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Patent US20030032673 - Therapeutic strategies for prevention and treatment of alzheimer's disease - Google PatentsSearch Images Maps Play YouTube News Gmail Drive More »Sign inPatentsThe invention relates to therapeutic agents for use in the prevention or treatment of Alzheimer's disease. In particular the invention relates to use of inhibitors of cell cycle re-entry and progression to the G1/S transition or inhibitors of progression of the cell cycle through the G1/S transition...http://www.google.com/patents/US20030032673?utm_source=gb-gplus-sharePatent US20030032673 - Therapeutic strategies for prevention and treatment of alzheimer's diseaseAdvanced Patent SearchPublication numberUS20030032673 A1Publication typeApplicationApplication numberUS 10/200,023Publication dateFeb 13, 2003Filing dateJul 19, 2002Priority dateJul 19, 2001Also published asDE60237725D1, EP1408938A1, EP1764092A2, EP1764092A3, EP1764092B1, EP1767197A2, EP1767197A3, EP1767197B1, EP1769791A2, EP1769791A3, EP1769791B1, EP2286803A1, EP2286875A1, EP2289511A1, EP2292238A1, EP2292240A1, US8343926, US8921321, US20130102553, WO2003007925A1Publication number10200023, 200023, US 2003/0032673 A1, US 2003/032673 A1, US 20030032673 A1, US 20030032673A1, US 2003032673 A1, US 2003032673A1, US-A1-20030032673, US-A1-2003032673, US2003/0032673A1, US2003/032673A1, US20030032673 A1, US20030032673A1, US2003032673 A1, US2003032673A1InventorsZsuzsanna NagyOriginal AssigneeIsis Innovation LimitedExport CitationBiBTeX, EndNote, RefManPatent Citations (5), Referenced by (48), Classifications (23), Legal Events (1) External Links: USPTO, USPTO Assignment, EspacenetTherapeutic strategies for prevention and treatment of alzheimer's disease
[0114] (Darzynkiewicz, Z. (1993) In Fantes P and Brooks R (eds) The cell cycle. Oxford University Press, Oxford, pp 43-68). [0115] Various techniques may be employed to obtain the values of TG1tr and TG1c. In a preferred embodiment TG1tr and TG1c may be obtained by determining the proportion of cells in the various phases of the cell cycle for both treated cells (non-neuronal cells from the test subject treated with the cell division inhibitor substance or stimulus that induces cell cycle arrest) and untreated control cells (non-neuronal cells from the same subject not exposed to the cell division inhibitor substance or stimulus that induces cell cycle arrest), in the presence of varying concentrations of candidate compound. The proportion of cells in the various phases of the cell cycle may be readily determined by incorporation of a labelled nucleotide analogue, preferably bromodeoxyuridine (BrdU), followed by fluorescence activated cell sorting (FACS analysis), or equivalent, as described in detail in Example 2. [0116] The presence of a cell cycle regulatory defect at the G1/S phase transition is indicated by a reduced relative lengthening of the G1 phase in the presence of the cell division inhibitor substance or stimulus in cells from the Alzheimer's subject, as compared to control cells not having a cell cycle regulatory defect at the G1/S phase transition (e.g. cells of the same type taken from an age-matched control individual not having Alzheimer's disease or any evidence of a regulatory defect at the G1/S transition or any genetic defect/allelic variation which might be expected to pre-dispose to Alzheimer's disease). The control cells not having a cell cycle regulatory defect at the G1/S phase transition are not to be confused with the “untreated control” cells used for calculation of RL, which are cells from the test subject which have not been exposed to inhibitory treatment. [0117] Candidate compounds which cause an increase in relative lengthening of the G1 phase in cells from a particular Alzheimer's patient in the presence of inhibitory treatment are scored as potentially useful in the treatment of Alzheimer's disease in that patient, because they are able to prevent progression through the cell cycle in cells of the Alzheimer's patient. [0118] (3) Assessment of Cell Cycle Regulatory Protein or mRNA Expression. [0119] Expression of cell cycle regulatory proteins may be assessed using standard techniques well known in the art such as, for example, immunoblotting, western blotting, ELISA or related methods. Assessment of expression of corresponding mRNAs encoding the cell cycle regulatory proteins may also be accomplished by means of standard methods such as, for example, hybridization techniques, “DNA chip” analysis or related