What is claimed is:
1. A method for identifying a compound that modulates aging, the method comprising the steps of:
(i) contacting the compound with a polypeptide, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C1 (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), LLW-4 (SEQ ID NO: 10) and HSF-1 (SEQ ID NO: 11), and human homologues thereof; and
(ii) determining the functional effect of the compound upon the polypeptide.
2. The method of claim 1, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C1 (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), and LLW-4, (SEQ ID NO: 10) or human homologues thereof.
3. The method of claim 1, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence of HSF-1 (SEQ ID NO: 11), or human homologues thereof.
4. The method of claim 1, wherein the functional effect is determined in vitro.
5. The method of claim 4, wherein the functional effect is determined by measuring enzymatic activity.
6. The method of claim 4, wherein the functional effect is determined by measuring ligand, substrate, or cofactor binding to the polypeptide.
7. The method of claim 4, wherein the functional effect is determined by measuring binding to a promoter sequence by the polypeptide.
8. The method of claim 1, wherein the polypeptide is expressed in a mitochondrial extract or an isolated mitochondrial membrane.
9. The method of claim 1, wherein the polypeptide is expressed in a eukaryotic host or host cell and the polypeptide is contacted with the compound in a living cell.
10. The method of claim 9, wherein the host cell is derived from C. elegans, mouse, rat, or human.
11. The method of claim 9, wherein the host is C. elegans, mouse, rat, or human.
12. The method of claim 9, wherein the functional effect is a determined by measuring ligand, substrate, or cofactor binding to the polypeptide.
13. The method of claim 9, wherein the functional effect is determined by measuring transcriptional activation.
14. The method of claim 9, wherein the functional effect is determined by evaluating age-associated parameters.
15. The method of claim 9, wherein the functional effect is determined by evaluating expression of an age-associated gene.
16. The method of claim 14, wherein the age-associated parameter is lifespan.
17. The method of claim 1, wherein the modulation is inhibition of aging.
18. The method of claim 17, wherein inhibition of aging occurs by activation of an HSF polypeptide encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence of HSF-1 (SEQ ID NO: 11), or human homologues thereof.
19. The method of claim 17, wherein inhibition of aging occurs by inhibition of a polypeptide encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C1 (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), and LLW-4 (SEQ ID NO: 10) or human homologues thereof.
20. The method of claim 1 wherein the polypeptide is recombinant.
21. The method of claim 1, wherein the compound is an antibody, an antisense molecule, or a small molecule.
22. A method for identifying a compound that modulates aging, the method comprising the steps of:
(i) contacting the compound with a polypeptide, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C1 (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), LLW-4 (SEQ ID NO: 10) and HSF-1 (SEQ ID NO: 11), and human homologues thereof,
(ii) determining the functional effect of the compound upon the polypeptide; and
(iii) contacting a host or host cell expressing the protein and evaluating an age-associated parameter in a host or host cell, thereby identifying a compound that modulates aging.
23. The method of claim 22, wherein the polypeptide is recombinant.
24. The method of claim 22, wherein the functional effect is a physical effect.
25. The method of claim 22, wherein the functional effect is a chemical effect.
26. The method of claim 22, wherein the functional effect is a phenotypic effect.
27. The method of claim 22, wherein the functional effect is determined in vitro.
28. The method of claim 22, wherein the functional effect is determined in a eukaryotic host organism or host cell.
29. The method of claim 22, wherein the age-associated parameter is lifespan.
30. A method for identifying a compound that modulates aging, the method comprising the steps of:
(i) contacting the compound with a polypeptide that is a component of the mitochondrial respiratory chain; and
(ii) determining the functional effect of the compound upon the polypeptide.
31. The method of claim 30, wherein the polypeptide is expressed in a mitochondrial extract or an isolated mitochondrial membrane.
32. A method for identifying a compound that modulates aging, the method comprising the steps of:
(i) contacting the compound with a polypeptide, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid listed in Table 5 or Table 6, or a nucleic acid encoding a polypeptide listed in Table 5 or Table 6, or human homologues thereof; and
(ii) determining the functional effect of the compound upon the polypeptide.
33. A method for identifying a compound that modulates aging, the method comprising the steps of:
(i) contacting the compound with a polypeptide, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid listed in Table 5 or Table 6, or a nucleic acid encoding a polypeptide listed in Table 5 or Table 6, or human homologues thereof
(ii) determining the functional effect of the compound upon the polypeptide; and
(iii) contacting a host or host cell expressing the protein and evaluating an age-associated parameter in a host or host cell, thereby identifying a compound that modulates aging.
34. A compound that modulates an aging process, wherein the compound is identified by the method of claim 1.
35. The compound of claim 34, wherein the compound is an antibody, an antisense molecule, or a small molecule.
36. The compound of claim 34, wherein the compound is antimycin or an analog thereof.
37. A method of increasing lifespan or treating premature aging in a subject, the method comprising the step of administering to the subject an effective amount of a compound identified using the method of claim 1.
