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Use of 14-3-3 proteins and a method for determining the same in the fluids or tissues of organisms |
The object of the present invention is to provide a method for the detection and/or quantification of the 14-3-3 proteins or their isoforms for early stage diagnosis of TSE-diseases, which method allows to perform the diagnosis in the living organism. It is furthermore an object, to detect a contamination of the sample by the parallel determination of a second antigen. This object according to the invention is solved by making use of the biochemical characteristics of the members of the 14-3-3 protein family, which bind to specific amino acid motifs like X(n)-XSXXSXXSX-X(n) or to the motif RSXpSXP (SEQ ID NO: 12) within peptides or proteins. For determining one or more isoforms or the entirety of the 14-3-3 protein(s) and for specific binding, one uses modified solid phases like e.g. microtiter plates, which are coated with a synthetic or natural peptide containing a binding motif for 14-3-3 proteins, e.g. a chemically synthesised peptide having the motif CAALPKINRSApSEPSLHR (SEQ ID NO: 1). After the addition of the extracts or bodily fluids to be investigated the detection and quantification of the generated peptide-14-3-3 protein complexes is accomplished by means of labeled antibodies. The use of the 14-3-3 protein family and/or of individual isoforms of the 14-3-3 proteins according to the invention can be employed as an effect monitor or biomonitor in aquatic invertebrates after environmental effects like the presence of polychlorinated biphenyls (PCBs), (xeno)estrogens etc. Moreover, the method can be used for early diagnosis of TSE-diseases like e.g. Creutzfeldt-Jakob disease (CJD) and its novel form (variant) in young persons (vCJD) and Bovine Spongiform Encephalopathy (BSE) or comparable diseases. Thus a diagnostic marker (surrogate marker) is available, which can be used in the living organism as a screening marker, confirmation marker or single marker. |
1. Method for the detection, determination and/or the quantification of an isoform and/or the entirety of the 14-3-3 protein isoforms from the 14-3-3 protein family in the living or dead organism in the human and in the veterinary field, characterised in that in a biological sample at least one isoform and/or the entirety of the 14-3-3 protein isoforms from the 14-3-3 protein family is/are contacted with synthetic or natural peptides comprising amino acid sequence motifs of the type X(n)-XSXXSXXSX-X(n), in particular CX(n)-XSXXSXXSX-X(n) (SEQ ID NO: 13), wherein X is a variable amino acid and S is serine or phosphoserine, and/or RSXpSXP (SEQ ID NO: 12) and/or a 14-3-3 specific antibody, and that the 14-3-3 protein isoform and/or the entirety of the 14-3-3 protein isoforms is detected, determined and/or quantified by means of affinity binding. 2. Method according to claim 1, characterised in that for the determination, for specific binding or concentration of the 14-3-3 proteins a solid phase is used, which is coated with the synthetic or natural peptides or antibodies being directed against purified or recombinant 14-3-3 proteins or 14-3-3 protein isoforms or peptides derived thereof. 3. Method according to claim 2, characterised in that a microtiter plate is used as the solid phase. 4. Method according to claim 2 or 3, whereat the synthetic or natural peptides used for the specific binding or concentration of the 14-3-3 proteins are coupled by means of the maleimide activated microtiter plates by the reaction of the sulfhydryl group of the N-terminal cysteine. 5. Method according to claim 2 or 3, whereat a streptavidin coated microtiter plate or solid phase is used, to which a biotinylated peptide with the above indicated binding motif of the 14-3-3 protein is coupled, and whereat, for the generation of the peptide-14,3,3 protein complexes, the incubation of a biotinylated peptide with the sample is performed either before or after the peptide has bound to the plate. 6. Method according to claim 2 or 3, whereat a carbodiimide or epoxide activated microtiter plate or solid phase is employed, to which a peptide with the above indicated binding motif(s) of the 14-3-3 protein binds, and whereat, for the generation of the peptide-14,3,3 protein complexes, the incubation of a peptide with the sample is performed either before or after the peptide has bound to the plate. 7. Method according to one of claims 1 to 6, characterised in that the determination of 14-3-3 protein is performed at a solid phase and a mobile phase, whereat the binding peptide and/or the antibody is coupled to the solid phase and the 14-3-3 protein to be determined is present in the mobile phase. 8. Method according to one of claims 2 to 7 in the form of a sandwich assay, characterised in that at least one isoform of the 14-3-3 protein family and/or its entirety is isolated by means of one or more specific capture antibodies being covalently or otherwise bound to solid phases, in particular to microtiter plates, followed by an immediate or delayed determination or quantification by means of a second, specific detection antibody. 9. Method according to one of claims 2 to 8, characterised in that the determination is accomplished in the form of a competition assay. 10. Method according to claim 9, wherein the competition takes place between the 14-3-3 protein to be determined in the sample and purified or recombinant 14-3-3 proteins or 14-3-3 protein isoforms or peptide fragments thereof, all of them competing for the same binding site at the binding peptide or antibody coupled to the solid phase, or whereat it is a competition between soluble 14-3-3 protein binding peptides in the mobile phase and the 14-3-3 protein binding peptide coupled to the solid phase, both competing for the 14-3-3 protein to be determined in the sample. 11. Method according to one of the preceding claims, characterised in that the method is combined in the same assay system or in the same kit with the determination of another biomarker (surrogate marker) or of a pathogenic agent in the same sample mixture by its binding to the same or to another solid phase in the form of a combination assay. 12. Method according to one of the preceding claims in the form of a capture assay, comprising the combination of binding the 14-3-3 protein to be determined to (a) a peptide recognition sequence and (b) to an antibody. 13. Method according to one of the preceding claims, characterised in that the determination of 14-3-3 protein is combined with the determination of a quality marker for the sample to be investigated. 14. Method according to one of the preceding claims, characterised in that the biological sample comprises cells, cell associations, tissues, organic fluid or bodily fluid like blood, serum, plasma, liquor, lacrimal secretion, milk secretion or urine. 15. Method according to one of the preceding claims, whereat the detection, the determination and/or the quantification of the generated peptide-14-3-3 protein complexes is accomplished by means of radionucleotide-, dye- or enzyme-labeled antibodies. 16. Use of the method according to one of the preceding claims for the determination of 14-3-3 binding proteins or 14-3-3 specific antibodies, whereat one measures the decrease of the binding of a pre-selected amount of 14-3-3 protein to the solid phase in the presence of these 14-3-3 binding proteins or 14-3-3 specific antibodies. 17. Use of the method according to one of the preceding claims for the detection and quantification of the 14-3-3 proteins or their isoforms in the diagnosis of TSE-diseases like Creutzfeldt-Jakob disease (CJD) and its novel form in young persons (vCJD), Gerstmann-Sträussler-Scheinker syndrome (GSS), Fatal familial insomnia (FFI), Kuru, Scrapie (Traberkrankheit; Gnubberkrankheit; tremblente de mouton), Bovine Spongiform Encephalopathy (BSE), Transmissible mink encephalopathy (TME), Chronic Wasting Disease of cervine animals (CWD), Spongiform encephalopathies in wild ruminants and Feline spongiform encephalopathy (FSE) in living and/or dead organisms and also of other diseases, which are associated with an alteration of the 14-3-3 protein concentration, and also for progression monitoring of therapies against diseases being associated with the 14-3-3 protein concentration. 18. Use of the method according to one of the preceding claims for the early stage diagnosis of BSE or Creutzfeldt-Jakob disease of the old or novel variant, in liquor, blood serum, plasma or other bodily fluids at the living or dead patient or organism. 19. Use of the 14-3-3 protein family or at least one isoform from the 14-3-3 protein family as a biomarker for determining the effects of xenobiotics of all kind or of natural environmental poisons in aquatic invertebrates and other organisms including the human. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The present invention relates to the use of one ore more isoforms from the 14-3-3 protein family for the universal, indirect detection of metabolic alterations in cells or complex cell systems. The use of isoforms of the 14-3-3 protein(s) as a biomarker can be employed for the detection of environmental stress both in environmental samples (water and soil samples) and in animals and cells, with this environmental stress being caused by natural and anthropogenic environmental pollutants, particularly by polychlorinated biphenyls (PCBs) and estrogens/xenoestrogens [(xeno)estrogens]. The present invention also relates to the development and employment of a novel method (an ELISA-method, called “14-3-3 Protein-Capture Assay”) for the quick qualitative and quantitative determination of the 14-3-3 proteins. The detection methods can also be employed for detecting the presence of 14-3-3 protein isoforms in bodily fluids from humans and animals being infected with pathogens of prion diseases. This is needed, since for these diseases there is currently no test-kit available for a surrogate marker, which allows to be determined in living organisms. Such a surrogate marker can be employed for early stage analytics and confirmation analytics of an infection with or a generation or an increased concentration of the pathogenic prion protein PrP Sc , which is regarded to be the pathogenic cause of the Transmissible Spongiform Encephalopathies (TSE). Prominent examples of TSE diseases, in which isoforms of the 14-3-3 proteins can be used as surrogate markers, include: Creutzfeldt-Jakob disease (CJD) and its novel form in young persons (variant CJD, vCJD) Gerstmann-Sträussler-Scheinker syndrome (GSS) Fatal familial insomnia (FFI) Kuru Scrapie (Traberkrankheit; Gnubberkrankheit; tremblente de mouton) Bovine Spongiform Encephalopathy (BSE) Transmissible mink encephalopathy (TME) Spongiform encephalopathies in wild ruminants Feline spongiform encephalopathy (FSE) Chronic Wasting Disease of Cervine Animals (CWD) At present, there exists no such surrogate marker, which would make it possible to obtain early information or perform early stage diagnostics for CJD and vCJD in patients, for BSE in living cattle, for Scrapie in the living sheep or for other TSE diseases in other animals. By employing specific antibodies directed against different isoforms of 14-3-3 protein, one is also allowed to achieve an improved distinction of prion diseases from other diseases, in which increased concentrations of 14-3-3 proteins in bodily fluids may be present. |
Stable pharmaceutical compositions containing pravastatin |
Stabilized pharmaceutical preparations containing a drug in which is sensitive to a low pH environment, such as pravastatin are disclosed in which pravastatin degradations is prevented with a buffering agent. The basic excipient enhances storage stability. |
1. A pharmaceutical composition comprising: pravastatin or a pharmaceutically acceptable salt, isomer, or derivitive thereof; and a buffering agent wherein said composition imparts a pH less than 9 when dissolved in 900 ml deionized water. 2. The dosage form of claim 1 wherein the composition further comprises filler. 3. The dosage form of claim 1 wherein the dosage form further comprises a binder. 4. The dosage form of claim 3 wherein said binder is microcrystalline cellulose, methyl cellulose, hydroxymethyl cellulose, hydroxypropyl methyl cellulose, acrylates, methacrylates, or polyvinylpyrrolidone. 5. The dosage form of claim 4 wherein said binder is microcrystalline cellulose. 6. The pharmaceutical composition as defined in claim 1 wherein the pravastatin is present in an amount from about 1 to 80% by weight of the composition. 7. The dosage form of claim 1 wherein said composition comprises about 1-6% of said buffering agent. 8. The pharmaceutical composition of claim 1 wherein the buffering agent is tromethamine. 9. The dosage form of claim 1 wherein the buffering agent is sodium phosphate dibasic. 10. The pharmaceutical composition as defined in claim 1 wherein the filler is present in an amount within the range from about 30 to about 95% by weight. 11. The pharmaceutical composition as defined in claim 5 wherein the filler is selected from the group consisting of sucrose, dextrose, lactose, cellulose derivatives, calcium carbonate, calcium sulfate, magnesium carbonate, corn starch, corn starch, mannitol, xylitol, fructose, sorbitol, or mixtures thereof. 12. The pharmaceutical composition as defined in claim 3 wherein the binder is present in an amount within a range from about 0 to 20% by weight. 13. The pharmaceutical composition as defined in claim 1 wherein the disintegrant is carboxymethyl cellulose sodium, carboxymethyl cellulose calcium, crospovidone, sodium starch glycolate, corn starch, insoluble cationic-exchange resins such as polyacrylin, microcrystalline cellulose, croscarmellose sodium. 14. The pharmaceutical composition as defined in claim 9 where the disintegrant is present in an amount within the range of from about 0.5 to about 10% by weight. 15. The pharmaceutical composition as defined in claim 1 having a pH of less than 8.5. 16. The pharmaceutical composition as defined in claim 1 having a pH of less than 8.0. 17. A pharmaceutical composition consisting essentially of: (a) 2-50 weight percent pravastatin or a pharmaceutically acceptable salt, isomer, or derivitive thereof; (b) 30-95 weight percent of a filler; (c) 0-20 weight percent of a binder, (d) 1-10 weight percent of a buffering agent; (e) 0.5-10 weight percent of a disintegrant; and (f) 0-6 weight percent of a lubricant; (g) wherein the pH is less than 9. 18. The pharmaceutical composition as defined in claim 15 wherein: (a) 4-25 weight percent pravastatin or a pharmaceutically acceptable salt, isomer, or derivitive thereof; (b) 60-80 weight percent of a filler; (c) 5-15 weight percent of a binder, (d) 2-6 weight percent of a buffering agent; (e) 2-6 weight percent of a disintegrate; and (f) 0.05-2 weight percent of a lubricant; (g) wherein the pH is less than 9. 19. The pharmaceutical composition as defined in claim 16 wherein the buffering agent is tromethamine. 20. The pharmaceutical composition as defined in claim 16 wherein the buffering agent is dibasic sodium phosphate. 21. The pharmaceutical composition as defined in claim 16 wherein the pH of the composition is less than 8.5. 22. The pharmaceutical composition as defined in claim 16 wherein the pH of the composition is less than 8.0. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention A pharmaceutical composition is provided for a medicament which is sensitive to a low pH environment of less than 3, such as pravastatin. Novel, stable oral dosage formulations of pravastatin are provided which include a buffering agent to stabilize and maintain the pH below 9 in an aqueous dispersion. As used in this application, the term pravastatin refers to the free base form of the drug as well as the pharmaceutically acceptable salts such as pravastatin sodium. 2. Description of the Related Art Pravastatin sodium, designated chemically as [1S-[1α (βS*,δs*), 2α, 6α, 8β-(R*),8aα]]-1,2,6,7,8,8a-hexahydro-β,δ,6-trihydroxy-2-methyl-8-(2-methyl-1-oxobutoxy)-1-naphthaleneheptanoic acid monosodium salt is a hydroxymethylglutaryl-CoA (HMG-CoA) reductase inhibitor antilipemic agent described in U.S. Pat. Nos. 4,346,227. It is indicated in hypercholesterolemic patients for primary prevention of coronary events including myocardial infarction (MI); to reduce the risk of undergoing myocardial revascularization procedures; to reduce the risk of cardiovascular mortality. It is indicated in hypercholesterolemic patients for secondary prevention of cardiovascular events, including MI and to slow the progression of coronary arteriosclerosis. Pravastatin is also used as an adjunct to diet for the reduction of elevated total- and LDL-cholesterol and triglyceride levels in patients with primary hypercholesterolemia and mixed dyslipidemia (Types IIa and IIb). The agent specifically competitively inhibits 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase, the enzyme that catalyzes the conversion of HMG-CoA to mevalonate, which is an early rate-limiting step in cholesterol biosynthesis. HMG-CoA reductase inhibitors increase HDL cholesterol and decrease LDL cholesterol, VLDL and plasma triglycerides. The usual dosing regimen is 10-40 mg once daily at bedtime. Certain drugs require an alkaline environment for the purposes of stability. Stability requirements are covered in the United States Pharmacopoeia (U.S.P.), in the Good Manufacturing Practices (GMPs) as well as in FDA Guidelines for stability studies. Buffers may be added to increase stability of certain pharmaceuticals. Buffers may also increase the thermo stability of drug in formulations that require drying during the process of producing the final dosage form. Pravastatin sodium is relatively polar hydrophilic, acid labile, and degrades to form its lactone and various isomers. Degradation results in lower bioavailability of pravastatin sodium. Pravastatin sodium requires a buffer to enhance storage stability. Strategies used in the prior art to stabilize pravastatin sodium formulations include: addition of a basifying agent to raise the pH to at least 9, and packaging of the product in a manner to decrease exposure to moisture. The stability of pravastatin sodium is affected by factors including formulation and storage conditions. Pravastatin sodium is known to be an acid labile compound. Labeling of pravastatin sodium tablets indicates storage at a temperature not to exceed 30° C. and protection from light and moisture. Stabilization is achieved by basification of the environment in which degradation occurs. Stabilized compositions in the prior art have a pH of 9 or over in an aqueous dispersion. The amounts of basifying agent range from 1 to 75%. Basifying agents used include magnesium oxide. Patents such as U.S. Pat. No. 5,180,589, U.S. Pat. No. 6,235,311, U.S. Pat. No. 5,225,202, U.S. Pat. No. 5,030,447 (which are incorporated herein by reference) describe aluminum oxide, all alkali metal hydroxides such as sodium hydroxide, potassium hydroxide or lithium hydroxide, or alkaline earth metal hydroxides such as calcium, magnesium, aluminum hydroxide, dihydroaluminum sodium carbonate, aluminum magnesium hydroxide sulfate, aluminum hydroxide magnesium carbonate co-dried gel, or ammonium hydroxides, calcium carbonate, magnesium carbonate, magnesium stearate, piperazine, sodium acetate, sodium citrate, sodium tartrate, sodium maleate, sodium succinate and mixtures thereof. Stabilization of the commercially available pravastatin sodium (Pravacol®) is achieved by basification by magnesium oxide which imparts a pH above 9, preferably about 10. Although the prior art basifying agents can prevent the degradation of pravastatin sodium, they are less desirable because some are strong bases which may have an adverse effect on excipients used with pravastatin sodium pharmaceutical compositions. For example, lactose discolors and emits a caramelized odor in the presence of certain basifying agents, for example piperazine. Additionally, the high alkalinity occurring at dissolution of these formulations may disrupt the acidic pH milieu of the gastrointestinal (GI) mucosa and is problematic for patients with pre-existing GI mucosal damage. The need exists for a stable pravastatin formulation prepared from an aqueous dispersion of hydrophobic polymers. Such formulations have a practical application, and represent a valuable contribution to the medical arts. The present invention provides such compositions, and offers efficient and cost effective methods of preparation. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention meets the unfulfilled needs of the pharmaceutical industry. The forgoing objectives are met by a pharmaceutical composition in the form of a tablet which has enhanced stability comprising: (a) pravastatin; (b) a filler; (c) a binder; (d) a buffering agent, and (e) a disintegrant wherein the pH of the composition is less than 9. The pharmaceutical formulation of the present invention is a solid dosage form of a medicament having a unique buffer system which results in excellent stability. Specifically, the current invention is directed to stable oral formulations of a medicament sensitive to a low pH environment, such as pravastatin, one or more fillers such as anhydrous lactose, mannitol, one or more binders, such as microcrystalline cellulose, polyvinylpyrrolidine (PVP), one or more disintegrants, such as croscarmellose sodium, one or more lubricants, such as magnesium stearate, one or more coloring agents, such as purple Lake, and one or more buffering agents, such as tromethamine or sodium phosphate dibasic to impart a pH to an aqueous dispersion of the composition of less than 9.0, preferably less than 8.5, and most preferably less than 8.0. A preferred formulation can be made by addition of tromethamine or sodium phosphate dibasic in a range of 1-10%, preferably 2-5%, as a buffering agent which imparts an increase in localized pH within the composition. This increase in localized pH prevents the pravastatin from degrading to form its lactone and various other isomers. The pH of the aqueous dispersion of the composition is about 8 but does not exceed 9 in an aqueous environment. The composition also exhibits excellent stability when stored under accelerated conditions of 40° C. and 75% relative humidity. Accordingly, it is an object of this invention to provide a novel and useful pravastatin formulation which is stable in a low pH environment. This represents an unexpected improvement in the art and substantially overcomes the disadvantages known to the prior art. It is also an object of the present invention to provide both a method of stabilizing pravastatin sodium to slow the degradation thereof and provide products that can be stored for long periods of time at room temperature, i.e. under humidity and temperature conditions usually encountered in pharmacies and medicine cabinets. It is a further object to provide solid oral pravastatin sodium dosage forms where the amount of active drug will be prevented from being reduced to less than 90% of its labeled strength, and more preferably not less than 95% of the labeled strength after one year of storage under controlled room temperature conditions. Other objects, features and advantages of the invention are not taught in the prior art but will be more apparent to those versed in the art from the following specification, taken in conjunction with the accompanying claims. detailed-description description="Detailed Description" end="lead"? |
Sphingosine kinase interacts with traf2 and modulates tumor necrosis factor-induced cellular activity |
The present invention relates generally to a method of modulating cytokine-mediated cellular activity and to agents useful for same. More particularly, the present invention contemplates a method of modulating tumor necrosis factor-mediated cellular activity by modulating an intracellular sphingosine kinase-dependent signalling mechanism. The method of the present invention is useful, inter alia, in the treatment and/or prophylaxis of conditions characterised by aberrant, unwanted or otherwise inappropriate cytokine-mediated cellular activity The present invention is further directed to methods for identifying and/or designing agents capable of modulating the subject sphingosine kinase-dependent signalling mechanism. |
1. A method of modulating cytokine-induced cellular activity, said method comprising contacting said cell with an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with a TRAF wherein inducing or otherwise agonising said association up-regulates said cellular activity and inhibiting or otherwise antagonising said association down-regulates said cellular activity. 2. The method of claim 1, wherein said cytokine comprises Tumor Necrosis Factor (TNF). 3. The method of claim 1 wherein said TRAF is TRAF2. 4. The method according to claim 2 wherein said TNF-induced cellular activity is the induction of anti-apoptotic characteristics and said modulation is down-regulation of the interaction of sphingosine kinase with TRAF. 5. The method according to claim 2 wherein said TNF-induced cellular activity is the induction of pro-inflammatory activity and said induction is down-regulation of the interaction of sphingosine kinase with TRAF. 6. The method according to claim 1 wherein said agent binds, links or otherwise associates with the C-terminal region of sphingosine kinase. 7. The method according to claim 6 wherein said C-terminal region is defined by the amino acid sequence PPEE. 8. The method according to claim 7 wherein said sphingosine kinase is human sphingosine kinase and said C-terminal region is defined by the amino acid sequence PPEE at amino acid residue numbers 379-382 of <400>1. 9. A method for the treatment and/or prophylaxis of a condition characterised by aberrant, unwanted or otherwise inappropriate cytokine induced cellular activity in a mammal, said method comprising administering to said mammal an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with a TRAF wherein inducing or otherwise agonising said interaction up-regulates said cellular activity and inhibiting or otherwise antagonising said interaction down-regulates said cellular activity. 10. The method of claim 9, wherein said cytokine comprises TNF. 11. The method of claim 9, wherein said TRAF is TRAF2. 12. The method according to claim 10 wherein said TNF-induced cellular activity is the induction of anti-apoptotic characteristics and said modulation is down-regulation of the interaction of sphingosine kinase with TRAF. 13. The method according to claim 12 wherein said condition is a neoplastic condition. 14. The method according to claim 10 wherein said TNF-induced cellular activity is the induction of pro-inflammatory activity and said induction is down-regulation of the interaction of sphingosine kinase with TRAF. 15. The method according to claim 9 wherein said agent binds, links or otherwise associates with the C-terminal region of sphingosine kinase. 16. The method according to claim 15 wherein said C-terminal region is defined by the amino acid sequence PPEE. 17. The method according to claim 16 wherein said sphingosine kinase is human sphingosine kinase and said C-terminal region is defined by the amino acid sequence PPEE at amino acid residue numbers 379-382 of <400>1. 18. The method according to claim 9 wherein said mammal is a human. 19-23. (canceled) 24. The method according to claim 10 wherein said TNF-induced cellular activity is the induction of pro-inflammatory activity and said induction is down-regulation of the interaction of sphingosine kinase with TRAF. 25-28. (canceled) 29. A pharmaceutical composition comprising the modulatory agent defined in accordance with the methods of claim 1 together with one or more pharmaceutically acceptable carriers and/or diluents. 30. (canceled) 31. A method for detecting an agent capable of modulating the interaction of TRAF with sphingosine kinase or its functional equivalent or derivative thereof said method comprising contacting a cell or extract thereof containing said sphingosine kinase and TRAF or its functional equivalent or derivative with a putative agent and detecting an altered expression phenotype associated with said interaction. 32. The method according to claim 31 wherein said TRAF is TRAF2. 33. The method according to claim 32 wherein said altered expression phenotype is an altered apoptosis profile. 34. The method according to claim 31 wherein said altered expression phenotype is modulation of the functional activity of sphingosine kinase. 35. A method for analysing, designing and/or modifying an agent capable of interacting with the TRAF binding site of sphingosine kinase or derivative thereof and modulating at least one functional activity associated with said sphingosine kinase said method comprising contacting said sphingosine kinase or derivative thereof with a putative agent and assessing the degree of interactive complementarity of said agent with said binding site. 36. The method according to claim 35 wherein said TRAF binding site is the C-terminal region of sphingosine kinase. 37. The method according to claim 36 wherein said C-terminal region is defined by the amino acid sequence PPEE. 38. The method according to claim 37 wherein said sphingosine kinase is human sphingosine kinase and said C-terminal region is defined by the amino acid sequence PPEE at amino acid residue numbers 379-382 of <400>1. 39. The agent identified in accordance with claim 35. 40. The agent of claim 39 when used in the method of claim 1. