device. In accordance with another aspect of the present invention, there is provided a method for securing and protecting a storage device for storing data to be handled by a computer from unauthorized access, the computer having a host central processing unit (CPU) and memory used by the host CPU to load programs in order to operate the computer and storage device, the method comprising: controlling access between the host CPU and the storage device independently of the host CPU; and unalterably storing computer programs for effecting said controlling access in a location separate from the memory and not addressable by the host CPU. Preferably, the method includes storing critical data and control elements associated with the basic operation of the computer and access to the storage device in a location separate from the memory and the storage device and not addressable by the host CPU. Preferably, the method includes independently supplying the host CPU with said critical data and control elements for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer. Preferably, the method includes authenticating a user of the computer having a prescribed profile of access to the storage device. Preferably, said authenticating includes enabling a user of the computer to enter a login identification and password and verifying the same to establish whether the user is an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. Preferably, said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and the verifying includes comparing the entered login identification and password with the login identification and passwords within said critical data and control elements and authenticating a user if there is match. Preferably, the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user to prescribed partitions or zones of the storage device. Preferably, the method includes blocking all data access by the host CPU to the data storage device during initialization of the computer and intercepting all said data access during the start up sequence after said initialization. Preferably, said critical data and control elements include identification data in respect of the storage device for enabling the computer to complete its peripheral check during said start up sequence. Preferably, said critical data and control elements include a custom boot sector for the computer that includes invoking the authentication step; and the method includes assuming operation of the computer during said start up sequence with the custom boot sector and authenticating the user of the computer at such time. Preferably, said authenticating includes enabling a particular prescribed level of authorized user to create and edit login identifications and passwords within the critical data and control elements for specifying authorized users having access to the storage device. Preferably, said authenticating includes enabling said particular prescribed level of authorized user to allocate and edit particular predetermined levels of access to said prescribed partitions or zones for all authorized users having access to the storage device within the critical data and storage elements. In accordance with a further aspect of the present invention, there is provided a security device for a computer having a host central processing unit (CPU), memory used by the host CPU to load programs in order to operate the computer and a storage device for storing data to be handled by the computer, the security device comprising: processing means independent of the host CPU for controlling access between the host CPU and the storage device; and; intercepting means to block all data access by the host CPU to the data storage device before initialization of the security device and intercept all said data access immediately after said initialization under the control of said processing means; wherein said processing means effects independent control of the host CPU and configuration of the computer in a manner so as to prevent unauthorized access to the storage device on said intercepting means intercepting said data access immediately after said initialization. Preferably, the security device includes program memory means independent of the memory of the computer and the storage device to unalterably store and provide computer programs for operating the processing means in a prescribed manner to control said access. Preferably, the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user of the computer to prescribed partitions or zones of the storage device. Preferably, the security device is adapted to be connected only in line with the data access channel between the host CPU and the storage device, and off the main data and control bus of the host CPU. In accordance with another aspect of the present invention, there is provided a method for securing and protecting a storage device for storing data to be handled by a computer from unauthorized access, the computer having a host central processing unit (CPU) and memory used by the host CPU to load programs in order to operate the computer and storage device, the method comprising: controlling access between the host CPU and the storage device independently of the host CPU; blocking all data access by the host CPU to the storage device during initialization of the computer; and intercepting all said data access during the start up sequence after said initialization to effect independent control of the host CPU and configuration of the computer in a manner so as to prevent unauthorized access to the storage device thereafter. Preferably, the method includes unalterably storing computer programs for effecting said controlling access in a location separate from the memory and not addressable by the host CPU. Preferably, said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and the verifying includes comparing the entered login identification and password with the login identification and passwords within said critical data and control elements and authenticating a user if there is match. Preferably, the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user to prescribed partitions or zones of the storage device. In accordance with another aspect of the present invention, there is provided a security device for a computer having a host central processing unit (CPU), memory used by the host CPU to load programs in order to operate the computer and a storage device for storing data to be handled by the computer, the security device comprising: blocking means for selectively blocking data access between the host CPU and the storage device; and authentication means to authenticate a user of the computer having a prescribed profile of access to the storage device; wherein said blocking means maintains said blocking data access until said authentication means completes correct authentication of the user of the computer. Preferably, the security device includes processing means independent of the host CPU for controlling the operation of said blocking means for blocking access between the host CPU and the storage device in response to said authentication means. Preferably, the blocking means blocks all data access by the host CPU to the data storage device before initialization of the security device and includes intercepting means to intercept all said data access immediately after said initialization under the control of said processing means. Preferably, said processing means effects independent control of the host CPU and configuration of the computer in a manner so as to prevent unauthorized access to the storage device, upon said intercepting means intercepting said data access immediately after said initialization and before loading of the operating system of the computer. Preferably, said authentication means enables a software boot of the computer to be effected after correct authentication of the user, and said processing means permits normal loading of the operating system during the start up sequence of the computer following said software boot. Preferably, said processing means controls said blocking means to effect blocking access to the storage device after correct authentication of the user in accordance with the prescribed profile of access of the user. Preferably, the security device includes program memory means independent of the memory of the computer and the storage device to unalterably store and provide computer programs for operating the processing means in a prescribed manner to control said access. Preferably, the security device includes memory store means independent of the memory means and the storage device of the computer to store critical data and control elements associated with the basic operation of the computer and access to the storage device. Preferably, said critical data and control elements are supplied to and used by the host CPU for verification of the storage device and operating the computer independently of the storage device during the start up sequence of the computer. Preferably, the authenitication means includes a login verifying means to enable a user of the computer to enter a login identification and password and have that login identification and password verified to authenticate said user being an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. Preferably, said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and said login verifying means accesses said critical data and control elements to effect authentication of a user. Preferably, the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user of the computer to prescribed partitions or zones of the storage device. Preferably, the security device is adapted to be connected only in line with the data access channel between the host CPU and the storage device, and off the main data and control bus of the host CPU. In accordance with another aspect of the present invention, there is provided a method for securing and protecting a storage device for storing data to be handled by a computer from unauthorized access, the computer having a host central processing unit (CPU) and memory used by the host CPU to load programs in order to operate the computer and storage device, the method comprising: selectively blocking all data access between the host CPU and the storage device; and authenticating a user of the computer having a prescribed profile of access to the storage device; wherein said blocking of data access is maintained until the user of the computer is correctly authenticated. Preferably, said selective blocking comprises controlling access between the host CPU and the storage device independently of the host CPU. Preferably, said selective blocking occurs during initialization of the computer and includes intercepting all said data access during the start up sequence immediately after said initialization and before loading of the operating system of the computer to