1. An isolated nucleic acid comprising a nucleotide sequence encoding a vascular endothelial cell growth factor-E (VEGF-E) polypeptide comprising amino acid residues 1 through 345 of FIG. 2 (SEQ ID NO:2).
2. The nucleic acid of claim 1 comprising the sequence of nucleotides 259 through 1293 of FIG. 1 (SEQ ID NO:1), or its complement.
3. An isolated nucleic acid comprising a nucleotide sequence which hybridizes under stringent conditions to the nucleotide sequence of claim 1.
4. A vector comprising the nucleic acid of claim 1.
5. A host cell comprising the nucleic acid of claim 1.
6. The host cell of claim 5, wherein said cell is a Chinese Hamster Ovary cell, an insect cell, an E. coli cell, or a yeast cell.
7. The host cell of claim 6 that is a baculovirus-infected insect cell.
8. A process for producing a vascular endothelial cell growth factor-E (VEGF-E) polypeptide comprising culturing the host cell of claim 5 under conditions suitable for expression of the VEGF-E polypeptide and recovering the VEGF-E polypeptide from the cell culture.
9. A polypeptide produced by the process of claim 8.
10. A VEGF-E polypeptide comprising amino acid residues 1 through 345 of FIG. 2 (SEQ ID NO:2).
11. A VEGF-E polypeptide selected from the group consisting of:
(a) a polypeptide comprising amino acid residues 1 through 345 of FIG. 2 (SEQ ID NO:2); and
(b) a polypeptide fragment of (a), wherein said fragment is biologically active.
12. A VEGF-E polypeptide encoded by the nucleotide sequence insert of ATCC deposit No. 209653.
13. A chimeric polypeptide comprising the polypeptide of claim 10 fused to a heterologous amino acid sequence.
14. The chimeric polypeptide of claim 13, wherein said heterologous amino acid sequence is an epitope tag sequence or a Fc region of an immunoglobulin.
15. A composition comprising the polypeptide of claim 10 in admixture with a carrier.
16. The composition of claim 15 comprising a therapeutically effective amount of the polypeptide, wherein the carrier is a pharmaceutically acceptable carrier.
17. A pharmaceutical product comprising:
(a) the composition of claim 15;
(b) a container containing said composition; and
(c) a label affixed to said container, or a package insert included in said pharmaceutical product, referring to the use of said VEGF-E polypeptide in the treatment of a cardiovascular or endothelial disorder.
18. A method for treating a cardiovascular or endothelial disorder in a mammal comprising administering to the mammal an effective amount of the composition of claim 15.
19. The method of claim 18 wherein the disorder is cardiac hypertrophy, trauma, or a bone-related disorder.
20. The method of claim 19 wherein said cardiac hypertrophy is characterized by the presence of an elevated level of PGF2.
21. The method of claim 19 wherein said cardiac hypertrophy has been induced by myocardial infarction.
22. The method of claim 21 wherein said VEGF-E polypeptide administration is initiated within 48 hours following myocardial infarction.
23. A method for diagnosing a disease or a susceptibility to a disease related to a mutation in a nucleic acid sequence encoding vascular endothelial cell growth factor-E (VEGF-E) comprising:
(a) isolating a nucleic acid sequence encoding VEGF-E from a sample derived from a host; and
(b) determining a mutation in the nucleic acid sequence encoding VEGF-E.
24. A method of diagnosing cardiovascular and endothelial disorders in a mammal comprising detecting the level of expression of a gene encoding a vascular endothelial cell growth factor-E (VEGF-E) polypeptide (a) in a test sample of tissue cells obtained from the mammal, and (b) in a control sample of known normal tissue cells of the same cell type, wherein a higher or lower expression level in the test sample indicates the presence of a cardiovascular or endothelial dysfunction in the mammal from which the test tissue cells were obtained.
25. A method for identifying an agonist to a vascular endothelial cell growth factor-E (VEGF-E) polypeptide comprising:
(a) contacting cells and a candidate compound under conditions that allow the polypeptide to stimulate proliferation of the cells; and
(b) measuring the extent to which cell proliferation is inhibited by the compound.
26. An agonist to a VEGF-E polypeptide identified by the method of claim 25.
27. A method for identifying a compound that inhibits the expression or activity of a vascular endothelial cell growth factor-E (VEGF-E) polypeptide, comprising:
(a) contacting a candidate compound with the polypeptide under conditions and for a time sufficient to allow the compound and polypeptide to interact; and
(b) measuring the extent to which the compound interacts with the polypeptide.
28. A compound identified by the method of claim 27.
29. An isolated antibody that binds a vascular endothelial cell growth factor-E (VEGF-E) polypeptide.
30. The antibody of claim 29 that is a monoclonal antibody.
31. A method for determining the presence of a vascular endothelial cell growth factor-E (VEGF-E) polypeptide comprising exposing a cell suspected of containing the VEGF-E polypeptide to the antibody of claim 29 and determining binding of said antibody to said cell.
32. A method of diagnosing cardiovascular, endothelial, or angiogenic disorders in a mammal comprising (a) contacting the antibody of claim 29 with a test sample of tissue cells obtained from the mammal, and (b) detecting the formation of a complex between the anti-VEGF-E antibody and the VEGF-E polypeptide in the test sample.
33. An article of manufacture, comprising:
a container;
a label on the container; and
a composition comprising the antibody of claim 29 contained within the container; wherein the label on the container indicates instructions for using the antibody in a therapeutic method or diagnostic method.
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 cutting insert for a milling cutter, the cutting insert comprising two polygonal main surfaces that are opposite each other and a plurality of side faces that are formed continuously with the sides of the main surfaces, the main surfaces being rake faces, the side faces being flank faces, and ridge lines at positions where the main surfaces and the side faces cross each other being used as cutting edges,
wherein the plurality of side faces include two major side surfaces that are opposite each other and a plurality of minor side surfaces that are formed continuously with the major side surfaces, a mounting hole is provided at the two major side surfaces in a direction in which the mounting hole extends therethrough, and areas of the two major side surfaces are greater than those of the main surfaces, and
wherein the major side surfaces and the minor side surfaces are provided with flutes along entire regions of the cutting edges, and flute faces of the flutes at a side along the cutting edges are tilted in a direction in which depths gradually increase with increasing distance from the cutting edges.
2. The cutting insert for the milling cutter according to claim 1, wherein a tilt angle of the flute face of each flute at the side along the cutting edge is greater than or equal to 2 degrees, and a flute width of each flute is greater than or equal to 0.5 mm and less than or equal to 5 mm.
3. The cutting insert for the milling cutter according to claim 1, wherein a central portion of each main surface is provided with a flat land that is seated on a support seat of a cutter body, and, with positions of the flat lands being lower than positions of the cutting edges, the rake faces that are tilted are provided between the cutting edges and the flat lands.
4. The cutting insert for the milling cutter according to either claim 2, wherein a central portion of each main surface is provided with a flat land that is seated on a support seat of a cutter body, and, with positions of the flat lands being lower than positions of the cutting edges, the rake faces that are tilted are provided between the cutting edges and the flat lands.