methods or amplification-based techniques such as reversed transcription (RT)-PCR, real time-PCR, nucleic acid sequence-based amplification (NASBA), etc. Suitable methods for the detection/quantitation of mRNAs that may be used in accordance to the invention will be well known to those skilled in the art. Certain of these methods, for example RT-PCR, rely on detection/quantitation of a cDNA copy of the relevant mRNA. [0120] The cell cycle regulatory defect present in Alzheimer's disease may result in changes in the pattern of expression of cell cycle regulatory proteins, and their corresponding mRNAs. Screening for changes in expression of particular cell cycle regulatory proteins and/or the corresponding mRNAs may therefore be used diagnostically to identify the presence of a cell cycle regulatory defect at G1/S. In addition, expression of cell cycle regulatory proteins may be used as a marker of progression through the cell cycle. Hence, the responsiveness of cells to inhibitory treatment (using method (A) or method (B), described above) may be assessed by looking at the expression of one or more cell cycle regulatory proteins, in order to determine the extent to which inhibitory treatment causes cell cycle arrest in cells from the test subject. Suitable cell cycle regulatory proteins include, but are not limited to, CDKN3, p15ink4B, p16ink4A, p19ink4D, p27kip1, p21cip1, p57kip2 and TP53. Full cDNA and amino acid sequences for these proteins are publicly available: a list of OMIM accession numbers for these proteins is provided below. OMIM is Online Mendelian Inheritance in Man, a database which is a catalog of human genes and genetic disorders. OMIM is accessible via the website of the National Center for Biotechnology Information (NCBI). Antibodies useful in the detection of each of these proteins are available commercially, or may be prepared according to standard techniques known in the art. [0121] Candidate compounds which cause the mRNA and/or protein expression profile for cell cycle regulatory proteins in cells from a particular Alzheimer's patient to resemble that of a “normal” cell with intact regulation at the G1/S transition, are scored as being potentially useful in the treatment of Alzheimer's disease in that patient, because they are able to prevent progression through the cell cycle in cells of the Alzheimer's patient. [0122] (4) Assessment of Cell Viability and Cell Death by any Method Known in the Art. [0123] When a proliferating cell is arrested in the G1/S transition one of two possible “downstream” phenomena may result, either differentiation or programmed cell death. These downstream phenomena may be used as an indication of the presence in a cell population of a regulatory defect at the G1/S transition, since if regulation of the G1/S transition is defective then the downstream effects of cell cycle arrest at G1/S will also be abnormal. A lower degree of cell death or higher degree of cell viability in response to inhibitory treatment (using method (A) or method (B)) in cells from a test subject, as compared to control cells, is taken as an indication that the subject has Alzheimer's disease. [0124] Candidate compounds which cause a higher degree of cell death or lower degree of cell viability in response to inhibitory treatment (using method (A) or method (B)) in cells from a particular Alzheimer's patient are scored as being potentially useful in the treatment of Alzheimer's disease in that patient. [0125] (5) Assessment of Cell Death Related (Inducing or Preventing) Protein or mRNA Expression Using Standard Techniques Known in the Art. [0126] In this approach, expression of cell death related proteins, or the corresponding mRNAs, is used as an indirect assessment of the downstream effects of inhibitory treatment with a cell division inhibitor (method (A)) or a stimulus inducing cell cycle arrest at the G1/S transition (method (B)). Suitable cell death related proteins include, for example, members of the bcl-2 family of proteins, of which there are many known in the art. Monoclonal antibody to Bcl-2 is commercially available from Sigma Corp. (St. Louis, Mo.), or may be prepared according to standard techniques known in the art. [0127] Candidate compounds which cause the mRNA and/or protein expression profile for cell death-related proteins in cells from a particular Alzheimer's patient to resemble that of a “normal” cell with intact regulation at the G1/S transition, are scored as being potentially useful in the treatment of Alzheimer's disease