enable independent control of the host CPU and configuration of the computer in a manner so as to prevent unauthorized access to the storage device. Preferably, the method includes performing a software boot of the computer after correct authentication of the user, and allowing normal loading of the operating system during the start up sequence of the computer thereafter. Preferably, the method includes controlling blocking access to the storage device after correct authentication of the user in accordance with the prescribed profile of access of the user. Preferably, the method includes unalterably storing computer programs for effecting said controlling access in a location separate from the memory and not addressable by the host CPU. Preferably, said authenticating includes enabling a user of the computer to enter a login identification and password and verifying the same to establish whether the user is an authorized user of the computer having a prescribed profile of access to the storage device before allowing the start up sequence of the computer to proceed further. Preferably, said login identification and passwords of authorized users and the prescribed profile of access thereof form part of said critical data and control elements and the verifying includes comparing the entered login identification and password with the login identification and passwords within said critical data and control elements and authenticating a user if there is match. Preferably, the prescribed profile of access comprises a prescribed allocation of predetermined levels of access permitted for an authorized user to prescribed partitions or zones of the storage device. |
Method of producing a light alloy wheel rim and an improved light alloy wheel rim for a motor vehicle |
The invention relates to a wheel rim made from a light alloy, such as an aluminium-based alloy, comprising a front face (10) and a tyre tread (20) which are welded together. The inventive method of producing the wheel rim (J) comprises the following steps: moulding of the front face (10) with a light alloy; provision of a weldable alloy strip with structural hardening; curving of said strip to produce a generally cylindrical shape; welding of the ends in relation to the curved strip with fusion of the strip material; forming of the strip by necking and/or spinning in order to form the tyre tread (20); and welding of the front face and the tyre tread to one another. |
1. A process for producing a rim (J) made of light alloy, such as an aluminum-based alloy, for a motor vehicle, the rim comprising a front face (10) and an outer rim piece (20) that are welded to each other, characterized in that it comprises the following: the step of casting the front face (10) with a light alloy; the step of providing an outer rim piece made of weldable, structural hardening alloy; the step of bending this outer rim piece in order to give it a cylindrical general shape; the step of welding the facing ends of the bent outer rim piece, with fusion of the material of the outer rim piece; the step of forming the outer rim piece, by swaging and/or flowturning, in order to form the outer rim piece (20); and the step of welding the front face and the outer rim piece to each other. 2. The process as claimed in claim 1, characterized in that the step of forming the outer rim piece is preceded by a step of expanding the outer rim piece, after its ends have been welded together. 3. The process as claimed in claim 1 or claim 2, characterized in that the forming step is preceded by an at least partial solution heat treatment and quenching operation. 4. The process as claimed in claim 1, characterized in that the weldable, structural hardening alloy is an aluminum alloy of the 6000 standard series. 5. The process as claimed in claim 1, characterized in that the step of welding the facing ends of the outer rim piece is carried out by a welding technique chosen from the group comprising electron-beam welding, vacuum electron-beam welding, MIG (metal inert gas) welding, tandem MIG welding, laser welding and combined MIG/laser welding. 6. The process as claimed in claim 1, characterized in that the operation of casting the front face (10) includes the formation of at least one weight-reducing hollow (11, 12), obtained by casting. 7. The process as claimed in claim 6, characterized in that at least one hollow (11, 12) is on an inner side of the front face (10). 8. The process as claimed in claim 6, characterized in that at least one hollow (11, 12) is near a connection region between the front face (10) and the outer rim piece (20). 9. The process as claimed in claim 6, characterized in that at least one hollow (11, 12) opens in a demolding direction of the front face. 10. A rim (J) made of light alloy, such as an aluminum-based alloy, for a motor vehicle, the rim comprising a front face (10) made of light alloy and an outer rim piece (20) also made of light alloy that are welded together, characterized in that the outer rim piece (20) is produced by bending and forming a part made of a weldable, structural hardening alloy, with the facing ends of the bent outer rim piece welded together with fusion of the material of the outer rim piece. 11. The rim as claimed in claim 10, characterized in that the weldable, structural hardening alloy is an aluminium alloy of the 6000 standard series. 12. The rim as claimed in claim 11, characterized in that the front face (10) includes at least one weight-reducing hollow (11, 12), obtained during the casting. 13. The rim as claimed in claim 12, characterized in that at least one hollow (11, 12) is on an inner side of the front face. 14. The rim as claimed in claim 12, characterized in that at least one hollow (11, 12) is near a connection region between the front face and the outer rim piece. 15. The rim as claimed in claim 12, characterized in that at least one hollow (11, 12) opens in a demolding direction of the front face. 16. The rim as claimed in claim 10, characterized in that it possesses two rim flanges (CJ1, CJ2), one (CJ1) of which is formed on the front face (10) and the other (CJ2) of which is formed on the outer rim piece (20). |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention aims to provide a process for producing a wheel made of light alloy that overcomes or at least alleviates the abovementioned problems and that makes it possible to obtain a light wheel for a minimal cost, while still maintaining suitable mechanical properties. In this regard, it was surprisingly found that, by using a light alloy that had undergone structural hardening for the rim and by forming this rim by bending, it is possible to use a welding technique involving the fusion of the parts (the facing ends) while still obtaining the final mechanical properties, especially in terms of the strength of the welded region, which are completely satisfactory. Thus, the present invention proposes, according to a first aspect, a process for producing a wheel made of light alloy, such as an aluminum-based alloy, for a motor vehicle, the wheel comprising a front face and a rim that are welded to each other, characterized in that it comprises the following: the step of casting the front face with a light alloy; the step of providing a strip made of weldable, structural hardening alloy; the step of bending this strip in order to give it a cylindrical general shape; the step of welding the facing ends of the bent strip, with fusion of the material of the strip; the step of forming the strip, by swaging and/or flowturning, in order to form the rim; and the step of welding the front face and the rim to each other. Preferred, but non-limiting, aspects of this process are the following: the step of forming the strip is preceded by a step of expanding the strip, after its ends have been welded together; the forming step is preceded by an at least partial solution-treatment and quenching operation; the weldable, structural hardening alloy is an aluminum alloy of the 6000 standard series; the step of welding the facing ends of the strip is carried out by a welding technique chosen from the group comprising electron-beam welding, vacuum electron-beam welding, MIG (metal inert gas) welding, tandem MIG welding, laser welding and combined MIG/laser welding; the operation of casting the front face includes the formation of at least one weight-reducing recess, obtained by casting; at least one recess is on an inner side of the front face; at least one recess is near a connection region between the front face and the rim; and at least one recess opens in a demolding direction of the front face. According to a second aspect, the present invention provides a wheel made of light alloy, such as an aluminum-based alloy, for a motor vehicle, the wheel comprising a front face made of light alloy and a rim also made of light alloy that are welded together, characterized in that the rim is produced by bending and forming a strip made of a weldable, structural hardening alloy, with the facing ends of the bent strip welded together with fusion of the strip material. Preferred, but non-limiting aspects of this wheel are the following: the weldable, structural hardening alloy is an aluminum alloy of the 6000 standard series; its front face includes at least one weight-reducing recess, obtained during the casting; at least one recess is on an inner side of the front face; at least one recess is near a connection region between the front face and the rim; at least one recess opens in a demolding direction of the front face; and the wheel possesses two wheel flanges, one of which is formed on the front face and the other of which is formed on the rim. |
Redox cell with non-selective permionic separator |
The ohmic losses in a redox cell, composed of a positive electrode inside a flow compartment of a positive halfcell acid electrolytic solution, a negative electrode inside a flow compartment of a negative halfcell acid electrolytic solution and a fluid impermeable membrane composed at least partially of an ion exchange resin separating said flow compartments of the respective halfcell electrolytic solution, may be reduced by using a membrane of mixed characteristics. The ion exchange resin of the membrane includes both a cation exchange resin and anion exchange resin allowing the migration through the membrane of anions as well as of protons (H+) of said acid electrolytic solutions. |