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Bibliographic details of the publications referred to by author in this specification are collected at the end of the description. The reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that that prior art forms part of the common general knowledge in Australia. Tumor necrosis factor-α (herein referred to as “TNF”), originally defined by its tumoricidal activity, is a pleiotropic cytokine which elicits a wide spectrum of organismal and cellular responses such as cell proliferation, apoptosis, and inflammatory and immunoregulatory responses. The different cellular responses to TNF are signalled through cell surface receptors (p55 TNF-R1 and p75 TNF-R2), and their adaptor proteins, initiating distinct and separate signalling pathways. These separate signals can lead to opposing cellular effects as best exemplified by TNF's both apoptotic and anti-apoptotic role (Locksley et al., Cell 104, 487, 2001). The strikingly different cellular responses to tumor necrosis factor, such as cell survival, activation and apoptosis, are signalled through the separate pathways. Discrete signalling is believed to be initiated by recruiting difference types of adaptor proteins to the TNF receptor superfamily complexes. For example, the recruitment of a complex including TRADD, FADD/MORT1 and RIP leads to the further recruitment and activation of various caspases and, subsequently, to cell death (Tartaglia et al., Cell 73, 213, 1993; Chinnaiyan et al., Science 274, 990, 1996). On the other hand, TNF induces the interaction of its receptor with a second class of adaptor protein, TNFR-associated factors (herein referred to as “TRAFs”) and recruits downstream signals such as NF-κB-inducing kinase (NIK) to activate NF-κB (Locksley et al., 2001 supra; Arch et al., Genes & Devel. 12, 2821, 1998). In addition, the recruitment of TRADD, FADD and FAN to TNF receptors enables activation of either acidic or neutral sphingomyelinase (Smase) yielding ceramide that has been implicated in the apoptotic signalling in several cell types. TNF activates another key enzyme, sphingosine kinase (SphK), in the sphingomyelin metabolic pathway resulting in production of sphingosine-1-phosphate (S1P). S1P is a potent antagonist of TNF mediated apoptosis. The TNF receptor-associated factors comprise six members, being TRAF1-6. TRAF2 is the prototypical member of the TRAF family. It can interact directly or indirectly with a number of TNF receptors to mediate the signal transduction of these receptors. TRAF2 can also interact with numerous intracellular proteins such as I-TRAF/TANK, RIP, NIK and the caspase inhibitors cIAPs, and thereby transduces signals required for the activation of the transcription factor NF-κB, the stress-activated protein kinase (SAPK or JNK) and anti-apoptosis (Hsu et al., Cell 81, 495, 1995; Rothe et al. Cell 83, 1243, 1995; Wang et al, Science 281, 1680, 1998). Structural studies have revealed the complexity and flexibility of TRAF2 as a signal junction (Park et al., Cell 101, 777, 2000). However, it is still not clear whether TRAF2 can differentially activate distinct downstream signals such as NF-κB and JNK, leading to different biological functions or how this may be achieved. Indeed, targeted gene deletion or truncated TRAF2 (ΔTRAF2) which lacks the N-terminal 86 amino acids that comprise the RING finger domain has showed that TRAF2 is essential for JNK activation and the suppression of TNF-induced apoptosis via NF-(B-independent pathways) (Yeh et al., Immunity 7:715, 1997; Lee et al., Immunity 7:703, 1997), but the precise mechanism of this function has not been established. Sphingolipids have recently emerged as signal molecules to mediate the pleiotropic activities of TNF (Hannun et al., Biochemistry 40:4893, 2001). The TNF signalling via sphingolipid turnover is exemplified by two distinct pathways: the formation of ceramide resulting from the activation of sphingomyelinase (SMase) and the production of sphingosine-1-phosphate (S1P) upon sphingosine kinase (SphK) activation. Despite the controversy regarding ceramide as an apoptotic inducer to mediate TNF-induced cell death (Hannun, Science 274:1855, 1996), S1P has been evident as an anti-apoptotic and mitogenic factor (Olivera and Spiegal, Nature 365:557 1993; Cuvillier et al., Nature 381:800, 1996; Xia et al., J. Biol. Chem. 27434499, 1999 and Cur. Biol. 10:1527, 2000). The signals which regulate TNF-mediated cellular activity have not been fully defined. Elucidation of these cellular signalling mechanisms is necessary for the development of therapeutic and/or prophylactic strategies directed to treating conditions characterised by aberrant or otherwise unwanted cellular activity, which cellular activity is directly or indirectly modulatable via a TNF-regulated signalling mechanism. Whilst structural studies have revealed the complexity and flexibility of TRAF2 (Park et al., 2000, supra) as a signal junction, it is still not clear whether and how TRAF2 can differentially activate its distinct downstream signals such as NF-κB and JNK, leading to different biological functions. In work leading up to the present invention, the inventors have identified a physical interaction between TRAF2 and sphingosine kinase (SphK), a lipid kinase that is responsible for the production of sphingosine-1-phosphate (S1P). The interaction of TRAF2 and SphK is capable of activating SphK, which is required for TRAF2 mediated activation of NF-(B but not JNK. In addition, it has been determined that TRAF2-promoted anti-apoptotic signal pathways critically involves SphK activation during TNF-mediated apoptosis. Accordingly, the interaction between the TNF receptor superfamily and sphingolipid signal pathways provides a mechanism of TRAF2-mediated signals specifically leading to cell activation and survival. The inventors have still further identified the TRAF2 binding motif of sphingosine kinase. The identification of these cellular signalling mechanisms and binding motif has now facilitated the development of methodology directed to modulating TNF mediated cellular activity by regulating the interaction of sphingosine kinase with TRAF. Identification of the sphingosine kinase/TRAF2 binding motif has still further facilitated the development of methodology directed to the identification, design and use of agents which interact with this binding motif thereby modulating TNF mediated cellular activity. |
<SOH> SUMMARY OF THE INVENTION <EOH>Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise”, and variations such as “comprises” and “comprising”, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. As used herein, the term “derived from” shall be taken to indicate that a specified integer may be obtained from a particular source albeit not necessarily directly from that source. The subject specification contains amino acid sequence information prepared using the programme PatentIn Version 3.1, presented herein after the bibliography. Each amino acid sequence is identified in the sequence listing by the numeric indicator <210> followed by the sequence identifier (e.g. <210>1, <210>2, etc). The length, type of sequence (protein (PRT), etc) and source organism for each amino acid sequence are indicated by information provided in the numeric indicator fields <211>, <212> and <213>, respectively. Amino acid sequences referred to in the specification are defined by the information provided in numeric indicator field <400> followed by the sequence identifier (eg. <400>1, <400>2, etc). One aspect of the present invention provides a method of modulating cytokine-induced cellular activity, said method comprising contacting said cell with an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with a TRAF wherein inducing or otherwise agonising said association up-regulates said cellular activity and inhibiting or otherwise antagonising said association down-regulates said cellular activity. In another aspect the present invention provides a method of modulating TNF-induced cellular activity, said method comprising contacting said cell with an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with a TRAF wherein inducing or otherwise agonising said association up-regulates said cellular activity and inhibiting or otherwise antagonising said association down-regulates said cellular activity. Yet another aspect of the present invention provides a method of modulating TNF induced cellular activity said method comprising contacting said cell with an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with TRAF2 wherein inducing or otherwise agonising said association up-regulates said cellular activity and inhibiting or otherwise antagonising said association down-regulates said cellular activity. Still another aspect of the present invention provides a method of down-regulating the TNF induced anti-apoptotic functional activity of a cell, said method comprising contacting said cell with an effective amount of an agent for a time and under conditions sufficient to down-regulate the interaction of sphingosine kinase with TRAF2 wherein inhibiting or otherwise antagonising said interaction up-regulates the apoptosis of said cell. In another aspect there is provided a method of down-regulating the TNF-induced pro-inflammatory functional activity of a cell, said method comprising Contacting said cell with an agent for a time and under conditions sufficient to down regulate the interaction of sphingosine kinase with TRAF2 wherein inhibiting or otherwise antagonising said interaction down-regulates one or more of said cells TNF induced pro-inflammatory activities. In still another aspect the present invention provides a method of modulating cytokine induced cellular activity said method comprising contacting said cell with an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with a TRAF, which agent binds, links or otherwise associates with the C-terminal region of sphingosine kinase, wherein inducing or otherwise agonising said interaction up-regulates said cellular activity and inhibiting or otherwise antagonising said interaction down-regulates said cellular activity. In yet still another aspect the present invention provides a method of modulating TNF induced cellular activity said method comprising contacting said cell with an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with TRAF2, which agent binds, links or otherwise associates with the C-terminal region of sphingosine kinase, wherein inducing or otherwise agonising said interaction up-regulates said cellular activity and inhibiting or otherwise antagonising said interaction down-regulates said cellular activity. A further aspect of the present invention provides a method of modulating TNF induced cellular activity said method comprising contacting said cell with an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with TRAF2, which agent binds, links or otherwise associates with the amino acid sequence PPEE at the C-terminal region of sphingosine kinase, wherein inducing or otherwise agonising said interaction up-regulates said cellular activity and inhibiting or otherwise antagonising said interaction down-regulates said cellular activity. Still another aspect of the present invention is directed to the method for the treatment and/or prophylaxis of a conditions characterised by aberrant, unwanted or otherwise inappropriate cytokine induced cellular activity in a mammal, said method comprising administering to said mammal an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with a TRAF wherein inducing or otherwise agonising said interaction up-regulates said cellular activity and inhibiting or otherwise antagonising said interaction down-regulates said cellular activity. Yet still a further aspect of the present invention provides a method for the treatment and/or prophylaxis of a condition characterised by aberrant, unwanted or otherwise inappropriate TNF induced cellular activity in a mammal, said method comprising administering to said mammal an effective amount of an agent for a time and under conditions sufficient to modulate the interaction of sphingosine kinase with a TRAF wherein inducing or otherwise agonising said interaction up-regulates said cellular activity and inhibiting or otherwise antagonising said interaction down-regulates said cellular activity. In another aspect there is provided a method for the treatment and/or prophylaxis of a neoplastic condition said method comprising administering to said mammal an effective amount of an agent for a time and under conditions sufficient to down-regulate the interaction of sphingosine kinase with TRAF2 wherein inhibiting or otherwise antagonising said interaction down-regulates TNF induced anti-apoptotic functional characteristics. In still another aspect there is provided a method for the treatment and/or prophylaxis of inflammation in a mammal, said method comprising administering to said mammal an effective amount of an agent for a time and under conditions sufficient to down-regulate the interaction of sphingosine kinase with TRAF2 wherein inhibiting or otherwise antagonising said interaction down-regulates the production of TNF induced inflammatory mediators by said mammal. Yet another aspect of the present invention contemplates the use of an agent, as hereinbefore defined, in the manufacture of medicament for the treatment of a condition in a mammal, which condition is characterised by aberrant, unwanted or otherwise inappropriate cytokine induced cellular activity, wherein said agent modulates the interaction of sphingosine kinase with a TRAF and wherein inducing or otherwise agonising said interaction up-regulates said cellular activity and inhibiting or otherwise antagonising said interaction down-regulates said cellular activity. In yet another further aspect, the present invention contemplates a pharmaceutical composition comprising the modulatory agent as hereinbefore defined together with one or more pharmaceutically acceptable carriers and/or diluents. Said agents are referred to as the active ingredients. Yet another aspect of the present invention relates to the agent as hereinbefore defined, when used in the method of the present invention. Another aspect of the present invention provides a method for detecting an agent capable of modulating the interaction of TRAF with sphingosine kinase or its functional equivalent or derivative thereof said method comprising contacting a cell or extract thereof containing said sphingosine kinase and TRAF or its functional equivalent or derivative with a putative agent and detecting an altered expression phenotype associated with said interaction. A further aspect of the present invention provides a method for detecting an agent capable of modulating the interaction of TRAF with sphingosine kinase or its functional equivalent or derivative thereof said method comprising contacting a cell containing said sphingosine kinase and TRAF or its functional equivalent or derivative with a putative agent and detecting an altered apoptosis profile associated with said interaction. In yet another aspect the present invention provides a method for detecting an agent capable of binding or otherwise associating with the TRAF2 binding site of sphingosine kinase or functional equivalent or derivative thereof said method comprising contacting a cell containing said binding site or functional equivalent or derivative thereof with a putative agent and detecting an altered expression phenotype associated with modulation of the function of sphingosine kinase or its functional equivalent or derivative. Still yet a further aspect of the present invention is directed to a method for analysing, designing and/or modifying an agent capable of interacting with the TRAF binding site of sphingosine kinase or derivative thereof and modulating at least one functional activity associated with said sphingosine kinase said method comprising contacting said sphingosine kinase or derivative thereof with a putative agent and assessing the degree of interactive complementarity of said agent with said binding site. In a related aspect, the present invention should be understood to extend to the agents identified utilising any of the methods hereinbefore defined. In this regard, reference to an agent should be understood as a reference to any proteinaceous or non-proteinaceous molecule which modulates at least one TRAF mediated functional activity. Single and three letter abbreviations used throughout the specification are defined in Table 1. TABLE 1 Single and three letter amino acid abbreviations Three-letter One-letter Amino Acid Abbreviation Symbol Alanine Ala A Arginine Arg R Asparagine Asn N Aspartic acid Asp D Cysteine Cys C Glutamine Gln Q Glutamic acid Glu E Glycine Gly G Histidine His H Isoleucine Ile I Leucine Leu L Lysine Lys K Methionine Met M Phenylalanine Phe F Proline Pro P Serine Ser S Threonine Thr T Tryptophan Trp W Tyrosine Tyr Y Valine Val V Any residue Xaa X |
Therapeutic protocols |
The present invention relates to the field of chemotherapy of diseases such as cell proliferation disorders including cancer. In particular, the present invention relates to the use of hyaluronan (HA) as a protective agent in the treatment of subjects. HA is administered in conjunction with a chemotherapeutic agent to facilitate the prolonged administration of a dose of the chemotherapeutic agent to be administered to a subject. Owing to the protective effects of the HA, the dose of chemotherapeutic agent may be substantially higher than a generally accepted effective dose, which would otherwise be expected to cause unacceptable side effects in the subject. |
1. A method which facilitates the prolonged administration of a dose of chemotherapeutic agent to a subject, wherein said dose is higher than a generally accepted effective dose, said method comprising the pre- and/or co-administration of an effective amount of HA. 2. The method of claim 1 wherein a single dose may be up to 200% higher and/or a cumulative dose may be up to 600% higher than generally accepted effective dose. 3. The method of claim 2 wherein the dose of chemotherapeutic agent is from about 10% to about 150% higher than the generally accepted effective dose. 4. The method of claim 3 wherein the dose of chemotherapeutic agent is from about 35% to about 100% higher than the generally accepted effective dose. 5. The method of of claim 1 wherein HA and the chemotherapeutic agent are simultaneously administered. 6. The method of of claim 1 wherein HA is administered from about 24 hours to about 5 minutes before the chemotherapeutic agent. 7. The method of of claim 1 wherein HA is administered from about 12 hours to about 10 minutes before the chemotherapeutic agent. 8. The method of of claim 1 wherein HA is administered about half an hour before the chemotherapeutic agent. 9. The method of of claim 1 wherein the effective amount of HA is from about 0.5 mg to about 20 mg per kilogram body weight per day. 10. The method of of claim 1 wherein the effective amount of HA is from about 5 mg to about 10 mg per kilogram body weight per day. 11. A method for the prolonged treatment of a subject with a dose of a chemotherapeutic wherein a single dose may be up to 200% higher and/or the cumulative dose may be up to 600% higher than a generally accepted effective dose, said method comprising pre- and/or co-administering an effective amount of HA with said chemotherapeutic agent. 12. The method of claim 12 wherein the dose of chemotherapeutic agent is from about 10% to about 150% higher than the generally accepted effective dose. 13. The method of claim 13 wherein the dose of chemotherapeutic agent is from about 35% to about 100% higher than the generally accepted effective dose. 14. The method of of claim 11 wherein HA and the chemotherapeutic agent are simultaneously administered. 15. The method of of claim 11 wherein HA is administered from about 24 hours to about 5 minutes before the chemotherapeutic agent. 16. The method of of claim 11 wherein HA is administered from about 12 hours to about 10 minutes before the chemotherapeutic agent. 17. The method of of claim 11 wherein HA is administered about half an hour before the chemotherapeutic agent. 18. The method of of claim 11 wherein the effective amount of HA is from about 0.5 mg to about 20 mg per kilogram body weight per day. 19. The method of of claim 11 wherein the effective amount of HA is from about 5 mg to about 10 mg per kilogram body weight per day. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Bibliographic details of references provided in the subject specification are listed at the end of the specification. Reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that this prior art forms part of the common general knowledge in any country. Many diseases that afflict animals, including humans, are treated with chemotherapeutic agents. For example, chemotherapeutic agents have proven valuable in the treatment of neoplastic disorders including connective or autoimmune diseases, metabolic disorders, and dermatological diseases, and some of these agents are highly effective (e.g. vincristine and bleomycin) and do not cause any toxic side effects problems, such as neutropenia Proper use of chemotherapeutic agents requires a thorough familiarity with the natural history and pathophysiology of the disease before selecting the chemotherapeutic agent, determining a dose, and undertaking therapy. Each subject must be carefully evaluated, with attention directed toward factors, which may potentiate toxicity, such as overt or occult infections, bleeding dyscrasias, poor nutritional status, and severe metabolic disturbances. In addition, the functional condition of certain major organs, such as liver, kidneys, and bone marrow, is extremely important. Therefore, the selection of the appropriate chemotherapeutic agent and devising an effective therapeutic regimen is influenced by the presentation of the subject. Unfortunately, many chemotherapeutics have severe side effects due to lack of selectivity between normal and malignant tissue. Unwanted toxic side effects may include cardiac toxicity, hair loss, gastrointestinal toxicity (including nausea and vomiting), neurotoxicity, lung toxicity, asthma and bone marrow suppression (including neutropenia). Bone marrow suppression associated with chemotherapy is the result, at least in part, of pronounced drug-induced depression of haematopoietic progenitor cells (HPCs) of the bone marrow (Shimamura et al., Exp. Hematol. 16: 681-685, 1988). The subsequent drop in feutrophil numbers leads to occurrences of secondary infections, the severity of which is directly related to the duration and severity of neutropenia (Bodey et al, Ann. Intern. Med. 64: 328, 1966). As a consequence of secondary infections patients are removed from their chemotherapy regime and placed on a treatment of broad-spectrum antibiotics resulting in limitations of the potential benefits of the cytotoxic treatment. However, despite the use of antibiotics, death from sepsis in severely neutropenic patients is not infrequent (Pettengel et al., Blood 80(6): 430-436, 1992). The level of neutropenia is generally dependent on the regenerative capacity of the bone marrow and/or the dose of the drug being administered. Indeed, neutropenia is often the main reason for decisions to reduce the drug dose being given to a subject (Dotti et al., Haeinatologica 80: 142-145, 1995). Since drug dose reduction is typically accompanied by a loss of effectiveness, or potential effectiveness, of the chemotherapy, drug dose reduction is undesirable. Therefore, there is a need to identify administration regimes, or co-administered agents, which may lessen the incidence or severity of neutropenia associated with chemotherapy, thereby allowing drug dose reduction to be avoided or even enabling the possibility of using higher than standard doses of a chemotherapeutic agent. In some early studies to reduce bone marrow suppression in a mouse model, accelerated recovery from neutropenia, by daily sub-cutaneous injections of human recombinant granulocyte colony-stimulating factor (rG-CSF) following initial injection of 5-fluorouracil (Shimamura et al., Blood 69: 353-355, 1987) and, more specifically, doxorubicin (Shimamura et al., 1988, supra), has been observed. Human rG-CSF exerts this effect by stimulating proliferation and differentiation of haematopoietic progenitor cells (Cohen et al., Proc. Natl. Acad. Sci. USA 84: 2484-2488, 1987). More recently, rG-CSF has been used as an adjunct, in patients undergoing cytotoxic chemotherapy, to enhance neutropenia recovery (Sheridan et al., Lancet 339: 640-644, 1992; Pettengell et al., 1992, supra; Anglin et al., Leuk. Lymphoma 11: 469-472, 1993; Kotaka et al., Int. J Urol. 6: 61-67, 1999). The application of rG-CSF, however, has to be optimally combined with repeated cycles of chemotherapy, due to the potential for the increased number of haematopoietic progenitor cells to become hypersensitive to cytotoxic drugs (Dotti et al., 1995, supra). There is a need to develop protocols where a high cancer or target cell-killing dose of chemotherapeutic agent can be used without or with reduced levels of toxic side effects. |
<SOH> SUMMARY OF THE INVENTION <EOH>Throughout this specification, unless the context requires otherwise, the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers. The present invention is predicated in part on the determination that hyaluronan (HA) may be used as a protective agent in subjects, when these subjects receive treatment with a generally cytotoxic drug. Situations wherein subjects may receive a cytotoxic drug include the treatment of life-threatening diseases such as cancer. In these circumstances, the primary therapeutic objective is to effect the regression of malignant cells. However, the presently available therapeutically effective agents are less specific than would be preferred, and their administration results, eventually, in the death of many of the subject's normal cells as well. Where cancer regression is not effected quickly enough, the concomitant unwanted effect on normal cells may be so high that the subject's condition deteriorates to the point where treatment must be curtailed or stopped. This can have disastrous consequences for the subject undergoing treatment. An agent that could moderate the need for cessation of treatment, for a period sufficiently long to facilitate the prolonged and increased therapeutic effect of the chemotherapeutic agent, would be a distinct advantage in the treatment of many forms of cancer and other diseases. While some of the beneficial effects of the inclusion of HA in treatment regimes have been known for some time, the protective effect of HA has not been appreciated until now. Accordingly, in one aspect, the present invention provides a method which facilitates the prolonged administration of a dose of chemotherapeutic agent to a subject, wherein said a single dose may be up to 200% higher and/or the cumulative dose may be up to 600% higher than a generally accepted effective dose, said method comprising the pre- and/or co-administration of an effective amount of HA. Under this regime, the pre- or co-administration of HA has the effect of ameliorating or even abolishing the otherwise unwanted concomitant deleterious effects on normal cells. As the patient's health is not being as adversely affected by the cytotoxic effects of the therapeutic agent, the said therapeutic agent may be administered at a higher than normal dose and allowed to act over a longer period of time. This increases the chances that the desirable cytotoxic effects of the administered drug, against unwanted malignant cells, will result in successful treatment. Accordingly, another aspect of the present invention contemplates a method for the prolonged treatment of a subject with a dose of a chemotherapeutic agent where a single dose may be up to 200% higher and/or the cumulative dose may be up to 600% higher than a generally accepted effective dose, said method comprising pre- and/or co-administering an effective amount of HA with said chemotherapeutic agent. The result of such treatment regimes is that the unacceptably severe side effects that are usually observed are obviated. As HA is a polymeric molecule, it may be formulated to comprise molecules of varying molecular weights. Although lower molecular weight formulations are also effective in the methods of the present invention, preferred formulations comprise HA having a modal molecular weight in the range 750,000 to 2 million Da. Higher molecular weight HA has the advantage of a tertiary structure whereby, at low concentrations, it self-aggregates forming a three-dimensional meshwork. This meshwork exhibits the characteristics of controllable porosity and molecular dimension, which enables the establishment of equilibrium between therapeutic molecules held within the volumetric domain of the polysaccharide and the external environment. The “loading” of the HA three-dimensional structure with therapeutic molecules results in a controlled release of the therapeutic agent at the pathological site, subsequently overcoming non-specific targeting of healthy tissue. A particularly preferred weight range is 750,000-1,000,000 Da. In a preferred embodiment, HA formulations may be administered to a subject simultaneously with or prior to administration of the chemotherapeutic agent. HA formulations may be generated in any number of ways, well known to those skilled in the art, including injectable solutions, powder formulations, tablets pills or capsules, or in any other convenient form. |
Method for production of the solid fuel from biomass |
The solid fuel formed into briquettes in the range of density by 400-1500 g/dm3, is produced from a perennial plant called Sida hermaphrodita Rusby as well as from a mixture of its stems with other chopped up elements of plant origin. Biomass made from stems of above-mentioned plant or from a mixture of its stems with other plants undergoes compressing in a briquetting press. The final product has energy output comparable to birch wood. Solid fuel made from biomass is mostly used in boiler-rooms for heating up hot-houses or for house fire-places. |
1. A method for production of the solid fuel from biomass, in the form of briquettes in the range of density by 400-1500 g/dm3 characterized in that biomass is prepared from stems of a perennial plant called Sida hennaphrodita Rusby chopped up into 20-100 mm long pieces, preferably 50 mm, next it is further chopped up to obtain average ratio of length to thickness between 4080, preferably 60 and participation of pieces longer than 10 mm from 2-20%, preferably 10%, while pieces smaller than 0.25 mm shoed constitute 5-15%, preferably 10%. 2. A method according to claims 1, characterized in that the chopped up stems of moisture contents by 3-18%, preferably 12% are compressed into briquettes of intended thickness and shape while lignin is naturally found in the raw material, 1 serves as a binder. 3. A method according to claim 1, characterized in that biomass is produced from stems of Sida hermaphrodita Rusby mixed with other chopped up substances of plant origin, preferably wood which constitutes less than 60% of dry matter. 4. A method according to claim 3, characterized in that a natural binder is added, preferably molasses equal to 0.3-2.0%, preferably 0.3% of dry matter, next it undergoes compressing. 5. A method according to claim 2, characterized in that biomass is produced from stems of Sida hermaphrodita Rusby mixed with other chopped up substances of plant origin, preferably wood which constitutes less than 60% of dry matter. |