in that patient. [0128] (6) Assessment of the Expression of DNA Damage Response Element Proteins or Corresponding mRNAs Using Standard Techniques. [0129] This approach may be used when the stimulus used to induce cell cycle arrest at G1/S is DNA damage, for example treatment with a chemical agent which causes DNA damage or exposure to UV radiation. Under normal circumstances the presence of DNA damage will induce a cell to arrest at the G1/S phase transition and attempt to repair the damaged DNA via activation of DNA damage response pathways. Alterations in the pattern of expression of proteins involved in the normal response to DNA damage, or the corresponding mRNAs, in response to the presence of damaged DNA may therefore be used as an indication of the presence of a cell cycle regulatory defect at the G1/S phase transition. Suitable DNA damage response elements include (OMIM accession numbers in parentheses), for example, TP53, Gadd34, Gadd45A(126335), Gadd45B(604948), Gadd45G(604949), Gadd153(126337) and PCNA(176740). A list of OMIM accession numbers for these DNA damage response elements is provided below. Antibodies specific for these proteins are available, or may be prepared according to standard techniques known in the art. [0130] Candidate compounds which cause the mRNA and/or protein expression profile for DNA damage response elements in cells from a particular Alzheimer's patient to resemble that of a “normal” cell with intact regulation at the G1/S transition, are scored as being potentially useful in the treatment of Alzheimer's disease in that patient. [0131] (7) Assessment of the DNA Content of the Non-Neuronal Cells, with or without Cell Cycle Analysis. [0132] In this approach, measurement of the DNA content of cells from the test subject treated with a cell division inhibitor (method (A)) or other stimulus inducing cell cycle arrest (method (B)) provides an indirect indication of the presence of a regulatory defect at the G1/S transition in such cells. The rationale behind this method is the difference in DNA content between cells in the G1 phase and cells in the G2 phase that have passed through the DNA replication stage of the cell cycle. When a population of normal cells (i.e. without a regulatory defect at G1/S) are treated to induce cell cycle arrest in G1 or at G1/S, the majority of the cells will remain in the G1 phase. However, if cells have a regulatory defect at G1/S, a proportion of the cells will pass through the G1/S checkpoint and undergo DNA replication. Thus an increased DNA content in cells from a test subject, as compared to control cells not having a regulatory defect at G1/S, following treatment to induce cell cycle arrest at GI is taken as an indication of the presence of a regulatory defect at G1/S. [0133] Candidate compounds which cause decreased DNA content in cells from a particular Alzheimer's patient exposed to inhibitory treatment (as compared to control cells exposed to no or zero control candidate compound), are scored as being potentially useful in the treatment of Alzheimer's disease in that patient, because they are able to prevent progression through the cell cycle in cells of the Alzheimer's patient. [0134] Online Mendelian Inheritance in Man (OMIM) Accession Numbers for Exemplary Cell Cycle Regulatory Proteins and DNA Damage Response Elements: Gene/protein OMIM Accession number CDKN3 123832 p15ink4B 600431 p16ink4A 600160 p19ink4D 600927 p27kip1 600778 p21cip1 116899 p57kip2 600856 TP53 191170 Gadd45A 126335 Gadd45B 604948 Gadd45G 604949 Gadd153 126337 PCNA 176740 Ku70 152690 KU80 194364 Ku86 604611 NDHII 603115 BLM 604610 RECQL 600537 RECQL4 603780 RECQL5 603781 [0135] Key references for these proteins are as follows: [0136] CDKN3: Cyclin-dependent kinase inhibitor 3, also known as cyclin-dependent kinase interactor 1 (CDI1), Gyuris, J. et al. Cell 75: 791-803, 1993; Hannon, G. J. et al. Proc Natl Acad Sci USA 91: 1731-1735, 1994. [0137] p15ink4B: cyclin dependent kinase inhibitor 2B, also known as CDKN2B, MTS2, TP15, Hannon, G. J. and Beach, D. Nature 371: 257-261, 1994; Quelle, D. E. et al. Oncogene 11: 635-645, 1995; Stone, S. et al. Oncogene 11: 987-991, 1995. Antibody to P15 is commercially available from BD Biosciences Clontech. [0138] p16ink4A: cyclin dependent kinase inhibitor 2A, also known as CDKN2A, MTS1, TP16, Quelle, D. E. et al. Oncogene 11: 635-645, 1995; Kamb, A. et al. Science 264: 436-440, 1994; Kamb, A. et al. Nature Genetics 8: 22-26, 1994. Monoclonal antibody to p16ink4A is commercially available