1 A redox cell composed of a positive electrode inside a flow compartment of a positive halfcell acid electrolytic solution, a negative electrode inside a flow compartment of a negative halfcell acid electrolytic solution and a fluid impermeable membrane composed at least partially of an ion exchange resin separating said flow compartments of the respective halfcell electrolytic solution, characterized in that said ion exchange resin has mixed ion exchanging properties supporting both the exchange of cations and of the anions for allowing the migration through the fluid impermeable membrane of anions as well as of protons (H+) of said acid electrolytic solutions. 2 The cell of claim 1, wherein said acid electrolytic solution belongs to the group composed of solutions of sulfuric acid, sulfonic acid, boric acid, ossalic acid, nitric acid and mixtures containing at least one of the acids. 3 The cell of claim 1, wherein said cation exchange resin belongs to the group composed of styrene, mono-divinylbenzene, polyvinyldene, polyethylene, polypropylene, polytetrafluoroethylene, polyvinylchloride, polyester, containing sulfonic or carboxylic groups and said anion exchange resin belongs to the group composed of styrene, mono-divinylbenzene, polyvinyldene, polyethylene, polypropylene, polytetrafluoroethylene, polyvinylchloride, polyester, containing aminic group. 4 The redox cell of claim 1, wherein said positive halfcell electrolytic solution contains a redox couple of V(V)/V(IV) and said negative halfcell electrolytic solution contains a redox couple of V(III)/V(II). |
Device for an optical system |
A device is part of an optical system (1) that works with an intermediate image and comprises stop(5) with associated aperture (6). The device comprises a light-reducing or light-absorbing arrangement (12) located at the side of the aperture. The arrangement is arranged to reduce considerably or eliminate completely light (11) incident to the side of the aperture, and thereby considerably reduce beam reflections. In this way, there is less likelihood of the optical system being detected when this is in use. |
1. Device for an optical system (1) that works with an intermediate image and comprises stop (5) with associated aperture (6), characterized in that it comprises a light-reducing or light-absorbing arrangement located at the side of the aperture (6), which arrangement is arranged to reduce considerably or eliminate completely light (11, 18) incident to the side of the aperture, and thereby reduce or eliminate stray light and reflections. 2. Device according to claim 1, characterized in that the light-reflecting or light-absorbing arrangement (12) comprises light-reflecting surface arrangements (13, 14) with surface coating(s) that give low reflection of the light (11, 18) incident to the side of the aperture. 3. Device according to claim 1, characterized in that in the case with the reflection of the light, the surface arrangement is arranged with its surfaces at an angle to each other that results in reflection in a controlled direction or directions. 4. Device according to claim 2, characterized in that the reflecting surfaces (13, 14) comprise or consist of mirrors provided with a coating that gives low reflection. 5. Device according to claim 3, characterized in that the surface arrangement comprises at least two different surfaces or surface arrangements arranged to produce at least two or three reflections of the light (11, 18) incident to the side of the aperture (6). 6. Device according to claim 5, characterized in that the light-reflecting or light-absorbing arrangement comprises a first part (20) that is arranged with the said aperture (6) and with one or more external first light-reflecting surfaces (14′) and a second part (21) that has one or more internal second light-reflecting surfaces (13′). 7. Device according to claim 6, characterized in that one or more first light-reflecting surfaces are in the shape of outer cones and one or more second light-reflecting surfaces are in the shape of inner cones, the half cone angles for the surfaces being unequal. 8. Device according to claim 6, characterized in that one or more reflecting first surfaces are located on a unit (20) in the shape of a truncated pyramid, and in that one or more second light-reflecting surfaces are located on an outer part (21) of the pyramidal unit, with the second light-reflecting surface(s) being opposite the first surface(s), the angles of inclination of the first and second light-reflecting surfaces being unequal. 9. Device according to claim 1, characterized in that an inner part (20) that has the first surface(s) (14′) is arranged with an opening (15′) or channel going right through it centrally, which opening widens out towards the lower section of the part, and in that the aperture (6) and the central opening (15′) have square or circular cross-sections. |
Female electrical contact |
The invention relates to a one-piece female electrical contact (1) which is cut and folded and which comprises, longitudinally, a rear area for connecting to an electrical conductor, a protective cage, which consists of a floor (4), two lateral walls (5, 6) and at least one ceiling element, and, on the front, a termination which is in contact with a complementary male contact. Said contact termination consists of at least one elastic contact blade (8, 9) comprising a blade which is connected to the rear of at least one ceiling element (22) and which extends towards the rear before folding. |
1. A cut and bent, one-piece, female electrical contact comprising, in the longitudinal direction, a rear zone of connection to an electrical conductor, a protective housing comprising a bottom (4), two side walls (5, 6) adjacent to the bottom (4) and bent back along a longitudinal axis, and at least one top element (22, 23), this electrical contact comprising, on the front, a contact terminal, which is situated in the housing, to a complementary male contact, this terminal contact terminal being formed by at least one spring contact tine (8) provided with a terminal part (10) pointed towards the front of the contact, characterized in that said spring contact tine is constructed from a small strip that is joined to the rear of the top element, this small strip extending, prior to bending, towards the rear and, after bending, towards the front perpendicularly to the bottom. 2. The female electrical contact according to claim 1, further characterized in that the bending of the small strip forming the tine consists of a twisting. 3. The female electrical contact according to claim 1, further characterized in that the tine has, in the region of its connection to the top element, a first bend (45) along an axis that is perpendicular to the longitudinal axis of the contact, a first section (46) extending away from the top element, a second biased bend (47) forming a second section (48), and a third bend (49) defining the rear part of a spring section (31) that is joined to a convex section (32) of contact to a complementary contact. 4. The electrical contact according to claim 3, further characterized in that the second section (48) includes an edge supported against the top element (23). 5. The electrical contact according to claim 1, further characterized in that the contact includes a retainer flap (20) for pretensioning the terminal part (10) of the tine, this flap being bent back from the front of the side wall (5) with respect to the tine, the tine comprising, on its part, in front of the contact zone, a section of support against the flap. 6. The electrical contact according to claim 1, further characterized in that the contact includes the means (25, 26) of retention in a contact carrier box. 7. The electrical contact according to claim 6, further characterized in that these means include at least one opening (25) introduced into a side wall of the contact. 8. The electrical contact according to claim 6, further characterized in that these means include at least one tab (26) constructed on one of the sides of the housing. 9. The electrical contact according to claim 1, further characterized in that it includes at lease one pair of tines (8, 9) and one pair of top elements (22, 23), each of the tines of the pair of tines being joined to one top element of the pair of top elements. 10. The electrical contact according to claim 5, further characterized in that the flap includes anchoring lugs (60, 61), the bottom and the top element containing receiving holes (62, 63) for these lugs. 11. The electrical contact according to claim 1, further characterized in that it includes a stiffening device in the linking region between the rear zone of connection and the housing, the stiffening device being formed by arches (70, 71) joining the top part of the side walls (5, 6) and an extension of the bottom adjoining the crimp tabs (50, 51) of an electrical wire. |
Electron beam exposure method and system therefor |
A method of leading an electron beam radiated from an electron emitter through openings provided in a stencil mask to a sensitive sample and exposing it includes placing the electron beam under a low field intensity where the electron beam progresses at a slow speed until reaching the openings of the stencil mask and thereafter placing the electron beam having passed through the openings of the stencil mask under a high field intensity where the electron beam progresses at a high speed. An apparatus for electron beam projection lithography comprises an electron emitter, a stencil mask having openings for permitting the electron beam radiated from the electron emitter to pass through, a base for supporting an exposure sample, and a device for placing the electron beam under a low field intensity as well as a high field generator for placing the electron beam under a high field intensity. |