Smart interactive billboard device |
A smart interactive billboard for allowing or other devices implemented with the smart billboard to display electronic/computer-based information and become interactive includes a transceiver adapted to communicate interactively with a server and at least one or more client devices to allow users to be interactive with the implemented billboards or other devices. A CPU including software controls the implemented billboards or other devices. A memory stores information received or uploaded to the implemented billboards or other devices from the server and client devices, and a display unit displays the information on the implemented billboards or other devices. |
1. A smart interactive billboard device, operable with stationary or mobile advertising platforms that display information to provide electronic/computer-based advertising, promotional and/or sales information, comprising: a transceiver operative to communicate with at least one client device by at least transmitting advertising, promotional and/or sales information; a CPU including software operative to control the transceiver and provide communication with the client device; and a memory adapted to store advertising, promotional and/or sales information for transmission by the transceiver to said at least one client device. 2. The smart interactive billboard device according to claim 1, further comprising a controller adapted to allow the billboard device to perform at least one of self-testing, reprogramming, information uploading, fault diagnostic examining or repairing. 3. The smart interactive billboard device according to claim 2, wherein the controller includes at least an operator input and an operator display. 4. The smart interactive billboard device according to claim 1, wherein said transceiver is operative to communicate interactively with at least one client device by further receiving advertising, promotional and/or sales information; said CPU including software operative to control the transceiver and provide interactive communication with the client device; and said memory further adapted to store advertising promotional and/or sales information received by the transceiver from said at least one client device. 5. The smart interactive billboard device according to claim 1, wherein the memory includes at least one of a flash memory, PROM or SRAM. 6. The smart interactive billboard device according to claim 1, further comprising an interface that is at least one of USB or UART. 7. The smart interactive billboard device according to claim 1, comprising means operable to enable a user via a client device to interactively download information from or upload information to the billboard device. 8. The smart interactive billboard device according to claim 1, comprising means operable to enable a user via the client device to perform electronic commerce transactions with respect to the information provided to the billboard device. 9. The smart interactive billboard device according to claim 1, comprising means operable to enable a user via the client device to exchange or transfer information amongst other users using the billboard device. 10. The smart interactive billboard device according to claim 1, wherein the information displayed on the stationary or mobile platform are advertisements. 11. The smart interactive billboard device according to claim 7, wherein the information uploaded to or download from the billboard device is at least one of electronic vouchers, electronic coupons, electronic tickets, electronic receipts, or electronic advertisements. 12. The smart interactive billboard device according to claim 1, wherein the smart interactive billboard device is adapted to be located and installed on stationary or mobile platforms. 13. The smart interactive billboard device according to claim 1, wherein the billboard device further comprises means for communicating interactively with a server through at least one of SMS, DVB, DAB, wireless modem or wireless dialup network. 14. The smart interactive billboard device according to claim 1, wherein the transceiver is adapted to communicate interactively with the client device through at least one of SMS, Bluetooth, infrared or IrDA. 15. A smart interactive billboard system, operable with stationary or mobile advertising platforms that display information to provide electronic/computer-based advertising, promotional and/or sales information, comprising: an advertising platform; a smart interactive billboard device mounted to said advertising platform and including: a transceiver operative to communicate interactively with at least one client device by at least transmitting advertising, promotional and/or sales information; a CPU including software operative to control the transceiver and enable interactive communication with the at least one client device; and a memory adapted to store information for transmission by or received by the transceiver, and a display unit, including a display screen, responsive to the smart interactive billboard device and operative to display on said screen advertising, promotional and/or sales information under control of said CPU. 16. The smart interactive billboard device according to claim 15, wherein the CPU is operative to update and change the displayed advertising, promotional and/or sales information. 17. The smart interactive billboard device according to claim 15, wherein the billboard device further comprises means for communicating interactively with a server through at least one of SMS, DVB, DAB, wireless modem or wireless dialup network. 18. The smart interactive billboard device according to claim 15, wherein the transceiver is adapted to communicate interactively with the client device through at least one of SMS, Bluetooth, infrared or IrDA. 19. The smart interactive billboard device according to claim 1S, wherein said transceiver is operative to communicate interactively with at least one client device by further receiving advertising, promotional and/or sales information; said CPU including software operative to control the transceiver and provide interactive communication with the client device; and said memory further adapted to store advertising, promotional and/or sales information received by the transceiver from said at least one client device. 20. A smart interactive billboard system for providing interactive communication with at least one client device comprising: a server; a plurality of advertising platforms; a smart interactive billboard device mounted to each said advertising platform and including: a transceiver means operative to communicate interactively with the client device and the server by at least one of transmitting and receiving information, a CPU including software operative to control the transceiver and the interactive communication with the server, and a memory adapted to store information for transmission by or received by the transceiver; and wherein the server is in communication with said plurality of smart interactive billboard devices and wherein said server includes a communication interface operative to at least request information from and deliver information to the plurality of interactive billboard devices. 21. The smart interactive billboard system according to claim 20, wherein at least one smart interactive billboard device is mounted to an advertising platform, wherein the platform further comprising a display unit operative to display advertising, promotional and/or sales information provided by the server. 22. The smart interactive billboard system according to claim 20, wherein the server is adapted to selectively transmit information to one or more interactive billboard devices. 23. The smart interactive billboard system according to claim 20, wherein the server and the smart interactive billboard devices are communicable through at least one of SMS, DVB, DAB, Wireless modem or wireless dialup network. 24. A smart interactive billboard system operative to enable interactive communication with a mobile user, comprising: an advertising platform; a smart interactive billboard device mounted to the advertising platform and including: a transceiver operative to communicate interactively with at least one user-operated client device by at least one of transmitting and receiving information, a CPU including software operative to control the transceiver and the interactive communication with the client device, and a memory adapted to store information for transmission by or received by the transceiver; and the at least one user-operated client device, including a client transceiver, operative to at least provide the user with the capability of at least one of uploading information to or receiving information from the smart interactive billboard device. 25. The smart interactive billboard system according to claim 24, wherein the advertising platform further comprises a display unit, including a display screen responsive to the smart interactive billboard device and operative to display information provided by the device. 26. The smart interactive billboard system according to claim 25, wherein the advertising platform is one of a stationary or a mobile platform. 27. The smart interactive billboard system according to claim 24, wherein the client device is at least-one of a personal computer, personal digital assistant, cellular or mobile phone, interactive television or radio system. 28. The smart interactive billboard system according to claim 24, wherein the client device is adapted to allow the user to perform electronic commerce transactions with respect to the information provided to the billboard device. 29. The smart interactive billboard system according to claim 24, wherein the client device is adapted to allow the user to exchange or transfer information amongst other users with client devices via the advertising platform. 30. The smart interactive billboard system according to claim 24, wherein the client device is a remote wired device. 31. The smart interactive billboard system according to claim 24, wherein the client device is a remote wireless device. 32. The smart interactive billboard system according to claim 24, wherein the client device and the smart interactive billboard device are responsive to at least one of SMS, Bluetooth, infrared or IrDA. 33. The smart interactive billboard system according to claim 25, wherein the advertising platform is a mobile platform and wherein the smart interactive billboard device includes a GSM receiver for receiving GSM signals containing GSM information; and the CPU is responsive to the received GSM signals and changes the display according to the GSM information. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1 Field of the Invention The present invention relates to a smart interactive billboard device that allows billboards or other devices, implemented with the smart billboard device, to display electronic/computer-based information and become interactive. More particularly, the present invention relates to a smart interactive billboard device that allows users to remotely upload information to, download information from, and perform electronic commerce transactions with the billboards, implemented with the smart billboard device. 2. Description of the Related Art Billboard advertising is a popular means for marketing a retailer's products since they are large, appealing and attractive. Products can include services, goods or the like. Generally, billboards are two-dimensional, is however, billboards can be three-dimensional. Advertisements that are posted on two-dimensional, static billboards can capture the attention of potential customers because of their size. The information provided on the billboards, however, is limited and their content is not readily changed. Moreover, they provide little or no opportunity for a customer to follow up when interested. For example, when a customer is driving pass the billboard, the customer may not be able to safely write down the information from the billboard while driving. Three-dimensional billboards can be more attractive than two-dimensional billboards since they usually contain animation or the like, and are visible in the daytime and nighttime. Also, the display may be changeable in short periods of time, thereby preventing a passing viewer from noting all relevant information. Similar to the two-dimensional billboards, the information provided on the three-dimensional billboards may be limited. Further, the information or advertisements provided on the conventional two- and three-dimensional billboards are difficult to update and maintain, as they require on-site attention. In addition, both types of billboards are non-interactive, i.e., the billboards do not support purchases or other electronic commerce transactions. Web-based and cell-phone based advertising is another popular marketing tool for retailers. The web-based and cell-phone based advertisements are capable of automatically updating according to a user's demographics, requirements and behavior. Further, the user can selectively choose or filter information from the advertisement, and perform electronic commerce transactions with respect to the displayed advertisement. However, there are restrictions depending on the user's device, i.e., wireless device such as a pager, mobile or cell phone or the like. U.S. Pat. No. 6,219,696 issued on Apr. 17, 2001 to Wynblatt et al., concerns a system for providing targeted Internet information to mobile devices is The system provides for the just-in-time distribution of information through mobile information terminals. The system involves the Internet as the primary source of information, and includes a mobile information terminal as the output device and a local agent that is locally operated. The mobile information terminal includes a receiver, a URL queue and a www renderer/browser. The local agent includes a short-range transmitter for distributing information pointers to mobile information terminals and a mechanism for transferring data into the transmitter. WO 00/62564 (Blow), System and Method for Distributing Advertising and Gathering Information in a Wireless Communication Network, published on Oct. 19, 2000, relates to a system and method for inserting and distributing commercial advertising to wireless phone subscribers A subscriber enters a user profile either when initially signing up for service or through an interactive menu on the phone handset. Through phone registration messages transmitted as overhead messages within the wireless network, the network determines the location of the phone. The system subsequently transmits advertising messages to the user at call origination based upon the user profile and cell location. The system uses the subscriber location to transmit messages to the user phone that will launch a browser like application within the phone. The browser displays textual or graphical advertisements on the handset display. In addition, the system initiates browser pop-up polling queries in which the user offers responses. The phone uses packetized data transmission to transmit the responses bark to the base station. The system accumulates and processes the various user responses, and uses location information to selectively transmit electronic coupon offers to local subscribers. There is a need for a smart interactive billboard device that may be on a fixed or mobile platform, and that allows billboards or other devices, implemented with the smart billboard device, to capture the attention of potential customers by displaying on the billboards electronic/computer-based information or advertisements. There is also a need for a compact, smart billboard device that allows users to become interactive with the billboards. i.e., download information to, upload information from, and perform electronic commerce transactions with respect to the, information displayed on the billboards. |
<SOH> SUMMARY OF THE INVENTION <EOH>Accordingly, it is an object of the present invention to overcome the disadvantages of the prior art, and to provide a smart interactive billboard device that can be installed on stationary and mobile platforms. It is another object of the present invention to provide a smart interactive billboard device that allows users or customers with client devices to download information from and upload information to billboards or other devices implemented with the smart billboard device. It is still a further object of the present invention to provide a smart interactive device that allows users or customers to perform electronic commerce transactions with respect to the information displayed on the billboards or other devices implemented with the smart billboard device. It is yet a further object of the present invention to provide a smart interactive device that allows: users or customers to exchange or transfer information amongst other users or customers using the billboards or other devices implemented with the smart billboard device. It is yet another object of the present invention to use the smart interactive device with a changeable display. Accordingly, to achieve the above objects, there is provided a smart interactive billboard device for allowing billboards or other devices implemented with the smart billboard device, such as a stationary or mobile platform with a display screen, to display electronic/computer-based information and become interactive. The smart billboard device includes a transceiver adapted to communicate interactively with a server and at least one or more client devices to allow users to be interactive with the implemented billboards or other devices. Also provided is a CPU including software for controlling the implemented billboards or other devices, and a memory adapted to store information uploaded to the implemented billboards or other devices, or information received from the server and client devices. A display unit for displaying the information on the implemented, billboards or other devices can also included. To further achieve the above objects, there is provided a smart interactive billboard system for allowing billboards or other devices implemented with a smart billboard device to display electronic/computer-based information and become interactive that includes a smart interactive billboard device and a server. The smart interactive billboard device includes a transceiver operative to communicate interactively with at least one user to allow the user to be interactive with the implemented billboards or other devices, a CPU including software operative to control the implemented billboards or other devices, and a memory adapted to store information received or uploaded to the implemented billboards or other devices. The server, which includes a server interface, is adapted to at least request information from and deliver information to the implemented billboards or other devices. To further achieve the above objects, there is provided a smart interactive billboard system for allowing billboards or other devices implemented with a smart billboard device to display electronic/computer-based information and become interactive that includes a smart interactive billboard device and at least one client device. The smart interactive billboard device includes a transceiver operative to communicate interactively with at least one user to allow the user to be interactive with the implemented billboards or other devices, a CPU including software operative to control the implemented billboards or other devices, and a memory adapted to store information received or uploaded to the implemented billboards or other devices. The client device, which includes a client interface, is adapted to at least provide the user with the capability to upload information to or receive information from the smart interactive billboard device. |
Cvd diamond cutting insert |
A blade, particularly a razor blade, comprises a layer of CVD diamond having a monolithic elongate cutting edge. |
1. A blade comprising a layer of CVD diamond have a monolithic elongate cutting edge. 2. A blade according to claim 1 wherein the cutting edge is a straight cutting edge. 3. A blade according to claim 1 or wherein the cutting edge is at least 10 mm in length. 4. A blade according to claim 1 or wherein the cutting edge is at least 20 mm in length. 5. A blade according to claim 1 or which is a razor blade. 6. A blade according to claim 5 wherein the length of the cutting edge is in the range 30 to 45 mm. 7. A blade according to claim 1 wherein the CVD diamond layer has a thickness in the range 50 μm-400 μm. 8. A blade according to claim 7 wherein the CVD diamond layer has a thickness in the range 150 μm-275 μm. 9. A blade according to claim 1 wherein the CVD diamond layer has a rectangular, square or triangular shape. 10. A blade according to claim 5 wherein the effective full angle of the blade at a distance of 40 μm from the cutting edge tip is in the range 12 to 28°. 11. A blade according to claim 5 wherein the effective full angle of the blade at a distance of 40 μm from the cutting edge tip is in the range 15 to 25°. 12. A blade according to claim 5 wherein the effective full angle of the blade at a distance of 40 μm from the cutting edge tip is in the range 17 to 23°. 13. A blade according to claims claim 1 wherein the cutting edge tip has a radius which is less than 60 nm. 14. A blade according to claim 1 wherein the cutting edge tip has a radius of less than 40 nm. 15. A blade according to claim 1 wherein the cutting edge tip has a radius of less than 20 nm. 16. A blade according to claim 1 which is a symmetric blade. 17. A blade according to claim 1 which is an asymmetric blade. 18. (Canceled). 19. (Canceled). |
<SOH> BACKGROUND OF THE INVENTION <EOH>This invention relates to a CVD diamond cutting insert and more particularly to a CVD diamond razor blade. Cutting human hair by shaving is a demanding and unique application. The hairs themselves do not cut easily, but can bend and then be pulled by an unsuitable or blunt blade causing pain. The most important region of the razor blade is generally accepted to be the first 40-150 μm from the cutting edge, which must cut cleanly, not show permanent deformation under cutting loads, and yet be as thin and frictionless as possible. Conventional razor blades are made from steel and go blunt during the hair shaving process. Techniques to improve the longevity of steel blades include the application of hard coatings and the treatment of the steel by, for example, ion implantation. Whilst these enhancement techniques do work, the improvement in longevity (the length of time the blade remains sharp) is only modest. In U.S. Pat. No. 4,720,918 it is stated that in conventional steel razor blades the effective full angle at a distance of about 40 μm back from the tip of the razor needs to be kept as low as possible consistent with a sufficiently strong edge, and in particular this angle is generally 20° or less. This reduces the drag on the hair being cut which otherwise results when using a wider blade angle, as the wider blade requires to open a wide wedge angle in the hair as it cuts through. This same U.S. patent suggests that for materials harder than steel, such as sapphire, titanium carbide and diamond, the blade can be made thinner with smaller angles at the cutting edge of the blade. No working examples of such blades are provided. It has been recognised that harder materials than steel, such as ceramics, generally make longer lasting blades. The harder the material the longer the blade will last, provided the edge does not damage or chip. FR 2 536 691 recognises this fact and shows several designs of diamond razor blades made from multiple, single crystals of diamond each of which has a sharpened edge. The use of multiple edges is however limited, as it requires careful fabrication and alignment to present a single continuous straight edge without steps in the regions where adjacent crystals are joined. WO 93/00204 describes a razor blade which includes a substrate with a wedge-shaped cutting edge and a layer of diamond or diamond-like material coating the substrate and more particularly coating the cutting edge. The substrate or core of the blade provides the rigidity and resilience required and the wear resistance is obtained from the diamond coating. In general these coatings are very thin, ensuring that the diamond is supported by the substrate material right to the cutting edge and that the coating does not adversely increase the cutting edge radius. |
<SOH> SUMMARY OF THE INVENTION <EOH>According to the present invention, a blade comprises a layer of CVD diamond having a monolithic elongate cutting edge. Essential to the blade of the invention is that the layer is made of CVD diamond and it has a monolithic elongate cutting edge. The cutting edge is monolithic and uninterrupted by bonding regions such as those in the blade of FR 2 536 691. The cutting edge will generally be at least 10 mm in length and preferably at least 20 mm in length. When the blade is a razor blade, the length will typically be in the range 30 to 45 mm. The shape of the layer of CVD diamond will depend on the application to which the blade is to be put. The shape, particularly in the case of razor blades, will generally be such as to provide at least one straight cutting edge and will typically be rectangular, square or triangular. The thickness of the CVD diamond layer will again depend on the use to which the blade is to be put. In the case of razor blades, the CVD diamond layer will typically have a thickness in the range 50-400 μm, preferably 150-275 μm CVD diamond is, as is known in the art, diamond produced by chemical vapour deposition. The diamond may be single crystal or polycrystalline. When the blade is a razor blade, it has been found that the profile of the cutting edge tip and region of the layer leading up to the cutting edge tip is of importance. In this regard, it is preferred that the effective full angle of the blade at a distance of 40 μm from the cutting edge tip is in the range 12 to 28°, preferably in the range 15 to 25°, and more preferably in the range 17 to 23°. The effective full angle and effective half angle of a razor blade is illustrated by the attached FIGS. 1 and 2 . Referring first to FIG. 1 , an asymmetric razor blade is shown and comprises a body 10 having a cutting edge region 12 . The cutting edge region 12 comprises inwardly sloping surfaces 14 , 16 . The surface 16 has an extra facet 18 formed in it. The cutting edge tip or extreme edge of the blade is at 20 . The effective half angle of the blade is the angle formed at the intersection of a line 22 passing through the centre of the blade and a line 26 drawn from a point 24 on one of the surfaces 14 , 16 . Point 24 is characterised in that it also lies on a line 34 drawn normal to line 22 typically at a distance of 40 μm from the tip 20 of the blade. For the surface 16 , the line 26 is not coincident with the surface 16 . The effective half angle is the angle 28 . For the surface 14 , the line 26 from the point 24 is coincident with the surface 14 and the effective half angle is the angle 30 . The effective full angle is the sum of the angles 28 and 30 . FIG. 2 illustrates the effective half angle and effective full angle for a symmetric blade. In this Figure, like parts to those of FIG. 1 carry like numerals. In this blade, the surface 14 also has a facet 18 formed on it. The lines 26 from the points 24 of both the surfaces 14 , 16 are not coincident with these surfaces and intersect line 22 at the cutting edge tip 20 . The angles 28 and 30 are the same. Thus, for a symmetric blade, the effective full angle is twice that of the effective half angles 28 , 30 . According to a further aspect of the invention, the cutting edge tip of the blade of the invention preferably has a radius which is less than 60 nm, preferably less than 40 nm, more preferably less than 20 nm. Tips with a radius of less than 5 nm can be produced and used. The tip radius of the blade is identified by the numeral 32 in FIG. 1 . The invention extends to the use of a blade as described above as a razor blade in the shaving of hair from a surface of a human or of an animal. Still further according to the invention, a razor comprises a cutting insert as described above, so mounted in a holder to present the monolithic elongate cutting edge suitable to carry out a shaving operation. |
Novel heterocyclic compounds, method for preparing same and use thereof as medicines, in particular as antibacterial agents |
The invention relates to new heterocyclic compounds of general formula (I), and their salts with a base or an acid: The invention also relates to a process for the preparation of these compounds, as well as their use as medicaments, in particular as anti-bacterial agents. |
