1. A method of identifying a compound that is useful in treating andor preventing dyslipidemia comprising, providing a cell comprising at least one expression construct comprising a DNA molecule encoding an XBP-1 polypeptide and a DNA molecule encoding an IRE-1 polypeptide;
a) contacting the cell with each member of a library of test compounds;
b) determining the activity of the XBP-1 polypeptide in the presence and absence of the test compound;
c) selecting from the library of test compounds a compound of interest that downmodulates the activity of the XBP-1 polypeptide;
d) determining the effect of the compound on de novo hepatic lipogenesis;
e) selecting a compound of interest that downmodulates de novo hepatic lipogenesis as compared to an appropriate control, thereby identifying a compound that is useful in treating andor preventing dyslipidemia.
2. The method of claim 1, wherein the effect of the compound on XBP-1 activity is determined by measuring the binding of XBP-1 to IRE-1.
3. The method of claim 1, wherein the effect of the compound on XBP-1 activity is determined by assaying the activity of IRE-1.
4. The method of claim 3, wherein the activity of IRE-1 is a kinase activity.
5. The method of claim 3, wherein the activity of IRE-1 is an endoribonuclease activity.
6. The method of claim 1, wherein the effect of the compound on XBP-1 activity is determined by assaying XBP-1 andor IRE-1 protein levels.
7. The method of claim 1, wherein the XBP-1 polypeptide comprises a transactivation domain and the cell further comprises a vector comprising a regulatory element responsive to the XBP-1 transactivation domain operatively linked to a reporter gene and the effect of the compound on the activity of the polypeptide is determined by evaluating the expression of the reporter gene in the presence and absence of the test compound.
8. The method of claim 1, wherein the cell comprises a recombinant expression vector encoding a spliced XBP-1 protein; and a vector comprising a regulatory element responsive to the spliced XBP-1 protein operatively linked a reporter gene and the effect of the compound on the activity of the polypeptide is determined by evaluating the expression of the reporter gene in the presence and absence of the test compound.
9. The method of claim 1, wherein the cell comprises a recombinant expression vector comprising a regulatory element responsive to XBP-1 spliced protein operatively linked a reporter gene, wherein the cell is contacted with an agent that induces ER stress, and the effect of the compound on the activity of the polypeptide is determined by evaluating the expression of the reporter gene in the presence and absence of the test compound.
10. The method of claim 7, 8, or 9, further comprising determining the effect of the identified compound on the ratio of unspliced XBP-1 mRNA to spliced XBP-1 mRNA.
11. The method of claim 7, 8, or 9, further comprising determining the effect of the identified compound on the production of XBP-1 protein.
12. The method of claim 1, further comprising providing an indicator cell comprising a glucose-dependent promoter operably linked to a reporter gene, wherein the cell in cultured in the presence of glucose, and the effect of the compound on de novo hepatic lipogenesis is determined by evaluating the expression of the reporter gene in the presence and absence of the test compound.
13. The method of claim 1, wherein the effect of the compound on de novo hepatic lipogenesis is determined by determining the effect of the compound on level of cholesterol andor triglyceride.
14. The method of claim 1, wherein determining the ability of the compound to modulate de novo hepatic lipogenesis comprises determining the expression of SREBP andor chREBP.
15. The method of claim 1, wherein the ability of the compound to modulate de novo lipogenesis comprises determining the expression of a lipogenic gene.
16. The method of claim 1, wherein:
the cell further comprises a DNA molecule to which XBP-1 binds, wherein the DNA molecule comprises a promoter selected from the group consisting of a Dgat2 promoter, a Scd1 promoter, an Acc2 promoter; and a PCSK9 promoter; and
the effect of the test compound on de novo hepatic lipogenesis is determined by evaluating the binding of XBP-1 to the DNA molecule in the presence and absence of the test compound.
17. The method of claim 1, further comprising administering the test compound identified as useful in modulating dyslipidemia to a non-human animal and determining the effect of the identified compound on obesity andor insulin resistance in the non-human animal.
18. The method of claim 1, further comprising administering the test compound identified as useful in modulating dyslipidemia to a non-human animal and determining the effect of the compound on at least one of aspartate aminotransferase levels, alanine aminotransferase levels, liver cell inflammation, and the presence of triglyceride vacuoles in liver cells.
19. The method of claim 1, further comprising determining the effect of the compound on at least one of serum triglycerides, total cholesterol and the distribution of cholesterol among LDL, IDL, and HDL.