38. The method of claim 37, wherein the compound is antimycin.
39. A method of increasing lifespan or treating premature aging in a subject, the method comprising the step of administering to the subject an effective amount of a compound that modulates the expression of a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C1 (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), LLW-4 (SEQ ID NO: 10) and HSF-1 (SEQ ID NO: 11), and human homologues thereof.
40. The method of claim 37 or 39, wherein the aging process is abnormal.
41. The method of claim 40, wherein the abnormal aging process is selected from Werner syndrome, Hutchinson-Guilford disease, Bloom’s syndrome, Cockayne’s syndrome, ataxia telangiectasia, and Down’s syndrome.
42. The method of claim 37 or 39, wherein the aging process is normal.
43. The method of claim 37 or 39, further comprising the step of evaluating an age-associated parameter of the subject.
44. A method of identifying a gene or gene product that modulates aging, the method comprising the steps of:
(i) providing a library of nucleic acids, each nucleic acid of the library comprising a segment of a gene of an organism; and
(ii) for each member of the library,
(a) generating double-stranded RNA (dsRNA) from the respective member of the library,
(b) providing the dsRNA to a cell or one or more cells of the organism to provide a ds-RNA treated cell or ds-RNA treated organism, and
(b) monitoring an age-associated parameter of the ds-RNA treated cell or the ds-RNA treated organism.
45. A method of identifying a gene or gene product that modulates aging, the method comprising the steps of:
(i) evaluating a relationship between presence or abundance for each species of a plurality of RNA or protein species with respect to age of a cell or organism;
(ii) producing or delivering a double-stranded RNA to a cell or one or more cells of an organism, the RNA corresponding to a species whose presence or abundance is correlated with age; and
(iii) monitoring an age-associated parameter of the cell or the one or more cells or the organism.
46. A method of identifying a compound that modulates aging, the method comprising the steps of:
(i) contacting a test compound to a living or biochemical system that comprising a C. elegans target protein selected from the group consisting of: a respiratory chain component, a heat shock promoter activator, LLW-1, LLW-2, LLW-3, LLW-4, a small GTPase, a protein in Table 5, and a protein in Table 6; and
(ii) evaluating a property associated with the target protein; and
(iii) evaluating an aging-associated parameter of a C. elegans organism contacted with the test compound.
47. A method of identifying a gene or gene product that modulates aging, the method comprising the steps of:
(i) providing a nematode in which activity of a target protein is reduced in the organism by RNA interference;
(i) expressing a gene encoding a candidate mammalian protein that is heterologous to the organism; and
(iii) evaluating an age associated parameter of the organism.
48. A C. elegans nematode that (1) expresses a heterologous gene in at least some cells, the heterologous gene encoding a respiratory chain component or glycolysis component that is non-identical to the corresponding endogenous respiratory chain component or glycolysis component, or a functional domain thereof; and (2) is deficient in at least some cells for an endogenous activity provided by the corresponding endogenous respiratory chain component or glycolysis component.
49. A method comprising: assessing an age-associated parameter of the nematode of claim 48.
50. A C. elegans nematode that (1) has a deficiency in at least some cells for an endogenous activity, the deficiency generated by dsRNA in the cells, and (2) has an average lifespan of at least 40% greater than an otherwise identical nematode without the deficiency.
51. A method of identifying a gene or gene product that modulates aging, the method comprising the steps of:
(i) providing the nematode of claim 50;
(ii) introducing a heterologous gene that encodes a heterologous polypeptide into the nematode;
(iii) expressing the heterologous gene in the nematode or a progeny of the nematode under conditions wherein the heterologous polypeptide is produced; and
(iv) monitoring an age-associated parameter of the nematode or the progeny of the nematode.
52. The method of claim 51, further comprising contacting a test compound to the nematode or the progeny prior to or during the monitoring.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1-16. (canceled)
17. Polymerisation process in which polyethylene is produced in slurry in a polymerisation reactor in the presence of a Ziegler Natta catalyst and an activator, and a stream or slurry containing the polymer is withdrawn from the reactor and transferred to a flash tank operating at a pressure and temperature such that at least 50 mol % of the liquid or non-polymer component of the stream entering the flash tank or slurry is withdrawn from the flash tank as a vapour and at least 98 mol %, more preferably at least 98.5 mol %, and most preferably at least 99.5 mol %, of the vapour withdrawn from the flash tank is capable of being condensed at a temperature of between 15 and 50\xb0 C., preferably 15 and 40\xb0 C., without compression, wherein a by-product suppressor, which reduces the amount of by-product formed per unit of polyethylene produced by at least 10%, and preferably the absolute amount of by-product formed in the reactor by at least 10%, compared with an identical polymerisation process where the by-product suppressor is not present, is used in the reactor.