1. A compound selected from the group consisting of a compound the formula: wherein: a) either R1 is selected from the group consisting of hydrogen, a —COOH, —CN, —COOR, —(CH2)n′R5, —CONR6R7 and R is selected from the group consisting of alkyl of 1 to 6 carbon atoms, optionally substituted by pyridyl, a —CH2-alkenyl of 3 to 9 carbon atoms, (poly)alkoxyalkyl of 1 to 4 oxygen atoms and 3 to 10 carbon atoms, aryl of 6 to 10 carbon atoms, aralkyl of 7 to 11 carbon atoms, the ring of the aryl or aralkyl optionally substituted by at least one member of the group consisting of —OH, —NH2, —NO2, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms and at least one halogen, R5 is selected from the group consisting of —COOH, —CN, —OH, —NH2, —CO—NR6R7, —COOR, —OR, —OCOH, —OCOR, —OCOOR, —OCONHR, —OCONH2, —OSO2R, —NHR, —NHCOR, —NHCOH, —NHSO2R, —NH—COOR, —NH—CO—NHR, —NH—CO—NH2 and —N3, R being defined as above, R6 and R7 are individually selected from the group consisting of hydrogen, alkyl of 1 to 6 carbon atoms, aryl 6 to 10 carbon atoms and aralkyl of 7 to 11 carbon atoms and alkyl of 1 to 6 carbon atoms substituted by pyridyl, n′ is 1 or 2, R3 and R4 together form a phenyl or a heterocycle of aromatic character with 5 or 6 vertices containing 1 to 4 heteroatoms chosen from nitrogen, oxygen and sulfur, and optionally substituted by at least one R′, R′ being selected from the group consisting of hydrogen, and the alkyl of 1 to 6 carbon atoms, optionally substituted by at least one member of the group consisting of hydroxy, oxo, halogen, cyano, nitro, alkenyl of 2 to 6 carbon atoms, halogen, amino, —OH, protected —OH, —OR, —NHCOH, —NHCOR, —NHCOOR, —COOH, —COOR, —C(C6H5)3 and —CH2—CH2.S(O)m-R, R being as defined previously and m being 0, 1 or 2, or b) R4 is hydrogen or —(CH2)n′1R5, n′1 being 0, 1 or 2 and R5 being as defined above, and R1 and R3 together form a phenyl or an optionally substituted heterocycle, as defined above, in both case a) and b) R2 is selected from the group consisting of hydrogen, halogen, R, —S(O)mR, —OR, —NHCOR, —NHCOOR and —NHSO2R, m and R being as defined previously, X is —C(O)—B— linked to the nitrogen atom by the carbon atom, B is selected from the group consisting of —O—(CH2)n″— linked with the carbonyl by the oxygen atom, —NR8—(CH2)n″— and —NR8—O— linked with the carbonyl by the nitrogen atom, n″ is 0 or 1 and R8 is selected from the group consisting of hydrogen, an —OH, —R, —OR, —Y —OY, —Y1, —OY1, —Y2, —OY2, —Y3, —O—CH2—CH2—S(O)m-R, —SiRaRbRc and —OSiRaRbRc, Ra, Rb and Rc are individually alkyl of 1 to 6 carbon atoms or aryl 6 to 10 carbon atoms and R and m are as defined previously, Y is selected from the group consisting of —COR, —COOR, —CONH2, —CONHR, —CONHOH, —CONHSO2R, —CH2COOH, —CH2COOR, —CH2CONHOH, —CH2CONHCN, —CH2tetrazole, protected —CH2tetrazole, —CH2SO3H, —CH2SO2R, —CH2PO(OR)2, —CH2PO(OR)(OH), —CH2PO(R)(OH) and —CH2PO(OH)2, Y1 is selected from the group consisting of —SO2R, —SO2NHCOH, —SO2NHCOR, —SO2NHCOOR, —SO2NHCONHR, —SO2NHCONH2 and —SO3H, Y2 is selected fro the group consisting of —PO(OH)2, —PO(OR)2, —PO(OH)(OR) and —PO(OH)(R), Y3 is selected from the group consisting of: tetrazole, tetrazole substituted by R, squarate, —NH or —NR tetrazole, —NH or —NR tetrazole, substituted by R, —NHSO2R and —NRSO2R, R being defined as above, n is 1 or 2 and its pharmaceutically acceptable salts with a base or acid. 2. A compound of claim 1, wherein n is 1. 3. A compound of claim 1 wherein R2 is hydrogen. 4. A compound of claim 1 wherein R3 and R4 together form a phenyl or heterocycle, optionally substituted, as defined in claim 1. 5. A compound of claim 1 wherein R3 and R4 together form phenyl or heterocycle selected from the group consisting of thienyl, furyl, pyrazolyl and triazolyl, optionally substituted. 6. A compound of claim 5 wherein R1 is selected from the group consisting of hydrogen, —COOCH3, —COOC2H5, —CONH2, —CONHCH3, —CONHCH2-phenyl and —CONHCH2-pyridyl. 7. A compound of claim 1 wherein B is —NR8—(CH2)n″ and n″ is 0. 8. selected from the group consisting of R8 is —Y1 or —OY1 which Y1 is selected from the group consisting of —SO2R, —SO2NHCOR, —SO2NHCOOR, —SO2NHCONHR and —SO3H and R is as defined in claim 1. 9. A compound of claim 7, wherein R8 is selected from the group consisting of hydrogen, hydroxyl, —CO-phenyl, —O-allyl, —OPO3H, —OPO3-benzyl, —OCH2COOH and —O-benzyl. 10. A compound of claim 1, selected from the group consisting of: the sodium salt of trans-3-oxo-4-(sulphooxy)-2,3,4,5-tetrahydro-2,5-methano-1H-2,4-benzodiazepine-1-carboxamide; the sodium salt of 3-methyl-5-(sulphooxy)-5,6,7,8-tetrahydro-4,7-methano-4H-[1,2,3]-triazolo[4,5-e][1,3]diazepine-6 (3H)-one; the sodium salt of trans-1-methyl-6-oxo-5-(sulphooxy)-4,5,6,8-tetrahydro-4,7-methano-1H-pyrazolo[3,4-e][1,3]diazepine-8(7H)-carboxamide; the sodium salt of trans-N-methyl-3-oxo-4-(sulphooxy)-2,3,4,5-tetrahydro-2,5-methano-1H-2,4-benzodiazepine-1-carboxamide; the sodium salt of 5-(sulphooxy)-5,6,7,8-tetrahydro-4,7-methano-4H-furo[2,3-e][1,3]diazepine-6-one; the pyridinium salt of 1-propyl-5-(sulphooxy)-4,5,7,8-tetrahydro-4,7-methano-imidazo[4,5-e][1,3]diazepine-6(1H)-one; the sodium salt of trans methyl 6-oxo-5-(sulphooxy)-5,6,7,8-tetrahydro-4,7-methano-4H-furo[2,3-e][1,3]diazepine-8-carboxylate; the sodium salt of 1-methyl-5-(sulphooxy)-5,6,7,8-tetrahydro-4,7-methano-4H-pyrazolo[3,4-e][1,3]diazepine-6(1H)-one; the sodium salt of trans-3-oxo-N-(4-pyridinylmethyl)-4-(sulphooxy)-2,3,4,5-tetrahydro-2,5-methano-1H-2,4-benzodiazepine-1-carboxamide; the sodium salt of trans-6-oxo-5-(sulphooxy)-5,6,7,8-tetrahydro-4,7-methano-4H-thieno[2,3-e][1,3]diazepine-8-carboxamide; the sodium salt of trans-6-oxo-5-(sulphooxy)-5,6,7,8-tetrahydro-4,7-methano-4H-furo[2,3-e][1,3]diazepine-8-carboxamide. the sodium salt of trans-1,2,3,5-tetrahydro-8-hydroxy-3-oxo-2-(sulphooxy)-1,4-methano-4H-2,4-benzodiazepine-5-carboxamide the sodium salt of trans-7-(acetylamino)-1,2,3,5-tetrahydro-8-hydroxy-3-oxo-2-(sulphooxy)-1,4-methano-4H-2,4-benzodiazepine-5-carboxamide the sodium salt of trans-1,5-dihydro-5-(hydroxymethyl)-2-(sulphooxy)-1,4-methano-4H-2,4-benzodiazepin-3(2H)-one the sodium salt of trans-4,5,6,8-tetrahydro-N-methyl-6-oxo-5-(sulphooxy)-4,7-methano-7H-thieno[2,3-e][1,3]diazepine-8-carboxamide the sodium salt of 7,8-dihydro-7-(sulphooxy)-5,8-methano-5H-thieno[2,3-e][1,3]diazepin-6(4H)-one and the triethylammonium salt of trans-2-bromo-4,5,6,8-tetrahydro-6-oxo-5-(sulphooxy)-4,7-methano-7H-thieno[2,3-e][1,3]diazepine-8-carboxamide. 11-19. (canceled) 20. An antibacterial composition comprising an antibacterial amount of a compound of claim 1 and a pharmaceutical carrier. 21. A compound of the formula: wherein a) either R′1 is selected from the group consisting of hydrogen, —CN, protected —COOH, —COOR″, —(CH2)n′R′5, —CONR6R7 and R″ is selected from the group consisting of alkyl of 1 to 6 carbon atoms, optionally substituted by pyridyl, —CH2-alkenyl of 3 to 9 carbon atoms, aryl of 6 to 10 carbon atoms and aralkyl of 7 to 11 carbon atoms, the ring of the aryl or aralkyl being optionally substituted by at least one member of the group consisting of —NO2, protected —OH, protected —NH2, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms or halogen, R′5 is selected from the group consisting of protected —OH, —CN, protected —NH2, —CO—NR6R7, protected —COOH, —COOR″, —OR″, —OCOH, —OCOR″, —OCOOR″, —OCONH2, —OCONHR″, protected —NHR″, —NHCOR″, —NHSO2R″, —NH—COOR″, —NH—CO—NHR″ and —NH—CONH2, R″ being as defined above in claim 1, n′ is 1 or 2, R6, R7 and R3 are defined as in claim 1, R′4 is R4 as defined in claim 1 or b) R′4 is hydrogen or —(CH2)n′, R′5, n′1 is 0 or 1 or 2 and R′5 is defined as above and R′1 and R′3 form phenyl or optionally substituted heterocycle as defined for R3 and R4 in claim 1 and in both a) and b) in both case a) and b) R′2 is selected from the group consisting of hydrogen, halogen, R″, —S(O)mR″, —OR″, —NHCOH, —NHCOR″, —NHCOOR″ and —NHSO2R″, R″ being as defined previously, ZH is selected from the group consisting of HO—(CH2)n″—, HNR′8—(CH2)n″— and HNR8—O—, n″ is as defined in claim 1 and R′8 is selected from the group consisting of hydrogen, R″, protected —OH, —OR″, —Y′, —OY, —Y′1, —OY′1, —Y′2, —OY′2, —Y′3, —O—CH2—CH2—S(O)m—R″, —SiRaRbRc and —OSiRaRbRc, Ra, Rb and Rc are individually alkyl of 1 to 6 carbon atoms or aryl of 6 to 10 carbon atoms and R″ and m being as defined in claim 1, ZH is not HO—(CH2)n— when R′1 is —(CH2)n′R′5; Y′ is selected from the group consisting of —COH, —COR″, —COOR″, —CONH2, —CONHR″, —CONHSO2R″, —CH2COOR″, protected —CH2tetrazole, —CH2SO2R″, —CH2PO(OR″)2, protected —CONHOH, protected —CH2COOH, protected —CH2CONHOH, protected —CH2SO3, protected —CH2PO(OR)(OH), protected —CH2PO(R)(OH), and protected —CH2PO(OH)2, Y′1 is selected from the group consisting of —SO2R″, —SO2NHCOH, —SO2NHCOR″, —SO2NHCOOR″, —SO2NHCONH2, —SO2NHCONHR″ and protected —SO3H, Y′2 is selected from the group consisting of —PO(OR″)2, protected —PO(OH)2, protected —PO(OH)(OR) and protected —PO(OH)(R), Y′3 is selected from the group consisting of protected tetrazole, tetrazole substituted by R″, protected squarate, protected —NH tetrazole, protected —NR″ tetrazole, protected —NH, —NR″ tetrazole substituted by R″—NHSO2R″ and —NSO2R″, R″ being defined as above, n is as defined in claim 1; X1 is a hydrogen and X2 is -Z-CO—X3, X3 is the remainder of the carbonylation agent, or X2 is -ZH and X1 is —CO—X3, X3 being defined as above. 22. A compound of the formula wherein a) either R′1 is selected from the group consisting of hydrogen, —CN, protected —COOH, —COOR″, —(CH2)n′R′5, —CONR6R7 and R″ is selected from the group consisting of alkyl of 1 to 6 carbon atoms, optionally substituted by pyridyl, —CH2-alkenyl of 3 to 9 carbon atoms, aryl of 6 to 10 carbon atoms and aralkyl of 7 to 11 carbon atoms, the ring of the aryl or aralkyl being optionally substituted by at least one member of the group consisting of —NO2, protected —OH, protected —NH2, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms or halogen, R′5 is selected from the group consisting of protected —OH, —CN, protected —NH2, —CO—NR6R7, protected —COOH, —COOR″, —OR″, —OCOH, —OCOR″, —OCOOR″, —OCONH2, —OCONHR″, protected —NHR″, —NHCOR″, —NHSO2R″, —NH—COOR″, —NH—CO—NHR″ and —NH—CONH2, R″ being as defined above in claim 1, n′ is 1 or 2, R6, R7 and R3 are defined as in claim 1, R′4 is R4 as defined in claim 1 or b) R′4 is hydrogen or —(CH2)n′, R′5, n′1 is 0 or 1 or 2 and R′5 is defined as above and R′1 and R′3 form phenyl or optionally substituted heterocycle as defined for R3 and R4 in claim 1 and in both a) and b) in both case a) and b) R′2 is selected from the group consisting of hydrogen, halogen, R″, —S(O)mR″, —OR″, —NHCOH, —NHCOR″, —NHCOOR″ and —NHSO2R″, R″ being as defined previously, ZH is selected from the group consisting of HO—(CH2)n″—, HNR′8—(CH2)n″— and HNR8—O—, n″ is as defined in claim 1 and R′8 is selected from the group consisting of hydrogen, R″, protected —OH, —OR″, —Y′, —OY′, —Y′1, —OY′1, —Y′2, —OY′2, —Y′3, —O—CH2—CH2—S(O)m—R″, —SiRaRbRc and —OSiRaRbRc, Ra, Rb and Rc are individually alkyl of 1 to 6 carbon atoms or aryl of 6 to 10 carbon atoms and R″ and in being as defined in claim 1; Y′ is selected from the group consisting of —COH, —COR″, —COOR″, —CONH2, —CONHR″, —CONHSO2R″, —CH2COOR″, protected —CH2tetrazole, —CH2SO2R″, —CH2PO(OR″)2, protected —CONHOH, protected —CH2COOH, protected —CH2CONHOH, protected —CH2SO3, protected —CH2PO(OR)(OH), protected —CH2PO(R)(OH), and protected —CH2PO(OH)2, Y′1 is selected from the group consisting of —SO2R″, —SO2NHCOH, —SO2NHCOR″, —SO2NHCOOR″, —SO2NHCONH2, —SO2NHCONHR″ and protected —SO3H, Y′2 is selected from the group consisting of —PO(OR″)2, protected —PO(OH)2, protected —PO(OH)(OR) and protected —PO(OH)(R), Y′3 is selected from the group consisting of protected tetrazole, tetrazole substituted by R″, protected squarate, protected —NH tetrazole, protected —NR″ tetrazole, protected —NH, —NR″ tetrazole substituted by R″—NHSO2R″ and —NSO2R″, R″ being defined as above, n is as defined in claim 1; and their derivatives in which the nitrogen is protected. 23. A compound of a formula selected from the group consisting of: wherein R′1, R′2, R3, R′4, n, R9 and A have the same meanings as in claim 16 and their non-toxic, pharmaceutically acceptable acid addition salts. 24. A compound of the formulae: in which R′1, R′2, R3, R′4, n, R′8 and A have the same meanings as in claim 17 and their non-toxic, pharmaceutically acceptable acid addition salts. 25. A compound of claim 21 or the form of its hydrochloride or trifloroacetate salt. 26. A compound of claim 22 or the form of its hydrochloride or trifloroacetate salt. 27. A compound of claim 23 or the form of its hydrochloride or trifloroacetate salt. 28. A compound of claim 24 or the form of its hydrochloride or trifloroacetate salt. 29. A method of combating bacterial infections in warm-blooded animals comprising administering to warm-blooded animals in need thereof an antibacterially effective amount of a compound of claim 1. |
Method of mycophenolate mofetil preparation |
Synthesis of mycophenolate mofetil (1), where R1=2-(-morpholinyl)ethyl and R2=hydrogen atom, includes reaction of mycophenolic acid with 4-(2-hydroxyethyl)morpholine in a suitable solvent under azeotropic separation of water. |
1. The process of preparation of mycophenolate mofetil by direct esterification of mycophenolic acid and 2-morpholinoethanol characterized with esterification carried out under boiling in ethers. 2. The process according to claim 1, characterized with the use of ethers as solvent of the general formula R3OR4, where R3 and R4 are independently alkyl or aryl. 3. The process according to claim 2, characterized with the use of ethers as solvent of boiling point above 120° C. 4. The process according to claim 1, characterized with the use of 1.01 up to 3.0 molar equivalents of 2-morpholinoethanol. 5. The process according to claim 3, characterized with the use of dibutylether as an inert solvent. 6. The process according to claim 5, characterized with the starting temperature of the reaction ranging between 130° C. and 138° C. and the final temperature of the reaction ranging between 140° C. and 145° C. 7. The process according to claim 5, characterized with the reflux time ranging from 30 to 80 hours. 8. The process according to claim 5, characterized with the ratio of mycophenolic acid to dibutylether ranging from 1 g/2 ml to 1 g/5 ml. |
<SOH> BACKGROUND ART <EOH>Synthesis of mycophenolate mofetil in accordance with the formula I (R 1 =2-morpholinoethyl, R 2 ═H) is described in the basic patent EP 281 713 B1 (1987) and several other patents: U.S. Pat. No. 4,808,592 (1989), U.S. Pat. No. 4,753,935 (1988), U.S. Pat. No. 4,952,579 (1990), U.S. Pat. No. 4,984,793 (1990), U.S. Pat. No. 4,786,637 (1988). In accordance with these patents mycophenolate mofetil may be prepared using two standard esterification methods (see Synthetic Organic Chemistry, R. B. Wagner and H. D. Zook (Wiley, New York), 1956, pages 479 to 532): reaction of mycophenolic chloride with excessive amount of 2-morpholinoethanol and condensation using dicyclohexylcarbodiimide (DDC). Esterification via the acid chloride is based on reaction of excessive amount of 2-morpholinoethanol with mycophenolic acid chloride that has been prepared from mycophenolic acid using suitable chlorinating agent (thionylchloride, oxalylchloride etc.). Use of the excessive amount of 2-morpholinoethanol (up to 3 equivalents), formation of dimmers (about 2%, R 1 =H or 2-morpholinoethyl, R 2 =mycophenolic acid) represents a disadvantage of the two-stage process, there are also problems with colour of the product. Formation of unjustifiable amount of impurities and dicyclohexylurea that may be eliminated from the reaction mixture only by a chromatography is a disadvantage of DCC use as an activating agent. The U.S. Pat. No. 5,247,083 dated 1993 describes preparation of mycophenolate mofetil by reflux of mycophenolic acid and 2-morpholinoethanol in a suitable solvent or a mixture of solvents under azeotropic water separation. Dichloromethane, benzene, toluene, xylene and higher hydrocarbons are given in the claims and examples. The most suitable solvents are toluene, xylene and their mixture in proportion 1:1. A long reaction period necessary to reach sufficient conversion (depending on the solvent used about 60 to 100 hours) and colour of the product (light violet crystal) are the disadvantages of this method. Object of the international application No. WO 00/34503 dated 2000 is mycophenolic acid esterification with 2-morpholinoethanol using enzyme catalysis. This way mycophenolate mofetil may be obtained in high yield and purity, however, the method may not be used in industry. Within this patent method of mycophenolic acid esterification by boiling in 2-morpholinoethanol without any solvent is described but considering price of 2-morpholinoethanol the method is not suitable either. |
Liver cancer prediction system for early detection and control method thereof |
The present invention relates to a liver cancer prediction system for early detection and control method thereof, which can perform hierarchical classification relating to a risk group for hepatocellular carcinoma, through an estimation of the incidence rate for the hepatocellular carcinoma and a relative risk of the incidence of hepatocellular carcinoma, both of which are found on an individual basis. General information on a patient, information depending on an ultrasonic test performed, clinical information including information on findings upon a first registration of a patient and information on findings upon a diagnosis of liver cancer, and information on a risk group, are stored in a database. A regression count which is an attributable ratio corresponding to each of risk factors is calculated based on the clinical information and risk group information stored in the database. An odds ratio of the incidence of liver cancer is measured by calculating risk probability of the incidence of liver cancer through a given operation process using the calculated regression count. It is thus possible to prevent the incidence of liver cancer per person depending on prediction of the incidence of liver cancer. Also hierarchical classification relating to a risk group for hepatocellular carcinomas is performed through the incidence rate for hepatocellular carcinoma and a relative risk of the incidence of liver cancer that are calculated on an individual basis. Therefore, a tailored model for prediction the incidence of liver cancer can be constructed. |
1. A liver cancer prediction system for early detection, comprising: a controller for controlling the entire operation of the system; a display unit for displaying information and a graphic user interface depending on the operation of the system under the control of the controller; an input unit for inputting initial set values, selecting a given menu based on the information displayed on the display unit and inputting information corresponding to the selected menu; a plurality of databases for storing general information on a patient, information depending on an ultrasonic test performed, clinical information including information on findings upon a first registration of a patient and information on findings upon a diagnosis of liver cancer, and information on a risk group; a regression counter for calculating a regression count which is an attributable ratio corresponding to each of risk factors based on the clinical information and risk group information stored in the database; and an odds ratio measurement unit for measuring an odds ratio of the incidence of liver cancer by calculating risk probability of the incidence of liver cancer through a given operation process using the regression count calculated in the regression counter. 2. The liver cancer prediction system as claimed in claim 1, wherein the database comprises a patient information database for storing/managing information on a patient, an ultrasonic information database for storing/managing ultrasonic information, a clinical information database for storing/managing clinical information on the patient, and a risk group information database for storing/managing information on the risk group. 3. The liver cancer prediction system as claimed in claim 1, wherein the controller receives, from the database, registration information on a corresponding patient that is stored by default upon entry of information depending on the ultrasonic test, and then displays such information on an activation window automatically. 4. The liver cancer prediction system as claimed in claim 1, wherein clinical information includes parameters such as a diagnosis subject, hepatitis, a diagnosis basis, a case history, examination findings and an odds ratio. 5. The liver cancer prediction system as claimed in claim 1, wherein the odds ratio measurement unit uses three kinds of core risk factors including a diagnosis subject, the cause of hepatitis and AFP in order to calculate the risk probability. 6. The liver cancer prediction system as claimed in claim 1, wherein the odds ratio measurement unit uses extended risk factors where other control factors including the three kinds of the core risk factors are taken into consideration in order to calculate the risk probability. 7. The liver cancer prediction system as claimed in claim 1, wherein the odds ratio measurement unit uses risk factors consisting of hepatitis, liver cirrhosis, hepatitis furuncle, ALT, α-FP (feto protein), age, sex (man/female), tolerance level to alcohol, where drinking history is not known, probability for liver cancer, an odds ratio, risk probability and a risk group. 8. The liver cancer prediction system as claimed in claim 1, wherein the odds ratio measurement unit finds an attributable ratio (regression count) by using the logistic regression that corresponds to the risk factor and then calculates risk probability. 9. The liver cancer prediction system as claimed in claim 1, wherein the odds ratio measurement unit comprises an odds ratio storage unit for storing risk probability and an odds ratio of the incidence of liver cancer that are previously made. 10. The liver cancer prediction system as claimed in claim 1, wherein the controller comprises a calculation-selecting unit for selecting whether to calculate risk probability using a core risk factor or perform calculation using an extended risk factor. 11. The liver cancer prediction system as claimed in claim 1, wherein the controller comprises a trace search unit for searching a trace monitoring item from risk group-assigning materials that are previously stored in the database, in case where an extended risk factor is selected. 12. The liver cancer prediction system as claimed in claim 1, further comprising an SMS management unit for generating a short message containing information on the result measured in the odds ratio measurement unit, and then providing the short message to a mobile communication terminal of a patient's attending physician that is previously registered through a mobile communication network. 13. The liver cancer prediction system as claimed in claim 1, further comprising an E-mail management unit for generating E-mail containing information on the result measured in the odds ratio measurement unit and transmitting the generated E-mail to an E-mail account of a patient's attending physician that is previously registered. 14. A liver cancer prediction system for early detection, comprising: a web server for providing a web service for predicting liver cancer to a user terminal through the Internet; a database for storing general information on a patient, information depending on an ultrasonic operation performed, clinical information including information on findings upon a first registration of a patient and information on findings upon a diagnosis of liver cancer, and information on a risk group; and a predicting server for performing liver cancer prediction based on information of the database. 15. The liver cancer prediction system as claimed in claim 14, wherein the web server comprises a controller for controlling the entire operation, a network connection unit for connection to the Internet, a web service unit for providing a web service for predicting liver cancer to the user terminal connected through the Internet, and a prediction server cooperation unit that cooperates with the prediction server to exchange data with the prediction server. 16. The liver cancer prediction system as claimed in claim 15, wherein the web server receives information on an odds ratio of the incidence of liver cancer calculated in the prediction server through the prediction server cooperation unit, and the web server further comprises an SMS management unit for generating a short message containing received information on the odds ratio of the incidence of liver cancer and then providing the short message to a mobile communication terminal of a patient's attending physician that is previously registered through a mobile communication network. 17. The liver cancer prediction system as claimed in claim 15, wherein the web server receives information on an odds ratio of the incidence of liver cancer calculated in the prediction server through the prediction server cooperation unit, and the web server further comprises an E-mail management unit for generating E-mail containing received information on the odds ratio of the incidence of liver cancer and providing the generated E-mail to an E-mail account of a patient's attending physician that is previously registered. 18. The liver cancer prediction system as claimed in claim 14, wherein the prediction server comprises: a controller for controlling the entire operation of the system, a display unit for displaying information depending on the operation of the system and a graphic user interface under the control of the controller, an input unit for inputting initial set values, selecting a given menu according to information displayed on the display unit and inputting information corresponding to the selected menu, a regression counter for calculating a regression count which is an attributable ratio corresponding to each of risk factors based on clinical information and risk group information stored in the database, and an odds ratio measurement unit for measuring an odds ratio of the incidence of liver cancer by calculating risk probability of the incidence of liver cancer through a given operation process using the regression count calculated in the regression counter. 19. The liver cancer prediction system as claimed in claim 18, wherein the controller receives, from the database, registration information on a corresponding patient that is stored by default when information depending on the ultrasonic test is inputted, and then displays such information on an activation window automatically. 20. The liver cancer prediction system as claimed in claim 18, wherein clinical information includes parameters such as a diagnosis subject, hepatitis, a diagnosis basis, a case history, examination findings and an odds ratio. 21. The liver cancer prediction system as claimed in claim 18, wherein the odds ratio measurement unit uses three kinds of core risk factors including a diagnosis subject, the cause of hepatitis and AFP in order to calculate the risk probability. 22. The liver cancer prediction system as claimed in claim 18, wherein the odds ratio measurement unit uses extended risk factors where other control factors including the three kinds of the core risk factors are taken into consideration in order to calculate the risk probability. 23. The liver cancer prediction system as claimed in claim 18, wherein the odds ratio measurement unit uses extended risk factors consisting of hepatitis, liver cirrhosis, hepatitis furuncle, ALT, α-FP (feto protein), age, sex (man/female), tolerance level to alcohol, where drinking history is not known, probability for liver cancer, an odds ratio, risk probability and a risk group. 24. The liver cancer prediction system as claimed in claim 18, wherein the odds ratio measurement unit finds an attributable ratio (regression count) corresponding to the risk factor and then calculates risk probability using the logistic regression. 25. The liver cancer prediction system as claimed in claim 18, wherein the odds ratio measurement unit comprises an odds ratio storage unit for storing risk probability and an odds ratio of the incidence of liver cancer that are previously made. 26. The liver cancer prediction system as claimed in claim 18, wherein the controller comprises a calculation-selecting unit for selecting whether to calculate risk probability using a core risk factor or using an extended risk factor. 27. The liver cancer prediction system as claimed in claim 18, wherein the controller comprises a trace search unit for searching a trace monitoring item from risk group-assigning materials that are previously stored in the database, in case where an extended risk factor is selected. 28. The liver cancer prediction system as claimed in claim 14, wherein the database comprises a patient information database for storing/managing information on a patient, an ultrasonic information database for storing/managing ultrasonic information, a clinical information database for storing/managing clinical information on the patient, and a risk group information database for storing/managing information on the risk group. 29. A method of controlling liver cancer prediction system including a controller for controlling the entire operation of the system; a display unit for displaying information depending on the operation of the system and a graphic user interface under the control of the controller; an input unit for inputting initial set values, selecting a given menu according to information displayed on the display unit and inputting information corresponding to the selected menu; a plurality of databases for storing general information on a patient, information depending on an ultrasonic operation performed, clinical information including information on findings upon a first registration of a patient and information on findings upon a diagnosis of liver cancer, and information on a risk group; a regression counter for calculating a regression count which is an attributable ratio corresponding to each of risk factors based on clinical information and risk group information stored in the database; and an odds ratio measurement unit for measuring an odds ratio of the incidence of liver cancer by calculating risk probability of the incidence of liver cancer through a given operation process using the regression count calculated in the regression counter, comprising: a patient information-managing step of displaying, on a display unit, a given menu wherein general information on a patient can be written, and storing information inputted through the input unit in the database; an ultrasonic information-managing step of displaying, on the display unit, a corresponding menu wherein information depending on an ultrasonic test performed can be written, and storing information inputted through the input unit in the database; a clinical information-managing step of displaying, on the display unit, a given menu wherein information on findings upon a first registration of a patient and information on finding upon a diagnosis of liver cancer can be written, and storing information inputted through the input unit in the database; a risk group-assigning step of displaying, on the display unit, a menu of a given format wherein additional risk groups can be assigned after the clinical information-managing step, and storing s risk group assigned according to information inputted through the input unit in the database; and an odds ratio measurement step of measuring an odds ratio of the incidence of liver cancer, by calculating probability of the incidence of liver cancer on the basis of clinical information stored in the clinical information-managing step and the risk group assigned in the risk group-assigning step. 30. The method as claimed in claim 29, wherein the ultrasonic information-managing step includes receiving, from the database, registration information on a corresponding patient that is stored by default when information depending on the ultrasonic test is inputted, and then displaying such information on the display unit. 31. The method as claimed in claim 29, wherein clinical information-managing step includes displaying clinical information including a diagnosis subject, hepatitis, a diagnosis basis, a case history, examination findings and an odds ratio, matching information selected or inputted through the input unit to respective factors of the clinical information and then storing the matched results in the database. 