from Sigma Corp. [0139] p19ink4D: cyclin-dependent kinase inhibitor 2D, also known as CDKN2D, Hirai, H. et al. Mol Cell Biol 15: 2672-2681, 1995; Okuda, T. et al. Genomics 29: 623-630, 1995. Antibody to p19 is commercially available from BD Biosciences Clontech. [0140] TP53: tumour protein p53, or p53, Levine, A. J. et al. Nature 351: 453-456, 1991; Levine, A. J. Cell 88: 323-331, 1997. Monoclonal antibody to TP53 is commercially available from Sigma Corp. [0141] p27kip1: cyclin dependent kinase inhibitor 1B, also known as CDKN1B, KIP1, Toyoshima, H. and Hunter, T. Cell 78: 67-74, 1994. Antibodies to p27 are commercially available from BD Biosciences Clontech and Sigma Corp. [0142] p21cip1: cyclin dependent kinase inhibitor 1A, also known as CDKN1A, WAF1, Harper, J. W. et al. Cell 75: 805-816, 1993. Antibody to p21cip1 is commercially available from BD Biosciences Clontech. [0143] p57kip2: cyclin dependent kinase inhibitor 1c, also known as CDKN1C, KIP2, Lee, M-H. et al. Genes Dev 9: 639-649, 1995. [0144] Gadd45A: growth arrest and DNA damage inducible gene alpha, also known as DDIT1, Carrier, F. et al. J Biol Chem 269: 32672-32677, 1994; Kearsey, J. M. et al. Oncogene 11: 1675-1683, 1995; Smith, M. L. et al. Science 266: 1376-1380, 1994. [0145] Gadd45B: growth arrest and DNA damage inducible gene beta, De Smeale, E. et al. Nature 414: 308-313, 2001. [0146] Gadd45C: growth arrest and DNA damage inducible gene gamma, Takekawa, M. and Saito, H. Cell 95: 521-530, 1998. [0147] Gadd153: DNA damage inducible-transcript, also known as CHOP, Ron, D. and Habener, J. F. Genes Dev 6: 439-453, 1992; Park, J. S. et al. Gene 116: 259-267, 1992. [0148] PCNA: proliferating cell nuclear antigen, Travali, S. et al. J Biol Chem 264: 7466-7472, 1989. Antibody to PCNA is commercially available from Sigma Corp. [0149] Ku70: thyroid autoantigen, also known as G22P1, p70, TLAA, Chan, J. Y. C. et al. J Biol Chem 264: 3651-3654, 1989. [0150] Ku80: X-ray repair, complementing defective, in chinese hamster, 5, also known as XRCC5. [0151] Ku86: Cooper, M. P. et al. Genes Dev 14: 907-912, 2000. [0152] NDHII: DEAD/H BOX 9 protein, also known as DDX9, RNA helicase A, nuclear DNA helicase II, Lee, C. G. and Hurwitz, J. J Biol Chem 268: 16822-16830 1992; Zhang, S. and Grosse, F. J Biol Chem 272: 11487-11494, 1993. [0153] BLM: Bloom syndrome gene, also known as RECQ protein-like 3, RECQL3, RECQ2, Ellis, N. A. et al. Cell 83: 655-666, 1995. [0154] RECQL: RECQ protein-like, also known as RECQL1, Puranam, K. L. and Blackshear, P. J. J Biol Chem 269: 29838-29845, 1994; Puranam, K. L. et al. Genomics 26: 595-598, 1995. [0155] RECQL4: RECQ protein-like 4, also known as RECQ4, Kitao, S. et al. Genomics 443-452, 1998. [0156] RECQL5: RECQ protein-like 5, also known as RECQ5, Kitao, S. et al. Genomics 443-452, 1998. Example 2 Protocols for Diagnostic Tests [0157] The following are specific protocols for separation and culture of lymphocytes, induction of cell division, induction of cell cycle arrest by either treatment with a cell division inhibitor or H2O2-induced hypoxia, BrdU incorporation/FACS analysis and MTT survival assay. These protocols, or minor adaptations thereof, may all be used diagnostically to test for the presence of a regulatory defect at the G1/S transition and in compound screening assays. [0158] Lymphocyte Separation [0159] Blood is collected in lithium heparin or EDTA vacutainers. Lymphocytes are isolated according to a standard protocol using Ficoll (Sigma). In order to standardize the culture methods for all patients, the separated lymphocytes are frozen and stored for further analysis. [0160] When lymphocytes are needed for culture, they are thawed in a 37� C. water bath and washed twice in RPMI (any medium or buffer which supports lymphocyte survival may be used to wash the cells with equivalent effect). Cell viability (e.g., by Trypan Blue exclusion) is typically approximately 80-90%. [0161] Lymphocyte Culture, Induction of Cell Division, Induction of Cell Cycle Arrest with Cell-Cycle Inhibitor, BrdU Incorporation and FACS Analysis [0162] Lymphocyte cultures are set up in duplicate in RPMI medium supplemented with 10% FCS at a concentration of 1�106 cells per 1 ml of culture media. Phytohaemaglutinin (PHA) is added to the cultures at a final concentration of 22 μg/ml to activate the lymphocytes. Cultures are incubated for 48 hours at 37� C. in a humidified atmosphere containing 5% CO2. After 48 hours incubation one culture is treated with 100 ng/ml rapamycin, while the other untreated