1. A method of electron beam projection lithography for leading and exposing an electron beam radiated from an electron emitter through openings provided in a stencil mask to an exposure sample, wherein said electron beam is placed under a comparatively low field intensity where the electron beam progresses at a comparatively slow speed until reaching the openings of said stencil mask, and wherein, thereafter, the electron beam having passed through the openings of said stencil mask is placed under a comparatively high field intensity where the electron beam progresses at a comparatively high speed. 2. A method according to claim 1, wherein said low field intensity V′1, said high field intensity V′2, and a voltage V applied to said stencil mask for generating said high field intensity are determined so as to satisfy a relationship (V′1−V′2)/4V=1/F, where F is a distance between the openings of said stencil mask and a convergent point of said electron beam having passed through said openings, so that said electron beam having passed through the openings of said stencil mask may converge. 3. An apparatus for electron beam projection lithography comprising: an electron emitter; a stencil mask having openings for permitting an electron beam radiated from said electron emitter to pass through; a base for supporting an exposure sample to be exposed upon receipt of the electron beam having passed through the openings of said stencil mask; a low field generator for placing said electron beam under a low field intensity so that said electron beam may progress at a slow speed until reaching the openings of said stencil mask; and a high field generator for placing said electron beam, having passed through the openings of said stencil mask, under a high field intensity where said electron beam progresses at a high speed. 4. An apparatus according to claim 3, wherein said low field generator includes an electrode disposed on the upstream side of said stencil mask relative to the progressing direction of said electron beam and a power source connected to said electrode; and wherein said high field generator includes a power source connected to said stencil mask as an electrode. 5. An apparatus according to claim 3, further comprising a second stencil mask disposed on the upstream side of said stencil mask relative to the moving direction of said electron beam and having openings larger than the corresponding openings of said stencil mask. 6. An apparatus according to claim 4, further comprising a second stencil mask disposed on the upstream side of said stencil mask relative to the moving direction of said electron beam and having openings larger than the corresponding openings of said stencil mask, wherein said low field generator further includes a power source connected to said second stencil mask as an electrode. 7. An apparatus according to claim 3, wherein said base for the exposure sample has a support-position-changing mechanism for changing positions to support said exposure sample to thereby change the distance between said exposure sample and said stencil mask. 8. An apparatus according to claim 5, wherein said base for the exposure sample has a support-position-changing mechanism for changing positions to support said exposure sample to thereby change the distance between said exposure sample and said stencil mask. |
<SOH> BACKGROUND ART <EOH>It is known, for transcribing a very fine geometric pattern such as a semiconductor integrated circuit on an electron exposure sample such as a silicon wafer, to use a mask (stencil mask) provided with a plurality of openings constituting the pattern and to expose the pattern on the exposure sample by an electron beam passed through the openings of the stencil mask. The stencil mask with the pattern formed is set as one with a very small thickness dimension so as to make its aspect ratio (the ratio of the width dimension to the depth dimension of an opening) appropriate. Such a thin stencil mask has poor mechanical strength and heat resistance. When an electron beam passes the openings of the stencil mask or crosses the openings, a part thereof hits the opening edges or portions between the openings of the stencil mask. The stencil mask absorbs the electron beam hitting the mask and thermally deforms, which sometimes caused deformation in the pattern formed in the stencil mask, spoiling the accuracy in transcribing the pattern into the exposure sample. Heretofore, in order to prevent such thermal deformation of a stencil mask, it has been proposed to pass an electron beam which is slow in travel speed, that is, of a low energy through openings of the stencil mask. See Japanese Patent Appln. Public Disclosure (KOKAI) No. 11-135424 and No. 9-274884. Japanese Patent Appln. Public Disclosure (KOKAI) No. 11-135424 discloses placing an electron beam radiated from an electron emitter, by applying a low voltage (2 KeV) to the electron emitter, under a low uniform motion and making the interval between a stencil mask and an exposure sample very small (50 μm). According to that, heating of the stencil mask is suppressed by the electron beam progressing at a slow speed, that is, in a low-energy state. Also, because of a narrow space between the stencil mask and the exposure sample, lateral diffusion of the electron beam due to mutual repulsive action of electrons under slow speed traveling (accordingly, long traveling) of the electron beam is minimized, and a so-called out-of-focus pattern on the exposure sample due to the lateral diffusion is prevented. However, even according to that, resolution is insufficient. Also, the electron beam, being under the low-energy state due to the slow speed, can only infiltrate into the surface layer of a resist of the exposure sample after passing the stencil mask. Therefore, the resist capable of forming the pattern is limited to a thin one (0.1 μm or less), and it is difficult to apply to a single-layer thick film resist (0.3 μm or over) which is practically required in a production process of a semiconductor. Further, because of a narrow space between the stencil mask and the exposure sample, use of an exposure sample with a coarse surface or an exposure sample having protrusion might damage the stencil mask. On the other hand, Japanese Patent Appln. Public Disclosure (KOKAI) No. 9-274884 discloses placing an electron beam radiated from an electron emitter in a high accelerated state (acceleration by application of a voltage (50 keV) for high acceleration), and then, just before the stencil mask, placing it in a low accelerated state (reduction by application of a low acceleration voltage (5 keV)), and placing the electron beam after passing through the openings of the stencil mask again in a high accelerated state (acceleration by application of a high acceleration voltage (50 keV)). According to this, by the reduction of the electron beam, a great heat load to the stencil mask is avoided, and by the re-acceleration of the electron beam having passed through the stencil mask, the arrival time of the electron beam at the exposure sample is shortened to avoid narrowing of the space between the exposure sample and the stencil mask, thereby enabling to transcribe on a single layer thick film resist. The re-acceleration of the electron beam, however, does not contribute to preventing the electron beam from diffusing laterally due to the reduction immediately before, thereby generating a so-called out-of-focus pattern. |
<SOH> SUMMARY OF THE INVENTION <EOH>It is an object of the present invention to provide a method, and an apparatus therefor, of electron beam projection lithography which improves resolution of the exposure on an exposure sample without increasing a heat load to a stencil mask. The method of electron beam projection lithography according to the present invention leads an electron beam radiated from an electron emitter through openings provided in a stencil mask to an exposure sample to expose it, and comprises placing the electron beam under a comparatively low electric field intensity for progressing at a comparatively slow speed until the electron beam reaches the openings of the stencil mask, and thereafter, placing the electron beam having passed through the openings of the stencil mask under a comparatively high electric field intensity where the electron beam progresses at a comparatively high speed. Preferably, the low field intensity (V′ 1 ), the high field intensity (V′ 2 ), and a voltage (V) to be applied to the stencil mask for generating the high field intensity are determined to satisfy the relationship: (V′ 1 −V′ 2 )/4V=1/F (where F is a distance between the openings of the stencil mask and an convergent point of the electron beam having passed through the openings) so that the electron beam having passed through the openings of the stencil mask may converge. The apparatus for electron beam projection lithography according to the present invention comprises: an electron emitter, a stencil mask having an openings which permit the electron beam radiated from the electron emitter to pass through; a base for supporting an exposure sample to be exposed upon receipt of the electron beam having passed through the openings of the stencil mask; a low field generator for placing the electron beam under a low field intensity so that the electron beam may progress at a slow speed until reaching the openings of the stencil mask; and a high field generator for placing the electron beam having passed through the openings of the stencil mask under a high field intensity where the electron beam progresses at a high speed. The low field generator may include an electrode disposed on the upstream side of the stencil mask relative to the progressing direction of the electron beam and a power source connected to the electrode, and the high field generator may include a power source connected to the electrode, using the stencil mask as an electrode. The apparatus for electron beam projection lithography can further comprise a second stencil mask disposed on the upstream side of the above-mentioned stencil mask in the progressing direction of the electron beam and having openings of a size the same as or larger than the corresponding openings of the above-mentioned stencil mask. In the exposure apparatus comprising the second stencil mask, the low field generator can further include a power source connected to the second stencil mask as an electrode. The base for the exposure sample may be one provided with a support-position-changing mechanism for changing a position supporting the exposure sample and thereby to change the distance between the exposure sample and the stencil mask. According to the present invention, the energy of the electron beam can be maintained at a comparatively low level by placing the electron beam radiated from the electron emitter and introduced to an exposure sample through the openings of the stencil mask under the low field intensity (under low acceleration) where the electron beam progresses at a slow speed up to immediately before reaching the openings of the stencil mask, thereby suppressing a temperature rise of the stencil mask due to a part of the electron beam which is incident on the stencil mask, hitting the peripheral edge of the openings to be absorbed thereby, and its incidental thermal deformation. Also, by placing the electron beam having passed through the stencil mask under a high field intensity (under high acceleration) where the electron beam progresses at a high speed, the traveling time of the electron beam required until reaching the exposure sample is reduced, avoiding a lateral diffusion of the electron beam during this time, thereby preventing a so-called out-of-focus pattern in the exposure sample from generating, which enables to attain high resolution exposure. Further, by preventing a lateral diffusion of the electron beam, the space between the exposure sample and the stencil mask can be comparatively wide, thereby reducing contamination of the stencil mask, that is, contamination by evaporation from the resist of the exposure sample accompanying the exposure as well as the number of cleaning times of the stencil mask, and to aim at reduction in breakage time of the stencil mask while cleaning and to aim at longevity of the stencil mask accompanying it. Further, it is possible to make an exposure sample with a comparatively coarse surface or an