32. The method as claimed in claim 29, wherein the clinical information of the clinical information-managing step comprises detail parameters such as a serial number, a diagnosis subject, hepatitis, a diagnosis basis, a case history, examination findings, findings upon a diagnosis of liver cancer, a diagnosis method. 33. The method as claimed in claim 29, wherein the clinical information-managing step comprises: first step of, after a serial number is inputted into a registration number inspection box and an enter key is then pressed, searching the serial number through a patient information table stored in the database; second step of, as a result of the search in the first step, if the serial number is not present in the table, displaying a message indicating that the serial number does not exist on the display unit and if the serial number is present in the table, displaying information on a name, an attending physician and reason on the display unit, and determining whether corresponding clinical information is stored in the database; and third step of, as a result of the determination in the second step, if corresponding clinical information does exist in the database, displaying contents stored in the database on the display unit, and if corresponding clinical information does not exist in the database, displaying a given guide message on the display unit. 34. The method as claimed in claim 29, wherein the odds ratio measurement step includes using three kinds of core risk factors including a diagnosis subject, the cause of hepatitis and AFP in order to calculate the risk probability. 35. The method as claimed in claim 29, wherein the odds ratio measurement step includes using an extended risk factor where other control factors including the three kinds of the core risk factors are taken into consideration in order to calculate the risk probability. 36. The method as claimed in claim 29, wherein the odds ratio measurement step includes using the risk factors consisting of hepatitis, liver cirrhosis, hepatitis furuncle, ALT, α-FP (feto protein), age, sex (man/female), tolerance level to alcohol, where drinking history is not known, probability for liver cancer, an odds ratio, risk probability and a risk group. 37. The method as claimed in claim 36, wherein the odds ratio measurement step includes finding an attributable ratio (regression count) corresponding to the risk factor and then calculates risk probability through logistic regression using the attributable ratio. 38. The method as claimed in claim 37, wherein the odds ratio measurement step comprises: a first step of defining three core risk factors for setting a logistic regression model as numerical type parameters; and a second step of inserting respective risk factor into a statistical prediction model depending in risk probability and an odds ratio of the incidence of liver cancer that are already made after being defined in the first step, thus displaying the odds ratio and a risk probability value based on the ratio on the display unit. 39. The method as claimed in claim 29, wherein the risk group-assigning step comprises: first step of searching a serial number through a patient information table stored in the database after the serial number is inputted through the input unit and the enter key is pressed; second step of, as a result of the search in the first step, if the serial number is not present in the table, displaying a message indicating that the serial number does not exist on the display unit, and if the serial number is present in the table, displaying information on a name and an attending physician and reason on the display unit and at the same time determining whether ‘risk group specification’ is stored in the database; and third step of, as a result of the determination in the second step, if corresponding clinical information is stored in the database, displaying contents stored in the database on the display unit, and if corresponding clinical information is not stored in the database, displaying a given guide message on the display unit. 40. The method as claimed in claim 29, wherein the risk group-assigning step comprises a step of selecting whether to calculate risk probability using a core risk factor or using an extended factor. 41. The method as claimed in claim 40, wherein the risk group-assigning step comprises a step of searching a trace monitoring item from risk group-assigning materials that are previously stored in the database, in case where an extended risk factor is selected. 42. The method as claimed in claim 29, wherein liver cancer prediction system further comprises an SMS management unit for managing a short message, and the method further comprises: a step of generating a short message containing the result calculated in the odds ratio measurement step; and a step of transmitting the short message to a mobile communication terminal of a patient's attending physician that is previously registered, through a mobile communication network. 43. The method as claimed in claim 29, wherein liver cancer prediction system further comprises an E-mail management unit for managing E-mail, and the method further comprises: a step of generating E-mail containing the result calculated in the odds ratio measurement step; and a step of transmitting the generated E-mail to an E-mail account of a patient's attending physician that is previously registered. |
<SOH> BACKGROUND ART <EOH>Liver cancer refers to a malignant tumor generated within the liver. It can be largely classified into hepatocellular carcinoma that is primarily generated within a hepatocyte, and metastatic liver cancer that is generated within extrahepatics and is then transferred to the inside of the liver. Liver cancer in this case means primary hepatocellular carcinoma. The hepatocellular carcinoma is one of the most common malignant tumors worldwide. The incidence rate of the hepatocellular carcinoma differs greatly from region to region. It is reported that hepatocellular carcinoma-prone regions are Africa and East Asia, where the incidence rate of hepatocellular carcinoma is 20 and above per 100,000 people. Whereas it is reported that the incidence rate of hepatocellular carcinoma is 10 or less per 100,000 people in U.S.A., North Europe, etc., with a relatively low incidence rate of this disease. Korea has a high incidence rate of hepatocellular carcinoma such as 30 per 100,000 male population and 7 per 100,000 female populations. Especially the incidence rate aged 40 to 60 is 74 in male and 15 in female, which is very high worldwide. Korea National Statistical Office reports that Korea has the second highest death rate in liver cancer next to Africa. According to the report on cancer death rate published by Korea National Statistical Office in 1996, about 10,000 persons died of this disease in a year, which shows a liver cancer death rate of 21.4%. This ratio is the second highest cancer after gastric cancer. The Office reports that the death rate of liver cancer in the forties to fifties is even higher than that of gastric cancer. In order to prevent liver cancer, it is required that we must exactly know the incidence carcinogenesis of liver cancer. If there are medicines for completely hindering carcinogenesis, it will be possible to easily prevent cancer. In recent years, an effort to prevent cancer with medicines has been actively made. In terms of liver cancer, however, significant advancements have not yet been reported so far. Although there has been proposed a method of administrating a medicine that changes aflatoxin within the body to non-carcinogens in some regions where people are severely exposed to aflatoxin, it is still only in a research stage. Therefore, even if it is uncertain to know carcinogenesis, the best alternative prevention method available has to be driven. The most efficient method to prevent a liver caner is to remove or avoid risk factors for hepatocellular carcinoma. The most widely used inspection methods for early detection of hepatocellular carcinoma are a liver ultrasonic inspection method and a serum alpha-feto protein level checking method. Computerized tomography (CT) is more accurate to detect the incidence of cancer than the ultrasonic inspection method, but it is impractical for a screening test due to the inconvenience and high cost. Meanwhile, the ultrasonic inspection method is easy to use and has a detection sensitiveness of about 75% or more for even a tumor of a size less than 3 cm. Accordingly, the ultrasonic inspection method has been widely used for the screening test for early detection of liver cancer. However, there is no method to analyze the long term individual ultrasonic inspection for prediction of the incidence of liver cancer. A patient has to suffer from an inconvenience of receiving diagnosis of liver cancer through the ultrasonic inspection method every time. |
<SOH> BRIEF DESCRIPTION OF DRAWINGS <EOH>Further objects and advantages of the invention can be more fully understood from the following detailed description taken in conjunction with the accompanying drawings in which: FIG. 1 is a diagram showing the connection of a prediction system based on the embodiment of the present invention, FIG. 2 is a block diagram illustrating the construction of a prediction system based on the embodiment of the present invention; FIG. 3 is a block diagram illustrating the construction of a web server based on the embodiment of the present invention; FIG. 4 is a block diagram illustrating the construction of a prediction server based on the embodiment of the present invention; FIG. 5 is a block diagram illustrating the entire configuration of a graphic user interface (GUI) of the prediction system based on the embodiment of the present invention; FIG. 6 is an exemplary view showing a GUI of an initial main menu based on the present invention; FIG. 7 is an exemplary view showing a GUI of a patient information-managing menu based on the present invention; FIG. 8 is an exemplary view showing a GUI of an ultrasonic information-managing menu based on the present invention; FIG. 9 is a table showing a list of information stored in a database based on the present invention; FIG. 10 is an exemplary view showing a GUI of a clinical information-managing menu based on the present invention; FIG. 11 is an exemplary view showing a GUI of a risk group-assigning menu based on the present invention; FIG. 12 is an exemplary view showing a GUI of a core risk factor menu based on the present invention; FIG. 13 is an exemplary view showing a GUI of an extended risk factor menu based on the present invention; FIG. 14 is an exemplary view showing a list of risk factors for measuring an odds ratio based on the present invention; FIG. 15 is the entire flowchart that explains a process of predicting liver cancer based on the present invention; and FIG. 16 is a flowchart that explains a process of a result notification based on the present invention. detailed-description description="Detailed Description" end="lead"? |
Dgks as modifiers of the p53 pathwha and methods of use |
Human DGK genes are identified as modulators of the p53 pathway, and thus are therapeutic targets for disorders associated with defective p53 function. Methods for identifying modulators of p53,comprising screening for agents that modulate the activity of DKG are provided. |
1. A method of identifying a candidate p53 pathway modulating agent, said method comprising the steps of: (a) providing an assay system comprising a purified DGK polypeptide or nucleic acid or a functionally active fragment or derivative thereof; (b) contacting the assay system with a test agent under conditions whereby, but for the presence of the test agent, the system provides a reference activity; and (c) detecting a test agent-biased activity of the assay system, wherein a difference between the test agent-biased activity and the reference activity identifies the test agent as a candidate p53 pathway modulating agent. 2. The method of claim 1 wherein the assay system comprises cultured cells that express the DGK polypeptide. 3. The method of claim 2 wherein the cultured cells additionally have defective p53 function. 4. The method of claim 1 wherein the assay system includes a screening assay comprising a DGK polypeptide, and the candidate test agent is a small molecule modulator. 5. The method of claim 4 wherein the assay is a kinase assay. 6. The method of claim 1 wherein the assay system is selected from the group consisting of an apoptosis assay system, a cell proliferation assay system, an angiogenesis assay system, and a hypoxic induction assay system. 7. The method of claim 1 wherein the assay system includes a binding assay comprising a DGK polypeptide and the candidate test agent is an antibody. 8. The method of claim 1 wherein the assay system includes an expression assay comprising a DGK nucleic acid and the candidate test agent is a nucleic acid modulator. 9. The method of claim 8 wherein the nucleic acid modulator is an antisense oligomer. 10. The method of claim 8 wherein the nucleic acid modulator is a PMO. 11. The method of claim 1 additionally comprising: (d) administering the candidate p53 pathway modulating agent identified in (c) to a model system comprising cells defective in p53 function and, detecting a phenotypic change in the model system that indicates that the p53 function is restored. 12. The method of claim 11 wherein the model system is a mouse model with defective p53 function. 13. A method for modulating a p53 pathway of a cell comprising contacting a cell defective in p53 function with a candidate modulator that specifically binds to a DGK polypeptide comprising an amino acid sequence selected from group consisting of SEQ ID NOs:21, 22, 23, 24, 25, 26, 27, 28, and 29, whereby p53 function is restored. 14. The method of claim 13 wherein the candidate modulator is administered to a vertebrate animal predetermined to have a disease or disorder resulting from a defect in p53 function. 15. The method of claim 13 wherein the candidate modulator is selected from the group consisting of an antibody and a small molecule. 16. The method of claim 1, comprising the additional steps of: (d) providing a secondary assay system comprising cultured cells or a non-human animal expressing DGK, (e) contacting the secondary assay system with the test agent of (b) or an agent derived therefrom under conditions whereby, but for the presence of the test agent or agent derived therefrom, the system provides a reference activity; and (f) detecting an agent-biased activity of the second assay system, wherein a difference between the agent-biased activity and the reference activity of the second assay system confirms the test agent or agent derived therefrom as a candidate p53 pathway modulating agent, and wherein the second assay detects an agent-biased change in the p53 pathway. 17. The method of claim 16 wherein the secondary assay system comprises cultured cells. 18. The method of claim 16 wherein the secondary assay system comprises a non-human animal. 19. The method of claim 18 wherein the non-human animal mis-expresses a p53 pathway gene. 20. A method of modulating p53 pathway in a mammalian cell comprising contacting the cell with an agent that specifically binds a DGK polypeptide or nucleic acid. 21. The method of claim 20 wherein the agent is administered to a mammalian animal predetermined to have a pathology associated with the p53 pathway. 22. The method of claim 20 wherein the agent is a small molecule modulator, a nucleic acid modulator, or an antibody. 23. A method for diagnosing a disease in a patient comprising: (a) obtaining a biological sample from the patient; (b) contacting the sample with a probe for DGK expression; (c) comparing results from step (b) with a control; (d) determining whether step (c) indicates a likelihood of disease. 24. The method of claim 23 wherein said disease is cancer. 25. The method according to claim 24, wherein said cancer is a cancer as shown in Table 1 as having >25% expression level. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The p53 gene is mutated in over 50 different types of human cancers, including familial and spontaneous cancers, and is believed to be the most commonly mutated gene in human cancer (Zambetti and Levine, FASEB (1993) 7:855-865; Hollstein, et al., Nucleic Acids Res. (1994) 22:3551-3555). Greater than 90% of mutations in the p53 gene are missense mutations that alter a single amino acid that inactivates p53 function. Aberrant forms of human p53 are associated with poor prognosis, more aggressive tumors, metastasis, and short survival rates (Mitsudomi et al., Clin Cancer Res 2000 October; 6(10):4055-63; Koshland, Science (1993) 262:1953). The human p53 protein normally functions as a central integrator of signals including DNA damage, hypoxia, nucleotide deprivation, and oncogene activation (Prives, Cell (1998) 95:5-8). In response to these signals, p53 protein levels are greatly increased with the result that the accumulated p53 activates cell cycle arrest or apoptosis depending on the nature and strength of these signals. Indeed, multiple lines of experimental evidence have pointed to a key role for p53 as a tumor suppressor (Levine, Cell (1997) 88:323-331). For example, homozygous p53 “knockout” mice are developmentally normal but exhibit nearly 100% incidence of neoplasia in the first year of life (Donehower et al., Nature (1992) 356:215-221). The biochemical mechanisms and pathways through which p53 functions in normal and cancerous cells are not fully understood, but one clearly important aspect of p53 function is its activity as a gene-specific transcriptional activator. Among the genes with known p53-response elements are several with well-characterized roles in either regulation of the cell cycle or apoptosis, including GADD45, p21/Waf1/Cip1, cyclin G, Bax, IGF-BP3, and MDM2 (Levine, Cell (1997) 88:323-331). Diacylglycerol (DAG) plays a role in intracellular signaling pathways as an allosteric activator of protein kinase C (PKC), which in turn is involved in the regulation of cellular differentiation and proliferation of diverse cell types. DAG also appears to be involved in regulating RAS and RHO family proteins by activating the guanine nucleotide exchange factors VAV and RASGRP1. DAG also occupies a central position in the synthesis of major phospholipids and triacylglycerols. Therefore, in order to maintain cellular homeostasis, intracellular DAG levels must be strictly regulated (Topham M. and Prescott, S. M.(1999) J. Biol. Chem. 274:11447-11450). DAG kinases (DGKs) phosphorylate DAG to phosphatidic acid, therefore removing DAG. DAGK is a modulator that competes with PKC for the second messenger DAG, in intracellular signaling pathway systems. Most DGKs contain structural motifs that may play regulatory roles, and form the basis for dividing the DGKs into 5 subtypes. Type I DGKs, such as DGK-alpha, beta, and gamma, have calcium-binding EF-hand motifs at their N termini. DGK-delta and DKG-eta contain N-terminal pleckstrin homology (PH) domains and are defined as type II. DGK-epsilon contains no identifiable regulatory domains and is a type III DGK The defining characteristic of type IV isozymes, such as DGK-zeta and iota is C-terminal ankyrin repeats. DGK-theta is placed into Group V, which contains 3 cysteine-rich domains and a PH domain. Diacylglycerol kinase alpha (DGKA) converts diacylglycerol to phosphatidic acid, thereby attenuating protein kinase C activity, and also contains an EF-hand domain. The identification and characterization of DGK-alpha or DAGK1, isoforms of DGK, (Schaap et al (1990) FEBS Lett. 275: 151-158) show that all DGKs have a conserved catalytic domain and at least 2 cysteine-rich regions homologous to the C1A and C1B motifs of PKCs (Topham and Prescott (1999) supra). In an expression profiling experiment using lung cancer cell line H1299 expressing temperature sensitive p53, DGKA was identified as one of many primary target genes regulated by p53. However, DGKA showed altered expression in control conditions as well (Kannan K et al. (2001) Oncogene 20:2225-2234). Diacylglycerol kinase delta (DGKD), has a pleckstrin homology domain and an EPH domain, preferentially phosphorylates the arachidonoyl type of diacylglycerol and is most abundant in skeletal muscle (Sakane et al (1996) Chem. 271: 8394-8401). Diacylglycerol kinase epsilon (DGKE), activates the preferential phosphorylation of arachidonoyl-containing diacylglycerols, regulates the cellular distribution of protein kinase C alpha and epsilon and polyunsaturated diacylglycerol turnover (Tang et al. (1996) J. Biol. 271: 10237-10241271). Diacylglycerol kinase gamma (DGKG), contains EF-hand motifs, zinc finger and ATP-binding site, and converts diacylglycerol to phosphatidic acid in a phosphatidylserine-dependent manner, and may regulate phospholipid turnover (Kai, M. et al. (1994) J. Biol. Chem. 269: 18492-18498). DGKG is expressed in the human retina, and mutations in this gene are known to cause retinal eye degeneration in Drosophlia (Masai, I. et al. (1993) Proc. Nat. Acad. Sci. 90: 11157-11161, 1993). Based on these findings, it was thought that mutations in this gene maybe involved in human disease, yet no evidence has been found to support this theory (Stohr, H. et al (1999) Proc. Nat. Acad. Sci. 90: 11157-11161, 1993). Diacylglycerol kinase theta (DGKQ) optimally phosphorylates substrates with an sn-2 unsaturated fatty acid, it is activated by thrombin, has catalytic activity that is lost by binding activated RhoA and may function in signal transduction (Houssa, B, et al. ( 1997) J. Biol. Chem. 272: 10422-10428) and is expressed in mammalian retina (Endele et al (1996) Genomics 33: 145-146). DGKs are found in a wide array of organisms ranging from yeast to man. Several homologs have been identified in rat (Houssa, B, et al. ( 1997) supra), mouse (Pilz, A. et al. (1995) supra), and Drosophila (Masai, I. et al. (1993) supra). The ability to manipulate the genomes of model organisms such as Drosophila provides a powerful means to analyze biochemical processes that, due to significant evolutionary conservation, has direct relevance to more complex vertebrate organisms. Due to a high level of gene and pathway conservation, the strong similarity of cellular processes, and the functional conservation of genes between these model organisms and mammals, identification of the involvement of novel genes in particular pathways and their functions in such model organisms can directly contribute to the understanding of the correlative pathways and methods of modulating them in mammals (see, for example, Mechler B M et al., 1985 EMBO J 4:1551-1557; Gateff E. 1982 Adv. Cancer Res. 37: 33-74; Watson K L., et al., 1994 J Cell Sci. 18: 19-33; Miklos G L, and Rubin G M. 1996 Cell 86:521-529; Wassarman D A, et al., 1995 Curr Opin Gen Dev 5: 44-50; and Booth D R. 1999 Cancer Metastasis Rev. 18: 261-284). For example, a genetic screen can be carried out in an invertebrate model organism having underexpression (e.g. knockout) or overexpression of a gene (referred to as a “genetic entry point”) that yields a visible phenotype. Additional genes are mutated in a random or targeted manner. When a gene mutation changes the original phenotype caused by the mutation in the genetic entry point, the gene is identified as a “modifier” involved in the same or overlapping pathway as the genetic entry point. When the genetic entry point is an ortholog of a human gene implicated in a disease pathway, such as p53, modifier genes can be identified that may be attractive candidate targets for novel therapeutics. All references cited herein, including sequence information in referenced Genbank identifier numbers and website references, are incorporated herein in their entireties. |
<SOH> SUMMARY OF THE INVENTION <EOH>We have discovered genes that modify the p53 pathway in Drosophila, and identified their human orthologs, hereinafter referred to as diacylglycerol kinases (DGKs). The invention provides methods for utilizing these p53 modifier genes and polypeptides to identify candidate therapeutic agents that can be used in the treatment of disorders associated with defective p53 function. Preferred DGK-modulating agents specifically bind to DGK polypeptides and restore p53 function. Other preferred DGK-modulating agents are nucleic acid modulators such as antisense oligomers and RNAi that repress DGK gene expression or product activity by, for example, binding to and inhibiting the respective nucleic acid (i.e. DNA or mRNA). DGK-specific modulating agents may be evaluated by any convenient in vitro or in vivo assay for molecular interaction with a DGK polypeptide or nucleic acid. In one embodiment, candidate p53 modulating agents are tested with an assay system comprising a DGK polypeptide or nucleic acid. Candidate agents that produce a change in the activity of the assay system relative to controls are identified as candidate p53 modulating agents. The assay system may be cell-based or cell-free. DGK-modulating agents include DGK related proteins (e.g. dominant negative mutants, and biotherapeutics); DGK-specific antibodies; DGK-specific antisense oligomers and other nucleic acid modulators; and chemical agents that specifically bind DGK or compete with DGK binding target. In one specific embodiment, a small molecule modulator is identified using a kinase assay. In specific embodiments, the screening assay system is selected from a binding assay, an apoptosis assay, a cell proliferation assay, an angiogenesis assay, and a hypoxic induction assay. In another embodiment, candidate p53 pathway modulating agents are further tested using a second assay system that detects changes in the p53 pathway, such as angiogenic, apoptotic, or cell proliferation changes produced by the originally identified candidate agent or an agent derived from the original agent. The second assay system may use cultured cells or non-human animals. In specific embodiments, the secondary assay system uses non-human animals, including animals predetermined to have a disease or disorder implicating the p53 pathway, such as an angiogenic, apoptotic, or cell proliferation disorder (e.g. cancer). The invention further provides methods for modulating the p53 pathway in a mammalian cell by contacting the mammalian cell with an agent that specifically binds a DGK polypeptide or nucleic acid. The agent may be a small molecule modulator, a nucleic acid modulator, or an antibody and may be administered to a mammalian animal predetermined to have a pathology associated the p53 pathway. detailed-description description="Detailed Description" end="lead"? |
Underwater sampling and mapping apparatus |
The present application provides a system, method and apparatus for measuring at least one physical and/or chemical variable at a plurality of different locations within a body of water. The method includes the steps of: providing dynamic measurement means configured to measure the at least one physical and/or chemical variable; towing said measurement means in said body of water at a predetermined depth, and simultaneously taking measurements of least one physical and/or chemical variable, at least periodically, and generating measurement data; and determining the location of the measurement means while taking said measurements of the at least one physical and/or chemical variable and generating corresponding location data. A method for mapping at least one physical and/or chemical variable for a body of water is also disclosed. |