culture is kept as a control. [0163] After a further 23 hours incubation BrdU is added at a concentration of 10 μg/ml to all cultures. After another hour cultures are “collected” and fixed in 70% ice-cold ethanol. [0164] BrdU incorporation is assessed using immunohistochemistry, followed by FACS analysis. The proportion of cells in various phases of the cell cycle is determined and data transformation performed to obtain the relative lengthening of the G1 phase. [0165] Calculations of relative lengthening of the G1 phase of the cell cycle in treated cultures relative to control cultures are based on the assumptions that cells are in the exponential phase of proliferation, and that the growth fraction in the cultures (ratio of dividing cells versus quiescent cells) is 1.0 (Darzynkiewicz Z (1993) In Fantes P and Brooks R (eds) The cell cycle. Oxford University Press, Oxford, pp 43-68). It is also assumed that rapamycin only alters the length of the G1 phase (Wagner E F, Hleb M, Hanna N and Sharma S (1998) J Immunol 161: 1123-31). Based on these assumptions, the relative lengthening of the G1 phase is calculated using the formula: RL=100f−100 (expressed in percent). The ratio of the G1 time in treated versus control cultures: f=TG1tr /TG1c=[ln 2−ln(2−G1tr)][ln(2−G1c)]/[ln(2−G1tr)][ln 2−ln(2−G1c)] (Darzynkiewicz, Z., 1993, ibid.) [0166] Lymphocyte Culture, Induction of Cell Division, Induction of Cell Cycle Arrest with H2O2-Induced Hypoxia, MTT Survival Assay [0167] Four sets of lymphocyte cultures are set up as above. Control cultures are left without any treatment, one set of cultures is treated with 100 ng/ml rapamycin, the third set is treated with 1 μM doxorubicin while the fourth set is treated with 120 μM H2O2. Doxorubicin induces DNA damage, leading to arrest at G2/M, rather than G1/S. H2O2 treatment produces oxidative stress, leading to a reversible and temporary cell cycle arrest at G1/S. [0168] After 20 hours of incubation a 4 hour long MTT survival assay (Chemicon International Ltd) is set up. Results are read using a microplate reader (570 filter, 630 reference filter). The ratio between cell numbers in treated cultures versus controls is expressed as percent. Example 3 Examining the Effects of Cell Cycle Inhibitor Drugs on Cell Division Kinetics and Alzheimer's-Type Protein Expression in the SH-SY5Y Neuroblastoma Cell Line [0169] The object of this study was to demonstrate that exemplary cell cycle inhibitor drugs are able to reduce indicators of Alzheimer's disease pathology in an in vitro model of Alzheimer's disease. There is currently no available animal model for sporadic Alzheimer's disease. However, the SH-SY5Y neuroblastoma cell line (ATCC accession number CRL-2266) is accepted as a suitable in vitro model. [0170] SH-SY5Y cells were treated with three candidate agents: rapamycin (Rapa) at 100 ng/ml, doxorubicin (Doxo) at 1 μM and Dexrazoxane (DexRaz) at 200 μM and then tested one day (d1) and two days (d2) after treatment for the following 7 parameters, which are indicative of cell cycle defects and “AD-type” pathology: [0171] Parameters Indicative of Changes in Cell Cycle/Cell Division Kinetics: [0172] 1) Cell survival and proliferation [0173] 2) Apoptosis [0174] 3) Relative lengthening of the G1 phase of the cell cycle [0175] 4) Relative lengthening of the G2 phase of the cell cycle [0176] Expression of AD-Related Proteins: [0177] 5) Expression of amyloid precursor protein (APP) [0178] 6) Expression of AD-type hyperphosphorylated tau protein [0179] 7) Expression of AD-type paired helical filament (PHF) tau protein [0180] The results are shown in FIGS. 1 to 7. [0181] In order to evaluate the effect of drug pre-treatment is protecting against the effects of oxidative stress cells were subjected to oxidative stress (by treatment with 120 μM H2O2) in the absence of drug treatment and after treatment with rapamycin or dexrazoxane, and then tested for the same parameters. Results are shown in FIGS. 1-7, panel b (FIG. 1b, FIG. 2b, FIG. 3b, FIG. 4b, FIG. 5b, FIG. 6b, FIG. 7b). [0182] Finally, cells were treated with doxorubicin after treatment with rapamycin or dexrazoxane and then tested for the same parameters, to evaluate the effects of combined treatment. Results are shown in FIGS. 1-7, panel c (FIG. 1c, FIG. 2c, FIG. 3c, FIG. 4c, FIG. 5c, FIG. 6c, FIG. 7c). [0183] Methods [0184] SH-SY5Y human neuroblastoma cells were cultured in Dulbecco's modified Eagle's medium (DMEM)/F12 supplemented with 10% fetal calf serum (FCS, PAA Laboratories, Austria), 2 mM