exposure sample with a curvature an object of exposure. The low and the high fields where the electron beam is to be placed can be generated respectively by the low field generator and the high field generator in the electron beam exposure apparatus according to the present invention. The low field generator includes an electrode disposed on the upstream side of the stencil mask relative to the progressing direction of the electron beam and a power source connected to the electrode, while the high field generator includes a power source connected to the stencil mask as an electrode. When fixing the low field intensity (V′ 1 ), the high field intensity (V′ 2 ), and the voltage (V) to be applied to the stencil mask for generating the high field intensity so as to satisfy (V′ 1 −V′ 2 )/4V=1/F (where F is a distance between the openings of the stencil mask and the convergent point of the electron beam having passed through the openings), a so-called lens effect can be given to an electron passing through the openings. This enables convergence of the electron beam against the exposure sample and a deep focal depth. As a result, the electron beam can be made to reach a deep portion of the comparatively thick resist, which is often provided in the exposure sample, thereby enabling it to transcribe an accurate fine pattern. Also, by this, it is possible to generate a gap enlarged by the resist of the exposure sample. When supporting the exposure sample with the base having a support-position-changing mechanism, a relative distance of the exposure sample with respect to the stencil mask can be properly set in accordance with the surface configuration and the general configuration of the exposure sample. Also, by changing the distance or the space between the exposure sample and the stencil mask, the high field intensity (V′ 2 ) can be changed without changing the applied voltage (V), thereby facilitating the field control for obtaining the lens effect. In exposing the exposure sample to the electron beam, it is also possible to dispose a second stencil mask having corresponding openings larger than the openings of the above-mentioned stencil mask. Since the second stencil mask is disposed on the upstream side of the original stencil mask (pattern-forming stencil mask) relative to the progressing direction of the electron beam, the electron beam radiated from the electron emitter, when successively passing through the openings of the second stencil mask and the openings of the mask for pattern formation, hits only the second stencil mask but does not hit the pattern-forming stencil mask located on the downstream side thereof. Therefore, the thermal expansion of the first stencil mask is further reduced, so that more accurate fine pattern transcription can be realized. Also, when providing the second stencil mask, by disposing a power source connected to the second stencil mask as an electrode, the electron beam moving at a slow speed can be further accelerated. |
Interface for removable storage devices |
An interface module and method to provide interfacing of removable digital storage devices (DSDs) to a host system, the interface module having multiple interfaces for the connection of digital storage devices, e.g. Compact Flash (86), a host communication means including a processor (110) for providing a communication channels to transport data (12) and other signals (18) between the digital storage device and the host system and means including a processor (110) for presenting data held on a connected DSD to the host system. Data contained in a DSD is presented to the host system in a transparent manner, that is the data appears as part of a local file or drive hierarchy without the intervention of the user. The invention enables constantly connecting and disconnecting different types of DSD which follow different protocols, without invoking installation routines, system error conditions, or other procedures involving heavy software or time overheads. |
1) An interface module for the transparent interfacing of removable digital storage devices (DSDs) to a host system, the module comprising: a connector interface means for the mounting and interconnection of at least one digital storage device; a host communication means for providing a communication channel to transport data and other signals between a digital storage device and the host system; and a data presenter means for presenting data held on a connected DSD to the host system. 2) The interface module of claim 1 wherein said connector interface means is adapted to read any data including low level data from the DSD. 3) The interface module of claim 1 wherein the host communication means provides a first communication channel to transport data between a digital storage device and the host system and a second communication channel to transport control signals between the interface module and the host system. 4) The interface module of claim 1 wherein the data presenter means further comprises a data buffer. 5) The interface module of claim 1 wherein the data presenter means presents data held on a connected DSD to the host system as part of a file hierarchy local to the host. 6) The interface module of claim 1 further comprising a data routing means for routing of data and signals from a temporarily connected DSD to the host system. 7) The interface module of claim 1 further comprising a detection means for detecting connection of an DSD to the interface module. 8) The interface module of claim 7 wherein the detection means detects the DSD without reading data from the DSD. 9) The interface module of claim 1 further comprising a connection control means for controlling the physical connection of DSDs. 10) The interface module of claim 1 further comprising a user interface control means for controlling the user interface of DSDs. 11) The interface module of claim 1 further comprising an indication means for indicating the status of DSDs to the interface module. 12) The interface module of claim 1 further comprising flash memory for storing application firmware. 13) The interface module of claim 1 wherein hardware control functions for the plurality of different DSD receptacles are integrated into a common control function within the interface module. 14) The interface module of claim 13 wherein the data access function and file system for said plurality of different DSD receptacles are integrated into a single storage device driver within the host computer system. 15) The interface module of claim 13 wherein hardware control functions are implemented via separate driver. 16) The interface module of claim 1 wherein said connector interface means has a modular architecture wherein different communication and/or device protocols are catered for by different connector interface modules. 17) The interface module of claim 1 wherein said communication channels comprise interconnection means arranged for high bandwidth transfer of data and other signal information between an externally connected DSDs and interface module. 18) The interface module of claim 1 wherein said communication channels between and externally connected DSD and the interface module are managed through the provision of proprietary interfaces and controllers. 19) The interface module of claim 1 further comprising a bridging connector to provide data and signal management by the interface module for providing communication channels between an externally connected DSD and the interface module. 20) The interface module of claim 1 wherein said communication channels further comprise interconnecting means arranged to provide for high bandwidth transfer of data and other signal information between the interface module and host. 21) The interface module of claim 1 wherein a PCMCIA controller is used to manage the flow of data to a host system. 22) The interface module of claim 1 wherein a universal serial bus (USB) is used to transfer data to a host system. 23) The interface module of claim 1 wherein IEEE 1394 is used to transfer data to a host system. 24) The interface module of claim 1 wherein an additional communication channel is provided between the interface module and the host system in the form of a serial RS232 bus. 25) The interface module of claim 1 wherein a separate input/output control block is used to control the connection and disconnection of DSDs to the interface module. 26) The interface module of claim 1 wherein the behavior of the I/O control block may be defined by application software executed on either the host system via the interface module or solely by the interface module. 27) The interface module of claim 1 further comprising an embedded OS that mediates interfacing of digital storage devices so as to present data stored on such a device as part of the host system local file/drive hierarchy wherein data stored on a DSD is mapped onto the file system of the host system and connecting or removing a device from the interface does not impact the normal operation of the host system. 28) The interface module of claim 1 wherein interface module is adapted to separately provide local bus interconnections for communication between other processing units. 29) The interface module of claim 1 wherein interface module is subdivided into processing units, each unit receiving a set of separately pluggable externally connecting modules and processing the external signal. 30) A method for the interfacing of removable digital storage devices (DSDs) to a host system, the method comprising the steps: connecting of at least one digital storage device; providing a communication channel to transport data and other signals between said digital storage device and the host system; and presenting data held on said connected DSD to the host system. 31) The method of claim 30 wherein the step of providing a communication channel further comprises providing a first communication channel to transport data between a digital storage device and the host system and a second communication channel to transport control signals between the interface module and the host system. 32) The method of method claim 30 wherein the step of presenting data comprises presenting data held on a connected DSD to the host system as part of a file hierarchy local to the host. 33) The method of method claim 30 further comprising routing of data and signals from a temporarily connected DSD to the host system. 34) The method of method claim 30 further comprising detecting connection of an external DSD to the interface module. 35) The method of method claim 30 further comprising detecting the DSD without reading data from the DSD. 36) The method of method claim 30 further comprising controlling the physical connection of DSDs. 37) The method of method claim 30 further comprising controlling the user interface of DSDs. 38) The method of method claim 30 further comprising indicating the status of DSDs to the interface module. 