1. A method for measuring at least one physical and/or chemical variable at a plurality of different locations within a body of water, said method including the steps of: providing dynamic measurement means configured to measure at least one physical and/or chemical variable; towing said measurement means in said body of water at a predetermined depth, and simultaneously taking measurements of least one physical and/or chemical variable, at least periodically, and generating measurement data; and determining the location of the measurement means while taking said measurements of the at least one physical and/or chemical variable and generating corresponding location data. 2. A method as claimed in claim 1 which includes the additional step of: controlling the depth of the measurement means in said body of water whilst towing said measurement means. 3. A method as claimed in claim 2 which includes the additional step of: detecting obstacles in a region adjacent to said measurement means; and wherein the step of controlling the depth of said measurement means is performed in response to detected obstacles. 4. A method as claimed in claim 3 in which the step of, detecting obstacles in a region adjacent to said measurement means, includes the sub-steps of: providing image capture means on said measurement means, adapted to generate a sequence of images of a region adjacent to said measurement means; and visually detecting said obstacles from said sequence of images. 5. A method as claimed in claim 1 wherein said measurement means includes water intake means configured to collect water from said body of water and wherein said method includes the additional step of: collecting at least one water sample from said body of water through said water intake means. 6. A method as claimed in claim 2 wherein the predetermined depth is determined relative to a bottom of the body of water. 7. A method as claimed in claim 1 wherein water depth and at least one other physical and/or chemical variable is measured by said measurement means. 8. A method as claimed in claim 1 wherein at least one physical and/or chemical variable measured by the measurement means is selected from a list including the following physical variables: water depth, water temperature, conductivity, and water turbidity. 9. A method as claimed in claim 1 wherein at least one physical and/or chemical variable measured by the measurement means is selected from a list including the following chemical variables: pH, dissolved oxygen, dissolved chloride, oxidation-reduction potential (ORP), soluble nitrate, ammonia, dissolved gases or chlorophyll A. 10. A method of mapping at least one physical and/or chemical variable in body of water, said method including the steps of: providing dynamic measurement means configured to measure the at least one physical and/or chemical variable; and towing said measurement means in said body of water at a predetermined depth, and simultaneously taking measurements of said at least one physical and/or chemical variable, at least periodically, and generating measurement data; determining the location of the measurement means while taking said measurements of the at least one physical and/or chemical variable and generating corresponding location data; and generating a map representative of the distribution of at least one physical and/or chemical variable within said body of water on the basis of the location and measurement data. 11. A method as claimed in claim 10 including the additional step of: determining the depth of the measurement means when measuring said at least one physical and/or chemical variable, and generating measurement depth data, and wherein said map is generated on the basis of the measurement depth data, measurement data and location data. 12. A method as claimed in claim 11 which includes the additional step of: controlling the depth of the measurement means in said body of water whilst towing said measurement means. 13. A method as claimed in claim 12 which includes the additional step of: detecting obstacles in a region adjacent to said measurement means; and wherein the step of controlling the depth of said measurement means is performed in response to detected obstacles. 14. A method as claimed in claim 13 in which the step of, detecting obstacles in a region adjacent to said measurement means, includes the sub-steps of: providing image capture means on said measurement means, adapted to generate a sequence of images of a region adjacent to said measurement means; and visually detecting said obstacles from said sequence of images. 15. A method as claimed in claim 10 in which said measurement means includes water intake means configured to collect water from said body of water and wherein said method includes the additional step of: collecting at least one water sample from said body of water through said water intake means. 16. A method as claimed in claim 10 wherein the predetermined depth is determined relative to a bottom of the body of water. 17. A method as claimed in claim 10 wherein water depth and at least one other physical and/or chemical variable is measured by said measurement means. 18. A method as claimed in claim 10 wherein at least one physical and/or chemical variable measured by the measurement means is selected from a list including the following physical variables: water depth, water temperature, conductivity, and water turbidity. 19. A method as claimed in claim 10 wherein at least one physical and/or chemical variable measured by the measurement means is selected from a list including the following chemical variables: pH, dissolved oxygen, dissolved chloride, oxidation-reduction potential (ORP), soluble nitrate, ammonia, dissolved gases or chlorophyll A. 20. A method as claimed in claim 10 which the map represents topographic contours of a body of water and the distribution of the at least one physical and/or chemical variable within said body of water. 21. A measurement means configured to measure at least one physical and/or chemical variable in a body of water, said measurement means including, a housing, and at least one sensor mounted at least partially within said housing, said sensor being configured to measure at least one physical and/or chemical variable, wherein said measurement means is configured to be towed in said body of water at a predetermined depth whilst simultaneously measuring, at least periodically, said least one physical and/or chemical variable. 22. A measurement means as claimed in claim 21 which further includes orientation means configured to orientate said measurement means relative to an apparent current experienced by said measurement means when said measurement means is being towed. 23. A measurement means as claimed in claim 21 wherein said orientation means includes at least one fin. 24. A measurement means as claimed in claim 21 which additionally includes water intake means to allow the collection of a sample of water from the body of water. 25. A measurement means as claimed in claim 21 which additionally includes image capture means adapted to generate a sequence of images of a region of said body of water adjacent to said measurement means, wherein in use the depth of the measurement means is controlled in response to the sequence of video images. 26. A measurement means as claimed in claim 21 wherein said at least one sensor is configured to measure one or more of the following physical variables: water depth, water temperature, conductivity, water turbidity. 27. A measurement means as claimed in claim 21 wherein said at least one sensor is configured to measure one or more of the following chemical variables: pH, dissolved oxygen, dissolved chloride, oxidation-reduction potential (ORP), soluble nitrate, ammonia, dissolved gases or chlorophyll A. 28. A measurement means as claimed in claim 21 wherein said housing includes a frame. 29. A measurement means as claimed in claim 21 wherein said housing includes a water permeable container configured to contain said at least one sensor. 30. A measurement means as claimed in claim 21 which further includes data storage means in communication with said at least one sensor, said data storage means being configured to store measurement data generated by said at least one sensor. 31. A system for taking a series of measurements of at least one physical and/or chemical variable in a body of water, said system including: measurement means configured to measure at least one physical and/or chemical variable in a body of water, wherein in use said measurement means is configured to be towed in said body of water at a predetermined depth whilst simultaneously measuring, at least periodically, said at least one physical and/or chemical variable to generate measurement data; and location means configured to determine the location of the measurement means while taking said measurements of the at least one physical and/or chemical variable to generate location data; and data storage means configured to store said measurement data and location data. 32. A system for taking a series of measurements as claimed in claim 31 which further includes depth control means configured to control the depth of the measurement means while said measurement means is being towed. 33. A system as claimed in claim 31 including image capture means adapted to provide a sequence of images of a region of the body of water adjacent the measurement means. 34. A system as claimed in claim 31 which further includes a water inlet means, mounted on said measurement means, configured to allow collection one or more water samples from the body of water. 35. A system as claimed in claim 31 wherein said at least one measurement means is configured to measure one or more of the following physical variables: water depth, water temperature, conductivity, water turbidity. 36. A system as claimed in claim 31 wherein said at least one sensor is configured to measure one or more of the following chemical variables: pH, dissolved oxygen, dissolved chloride, oxidation-reduction potential (ORP), soluble nitrate, ammonia, dissolved gases or chlorophyll A. 37. A dataset including a plurality of measurements obtained according to claim 1. 38. A map representing a dataset as claimed in claim 37. 39. A map generated according to the method of claim 10. |
<SOH> BACKGROUND OF THE INVENTION <EOH>It is now clear that human activity can have a great impact on the environment. As a result of this realisation, an environmental impact statement or environmental assessments are often made before a development is begun. The appreciation of the potential for human impact on the environment may also lead to an increase in research and environmental modelling in order to determine the, mechanisms for, and effects of, human activity on the environment. In order to increase the speed, quality, accuracy and cost effectiveness of environmental assessment, environmental monitoring and base line surveying improved techniques and systems for measuring physical variable in the environment are desired. |
<SOH> SUMMARY OF THE INVENTION <EOH>According to a first aspect of the present invention there is provided a method for measuring at least one physical and/or chemical variable at a plurality of different locations within a body of water, said method including the steps of: providing dynamic measurement means configured to measure the at least one physical and/or chemical variable; towing said measurement means in said body of water at a predetermined depth, and simultaneously taking measurements of least one physical and/or chemical variable, at least periodically, and generating measurement data; and determining the location of the measurement means while taking said measurements of the at least one physical and/or chemical variable and generating corresponding location data. Preferably the method includes the additional the step of: controlling the depth of the measurement means in said body of water whilst towing said measurement means. Preferably the method includes the additional the step of: detecting obstacles in a region adjacent to said measurement means; and wherein the step of controlling the depth of said measurement means is performed in response to detected obstacles. Preferably the step of, detecting obstacles in a region adjacent to said measurement means, includes the sub-steps of: providing image capture means on said measurement means, adapted to generate a sequence of images of a region adjacent to said measurement means; and visually detecting said obstacles from said sequence of images. Preferably said measurement means includes water intake means configured to collect water from said body of water and wherein said method includes the additional step of: collecting at least one water sample from said body of water through said water intake means. Preferably the predetermined depth is determined relative to a bottom of the body of water. Preferably water depth and at least one other physical and/or chemical variable is measured by said measurement means. Preferably the at least one physical and/or chemical variable measured by the measurement means is selected from a list including the following physical and chemical variables: water depth, water temperature, conductivity, and water turbidity, pH, dissolved oxygen, dissolved chloride, oxidation-reduction potential (ORP), soluble nitrate, ammonia, dissolved gases or chlorophyll A. According to a second aspect of the present invention there is provided a method of mapping at least one physical and/or chemical variable in body of water, said method including the steps of: providing dynamic measurement means configured to measure the at least one physical and/or chemical variable; and towing said measurement means in said body of water at a predetermined depth, and simultaneously taking measurements of said at least one physical and/or chemical variable, at least periodically, and generating measurement data; determining the location of the measurement means while taking said measurements of the at least one physical and/or chemical variable and generating corresponding location data; and generating a map representative of the distribution of the at least one physical and/or chemical variable within said body of water on the basis of the location and measurement data. Preferably the method additionally includes the additional the step of: determining the depth of the measurement means when measuring said at least one physical and/or chemical variable, and generating measurement depth data, and wherein said map is generated on the basis of the measurement depth data, measurement data and location data. Preferably the method includes the additional the step of: controlling the depth of the measurement means in said body of water whilst towing said measurement means. Preferably the method includes the additional the step of: detecting obstacles in a region adjacent to said measurement means; and wherein the step of controlling the depth of said measurement means is performed in response to detected obstacles. Preferably the step of, detecting obstacles in a region adjacent to said measurement means, includes the sub-steps of: providing image capture means on said measurement means, adapted to generate a sequence of images of a region adjacent to said measurement means; and visually detecting said obstacles from said sequence of images. Preferably said measurement means includes water intake means configured to collect water from said body of water and wherein said method includes the additional step of: collecting at least one water sample from said body of water through said water intake means. Preferably the predetermined depth is determined relative to a bottom of the body of water. Preferably water depth and at least one other physical and/or chemical variable is measured by said measurement means. Preferably the at least one physical and/or chemical variable measured by the measurement means is selected from a list including the following physical and chemical variables: water depth, water temperature, conductivity, and water turbidity, pH, dissolved oxygen, dissolved chloride, oxidation-reduction potential (ORP), soluble nitrate, ammonia, dissolved gases or chlorophyll A. Preferably the map represents topographic contours of a bottom of the body of water and the distribution of the at least one physical and/or chemical variable within said body of water. According to a third aspect of the present invention there is provided measurement means configured to measure at least one physical and/or chemical variable in a body of water, said measurement means including, a housing, and at least one sensor mounted at least partially within said housing, said sensor being configured to measure at least one physical and/or chemical variable, wherein said measurement means is configured to be towed in said body of water at a predetermined depth whilst simultaneously measuring, at least periodically, said least one physical and/or chemical variable. The measurement means can further include orientation means configured to orientate said measurement means relative to an apparent current experienced by said measurement means when said measurement means is being towed. Preferably said orientation means includes at least one fin. The measurement means can further include water intake means to allow the collection of a sample of water from the body of water. The measurement means can further include image capture means adapted to generate a sequence of images of a region of said body of water adjacent to said measurement means, wherein in use the depth of the measurement means is controlled in response to the sequence of video images. Preferably the at least one sensor is configured to measure one or more of the following physical and chemical variables: water depth, water temperature, conductivity, water turbidity, pH, dissolved oxygen, dissolved chloride, oxidation-reduction potential (ORP), soluble nitrate, ammonia, dissolved gases or chlorophyll A. Preferably there is more than one sensor. Each sensor can measure one or more physical and/or chemical variables. Preferably said housing includes a frame. Preferably said housing includes a water permeable container configured to contain said at least one sensor. The measurement means can further include data storage means in communication with said at least one sensor, said data storage means being configured to store measurement data generated by said at least one sensor. According to a fourth aspect of the present invention there is provided a system for taking a series of measurements of at least one physical and/or chemical variable in a body of water, said system including: measurement means configured to measure at least one physical and/or chemical variable in a body of water, wherein in use said measurement means is configured to be towed in said body of water at a predetermined depth whilst simultaneously measuring, at least periodically, said at least one physical and/or chemical variable to generate measurement data; and location means configured to determine the location of the measurement means while taking said measurements of the at least one physical and/or chemical variable to generate location data; and data storage means configured to store said measurement data and location data. Preferably the system includes depth control means configured to control the depth of the measurement means while said measurement means is being towed. Preferably the system includes image capture means adapted to provide a sequence of images of a region of the body of water adjacent the measurement means. Preferably the system includes water inlet means, mounted on said measurement means, configured to allow collection one or more water samples from the body of water. Preferably the at least one measurement means is configured to measure one or more of the following physical or chemical variables: water depth, water temperature, conductivity, water turbidity, pH, dissolved oxygen, dissolved chloride, oxidation-reduction potential (ORP), soluble nitrate, ammonia, dissolved gases or chlorophyll A. The present invention also provides a dataset including a plurality of measurements obtained according to the method described above, and a map representing such a dataset. The present invention additionally provides a map generated according to the mapping method of described above. |
Apparatus for preparing a beverage suitable for consumption, such as coffee |
An apparatus for preparing a beverage suitable for consumption such as coffee on the basis of a hot liquid such as water provided with a storage tank, a heater and a three-way valve. The three-way valve is provided with in inlet, a first outlet and a second outlet. The pump is in fluid communication with the storage tank and an inlet of the heater. There is a fluid communication between the outlet of the heater and the inlet of the three-way valve, between the first outlet and the storage tank and/or an inlet of the pump and between the second outlet and a beverage preparing unit for preparing the beverage suitable for consumption on the basis of the hot liquid. The three-way valve is provided with a first member which is freely movable between a first and second extreme position wherein the first member can move under the influence of the liquid flow from the inlet to the first outlet and/or the second outlet from the first extreme position towards the second extreme position and in the second extreme position closes off the first outlet so that the liquid flow then extends from the inlet to the second outlet. The three-way valve is further provided with a second member which is freely moveable under the influence of a liquid flow from the inlet to the second outlet between a third and fourth extreme position wherein the second member, under the influence of the liquid flow from the inlet to the second outlet, moves from the third extreme position to the fourth extreme position and in the third extreme position blocks a liquid flow from the second outlet in the direction of the inlet. |
1. An apparatus for preparing at least one type of beverage suitable for consumption such as coffee or cocoa on the basis of a hot liquid such as water, provided with a storage tank which, in use, is filled with a liquid such as water, a heater and a three-way valve, wherein the three-way valve is provided with an inlet, a first outlet and a second outlet, a pump being in fluid communication with the storage tank and an inlet of the heater for pumping the liquid from the storage tank to the heater and wherein an outlet of the heater is in fluid communication with the inlet of the three-way valve, the first outlet of the three-way valve being in fluid communication via a first outflow path with the storage tank and/or an inlet of the pump for returning liquid from the three-way valve to the storage tank and/or the pump and wherein the second outlet of the three-way valve is in fluid communication via a second outflow path with a beverage preparing unit for preparing, on the basis of the hot liquid, the beverage suitable for consumption, the three-way valve being provided with a first member which is freely moveable, under the influence of a liquid flow from the inlet to the first outlet and/or the second outlet, between a first and second extreme position, wherein the first member can move, under the influence of the liquid flow from the inlet to the first outlet and/or the second outlet from the first extreme position towards the second extreme position and in the second extreme position closes off the first outlet, so that the liquid flow then extends from the inlet to the second outlet, characterized in that the three-way valve is further provided with a second member which is freely moveable, under the influence of a liquid flow from the inlet to the second outlet, between a third and a fourth extreme position, wherein the second member can move, under the influence of the liquid flow from the inlet to the second outlet, from the third extreme position towards the fourth extreme position, and in the third extreme position blocks a liquid flow from the second outlet in the direction of the inlet. 2. An apparatus according to claim 1, characterized in that the three-way valve is disposed in one single housing. 3. An apparatus according to claim 1 or 2, characterized in that the three-way valve is provided with a first compartment in which the first member is situated, a second compartment in which the second member is situated and a fluid communication between the first and second compartment. 4. An apparatus according to claim 3, characterized in that the first compartment is provided with the inlet and the first outlet. 5. An apparatus according to claim 3 or 4, characterized in that the second compartment is provided with the second outlet. 6. An apparatus according to claims 4 and 5, characterized in that an inflow opening of the fluid communication in the first compartment is located between the inlet and the first outlet. 7. An apparatus according to claim 6, characterized in that the first member and the first compartment are configured such that when the first member is in the second extreme position with the first outlet closed off, a liquid flow from the inlet to the second outlet extends through a part of the first compartment situated between the first member and the inlet. 8. An apparatus according to any one of claims 3-7, characterized in that the first and second compartment are separated from each other by means of a dividing wall while the fluid communication extends through the dividing wall. 9. An apparatus according to claims 7 and 8, characterized in that said part of the first compartment comprises the inflow opening of the fluid communication. 10. An apparatus according to any one of the preceding claims, characterized in that the three-way valve allows a liquid flow from the inlet to the first outlet when the first member is not in the second extreme position. 11. An apparatus according to any one of the preceding claims, characterized in that the three-way valve allows a liquid flow from the inlet to the second outlet when the second member is not in the third extreme position. 12. An apparatus according to any one of the preceding claims, characterized in that the first member is of cylindrical design. 13. An apparatus according to any one of the preceding claims, characterized in that the second member is of convex design. 14. An apparatus according to any one of the preceding claims, characterized in that the first member is heavier than the second member. 15. An apparatus according to any one of the preceding claims, characterized in that a flow resistance of the first outflow is smaller than a flow resistance of the second outflow in combination with the beverage preparing unit. 16. An apparatus according to any one of the preceding claims 1-11, characterized in that the first member is designed as a freely moveable circumferential edge of a first flexible umbrella valve and the three-way valve is provided with a first seat for the first umbrella valve wherein the first umbrella valve is connected to the first seat and the first seat is provided with at least one groove wherein in the first extreme position the circumferential edge of the first umbrella valve abuts against the first seat and extends over the groove so that a space is present between the circumferential edge of the first umbrella valve and the groove, which space releases a liquid flow from the inlet to the first outlet and wherein in the second extreme position the circumferential edge of the first umbrella valve abuts against the seat and extends into the groove so that the circumferential edge closes off the first outlet. 17. An apparatus according to claim 16, characterized in that in the second extreme position the circumferential edge of the first umbrella valve abuts against the seat and the bottom of the groove. 18. An apparatus according to claim 16 or 17, characterized in that the first umbrella valve is situated in a liquid flow path extending between the inlet and the first outlet. 19. An apparatus according to claim 18, characterized in that the first umbrella valve has a convex outer surface facing away from the first outlet. 20. An apparatus according to any one of the preceding claims 1-12 or 16-19, characterized in that the second member is designed as a freely moveable circumferential edge of a second flexible umbrella valve and the three-way valve is provided with a second seat for the second umbrella valve wherein the second umbrella valve is connected to the second seat, while in the third extreme position the circumferential edge of the second umbrella valve abuts against the second seat and blocks a liquid flow from the second outlet in the direction of the inlet and wherein in the fourth extreme position an intermediate space is present between the circumferential edge of the second umbrella valve and the second seat. 21. An apparatus according to claim 20, characterized in that the second umbrella valve is situated in a liquid flow path extending between the inlet and the second outlet. 22. An apparatus according to claim 21, characterized in that the second umbrella valve has a convex outer surface facing towards the second outlet. 23. An apparatus according to any one of the preceding claims, characterized in that from the inlet to the first member and from the inlet to the second member non-closable fluid flow paths are present. 24. A three-way valve provided with an inlet, a first outlet and a second outlet wherein the three-way valve is provided with a first member which is freely moveable, under the influence of a liquid flow from the inlet to the first outlet and/or the second outlet, between a first and second extreme position wherein the first member can move, under the influence of the liquid flow from the inlet to the first outlet and/or the second outlet, from the first extreme position towards the second extreme position and in the second extreme position closes off the first outlet so that the liquid flow then extends from the inlet to the second outlet, characterized in that the three-way valve is further provided with a second member which is freely moveable, under the influence of a liquid flow from the inlet to the second outlet, between a third and fourth extreme position wherein the second member can move, under the influence of the liquid flow from the inlet to the second outlet, from the third extreme position in the direction of the fourth extreme position and in the third extreme position blocks a liquid flow from the second outlet in the direction of the inlet. 25. A three-way valve according to claim 24, characterized in that the three-way valve is disposed in one single housing. 26. A three-way valve according to claim 24 or 25, characterized in that the three-way valve is provided with a first compartment in which the first member is situated, a second compartment in which the second member is situated, and a fluid communication between the first and second compartment. 27. A three-way valve acclaim 26, characterized in that the first compartment is provided with the inlet and the first outlet. 28. A three-way valve according to claim 26 or 27, characterized in that the second compartment is provided with the second outlet. 29. A three-way valve according to claims 27 and 28, characterized in that an inflow opening of the fluid communication in the first compartment is situated between the inlet and the first outlet. 30. A three-way valve according to claim 29, characterized in that the first member and the first compartment are configured such that when the first member is in the second extreme position with the first outlet closed off, a liquid flow from the inlet to the second outlet extends through a part of the first compartment situated between the first member and the inlet. 31. A three-way valve according to any one of claims 26-30, characterized in that the first and second compartment are separated from each other by means of a dividing wall while the fluid communication extends through the dividing wall. 32. A three-way valve according to claims 30 and 31, characterized in that said part of the first compartment comprises the inflow opening of the fluid communication. 33. A three-way valve according to any one of claims 24-32, characterized in that the three-way valve allows a liquid flow from the inlet to the first outlet when the first member is not in the second extreme position. 34. A three-way valve according to any of claims 24-33, characterized in that the three-way valve allows a liquid flow from the inlet to the second outlet when the second member is not in the third extreme position. 35. A three-way valve according to any one of the preceding claims 24-34, characterized in that the first member is of cylindrical design. 36. A three-way valve according to any one of the preceding claims 24-35, characterized in that the second member is of convex design. 37. A three-way valve according to any one of the preceding claims 24-36, characterized in that the first member is heavier than the second member. 38. A three-way valve according to any one of the preceding claims 24-34, characterized in that the first member is designed as a freely moveable circumferential edge of a first flexible umbrella valve and the first outlet is provided with a first seat for the first umbrella valve, the first umbrella valve being connected with the first outlet and the first seat being provided with a groove while in the first extreme position the circumferential edge of the first umbrella valve abuts against the first seat and extends over the groove so that a space is present between the circumferential edge of the first umbrella valve and the groove, which space releases a liquid flow from the inlet to the first outlet and wherein in the second extreme position the circumferential edge of the first umbrella valve abuts against the seat and extends in the groove so that the circumferential edge closes off the first outlet. 39. A three-way valve according to claim 38, characterized in that in the second extreme position the circumferential edge of the first umbrella valve abuts against the seat and the bottom of the groove. 40. A three-way valve according to claim 38 or 39, characterized in that the first umbrella valve is in a liquid flow path extending between the inlet and the first outlet. 41. A three-way valve according to claim 40, characterized in that the first umbrella valve has a convex outer surface facing away from the first outlet. 42. A three-way valve according to any one of the preceding claims 24-34 or 38-41, characterized in that the second member is designed as a freely moveable circumferential edge of a second flexible umbrella valve and the second outlet is provided with a second seat for the second umbrella valve while the second umbrella valve is connected to the second seat wherein in the third extreme position the circumferential edge of the second umbrella valve abuts against the second seat and blocks a liquid flow from the second outlet in the direction of the inlet and wherein in the fourth extreme position an intermediate space is present between the circumferential edge of the second umbrella valve and the second seat. 43. A three-way valve according to claim 42, characterized in that the second umbrella valve is located in a liquid flow path extending between the inlet and the second outlet. 44. A three-way valve according to claim 43, characterized in that the second umbrella valve has a convex outer surface facing towards the second outlet. 45. A three-way valve according to any one of the preceding claims 24-44, characterized in that from the inlet to the first member and from the inlet to the second member non-closable fluid flow paths are present. |