L-glutamine, 100 IU/ml penicillin and 100 μg/ml streptomycin (Gibco) in a humidified incubator at 37� C. and 5% CO2. [0185] After 24 h incubation the culture medium was replaced by DMEM/F12 with 10% FCS containing various concentrations of drug. The cells were incubated for another 24 or 48 hours. [0186] Cell survival and proliferation was assessed by MTT proliferation and cell survival assay, performed according to the manufacturers recommendations (Chemicon International, Ltd., Hampshire, UK) in the final 4 hours of the 24 hours period. The results of the assay were evaluated using an Opsys MR ELISA plate reader (Dynex Technologies, Middlesex, UK) at 570 nm with a reference wavelength of 630 nm. [0187] The cell cultures were repeated in identical conditions in order to analyze cell cycle kinetics by flow cytometry and to study protein expression. [0188] The cultures prepared for flow cytometry were harvested after a short incubation with Trypsin EDTA (Sigma) in PBS. Cells were fixed in 70% ice-cold alcohol. Prior to staining with propidium iodide (PI) the samples were washed twice in PBS containing Triton-X at 0.1%. On flow cytometry the ratio of G1 and G2 cells was measured. Cells with sub-G1 DNA content were regarded as the apoptotic fraction. [0189] The cultures prepared for protein expression assays were harvested after a short wash with cold PBS by scraping. The collected cells were lysed in 150 μl of lysis buffer containing 50 mM Tris-HCl (pH 7.4), 100 mM NaCl, 1 mM EDTA, 0.5% (v/v) Nonidet P-40, 50 mM NaF, 0.2 mM Na3VO4, 20 μg/ml phenylmethylsulfonyl fluoride, and 5 μg/ml leupeptin, and incubated on ice for 30 min. The cell lysate was centrifuged at 14,000 g for 20 min at 4� C. to remove insoluble pellets. Protein concentration was determined by the Bradford assay. ELISA assays were performed using a panel of antibodies. Secondary antibodies (anti rabbit or anti mouse as appropriate) conjugated to horseradish peroxidase were used. [0190] Antibodies used in this study: Antigen Antibody Dilution Source Phosphorylated tau AT8 1:1000 Insight Biotechnology PHF tau Tau2 1:1000 Novocastra APP APP 1:1000 Novocastra [0191] Results [0192] The addition of 100 ng/ml rapamycin to the SH-SY5Y neuroblastoma culture resulted in significantly lower cell numbers in the culture than in control cultures (FIG. 1a). This cell number reduction was not due to apoptosis, since rapamycin also protected the neuroblastoma cells against apoptosis (FIG. 2a). The reduced cell numbers were due to lengthening of the cell division process, primarily the G2 phase of the cell cycle (FIG. 4a). This was associated with a small increase in APP production (FIG. 5a), and PHF-tau formation (FIG. 7a). The treatment also increased the amount of hyperphosphorylated tau after one day (FIG. 6a). The cell cycle effect of the rapamycin disappeared after 1 day. Although the cell cycle was accelerated after cells escaped from the rapamycin inhibition after 1 day, the reduction of AD-related proteins was maintained. [0193] Following rapamycin treatment, oxidative stress-induced apoptotic cell death in the cell line increased significantly (FIG. 2b). However, this was also accompanied by a significant lengthening of the G1 phase of the cell cycle (FIG. 3b). This effect was also short lasting, and the cells escaped inhibition after 1 day (FIG. 3b, compare d1 with d2). The rapamycin pre-treated cultures also showed increased tau phosphorylation and PHF formation under oxidative stress (FIG. 6b; FIG. 7b). Interestingly, this increase was followed by a decrease of these proteins, while the initial APP reduction was followed by a slight increase by day 2. [0194] The addition of dexrazoxane to the cell cultures resulted in reduced cell numbers only after 2 days of treatment FIG. 1a). This delayed effect was more interesting since the dexrazoxane did not seem to protect cells against apoptotic cell death as efficiently as rapamycin did (FIG. 2a). The effects of the drug were due to an acceleration of the G1 phase of the cell cycle that was accompanied by little change in G2 time on day one and massive G2 lengthening on day 2 (FIGS. 3a and 4 a). Although the drug increased the expression of all three AD-related proteins on the first day, by day two all three proteins were reduced in the cell lysates (FIGS. 5a, 6 a, 7 a). [0195] Doxorubicin treatment on its own caused reduced cell numbers (FIG. 1a) mainly due to its cell cycle inhibitory effect, since it appeared to be protective against apoptosis (FIG. 2a). This cell cycle