39) The method of method claim 30 further comprising accessing hardware control functions for a plurality of different DSD receptacles using a common control function. 40) The method of method claim 30 further comprising accessing data for the plurality of different DSD receptacles by accessing a single storage device driver within the host computer system. 41) The method of method claim 30 further comprising accessing hardware control functions for the plurality of different DSD receptacles by accessing a separate device driver within the host computer system. 42) A method of and apparatus for interfacing removable digital storage devices to a host system wherein data held on said devices is presented transparently as part of the host system local file or drive hierarchy, wherein a driver stack interfaces said devices to the host system, the driver stack providing a mirror of the host system replicating commands and data across a driver stack. 43) A method of and apparatus for interfacing a variety of removable digital storage devices (DSDs) to a host computer system wherein specific mechanical and electrical interfaces are provided for each type of DSD and data held on said devices is presented transparently as part of a host system file system, wherein a driver stack interfaces said different devices to the application level software in the host system via a common application programming interface. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>Embodiments of the invention will now be described, by way of example only, with reference to the accompanying drawings, in which: FIG. 1 shows a kiosk fascia including interface means for multiple digital storage devices in accordance with the present invention; FIG. 2 is a system application diagram for the interfacing of said digital storage devices to a host system in accordance with an embodiment of the invention; FIG. 3 is block diagram of an interface module with the embodiment of FIGS. 1 and 2 ; FIG. 4 is a block diagram of the interface module for Within another embodiment of the invention; FIG. 5 is a block diagram of yet another embodiment of the interface module; FIG. 6 is a block diagram of yet another embodiment of the interface module; FIG. 7 is a software component diagram for the embodiments of FIGS. 3, 4 or 6 ; FIG. 8 illustrates a data communications stack within the embodiment of FIG. 7 . detailed-description description="Detailed Description" end="lead"? |
Homeotic genes and proteins, and uses thereof |
The homeotic gene AGL42 in Arabidopsis has been cloned and characterized and is presented herein, both alone and in association with expression control sequences and in vectors which may be used e.g. for transgenic expression. The gene exhibits the role of controlling meristem maintenance. A labeled construct is shown to serve as a marker for identity of the quiescent cells within the meristem. Both the DNA and protein are disclosed. |
1. A method for controlling the morphogenesis of at least one plant organ in the plant Arabidopsis comprising controlling the expression of the homeotic gene AGL42. 2. A DNA sequence or degenerate variant thereof, which encodes the homeotic gene AGL42 in Arabidopsis, or a fragment thereof, selected from the group consisting of: (A) the DNA sequence of FIG. 5; (B) DNA sequences that hybridize to the DNA sequence of FIG. 1 under standard hybridization conditions; (C) DNA sequences that code on expression for an amino acid sequence encoded by the foregoing DNA sequence of FIG. 5; (D) degenerate variants thereof; (E) alleles thereof; and (F) hybridizable fragments thereof. 3. A recombinant DNA molecule comprising a DNA sequence or degenerate variant thereof, which encodes the homeotic gene AGL42 in Arabidopsis, or a fragment thereof, selected from the group consisting of: (A) the DNA sequence of FIG. 5; (B) DNA sequences that hybridize to the foregoing DNA sequence under standard hybridization conditions; (C) DNA sequences that code on expression for an amino acid sequence encoded by the foregoing DNA sequence; (D) degenerate variants thereof; (E) alleles thereof; and (F) hybridizable fragments thereof. 4. The DNA sequence of either of claims 2 or 3, operatively linked to an expression control sequence. 5. The DNA sequence of claim 4, wherein said expression control sequence is selected from the group consisting of the CaMV 19S promoter, the CaMV 35S promoter, the rbcs E9 gene from pea, the wheat histone H3 gene, the nopaline synthase gene of the Ti plasmid of Agrobacterium tumefaciens, the octopine synthase gene of the Ti plasmid of Agrobacterium tumefaciens, the mannopine synthase promoter, the light-inducible promoter from the small subunit of ribulose bis-phosphate carboxylase (ssRUBISCO), and combinations thereof. 6. A plant transformation vector containing a DNA sequence or degenerate variant thereof, which encodes the homeotic gene AGL42 in Arabidopsis, or a fragment thereof, or a fragment thereof, selected from the group consisting of: (A) the DNA sequence of FIG. 5; (13) DNA sequences that hybridize to the foregoing DNA sequence under standard hybridization conditions; (C) DNA sequences that code on expression for an amino acid sequence encoded by the foregoing DNA sequence; (D) degenerate variants thereof; (E) alleles thereof; and (F) hybridizable fragments thereof. 7. The DNA sequence of either of claims 2 or 3, operatively linked to a label. 8. The DNA sequence of claim 7, wherein said label comprises Green Fluorescent Protein. 9. The construct pAGL42::GFP. 10. A polypeptide expressed by the homeotic gene AGL42, which polypeptide is expressed by the DNA molecule of claim 3. 11. A polypeptide capable participating in meristem maintenance in the plant Arabidopsis, which has the amino acid sequence shown in FIG. 6. 12. A probe capable of screening for a nucleic acid encoding the polypeptide AGL42 in alternate plant species, which probe is a labeled DNA molecule of claim 3. 13. Use of the nucleic acid sequence of claim 2, the DNA molecule of claim 3 or the polypeptide of claim 11 for the regulation and/or maintenance and/or promotion of root growth and stability. |
<SOH> BACKGROUND OF THE INVENTION <EOH>I The study of plant morphology and morphogenesis has developed extensively with the advent of recombinant technology. Investigators have identified individual homeotic genes that act either alone or in concert with each other to determine structures in the developing plant. Variations in plant morphology have been noted through the ages, and studies have revealed that mutations of critical homeotic genes may account for such structural aberrations. In many instances such changes may be prompted by response of the plant to environmental stress. The bulk of these investigations have sought to determine both the manner in which such aberrations result and possible avenues for application of such aberrations either to develop new plant strains and structures, or alternatively, to correct the defect caused by the mutation and to assure uniform and normal plant growth. More particularly, plants adapt to environmental stresses through altered growth patterns and physiological changes. These changes occur primarily at the tips of the plant, through the regulated division of cells in regions termed apical meristems. Meristems are first laid down in the embryo, but actively and continuously produce new organs in the germinated seedling and adult plant. The root apical meristem is a particularly suitable system for studying organogenesis because of its relatively simple organization and well characterized, predictable cell divisions (Dolan et al., 1993). Most of the root is a cylinder composed of concentric layers of four basic tissues (epidermis, ground tissue [cortex and endodermis], pericycle and vascular bundle from outside to in; FIG. 1 ). Anatomical studies and fate mapping trace the origin of each tissue type to a ring of cells at the root apex known as initials (Scheres et al., 1994). Initials produce two cells in every stem-cell division: the regenerated initial, and a daughter cell which differentiates as it is displaced from the initial by further rounds of division. Thus, growth in the root occurs by addition of cells at the tip. Surrounding the root apex is a set of protective cells which make up the root cap. The columella (central) root cap has its own set of initials which produce daughters downwards in the direction of the root tip. Lateral root cap shares a common initial with the epidermis. Together, the initials surround a set of approximately four central cells in Arabidopsis termed central cells, or the quiescent center (QC). The meristem is considered to comprise the QC, initials, and their rapidly dividing derivatives in the root tip. The QC, at the heart of the mersitem, has been the focus of intense investigation for nearly two centuries. Early theories posited that central cells must divide rapidly to give rise to all tissues of the root (see Barlow, 1976). This idea was debunked by the English investigator Frederick Clowes, who fed 3 H-thymidine to maize seedlings, and discovered that the population of central cells rarely enter S phase of the cell cycle (Clowes, 1961). Slow cycling time has since been described as an almost universal aspect of stem cells, which often accompanies self-renewal as a defining character (Morrison et al., 1997). Soon after the discovery of mitotic quiescence, levels of RNA and protein synthesis were determined to be very low in the QC (Clowes, 1961). Despite these properties, it was shown that occassional divisions could give rise to any cell of the root, and damaged meristems were regenerated by inducing QC proliferation (Feldman, 1976). This led Barlow to propose his Founder Cell Theory, in which QC cells were described as the ultimate source of all cells in the root (Feldman, 1984). Totipotency of QC cells was demonstrated through in vitro experiments, which proved that the QC is sufficient for regenerating an organized root in culture (Feldman & Torrey, 1976). Torrey (1972) looked beyond the role of the QC as a stem cell population by proposing a function in patterning. He argued that the QC served as a “template”, by which the meristem and root was patterned. Although a quiescent center has been detected in all plant species analysed to date (including Arabidopsis ; Dolan et al., 1993), its function remains poorly characterized. More recent insights into QC function have come from laser ablation studies. Ablation of a single QC cell results in the differentiation of contacting initials, as evidenced by accumulation of starch in the columella initial and a premature asymmetric division in the ground tissue (van den Berg et al., 1995). Thus, the QC must be responsible for a short-range signal that prevents the differentiation of surrounding stem cells. An additional, long-range “top-down” signal for differentiation was also detected in roots by laser ablation (van den Berg, 1997). In order to maintain its undifferentiated state, the QC must be refractile to this signaling from more mature cells. Given their function in continuous organogenesis, meristems must be able to balance stem-cell maintenance with differentiation. In the shoot apical meristem, stem cells in a central zone undergo cell divisions to produce daughters in a peripheral zone where organ primordia are initiated. Balance between these processes in the shoot has recently been shown to result from an autoregulatory loop between signaling molecules (Schoof et al., 2000). The homeodomain protein WUSCHEL is expressed in an organizing region just below the central zone, and is required for maintenance of stem cell fate (Mayer et al., 1998). WUS activates transcription of CLV3, a ligand which signals the CLVI receptor kinase in the central and peripheral zones to inhibit transcription of WUS. Thus, as stem cells leave the central zone, they lose their ability to repress WUS, an activator of stem cell fate; these cells are replenished by the activity of WUS in the central zone. Superficial similarities between the root and shoot meristems, in combination with the results of laser ablation, hint that a similar feedback regulation may occur between the QC and surrounding cells. Along these lines, the FASCIATA1 and 2 genes have recently been shown to affect both shoot and root apical meristem organization as a double mutant (Kaya et al., 2001). FAS genes encode proteins involved in replication-fork dependent nucleosome assembly, and are thus hypothesized to maintain stable epigenetic transcriptional states in the meristems. In fas double mutants, the WUS expression domain is expanded, leading to stem cell overproliferation and stem fasciation. Candidate targets in the root are few. Recently, the root-tip specific expression of CakAt1, a CDK-activating kinase, has been shown to be necessary for preventing differentiation of initials (Umeda et al., 2000). This is likely to be a downstream player of signaling or morphogenic processes. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is predicated on the discovery that a gene named herein AGL42, a MADS box transcription factor, expresses specifically in the quiescent center and is necessary for the maintenance of meristem organization. The gene was identified by an enhancer trap strategy. DS box genes are well known for their roles in the flower, but among the root-specific MADS genes, only ANR1 has been assigned a function (in nitrate response; Zhang and Forde, 1998). This family has played key developmental roles in the plant. The availability of this new gene with remarkable expression and phenotype in the QC will finally allow for a molecular characterization of this important but enigmatic stem cell population. The invention also extends to the homeotic gene AGL42 in Arabidopsis having the DNA sequence set forth in FIG. 5 . The invention extends further to DNA sequences that code the expression of the amino acid sequence set forth in FIG. 6 as well as DNA sequences that would hybridize thereto. The invention further extends to the operative linkage of said DNA sequence to an expression control sequence, including promoters such as the promoter cauliflower mosaic virus (CaMV) 35 S. Naturally, other promoters such as nopaline synthetase and the rbcs E9 gene from pea are representative of other nonlimiting examples thereof. The CaMV 35 S promoter is preferred as it is constitutively expressed and is particularly well-suited for the method illustrated herein. The nucleic acid sequence of the AGL42 cDNA, presented as exons I through VII and the 3′-UTR untranslated sequence is depicted in FIG. 5 . The amino acid sequence of the AGL42 polypeptide is depicted in FIG. 6 . In accordance with an aspect of the invention, the present gene or regulatory elements thereof may be e.g. fused to a marker such as Green Fluorescent Protein (GFP), and used to identify and monitor expression of the central cells (quiescent center) in the meristem. Such a construct was prepared in accordance herewith and its utility has been established as set forth infra. Other objects and advantages will become apparent to those skilled in the art from a consideration of the ensuing description taken in conjunction with the following illustrative drawings. |
Internet security |
In a communications system in which an incoming email is received at an email server within a secure domain, the incoming email is copied to a secondary server outside that secure domain. The copy email message can then be retrieved from the secondary server from a remote device outside the secure domain. |
1. A communication system in which an incoming email received at an email server within a secure domain is copied to a secondary server outside that secure domain if the end user is Present so that the copy email message can be retrieved therefrom from a remote device outside the secure domain. 2. A system according to claim 1 in which the email copy sent to the secondary server contains parameters which allow an application at the secondary server to use changes in the end-user's Presence parameters to activate email availability limitations or to delete the email. 3. A system according to claim 1 in which the copy email message is encrypted using the public key of a public/private key pair and the remote device contains the private key thereof to enable to retrieved message to be decrypted. 4. A system according to claim 3 in which the choice of public/private key pair used is related to Presence parameters and the remote device contains a private key related to the end user's Presence to enable the message to be decrypted. 5. A system according to claim 1 including means for copying a part of the incoming email message and sending it to the secondary email server so that the copied part of the message acts as a prompt to alert the user of the remote device that the full message is awaiting retrieval. 6. A system according to claim 5 including means for using the Presence parameters to determine which part or parts of the email message should be copied and sent to the secondary server. 7. A system according to claim 1 in which a record of the copied email is kept at a PC client associated with the email server so that changes in the end-user's Presence can be used as basis for sending a request for deletion of the email at the secondary server. 8. A system according to claim 1 in which a screensaver application at the remote device or at the PC client is used as input to the Presence server so that the screensaver status forms part of the Presence parameters. 9. A system according to claim 2 in which a key used to encrypt the message or a part of the message is created dynamically using the Presence parameters of the remote device for which it is intended so that the email message or part thereof can only be decrypted by a remote device having the same Presence parameters. 10. A system according to claim 1 in which an email message can be decrypted or retrieved by a remote access device only when the Presence parameters of the remote device have been associated with the Presence parameters of at least one other device; a key used to encrypt the data being dependent on the Presence parameters of both the remote device and the said at least one other device. 11. A system according to claim 9 in which the decryption of the email message at the remote device is used to activate a notification application which notifies other devices or servers about the Presence parameters of the decrypting remote device at the time of decryption. 12. A system according to claim 1 in which an email message can be retrieved only by a remote access device when it is associated with a second device; the key used to encrypt the data being dependent on information from or relating to both devices. 13. A system according to claim 1 in which the key used for decryption of the email message carries information relating to interfaces available at the remote access device and only permits decryption of messages intended only for a predetermined interface or interfaces. 14. A communication system in which an incoming email received at an email server within a secure domain is copied to a secondary server outside that secure domain so that the copy email message can be retrieved therefrom from a remote device outside the secure domain. 15. Computer software recorded in machine readable form for implementing the system of claim 1. |
Red fluorescent material and composition containing the same |
A red fluorescent material having improved fluorescent intensity and an ink composition containing the material. The red fluorescent material is represented by the general formula (1): wherein X1 and X2 each independently represents hydrogen, halogeno, alkyl, alkoxyl, hydroxyl, amino, alkylamino, dialkylamino, aryl, or aralkyl; Y represents a C1-10 fluorohydrocarbon group; and Z1, Z2, and Z3 each independently represents hydrogen, halogeno, alkyl, alkoxyl, hydroxyl, amino, alkylamino, dialkylamino, aryl, or aralkyl. |
1. A red fluorescent material represented by the following general formula (1), (wherein, X1 and X2 each independently represents a hydrogen atom, halogen atom, alkyl group, alkoxyl group, hydroxyl group, amino group, alkylamino group, dialkylamino group, aryl group or aralkyl group; Y represents a fluorohydrocarbon group having from 1 to 10 carbon atoms; and Z1, Z2 and Z3 each independently represents a hydrogen atom, halogen atom, alkyl group, alkoxyl group, hydroxyl group, amino group, alkylamino group, dialkylamino group, aryl group or aralkyl group.) 2. A red fluorescent material according to claim 1, wherein X1 and X2 each is a hydrogen atom. 3. A red fluorescent material according to claim 1, wherein Y is a trifluoromethyl group. 4. A red fluorescent material according to claim 1, wherein Z1, Z2 and Z3 each is a hydrogen atom. 5. An ink composition including a red fluorescent material according to any one of claims 1 to 4. |
<SOH> BACKGROUND ART <EOH>Compounds that are colorless under visible light but emit color in red under UV light irradiation such as a tris(thenoyl trifluoroacetonate)europium complex, tris(benzoyl trifluoroacetonate)europium complex and so on are known. These are intended to apply to various inks and so on. These compounds are relatively high in the emission intensity; however, red fluorescent materials higher in the quantum efficiency of the emission have been in demand. The present invention intends to provide a red fluorescent material that emits in red under UV light irradiation, is excellent in the stability, and has an emission intensity of fluorescence improved more than the conventional one, and an ink composition containing the same. |
Capacitive measuring system |