Map of a property |
The present invention provides a method of mapping a property of a three dimensional object. The method comprises the steps of mapping the property for at least a portion of the object, providing a line or region which defines intersections with part of the mapped portion, and displaying the property for the intersections. The present invention also provides another method of mapping a property of an object. The method comprises the steps of mapping the property for a slice within the object, providing a line within the slice, and displaying the property for the line. |
1. A method of mapping a property of a three dimensional object, said method comprising the steps of mapping the property for at least a portion of the object, providing a line or region which defines intersections with part of said mapped portion, and displaying the property for the intersections. 2. A method of mapping a property of a three dimensional object, said method comprising the steps of mapping the property for a plurality of slices within the object, providing a line or region which defines intersections with part of at least some of the plurality of slices, and displaying the property for the intersections. 3. A method as defined in claim 1 wherein the property relates to nuclear magnetic resonances. 4. A method as defined in claim 2 wherein mapping of the property for the plurality of slices results in a plurality of nuclear magnetic resonance images. 5. A method as defined in claim 1 wherein the line or region is a line. 6. A method as defined in claim 5 wherein the line is a plurality of lines. 7. A method as defined in claim 6 wherein each of the lines is substantially perpendicular to the surface of the three-dimensional object. 8. A method as defined in claim 1 which further comprises the step of approximating the shape of the object by a three dimensional lattice of two-dimensional forms created by a first computer routine. 9. A method as defined in claim 8 wherein the lattice is a mesh. 10. A method as defined in claim 8 wherein each of the lines is a line substantially perpendicular to one of the two-dimensional forms. 11. A method as defined in claim 2 wherein the slices are selected using a second computer routine. 12. A method as defined in claim 3 wherein a third computer routine maps nuclear magnetic resonance values for the intersections by displaying an array of values. 13. A method as defined in claim 12 which further comprises the step of analysing the array of values using a fourth computer routine. 14. A method as defined in claim 13 wherein the fourth computer routine analyses numerical properties. 15. A method as defined in claim 12 wherein the step of analysing the array of values comprises differentiation. 16. A method as defined in claim 15 wherein analysing the array of values comprises determining the positions of the maxima, minima and/or zero-points of the array of values and the derivatives of the array of values. 17. A map produced by a method of mapping a property of a three dimensional object, said method comprising the steps of mapping the property for at least a portion of the object, providing a line or region which defines intersections with part of said mapped portion, and displaying the property for the intersections. 18. A map produced by a method of mapping a property of a three dimensional object, said method comprising the steps of mapping the property for a plurality of slices within the object, providing a line or region which defines intersections with part of at least some of the plurality of slices, and displaying the property for the intersections. 19. A method of mapping a property of an object, said method comprising the steps of mapping the property for a slice within the object, providing a line within the slice, and displaying the property for the line. 20. A method as defined in claim 19 wherein the property wherein relates to nuclear magnetic resonances. 21. A method as defined in claim 19 wherein mapping of the property for the slice results in a nuclear magnetic resonance image. 22. A method as defined in claim 19, wherein the line is a plurality of lines. 23. A method as defined in claim 19 wherein a fifth computer routine maps nuclear magnetic resonance values for the intersections by displaying an array of values. 24. A method as defined in claim 23 which further comprises the step of analysing the array of values using a sixth computer routine. 25. A method as defined in claim 24 wherein the fifth computer routine analyses numerical properties. 26. A method as defined in claim 23 wherein the step of analysing the array of values comprises differentiation. 27. A method as defined in claim 26 wherein the step of analysing the array of values comprises determining the positions of the maxima, minima and/or zero-points of the array of values and the derivatives of the array of values. 28. A map produced by a method of mapping a property of an object, said method comprising the steps of mapping the property for a slice within the object, providing a line within the slice, and displaying the property for the line. |
<SOH> BACKGROUND OF THE INVENTION <EOH>It is currently of significant interest in different areas of research and analysis, in particular in medicine, to obtain non-destructively images or line-scans related to regions within objects. Typical objects may include biological specimen such as live human brains. The analysis of such images or line-scans may reveal information that is essential for understanding the functional organisation of the human brain. In order to gain a more detailed understanding of the activation of the cerebral cortex of the brain, the anatomical organisation of the brain has to be known. With regard to the cerebral cortex, which is of particular interest in studies of cognition, a key question is how one relates the results of a functional neuro-imaging study with particular cortical areas in which changes in neural activity occur. An enormous and costly effort is expended in carrying out functional neuro-imaging of ever increasing resolution, but in the end the researcher is left with only a crude anatomical estimate of the activated cortical areas. If this situation is to be resolved, better human brain atlases are required. However, these suffer from one major problem; namely inter-subject variations in brain micro- and macrostructure. Histological examination of the cyto-architecture, myelo-architecture and chemo-architecture of the brain remains the best method of accurately defining anatomically distinct regions. Ideally, if this can be performed in every subject used in functional neuroimaging studies, each individual's functional results can be precisely and accurately related with their own anatomical brain map. Furthermore, if a functionally active cortical focus were consistently found to be correlated with a well defined anatomical region across a population of subjects, this would significantly advance understanding of human brain organisation. Obviously, this is not practical except in a few rare cases. Instead, the results from postmortem brains of “non-subjects” in functional studies are used which again results in problems of inter-subject variability. |
<SOH> SUMMARY OF THE INVENTION <EOH>According to a first aspect of the invention there is provided a method of mapping a property of a three dimensional object, said method comprising the steps of mapping the property for at least a portion of the object, providing a line or region which defines intersections with part of said mapped portion, and displaying the property for the intersections. According to a second aspect of the invention there is provided a method of mapping a property of a three dimensional object, said method comprising the steps of: mapping the property for a plurality of slices within the object, providing a line or region which defines intersections with part of at least some of the plurality of slices, and displaying the property for the intersections. Preferably the property relates to nuclear magnetic resonances. More preferably, mapping of the property for the slices results in a plurality of nuclear magnetic resonance images. It should be understood that nuclear magnetic resonances and nuclear magnetic resonance images are terms usually used by physicists and chemists. It is common practice to abbreviate these terms in a clinical environment to magnetic resonances and magnetic resonance images, respectively. Preferably the line or region is a line. More preferably, the line is a plurality of lines. Preferably, each of the lines is substantially perpendicular to the surface of the three-dimensional object. In one embodiment the object is a biological object such as the brain of a subject (or another organ of a human or animal, or a semi-biological or inanimate object) and each of the lines is a line through the cortex of the brain. The cortex of the human brain is convoluted and contains a plurality of grooves. Therefore, a line substantially perpendicular to the surface, for example within a groove, would typically intersect not only one slice, but can intersect a plurality of slices which are related to nuclear magnetic resonance images taken from adjacent areas of the brain. Preferably, the method further comprises the steps of approximating the shape of the object by a three dimensional lattice of two-dimensional forms created by a first computer routine. More preferably, the lattice is a mesh. Preferably, the lines are each substantially perpendicular to a respective one of the two-dimensional forms. Preferably the intersections of each of the lines with each of the slices are selected using a second computer routine. More preferably a third computer routine maps nuclear magnetic resonance values for the intersections by displaying an array of values. Preferably the method further comprises the step of analysing the array of values using a fourth computer routine. More preferably the fourth computer routine analyses quantitative properties. Preferably analysing the array of values using the fourth computer routine comprises differentiation. More preferably analysing the array of values comprises determining the positions of the maxima, minima and/or zero-points of the array of values and the derivatives of the array of values. In one embodiment, the positions of the inner and outer boundaries of the grey matter of the brain may be approximated by determining the positions of the maxima of the first derivative of the array of values. Two closely followed zero-points of the first derivative may indicate the presence of lamination. By measuring the relative positions of the zero points the thickness of the lamination can be determined. According to a third aspect of the invention there is provided a map produced by a method of mapping a property of a three dimensional object, said method comprising the steps of: mapping the property for at least a portion of the object, providing a line or region which defines intersections with part of said mapped portion, and displaying the property for the intersections. According to a fourth aspect of the invention there is provided a map produced by a method of mapping a property of a three dimensional object, said method comprising the steps of: mapping the property for a plurality of slices within the object, providing a line or region which defines intersections with part of at least some of the plurality of slices, and displaying the property for the intersections. According to a fifth aspect of the invention there is provided a method of mapping a property of an object, said method comprising the steps of: mapping the property for a slice within the object, providing a line within the slice, and displaying the property for the line. Preferably the property relates to nuclear magnetic resonances. More preferably, mapping of the property for the slice results in a nuclear magnetic resonance image. Preferably, the line is a plurality of lines. More preferably in the fourth aspect a computer routine maps nuclear magnetic resonance values for each of the lines by displaying an array of values. In one embodiment the object is a biological object such as a brain and each of the lines is a line through the cortex of the brain. Preferably the method further comprises the step of analysing the array of values using a fifth computer routine. More preferably the fifth computer routine analyses numerical properties. Preferably analysing the array of values using the fifth computer routine comprises differentiation. More preferably analysing the array of values comprises determining the positions of the maxima, minima and/or zero-points of the array of values and the derivatives of the array of values. In one embodiment, the positions of the inner and outer boundaries of the grey matter of the brain may be approximated by determining the positions of the maxima of the first derivative of the array of values. Two closely followed zero-points of the first derivative may indicate the presence of lamination. By measuring the relative positions of the zero points the thickness of the lamination can be determined. According to a sixth aspect of the invention there is provided a map produced by a method of mapping a property of an object, said method comprising the steps of: mapping the property for a slice within the object, providing a line within the slice, and displaying the property for the line. |
Device and method for determining a physical address from a virtual address, using a hierarchical mapping rule comprising compressed nodes |
A method for determining a physical address from a virtual address, wherein a mapping regulation between the virtual address and the physical address is implemented as hierarchical tree structure with compressed nodes. First, a compression indicator included in the mapping regulation is read, and a portion of the virtual address associated with the considered node level is read. Using the compression indicator and the portion of the virtual address, an entry in the node list of the considered node is determined. The determined entry is read, whereupon the physical address can be determined directly, if the considered node level has been the hierarchically lowest node level. If higher node levels to be processed are present, the previous steps in determining the physical address for compressed nodes of lower hierarchy level are repeated until the hierarchically lowest node level is reached. |
1. A method for determining a physical address associated with a corresponding storage location in a physical memory space from a virtual address by using a hierarchical mapping regulation with a plurality of hierarchically arranged node levels, wherein a virtual address space associated with the virtual address is larger than the physical address space, the virtual address having a plurality of portions, each portion of the virtual address being associated to a corresponding node level of the virtual address space, each node level having at least one node, each node having a node list with corresponding list entries, each list entry of an associated node list referring to one of a corresponding node of a hierarchically lower node level and an associated physical address, wherein a length of each portion of the virtual address defines a maximum number of list entries included in each node belonging to the node level associated to the portion, wherein at least one node of the plurality of node levels is a compressed node including an associated node list having fewer than the maximum number of list entries, wherein the hierarchical mapping regulation includes a compression indicator associated to said at least one compressed node, and wherein the method comprises: reading the compression indicator; reading a portion of the virtual address; determining an entry in a node list of a node of a node level associated to the read portion of the virtual address, by using the read portion of the virtual address and the compression indicator; reading the determined entry of the node list to identify a node of a hierarchically lower node level referenced by the determined entry; and determining the physical address by using the node of the hierarchically lower node level referenced by the determined entry. 2. The method according to claim 1, wherein the compression indicator is stored at a predetermined position in the node list of the compressed node, and wherein reading the compression indicator comprises jumping to the predetermined portion to read the compression indicator. 3. The method according to claim 1, wherein the compression indicator for a compressed node is stored in the list entry of a node list of a node of a hierarchically higher node level, and wherein reading the compression indicator comprises obtaining the compression indicator by reading a list entry of a node list of the node of the hierarchically higher node level. 4. The method according to claim 1, wherein a node list of a compressed node is compressed such that the number of list entries in comparison to the maximum number is halved, if the most significant bit of each list index for all list entries is non-zero, and wherein in the step of determining the compression indicator is interpreted such that the most significant bit of the portion of the virtual address is ignored in determining the entry. 5. The method according to claim 1, wherein a node list of a compressed node is compressed such that the number of list entries is quartered in comparison to the maximum number, if the two most significant bits of each list index for all list entries are non-zero, and wherein in the step of determining the compression indicator is interpreted such that the two most significant bits of the virtual address are ignored in determining the entry. 6. The method according to claim 1, wherein an addressing by pages of the physical memory is provided, wherein a memory page is addressable via a base address and an offset value, wherein the node lists of at least two compressed nodes are stored in the same physical memory page, and wherein a list entry of a node list of a node of a hierarchically higher node level comprises both a base address and an offset value for the memory page, where the node list for the node is stored, which is referenced by the entry, and wherein in the step of determining the entry the determination is carried out by using the base address and the offset value. 7. The method according to claim 1, wherein, in determining the physical address, the steps reading the compression indicator, reading a portion of the virtual address, determining an entry of the node list and reading the determined entry, are carried out until the hierarchically lowest node level is reached. 8. The method according to claim 1, wherein a predetermined value of a portion of the virtual address shows that a node level is skipped. 9. An apparatus for determining a physical address associated with a corresponding storage location in a physical memory space from a virtual address by using a hierarchical mapping regulation with a plurality of hierarchically arranged node levels, wherein a virtual address space associated with the virtual address is larger than a physical address space, the virtual address having a plurality of portions, each portion of the virtual address being associated to a corresponding node level of the virtual address space, each node level having at least one node, each node having a node list with corresponding list entries, each list entry of an associated node list referring to one of a corresponding node of a hierarchically lower node level and an associated physical address, wherein a length of each portion of the virtual address defines a maximum number of list entries included in each node belonging to the node level associated to said each portion, wherein at least one node of the plurality of node levels is a compressed node including an associated node list having fewer than the maximum number of list entries, wherein the hierarchical mapping regulation includes a compression indicator associated to said at least one compressed node, and wherein the apparatus comprises: means for reading the compression indicator; means for reading a portion of the virtual address; means for determining an entry in a node list of a node of the node level associated to the read portion of the virtual address, by using the virtual address, by using the read portion of the virtual address and the compression indicator; means for reading the determined entry of the node list to identify a node of a hierarchically lower node level which is referenced by the determined entry; and means for determining the physical address by using the node of the hierarchically lower node level which is referenced by the determined entry. 10. A computer system comprising: a physical memory including a plurality of storage locations, each storage location having an associated physical address; and an apparatus for determining a selected physical address from a virtual address by using a hierarchical mapping regulation having a plurality of hierarchically arranged node levels, wherein the virtual address includes a plurality of portions, each portion of the virtual address being associated to a corresponding node level of the virtual address space, each node level having at least one node, each node having a node list with corresponding list entries, each list entry of an associated node list referring to one of a corresponding node of a hierarchically lower node level and an associated physical address, wherein a length of each portion of the virtual address defines a maximum number of list entries included in each node belonging to the node level associated to said each portion, wherein at least one node of the plurality of node levels is a compressed node including an associated node list having fewer than the maximum number of list entries, wherein the hierarchical mapping regulation includes a compression indicator associated to said at least one compressed node, and wherein the apparatus comprises: means for reading the compression indicator; means for reading a portion of the virtual address; means for determining an entry in a node list of a node of the node level associated to the read portion of the virtual address, by using the virtual address, by using the read portion of the virtual address and the compression indicator; means for reading the determined entry of the node list to identify a node of a hierarchically lower node level which is referenced by the determined entry; and means for determining the physical address by using the node of the hierarchically lower node level which is referenced by the determined entry. 11. The computer system according to claim 10, laid out as a chip card, smart card or security IC. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention The present invention relates to computer systems and especially to computer systems with virtual addressing. 2. Description of the Related Art Virtual addressing has been known for a while from the field of workstations. By using a certain mapping regulation virtual addresses are mapped to physical addresses. The physical addresses address memory cells of a physical memory, such as the working memory, a hard drive, a tape memory, etc. Virtual addresses however, do not reference directly to physical memory cells but merely indirectly via the mapping regulation. It is the advantage of this type of addressing that the programmer of an application does not have to take care of the different physical memories present in a computer system. The programmer has a virtual address space at his hands that he can use for his program as required. The mapping to the physical address space that a special computer system makes available is generated separately from the program code so that by making available different mapping regulations a program programmed with virtual addresses can run on different computer systems. In a processor with a virtual memory system an application runs in a so-called virtual address space. Each address of the virtual memory where data read/writable by the application or executable code exist is mapped to an address on the physical memory where those data or this code is actually stored. The virtual address (VA) and the physical address (PA) associated via the mapping regulation do not need to have any relation at all. Further, the virtual address space can be significantly larger than the physical address space. Virtual addresses without read/writable data or executable code are normally not mapped to a physical memory. This mapping is totally transparent for the processed application. When organizing the memory in pages, the virtual address space is divided into equal overlapping-free memory areas. One page in the physical address space is associated to a page in the virtual address space via the mapping regulation; the page in the physical address space is also referred to as page frame. The payload data memory of a page frame of the physical address space has the same size as a page of the virtual address space. The allocation of a virtual page to a physical page is usually achieved by the so-called page table, comprising address pairs of respective start addresses of the virtual pages and the associated physical pages. In workstations, part of the page table is in a cache that is also referred to as “Translation Look Aside Buffer (TLB)”. If the start address pair for a virtual page and the associated physical page is in the TLB, then the calculation of the address mapping into the virtual memory area happens accelerated since only one access to a table is necessary in order to obtain the physical address associated to a virtual address. If the start address pair, i.e. the virtual address and its associated physical address is in the TLB, then a TLB miss takes place, which usually leads to a trap to the operating system which has to add the address tupel to the TLB. In the area of workstations, the mapping regulation between virtual address space and physical address space that can, for example, be implemented as a single page table, is held in the volatile working memory. When a workstation is booted up, it first starts in the real addressing mode. This means that the operating system of the workstation causes the CPU of the workstation to gradually set up a page table in the volatile working memory in the workstation in the real, i.e. physical addressing mode. Only when a page table is constructed, the workstation can switch to the virtual addressing mode. If the CPU asks for data at a virtual address, then the associated physical address is determined in the volatile working memory of the CPU in order to be able to fetch data from the memory. Common workstations are therefore distinguished by the fact that they boot up in a real addressing mode and then switch to the virtual addressing mode when the mapping regulation from the virtual address space to the physical address space in the volatile memory is set up. It is one disadvantage of this concept that a relatively large working memory area is necessary in order to store a page table. This disadvantage is not of high importance for workstations, since they have large amounts of working memory available. For other applications, such as for security relevant computer systems, such as implemented in chip card ICs, the memory resources are limited due to the small available space. The provision of an amount of volatile working memory to store a page table leads to the fact that the application carried out on the chip card might have too little working memory and therefore experience performance losses. It is another disadvantage of the known concept that a significant management effort is necessary to, at first, set up the page table when booting up the computer system, i.e. to gradually calculate the address allocations from the stored information and to store them. Besides the fact that computer resources are necessary for that, respective programs also have to be made available on a chip card in order to take the necessary precautions for the virtual addressing mode. Such programs also need memory space that is a limited resource, especially in chip cards or other security ICs, due to reasons of space. In the specialist book “Computer Architecture: a quantitative approach” by David A. Patterson, second edition, 1996, Morgan Kaufmann Publishers, Inc., pages 449-453 the memory management of the alpha AXP processor is described. A hierarchical tree structure with three levels is used here as mapping regulation for determining a physical address from a virtual address. One node level comprises at least one node with a page table. The virtual address is divided into portions, wherein each portion of the virtual address is associated to a node level. The address translation begins with adding the highest-level address field to a page table base register, whereupon the memory is read out at this position to obtain the basis of the page table of the second level. The address field of the virtual address for the second level is then again added to this newly fetched address. Then the memory is accessed again to obtain the basis of the page table of the third level. The address field of the third level is added to this base address, whereupon the memory is read out by using this sum to obtain the physical address of the page that is referenced. A page offset is added to this address to obtain the physical address associated to the virtual address. Each page table in the alpha AXP architecture fits into a single page, so that all page table addresses are physical addresses that do not need any further translation. Each page table for each node level is therefore exactly one physical page long. If the virtual address space is larger than the physical address space, which is especially an advantage when flexibility for future expansions of the physical address space is wanted, the page tables, i.e. the node lists of the nodes, contain a large number of zero entries, i.e. of entries that do not reference to a physical address or to no node of a lower node level. Due to the fact that each page table is exactly one physical memory page long, the known hierarchical addressing wastes a lot of memory space. The amount of wasted memory space is getting larger the larger the virtual address space is in comparison to the physical address space. The memory taken up by zero entries in the node lists leads to the fact that more memory space, for example in form of an external RAM has to be made available, especially in computer systems with limited memory resources, as they can be found on chip cards, smart cards or security ICs, which makes a chip card more expensive. If, however, due to reasons of chip space no further memory can be made available especially the working memory resources of the chip cards are more limited than absolutely necessary, which leads to the fact that less expensive programs can be computed in the chip card or that the performance of the chip card suffers. |