inhibitory effect was due mainly to the lengthening of the G2 phase of the cell cycle (FIG. 4a). The effect on G1 was reversed after 1 day (FIG. 3a), while the G2 inhibitory effect diminished (FIG. 4a). This phenomenon led to the elevated expression of APP (FIG. 5a) and PHF tau (FIG. 7a) despite the reduction of phospho-tau (FIG. 6a). [0196] Rapamycin pre-treatment altered the effects of doxorubicin, in that it caused more apoptosis, and lead to a significant lengthening of both the G1 and G2 phases of the cycle after 1 day (FIG. 3c; FIG. 4c). Both effects were reversible and disappeared by day 2. The rapamycin pre-treatment also led to reduced APP and PHF tau formation that lasted after the cell cycle effects disappeared (FIG. 5c; FIG. 7c); the reduction of phospho tau by rapamycin pre-treatment disappeared after 1 day. [0197] Dexrazoxane pre-treatment increased the G2 lengthening effect of doxorubicin (FIG. 4c), and led to the reduction of the APP and PHF-tau production while it did not significantly effect the phospho-tau production. [0198] In summary, the lengthening of the G2 phase under the effect of doxorubicin, although it seems to be protective against apoptotic cell death, also leads to the elevation of AD-related protein expression. [0199] Rapamycin does not seem to be an effective G1 inhibitor in this cell line, causing only a slight change in G1 time on its own. It causes a G2 delay more readily. This in turn is associated with protection against apoptosis and slight changes in the AD-related protein expression, reducing the hyperphosphorylated tau and slightly increasing APP production. Rapamycin pre-treatment is not protective against oxidative stress-induced damage but it accelerated it. However, in combination with doxorubicin it had strong protective effect against the accumulation of AD-related proteins, although the GI inhibitory effects of such treatment were short-lived. [0200] Dexrazoxane effectively reduced AD-related protein expression after two days in culture, however it did not confer protection against cell death on its own. Additionally, when followed by a doxorubicin-induced G2 inhibition, its protective effects against AD-related protein expression became more robust. CONCLUSION [0201] The results of this study provide evidence of the effectiveness of cell-cycle regulatory drugs, or combinations thereof, in reducing parameters of Alzheimer's disease pathology and protecting against oxidative stress-induced damage in a cell culture model of AD. [0202] Each of the patents, patent applications and references that are recited in this application are herein incorporated in their entirety by reference. Having described the presently preferred embodiments, and in accordance with the present invention, it is believed that other modifications, variations and changes will be suggested to those skilled in the art in view of the teachings set forth herein. It is, therefore, to be understood that all such variations, modifications, and changes are believed to fall within the scope of the present invention as defined by the appended claims. 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Inc.Trisubstituted boron-containing moleculesUS20110152217 *Dec 17, 2009Jun 23, 2011Anacor Pharmaceuticals, Inc.Polymorphs of (s)-3-aminomethyl-7-(3-hydroxy-propoxy)-3h-benzo[c][1,2]oxaborol-1-olUS20110166103 *Sep 1, 2009Jul 7, 2011Anacor Pharmaceuticals, Inc.Boron-containing small moleculesUS20110172187 *Jul 14, 2011Anacor Pharmaceuticals, Inc.Boron-containing small moleculesUS20110190235 *Aug 6, 2010Aug 4, 2011Anacor Pharmaceuticals, Inc.Boron-containing small molecules as antiprotozoal agentsUS20110207701 *Aug 16, 2010Aug 25, 2011Anacor Pharmaceuticals, Inc.Boron-containing small molecules as antiprotozoal agentsUS20110207702 *Oct 15, 2009Aug 25, 2011Anacor Pharmaceuticals, Inc.Boron-containing small molecules as anti-protozoal agentsUS20110212918 *Nov 11, 2010Sep 1, 2011Anacor Pharmaceuticals, Inc.Boron-containing small moleculesWO2007095638A2 *Feb 16, 2007Aug 23, 2007Anacor Pharmaceuticals, Inc.Boron-containing small molecules as anti-inflammatory agentsWO2007095638A3 *Feb 16, 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text: Hot Wheels Toys > Refine Price $ to $ Go Top brands LEGO Disney Fisher-Price Paw Patrol Little Tikes Store availability Sort Best sellers Price: low to high Price: high to low Highest rating New As Advertised $9.47 Hot Wheels, 9-Pack 4.5 stars (77) ratings Free shipping on orders over $50 Free store pickup $19.99 Hot Wheels 20-Piece