The invention relates to a measuring device comprising at least one measuring probe (10), means for sequentially applying a controlled supply voltage between the measuring probe (10) and a reference element, and means for integrating accumulated electric charges on the measuring probe (10), characterized in that said device also comprises at least one auxiliary measuring probe (100) which is also sequentially linked to controlled electric supply means and to charge integrating means, said auxiliary measuring probe (100) having a capacity, in relation to a potential detection zone, which is different from the main measuring probe (10), whereby comparative utilization of signals respectively emitted by the two measuring probes (10,100) enables the influence of the main measuring probe to be determined. |
1. A measuring device comprising at least one measurement probe (10), means (30) capable of sequentially applying a controlled supply voltage between the measurement probe (10) and a reference element (20) and means (50) capable of integrating the electrical charges accumulated on the measurement probe (10), characterized in that it furthermore includes at least one auxiliary measurement probe (100) connected, also sequentially, to controlled power supply means (30) and to charge integration means (50), said auxiliary measurement probe (100) having, with respect to a potential detection region, a different capacitance from the main measurement probe (10), in such a way that it is possible, by comparing the signals emanating from the two measurement probes (10, 100) respectively, to determine the influence of the main measurement probe. 2. The device as claimed in claim 1, characterized in that the auxiliary measurement probe (100) has a controlled area that is small compared to the main measurement probe (10). 3. The device as claimed in claim 2, characterized in that the auxiliary measurement probe (100) lies at the same distance from the potential detection region, for example the upper surface of a seat, as the main measurement probe (10). 4. The device as claimed in either of claims 2 and 3, characterized in that the area and the location of the auxiliary measurement probe (100) are such that the latter always experiences the same external influence when a transient external phenomenon occurs, irrespective of the magnitude of this phenomenon. 5. The device as claimed in one of claims 2 to 4, characterized in that the area and the location of the main measurement probe (10) are such that the area of this main measurement probe (10) influenced by the transient external phenomenon depends on the magnitude of this phenomenon. 6. The device as claimed in one of claims 2 to 5, characterized in that the auxiliary measurement probe (100) is centered on the detection region. 7. The device as claimed in one of claims 2 to 6, characterized in that the auxiliary measurement probe (100) has a larger transverse dimension, of around a few centimeters, for example less than 3 cm and preferably less than 1 cm. 8. The device as claimed in one of claims 2 to 7, characterized in that the auxiliary measurement probe (100) has a total area of less than a few square centimeters, for example less than 9 cm2 and preferably less than 4 cm2. 9. The device as claimed in one of claims 2 to 8, characterized in that it includes means capable of using the signal emanating from the integration of the charges accumulated on the auxiliary measurement probe (100) to determine the distance separating a body (P) from the auxiliary measurement probe (100) and then consequently to normalize the measurement obtained from the main probe (10). 10. The device as claimed in claim 1, characterized in that the auxiliary measurement probe (100) is located at a different distance from the potential detection region than the main measurement probe (10). 11. The device as claimed in claim 10, characterized in that the auxiliary measurement probe (100) has an area identical to that of the main measurement probe (10). 12. The device as claimed in claim 10, characterized in that the auxiliary measurement probe (100) has a different area from that of the main measurement probe (10), but in a known ratio relative to the latter. 13. The device as claimed in one of claims 10 to 12, characterized in that the auxiliary measurement probe (100) is close to the main measurement probe (10), so that the difference in influence of an external body (P) on the respective two probes (10, 100) depends only on the difference in distance between the body (P) and these probes (10, 100). 14. The device as claimed in one of claims 10 to 13, characterized in that it includes means capable of combining the signals detected on the two measurement probes (10, 100) in order to determine the distance (e) between the main probe (10) and a body (P) to be detected and subsequently to factor out said distance in the measurement. 15. The device as claimed in claim 1, characterized in that the auxiliary measurement probe (100) and the main measurement probe (10) are asymmetric with respect to a reference element (110). 16. The device as claimed in claim 15, characterized in that the auxiliary measurement probe (100) is located at a different distance from a reference element (110) than the main measurement probe (10). 17. The device as claimed in either of claims 15 and 16, characterized in that it includes means capable of determining the distance (e) separating a body (P) to be detected from the measurement probes (10, 100) by combined use of the signals emanating from these two measurement probes (10, 100), for example by a ratio of the two capacitances measured on these two probes. 18. The device as claimed in one of claims 15 to 17, characterized in that the two, main (10) and auxiliary (100), measurement probes are coplanar with the reference element (110). 19. The device as claimed in one of claims 15 to 18, characterized in that the main (10) and auxiliary (100) measurement probes have identical, for example rectangular, areas. 20. The device as claimed in one of claims 15 to 19, characterized in that the reference element (110) has an area identical to the main (10) and auxiliary (100) measurement probes. 21. The device as claimed in one of claims 15 to 18, characterized in that the main (10) and auxiliary (100) measurement probes have different areas in a known ratio. 22. The device as claimed in one of claims 15 to 21, characterized in that, when one of the two measurement probes (10, 100) is active, the other measurement probe (100, 10) itself serves as auxiliary reference element. 23. The device as claimed in one of claims 1 to 22, characterized in that the main measurement probe (10) has at least one dimension greater than the largest possible dimension of the body (P) that can be detected. 24. The device as claimed in one of claims 1 to 23, characterized in that it comprises a probe (100) of U-shaped configuration comprising two mutually parallel main strands (100, 204) placed respectively on either side of another probe (10). 25. The device as claimed in one of claims 1 to 24, characterized in that it comprises at least one probe (10, 100) having a width that varies along its length. 26. The device as claimed in claim 25, characterized in that it comprises at least one probe (10, 100) having three portions, namely a central portion (16, 106) and two end portions (18, 19; 108, 109). 27. The device as claimed in either of claims 25 and 26, characterized in that it comprises at least one probe (10) having a central portion (16) of large width (l16) and two end portions (18, 19) of small width (l18, l19). 28. The device as claimed in one of claims 25 to 27, characterized in that it comprises at least one probe (100) having a central portion (106) of small width (l106) and two end portions (108, 109) of large width (l108, l109). 29. The device as claimed in one of claims 1 to 28, characterized in that it comprises at least one nonrectilinear probe (10, 100, 110). 30. The device as claimed in claim 29, characterized in that it comprises at least one probe (10, 100, 110) formed from various rectilinear segments joined together in pairs via their ends by transition elements of the concave dihedron type with alternating concavities. 31. The device as claimed in one of claims 1 to 30, characterized in that it comprises a first set of two probes (10, 100) that are located at a defined distance from each other and a third probe (110) located at a greater distance from the two first mentioned probes (10, 100) than the gap that exists between said two probes. 32. The device as claimed in claim 31, characterized in that it includes means capable of sequentially applying a voltage between, on the one hand, the two first mentioned probes (10, 100) and, on the other hand, between the third probe (110) and each of the two first mentioned probes (10, 100), respectively. 33. The device as claimed in one of claims 1 to 32, characterized in that it comprises two electrically conducting bodies, constituting the measurement probe (10) and the reference element (20) respectively, power supply means (30) capable of delivering a DC voltage of controlled amplitude, an integrating stage (50) comprising a capacitor switch system (53) and control means (40) suitable for cyclically defining, at a controlled frequency, a series of two sequences (T1, T2), namely a first sequence during which the power supply means (30) are connected to the measurement probe (10) in order to apply an electric field between the measurement probe (10) and the reference element (20) and to accumulate electric charges on the measurement probe (10), then a second sequence during which the power supply means (30) are disconnected from the measurement probe (10) and the latter is connected to a summing point of the integrating stage (50) in order to transfer charges into the integrating stage (50) and obtain, as output by the latter, a signal representative of the permittivity that exists between the measurement probe (10) and the reference element (20), the integrating stage (50) furthermore including an operational amplifier (51), a first integrating capacitor (52) connected in a feedback loop onto this amplifier (51), and a second capacitor (53) switched between the output and the input of the operational amplifier (51) at the rate of the sequences (T1, T2) controlled by the control means (40), in such a way that, in the steady equilibrium state, the operational amplifier (51) delivers, as output, a voltage “Vsequilibrium” equal to: −Ecs/C53, in which relationship −E denotes the amplitude of the voltage at the terminals of the power supply means (30), and Cs and C53 denote the values of the capacitances defined, on the one hand, between the measurement probe (10) and the reference element and, on the other hand, the switched second capacitor (53), respectively. |
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