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. A bucket-mounted concrete tool-cleaning device, further comprising:
a cleaner head base that clips onto a bucket such that an angled arm is presented into an interior of said bucket at an acute angle and whereby at least one snap-on cleaner head attaches onto said angled arm and includes an open, inner channel adapted to receive a concrete tool or float to be cleaned and further including inwardly directed bristles for cleaning residual concrete from a concrete tool or float.
2. The bucket-mounted concrete tool-cleaning device as described in claim 1 wherein the cleaner head base is further defined by a base arm and a top lip arm both of which are attached to the angled arm.
3. The bucket-mounted concrete tool-cleaning device as described in claim 2 wherein the top lip arm features a lip that engages a top lip of said bucket.
4. The bucket-mounted concrete tool-cleaning device as described in claim 3 wherein a lip slides underneath the top lip of said bucket.
5. The bucket-mounted concrete tool-cleaning device as described in claim 4 wherein the lip includes a curved recess, which accommodates a curvature of the bucket.
6. The bucket-mounted concrete tool-cleaning device as described in claim 2 wherein the base arm is composed of a horizontally oriented section that abuts against an inner surface of the bucket.
7. The bucket-mounted concrete tool-cleaning device as described in claim 6 wherein the base arm includes a base plate that rests up against the inner surface of the bucket.
8. The bucket-mounted concrete tool-cleaning device as described in claim 1 wherein the angled arm includes a plurality of slots, which are equally spaced along the length of the angled arm.
9. The bucket-mounted concrete tool-cleaning device as described in claim 8 wherein the snap-on cleaner head includes a clip protruding from a rear surface of the snap-on cleaner head, which enables the snap-on cleaner head to attach to the angled arm at one of the slots.
10. The bucket-mounted concrete tool-cleaning device as described in claim 1 wherein the plurality of bristles adorn an inner surface of the open, inner channel such that upon insertion of the tool or float, residual concrete is scrubbed off; wherein the open, inner channel includes an opening that enables a tool handle to pass there through when engaging the tool against the bristles.
11. The bucket-mounted concrete tool-cleaning device as described in claim 1 where in the cleaner head base includes a screw that is threadably engaged therewith and which extends and retracts laterally thereto; wherein the screw is rotated to tighten and lock the cleaner head base to the bucket.
12. A bucket-mounted concrete tool-cleaning device, further comprising:
a cleaner head base that clips onto a bucket such that an angled arm is presented into an interior of said bucket at an angle and whereby at least one snap-on cleaner head attaches onto said angled arm and includes an open, inner channel adapted to receive a concrete tool or float to be cleaned and further including inwardly directed bristles for cleaning residual concrete from a concrete tool or float;
wherein the cleaner head base is further defined by a base arm and a top lip arm both of which are attached to the angled arm;
wherein the top lip arm features a lip that engages a top lip of said bucket;
wherein a lip slides underneath the top lip of said bucket;
wherein the base arm is composed of a horizontally oriented section that abuts against an inner surface of the bucket;
wherein the base arm includes a base plate that rests up against the inner surface of the bucket.
13. The bucket-mounted concrete tool-cleaning device as described in claim 12 wherein the lip includes a curved recess, which accommodates a curvature of the bucket.
14. The bucket-mounted concrete tool-cleaning device as described in claim 12 wherein the angled arm includes a plurality of slots, which are equally spaced along the length of the angled arm.
15. The bucket-mounted concrete tool-cleaning device as described in claim 14 wherein the snap-on cleaner head includes a clip protruding from a rear surface of the snap-on cleaner head, which enables the snap-on cleaner head to attach to the angled arm at one of the slots.
16. The bucket-mounted concrete tool-cleaning device as described in claim 12 wherein the plurality of bristles adorn an inner surface of the open, inner channel such that upon insertion of the tool or float, residual concrete is scrubbed off; wherein the open, inner channel includes an opening that enables a tool handle to pass there through when engaging the tool against the bristles.
17. The bucket-mounted concrete tool-cleaning device as described in claim 16 wherein the open, inner channel may include a small recess, which is suited for cleaning off a jointer edge of jointer concrete tools.
18. The bucket-mounted concrete tool-cleaning device as described in claim 12 wherein the cleaner head base includes a screw that is threadably engaged therewith and which extends and retracts laterally thereto; wherein the screw is rotated to tighten and lock the cleaner head base to the bucket.