<SOH> SUMMARY OF THE INVENTION <EOH>It is the object of the present invention to provide a more efficient concept for determining a physical address from a virtual address as well as a more efficient computer system. In accordance with a first aspect, the present invention provides a method for determining a physical address from a virtual address by using a hierarchical mapping regulation with hierarchically arranged node levels, wherein a virtual address space is larger than a physical address space, the virtual address having portions, one portion of the virtual address being associated to a node level, the node level having at least one node, each node having a node list with list entries, a list entry of a node list referring to a node of a hierarchically lower node level, wherein by using a list entry of a node list of a hierarchically lower node level the physical address can be determined, a length of the portion of the virtual address defining a maximum number of entries of a node list of a node belonging to the node level associated to the portion, a node, whose node list has a lower than the maximum number of entries, being a compressed node, and the mapping regulation having a compression indicator associated to a compressed node, having reading the compression indicator; reading a portion of the virtual address; determining an entry in a node list of a node of a node level associated to the read portion of the virtual address, by using the read portion of the virtual address and the compression indicator; reading the determined entry of the node list to identify a node of a hierarchically lower node level referenced by the determined entry; and determining the physical address by using the node of the hierarchically lower node level referenced by the determined entry. In accordance with a second aspect, the present invention provides an apparatus for determining a physical address from a virtual address by using a hierarchical mapping regulation with hierarchically arranged node levels, a virtual address space being larger than a physical address space, the virtual address having portions, one portion of the virtual address being associated to a node level, the node level having at least one node, each node having a node list with list entries, a list entry of a node list referring to a node of a hierarchically lower node level, wherein by using a list entry of a node list of a hierarchically lower node level the physical address can be determined, a length of the portion of the virtual address defining a maximum number of entries of a node list of a node belonging to the node level associated to the portion, a node, whose node list has a lower than the maximum number of entries, being a compressed node, and the mapping regulation having a compression indicator associated to a compressed node, having means for reading the compression indicator; means for reading a portion of the virtual address; means for determining an entry in a node list of a node of the node level associated to the read portion of the virtual address, by using the virtual address, by using the read portion of the virtual address and the compression indicator; means for reading the determined entry of the node list to identify a node of a hierarchically lower node level which is referenced by the determined entry; and means for determining the physical address by using the node of the hierarchically lower node level which is referenced by the determined entry. The present invention is based on the knowledge that a compression of node lists has to be carried out to reduce the memory space taken up by the hierarchical mapping regulation between virtual address and physical address. According to the invention, a compressing indicator that is used in determining an entry in a node list of a node together with a respective portion of the virtual address to read the determined entry and then to find out the physical address by using the determined entry is stored together with a mapping regulation in an addressing structure where the virtual address space is larger than the physical address space and where hierarchical disposed node levels are present. The hierarchical mapping regulation comprises hierarchically disposed node levels, one node level having at least one node. A node list of list entries is associated to each node. Further, the virtual address has portions, always one portion of the virtual address being associated to one node level. The node list of a node level comprises list entries, each list entry identified by an index, and each list entry comprising a reference to a node of a hierarchically lower node level. The physical address is determinable by using a list entry of a node of a hierarchically lowest node level, either as a direct physical address or, if an addressing by pages is used, by using the base address and the offset value of the physical memory page. The length of a portion of the virtual address defines a maximum number of entries of a node list of a node. If one portion of the virtual address has a length of, for example, four bits, a node of a node level associated to this portion of the virtual address, for example a maximum of 16 list entries, if node compression is present, since 16 different indices for 16 different node entries can be signaled. According to the invention, a node whose node list has a lesser than maximum number of entries is a compressed node. This fact and the way in which a node is compressed are given by a compressing indicator indicating a compressed node. In a preferred embodiment of the present invention compressions of nodes are preferred, which comprise a compression to one half of the list entries compared to the maximum number, to one quarter of list entries compared to the maximum number, to one eighth of list entries in comparison to the maximum number, etc. Preferably, compression is carried out when the most significant bits of all list entries of an uncompressed node list are equal. Then, compression by one half can be achieved and the most significant bit of the respective portion of the virtual address is ignored in indexing the node list, due to the node indicator of the mapping regulation. Further, if the two most significant bits of all non-zero entries in an uncompressed node list are equal, a compression to a quarter of the uncompressed node list is achieved, and the two most significant bits of the respective portion of the virtual address are ignored in the indexing of a node list due to the node indicator. If the three most significant bits of all non-zero entries in an uncompressed node list are equal, a compression to one eighth of the maximum number of node list entries can be achieved, etc. Different possibilities for storing the compression indicators exist. Generally, it has to be made sure that in address translating the compression indicator for a node list is read before the node list entry is indexed. A preferred form for storing the compression indicator is to write it into a list entry of a node of a higher node level so that in address translating it can be found out already when reading the list entry of a higher node level, whether the node list referenced by the read list entry is compressed and if the compression type is not standard, how it is compressed. Alternatively, the compression indicator can also be, for example, in a default position, even in the compressed node list itself, which is always accessed before a special entry of the node list is indexed. It is one advantage of the present invention that compressed node lists need less physical memory space than the non-compressed node lists so that this gained memory space is available. In a preferred embodiment of the present invention where an addressing by pages structure is used the node lists are no longer stored in individual physical memory pages, but as many node lists as possible are stored in the same physical memory page so that the case occurs that several node lists of nodes of the same node level are arranged in the same physical memory page or even several node lists of nodes of different node levels are stored in the same physical memory page. This has the advantage that the physical memory is “tidy”, so that not only parts of memory pages but whole memory pages, which are not taken up by the mapping regulation, are available for other data. Thereby, a fragmentation of the physical address space is counteracted. In a preferred embodiment of the present invention at least part of the mapping regulation and especially merely the root node itself or even only a reference to it and a generating regulation for the mapping regulation is stored by using the reference in a non-volatile memory, so that the computer system with the inventive addressing can already boot up in the virtual mode. According to a preferred embodiment, the security of a computer system with virtual addressing is improved by the fact that the computer system does not use a physical addressing mode, but works in the virtual addressing mode from the beginning. To achieve that, the hierarchical mapping regulation by which a physical address can be determined from a virtual address is stored in a non-volatile memory of the computer system. The mapping regulation is stored in the non-volatile memory so that it is immediately present in booting up the computer system and does not have to be generated in a real mode first and then stored in a volatile memory as in the prior art. Thereby the complete management effort and the required programs to generate a mapping regulation, for example in the shape of a hash table, becomes obsolete, since the mapping regulation is already fully available in the non-volatile memory. For the computer system to boot up in the virtual addressing mode, apart from the non-volatile memory where the mapping regulation is stored, a hardware state machine is provided that can access the non-volatile memory and is designed to determine the physical address associated with the virtual address by using the virtual address and the mapping regulation. The hardware state machine executes a predetermined algorithm automatically, as it is common for state machines, wherein the algorithm executed by the hardware state machine obtains, on the one hand, input data from the non-volatile memory, and on the other hand the virtual address as input data to output the physical address as output data. Inventively, however, it is preferred to store the mapping regulation in the shape of a hierarchical tree with one root not for a root level, at least one intermediate node for at least one intermediate level and an end node for an end level, so that the hardware state machine controlled by the virtual address and the lists for the individual node stored in the non-volatile memory performs a so-called page table walk to output the physical address corresponding to the virtual address after passing through the tree. In a preferred embodiment of the present invention, where the virtual address space is significantly larger than the physical address space, and lists for nodes therefore have relatively few used entries and relatively many zero entries, the lists for nodes of the hierarchical tree are compressed to save memory space in the non-volatile memory. In the case of the hierarchically organized mapping regulation between virtual and physical address not the whole mapping regulation has to be stored in the non-volatile memory, but at least part of the mapping regulation by which it is possible to start a boot up process of the system in the virtual mode. In appropriate list entries in the virtual mode it is then possible to generate the remaining part of the mapping regulation in the volatile memory already when retrieving the needed data from the physical memory and use it for further address translation from virtual to physical. The hardware state machine can therefore also access data programmed in the volatile memory during runtime after boot up of the system in the virtual mode. Further, in a preferred embodiment of the present invention, addressing by pages is preferred. To avoid memory fragmentation in this case as many combined lists as possible are stored in the same physical page. It is another advantage of the present invention that in the case of the mapping regulation as hierarchical tree structure a differentiated access right placing with an adjustable granularity can be achieved by implementing access rights at the node of the tree. It is another advantage of the present invention that by storing the mapping regulation in non-volatile memory in a form as it can be used by the hardware state machine without intervention of the CPU, CPU resources will be saved. The mapping regulation, for example the page table or the hierarchical structure of node lists has of course to be generated. This, however, can be performed outside the operation of the computer system, so to speak “off line” or in the case of a chip card “off card”. Generating the mapping regulation and storing the mapping regulation in non-volatile memory does therefore not need any valuable online resource of memory or CPU but can be performed when there is enough time, for example in the production of the chip card or in the case of a dynamic modification of the mapping regulation when the computer system does not carry out any security relevant application. Inventively, therefore, processing steps can be transferred out of the online operation in order to make valuable online resources, such as computer performance, memory space, energy consumption etc. available, or to save it. |
Display system |
A display system can accommodate a number of products. The system includes one or more modules. Each module may be mounted on a support wall and includes back and shelf portions. Advantageously, a number of modules are arranged in a number of rows, one row above another. A number of dividers are removably secured to at least one of the walls and may be moved in stepwise or continuous fashion to separate groups of the products within each row. Various of the modules may include hook bars secured to the back wall portions for hanging certain of the products and/or space thieves removably secured to the shelf portions for forwardly offsetting stacked or hung products. |
1. A combination comprising: a substantially vertical support wall having at least one array of mounting apertures; a plurality of products; and a device supporting said products in at least one condition and, in turn, supported by said support wall in an installed condition and comprising: a back portion secured to the support wall in said installed condition via said mounting apertures and having front and back sides, left and right sides, and upper and lower ends; and a shelf portion hinged to a lower portion of the back portion for articulation between stowed and deployed conditions and having a first surface which is a substantially upper surface in said deployed condition and in close facing or contacting proximity to the back portion front side in said stowed condition. 2. The combination of claim 1 further comprising: a plurality of corrugated trays supported by the upper surface, at least some of the products being in a plurality of stacks, each tray containing at least one such stack. 3. The combination of claim 1 further comprising: a plurality of dividers, removeably secured to at least one of the back and shelf portions in said deployed condition. 4. The combination of claim 3 wherein: the dividers each comprise the unitary plastic molding of: a base from which a pair of front and rear rails depend, the rails having a depending leg portion and a foot portion; and an upstanding web connected to the base; and the shelf portion includes a first elongate transverse slot receiving the rails. 5. The combination of claim 1 further comprising: a signage carrier comprising the unitary plastic molding of: a base from which a pair of front and rear rails depend, the rails having a depending leg portion and a foot portion; an upstanding web connected to the base; and a signage receiving portion extending forward from the web and having left and right side surfaces and a rim and inward projections from said rim spaced outboard of said left and right side surfaces; and wherein: the shelf portion includes a first elongate transverse slot receiving the rails; and the signage receiving portion carries a pair of left and right signage cards along said left and right surfaces and retained by the projections. 6. The device of claim 1 wherein the back portion is so secured to the support wall in said installed condition by: a plurality of mounting brackets, each comprising a unitary plastic molding of: a pair of mounting prongs dimensioned and positioned to be inserted into a pair of the mounting apertures; and an upwardly open channel receiving an associated portion of the back portion. 7. The device of claim 6 wherein: the back portion has a plurality of apertures and said associated portions are at upper portions of said apertures. 8. The device of claim 7 wherein: there are at least three such apertures being left, right, and center such apertures and three such mounting brackets, engaging the upper portions of the left, right, and center apertures respectively. 9. The device of claim 1 wherein the shelf portion upper surface has a primary substantially flat portion and a raised lip forward thereof. 10. The device of claim 9 wherein: from a front extreme of said shelf portion a first lip surface extends upward and rearward within 30 degrees of vertical; and a second lip surface extends rearward and upward from the first lip surface within 30 degrees of horizontal. 11. The device of claim 10 wherein an at least partially transparent extrusion is secured to at least one of the first and second lip surfaces and defines a signage receiving channel. 12. A display module for mounting to a support surface and comprising: a molded plastic member having: a bottom wall having an upper surface and a rear wall having a front surface; apertures on the bottom and rear walls for receiving mounting projections of one or more dividers. 13. The device of claim 12 wherein the bottom and rear walls are hinged relative to each other. 14. The device of claim 12 wherein the bottom wall comprises a plurality of first mounting apertures and the device further comprises a removable and reinstallable space thief having a pair of projections at its bottom end for engaging the first mounting apertures. 15. The device of claim 12 wherein a principal portion of the upper surface of the bottom wall has a front to back decline of 5-20°. 16. The device of claim 12 wherein a front face of the bottom wall extends above the remaining upper surface to provide a retaining lip. 17. The device of claim 14 wherein the plurality of first mounting apertures are arrayed to provide multiple front to back positions for the space thief. 18. The device of claim 12 further comprising: at least one such divider so mounted to the member; and a pair of mounting prongs for engaging holes of the support surface. 19. The device of claim 18 wherein: said prongs are formed by opposite end portions of a metal wire; a central portion of the wire is captured by the member to support the member; and the ends have a separation from each other and from the central portion effective to permit the ends to be inserted into holes on either side of a one inch square section tubing along the front of the support surface. 20. The device of claim 12 further comprising: a hook mounting fixture consisting essentially of an elongate body extending generally vertically along the rear wall front surface and a plurality of mounting projections extending rearward from the body and engaging the rear wall to removeably secure the hook mounting fixture to the rear wall; and a plurality of hooks, each having a base portion removeably mounted to the body and a shaft extending generally forward from the base portion. 21. A system for displaying product comprising: a gondola unit extending from a first end to a second end, the first and second ends being referenced as right and left ends, comprising: a gondola base having a front edge and an upper surface; a mounting wall extending upward from the base and having a front surface and an array of mounting holes; a plurality of display modules, each comprising: a back portion; and a bottom portion extending forward from the back portion, wherein: the plurality of display modules are mounted to the mounting wall and arranged on a plurality of rows, each row comprising at least two such modules; one or more divider walls, each mounted to an associated module, separate groups of the product; one or more of the modules have product-carrying hooks mounted to and extending forward from the back portion of such module; one or more of said product-carrying hooks are positioned one above another on a given mounting fixture mounting such hooks to a common module back portion; and each of one or more space thief members has a bottom portion removably mounted to an associated one of the mounting fixtures and a front surface forwardly offsetting product relative to the back portion. 22. The display system of claim 21 further comprising: a signage-carrying header mounted to the mounting wall and extending at a height above an upper row of said plurality of rows; at least one elongate, vertically extending sign member mounted to two associated modules of two adjacent rows and carrying signage on two opposite left and right facing sides; a base cover having a first portion atop the base upper surface and a second portion depending in front of a front edge portion of the base; and wherein: at least one of the space thief members is associated with one of the mounting fixtures with the product-carrying hooks positioned on such mounting fixture extending through an aperture on such space thief member. 23. A product display module comprising: a molded plastic back portion having: means for receiving product-carrying elements; means for receiving hangers for mounting the module to a product support structure; and means for receiving divider walls; and a molded plastic shelf portion extending forward from the back portion in at least a deployed condition and having: an upper surface; means formed in said upper surface for receiving said divider walls; and means formed in said upper surface for receiving signage carriers. 24. The display module of claim 23 further comprising: means for mounting the shelf portion to the back portion permitting the shelf portion to be alternatively position in said deployed condition and a relatively compact stowed condition. 25. A product display system comprising: one or more display modules, each comprising: molded plastic back portion; and a molded plastic shelf portion extending forward from the back portion, means for mounting the display modules to a support structure; divider means for separating groups of the product; product-carrying hooks mounted to and extending forward from the back portion of at least one such module; and one or more space thief members having a bottom portion removably mounted to the shelf portion of at least one such module. 26. The display system of claim 25 comprising a plurality of said modules mounted to a mounting wall in a plurality of rows and further comprising: a signage-carrying header mounted to the mounting wall and extending at a height above an upper row of said plurality of rows; and at least one elongate, vertically extending sign member mounted to two associated modules of two adjacent rows and carrying signage on two opposite left and right facing sides. |
<SOH> BACKGROUND OF THE INVENTION <EOH>(1) Field of the Invention This invention relates to displays, and more particularly to displays mountable on vertical support surfaces such as pegboard walls and in-store shelving gondolas. (2) Description of the Related Art A wide variety of systems exist for displaying goods in the retail environment. Many such systems are used to display many small product packages. These various systems make use of many product holding means including hooks, trays, chutes, and the like. Some displays are freestanding while others are mounted to a support structure such as a shelving gondola. Common gondola configurations feature long rows of shelving facing aisles on either side of the gondola. At the gondola's ends, additional shelving or other display areas define end caps. One common auxiliary display system is known as the power wing, typically secured at the side of an end cap and protruding slightly into the adjacent aisle. Smaller displays may be secured to the sides of the power wing and may face the longitudinal direction of the aisle. Such smaller displays are often identified as mini wings. Mini wing-type displays may also be mounted to shelving fronts to protrude into an aisle. These may include portions facing the aisle or facing the longitudinal direction of the aisle. |
<SOH> BRIEF SUMMARY OF INVENTION <EOH>A display system can accommodate a number of products. The system includes a display module having a back portion and a shelf or bottom portion, preferably hinged relative to each other. The modules are preferably hangable, and are preferably mounted on a vertical support surface such as a gondola pegboard wall. The modules may be arrayed in one or more rows. The products may be supported by the shelves individually or stacked on the shelves thieves may forwardly offset product relative to the module back. Dividers may be provided to separate groups of products. Mounting features may provide stepwise or continuous transverse positioning of the dividers, hooks, space thieves and other elements such as signage. Longitudinal adjustment is particularly advantageous for the space thieves. The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims. |
Methods, configuration and computer program having program code means and computer program product for determining a position of a mobile communications device within a communications network |
To determine a position of a mobile communications device within a communications network (localization). possible location areas for the mobile communications device are determined from communications signals of the mobile communications device by using base stations located within the communication network, and these possible location areas are superimposed to form a common location area while using a non-linear quantity-based filter. The position of the mobile communications device is then determined while using the common locations area. |
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