18. Process according to claim 17, wherein the by-product suppressor reduces the concentration in the reactor of at least one of the components in the slurry which has a molecular weight below 50 by at least 5%, preferably at least 10% compared with an identical process where the by-product suppressor is not present.
19. Process according to claim 17, wherein the by-product suppressor is a halogenated hydrocarbon, preferably a chloromethane of the formula CHxCl4-x where x is an integer from 1 to 3, and most preferably chloroform, CHCl3.
20. Polymerisation process in which polyethylene is produced in slurry in a polymerisation reactor in the presence of a Ziegler Natta catalyst and an activator, and a stream or slurry containing the polymer is withdrawn from the reactor and transferred to a flash tank operating at a pressure and temperature such that at least 50 mol % of the liquid or non-polymer component of the stream entering the flash tank or slurry is withdrawn from the flash tank as a vapour and at least 98 mol %, more preferably at least 98.5 mol %, and most preferably at least 99.5 mol %, of the vapour withdrawn from the flash tank is capable of being condensed at a temperature of between 15 and 50\xb0 C., preferably 15 and 40\xb0 C., without compression, wherein a halogenated hydrocarbon, preferably a chloromethane of the formula CHxCl4-x where x is an integer from 1 to 3, is present in the slurry.
21. Process according claim 17, wherein a stream containing the polymer is withdrawn from the reactor and transferred to a flash tank operating at a pressure and temperature such that at least 50 mol % of the non-polymer component of the stream entering the flash tank is withdrawn from the flash tank as a vapour.
22. Process according to claim 17, wherein the molar ratio of the by-product suppressor or halogenated hydrocarbon added to the reactor to titanium added to the reactor is greater than 0.1, preferably between 0.2 and 1.
23. Process according to claim 17, wherein the concentration in the stream entering the flash tank of components having a molecular weight below 50 gmol, Clights (mol %), satisfies the equation Clights<7+0.07(40\u2212Tc)+4.4(Pc\u22120.8)\u22127(CH2CEt) where Tc and Pc are respectively the temperature (in \xb0 C.) and pressure (MPa g) at the location where the vapour withdrawn from the flash tank is condensed, and CH2 and CEt are the molar concentrations in the flash tank of hydrogen and ethylene respectively.
24. Process according to claim 23, wherein the concentration in the reactor of components having a molecular weight below 50 also satisfies the equation Clights<7+0.07(40\u2212Tc)+4.4(Pc\u22120.8)\u22127(CH2CEt) where Clights, CH2, and CEt in this case are the concentrations of components having a molecular weight below 50, hydrogen and ethylene respectively in the reactor.
25. Process according to claim 23, wherein the concentration in the slurry entering the flash tank of hydrogen, ethylene and components having a molecular weight below 50 is the same as the concentration of hydrogen, ethylene and components having a molecular weight below 50 in the second reactor.
26. Process according to claim 17, wherein at least 80 mol %, more preferably 90 mol %, most preferably 95 mol % of the liquid component of the slurry is withdrawn from the flash tank as a vapour.
27. Process according to claim 17, wherein the polyethylene is a bimodal polymer made in two reactors in series, and the by-product suppressor is added to at least one of the reactors.
28. Process according to claim 27, wherein a high molecular weight (HMW) polymer is made in suspension in the first reactor and a low molecular weight (LMW) polymer is made in suspension in the second reactor in the presence of the first polymer.
29. Process according to claim 27, wherein a high molecular weight (HMW) polymer is made in suspension in the first reactor and a low molecular weight (LMW) polymer is made in suspension in the second reactor in the presence of the first polymer, and the ratio of the average activity in the second LMW reactor to the average activity in the first HMW reactor is from 0.25 and 1.5, where average activity in each reactor is defined as the rate of polyethylene produced in the reactor (kgPEhr)ethylene concentration in the reactor (mol %)\xd7residence time in the reactor (hours)\xd7feed rate of catalyst into the reactor (ghr).
30. Process according to claim 27, wherein a high molecular weight (HMW) polymer is made in suspension in the first reactor and a low molecular weight (LMW) polymer is made in suspension in the second reactor in the presence of the first polymer, and the ratio of ethylene concentration (in mol %) in the second reactor to that in the first reactor is 5 or less, preferably 3 or less, and more preferably 2.5 or less.
31. Process according to claim 27, wherein a high molecular weight (HMW) polymer is made in suspension in the first reactor and a low molecular weight (LMW) polymer is made in suspension in the second reactor in the presence of the first polymer, and the concentration of ethylene in the second reactor is less than 8 mol %, preferably between 1.5 mol % and 8 mol %.
32. Process according to claim 17, wherein the polyethylene is a multimodal polyethylene having a shear ratio of at least 15, preferably between 21 and 29, where \u201cshear ratio\u201d is the ratio of the high load melt index HLMI of the polyethylene to the MI5 of the polyethylene, both being measured according to ISO Standard 1133 at a temperature of 190\xb0 C.