Die-Cast Vehicles Gift Pack 4.5 stars (37) ratings Free shipping on orders over $50 Free store pickup $24.88 Hot Wheels Super Loop Raceway 4.0 stars (21) ratings Free shipping on orders over $50 Free store pickup today $65.50 List price $89.97 Save $24.47 14" Hot Wheels Boy's Bike 4.0 stars (16) ratings Free shipping Free store pickup New $69.00 16" Dynacraft Boy's Hot Wheels Bike 4.0 stars (1) ratings Free shipping Free store pickup today $19.88 Hot Wheels 100 Car Case 3.5 stars (38) ratings Free store pickup today Rollback $17.95 Hot Wheels Spinshotz Hyper Speed Showdown Play Set 5.0 stars (1) ratings Free shipping on orders over $50 Free store pickup $9.99 Hot Wheels Track Builder Digital Speedometer 5.0 stars (1) ratings Free shipping on orders over $50 Free store pickup $6.70 Hot Wheels Track Builder Sideshot Launcher 4.5 stars (3) ratings Free shipping on orders over $50 Free store pickup $5.99 Hot Wheels Track Builder Flip Tower 3.5 stars (2) ratings Free shipping on orders over $50 Free store pickup Reduced Price $25.99 Was $31.19 Save $5.20 Hot Wheels Airbrush Auto Designer 2.0 stars (12) ratings Sold & Shipped by Hobby Warehouse Free shipping $9.47 Hot Wheels Basic Car 9pk Assortment 4.5 stars (125) ratings Free shipping on orders over $50 Free store pickup today $21.99 Angry Birds Hot Wheels Slingshot Launch Track Set 3.5 stars (16) ratings Sold & Shipped by Toywiz $11.32 Was $13.05 Save $1.73 Hot Wheels Scorpion Gravity Track Set Free shipping on orders over $50 Free store pickup New $59.00 12" Dynacraft Boy's Hot Wheels Bike Free shipping Free store pickup today $8.40 Was $10.23 Save $1.83 Hot Wheels Track Builder Rocket Launcher 4.5 stars (2) ratings Free shipping on orders over $50 Free store pickup Rollback $10.10 Hot Wheels Marvel Guardians of the Galaxy Rocket's Tailspin Takedown 5.0 stars (2) ratings Free shipping on orders over $50 Free store pickup $9.46 List price $12.99 Save $3.53 Shakespeare Disney Cars Kit 4.0 stars (25) ratings Free shipping on orders over $50 Free store pickup today $9.38 Hot Wheels Crainiac Track Set 5.0 stars (1) ratings Free shipping on orders over $50 Free store pickup $10.20 Hot Wheels Track Essentials Fan Man Free shipping on orders over $50 Free store pickup $15.59 HOT WHEELS TRACK BUILDER (Assorted Sets) Sold & Shipped by Shippped Free shipping $14.99 Shakespeare Barbie, Tacklebox Kit 182320 4.0 stars (15) ratings Free shipping on orders over $50 Free store pickup $14.03 Hot Wheels Monster Jam Grave Digger Sound Smashers Truck 4.5 stars (4) ratings Free shipping on orders over $50 Free store pickup $14.99 Sakar Hot Wheel Molded Walkie-Talkie Free shipping on orders over $50 Free store pickup Reduced Price $14.99 Was $39.99 Save $25.00 Star Wars Hot Wheels Disney Blast & Battle Lightsaber Car Luncher - 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My model greyhound LOVES when the hats come out! He'll do anything for extra attn. & cookies. : dogswearinghatsjump to contentmy subredditsannouncementsArtAskRedditaskscienceawwblogbookscreepydataisbeautifulDIYDocumentariesEarthPornexplainlikeimfiveFitnessfoodfunnyFuturologygadgetsgamingGetMotivatedgifshistoryIAmAInternetIsBeautifulJokesLifeProTipslistentothismildlyinterestingmoviesMusicnewsnosleepnottheonionOldSchoolCoolpersonalfinancephilosophyphotoshopbattlespicsscienceShowerthoughtsspacesportstelevisiontifutodayilearnedTwoXChromosomesUpliftingNewsvideosworldnewsWritingPromptsedit subscriptionsfront-all-random | funny-Fitness-photoshopbattles-food-WritingPrompts-AskReddit-explainlikeimfive-gifs-books-nosleep-Music-listentothis-aww-DIY-personalfinance-Jokes-pics-creepy-sports-askscience-todayilearned-videos-IAmA-gaming-Documentaries-mildlyinteresting-television-GetMotivated-gadgets-tifu-Futurology-EarthPorn-InternetIsBeautiful-worldnews-movies-OldSchoolCool-science-news-philosophy-Showerthoughts-UpliftingNews-space-TwoXChromosomes-nottheonion-Art-history-LifeProTips-dataisbeautifulmore »reddit.com dogswearinghatscommentsrelatedwant to join? sign in or create an account in seconds|Englishlimit my search to /r/dogswearinghatsuse the following search parameters to narrow your results:subreddit:subredditfind submissions in "subreddit"author:usernamefind submissions by "username"site:example.comfind submissions from "example.com"url:textsearch for "text" in urlselftext:textsearch for "text" in self post contentsself:yes (or self:no)include (or exclude) self postsnsfw:yes (or nsfw:no)include (or exclude) results marked as NSFWe.g. subreddit:aww site:imgur.com dogsee the search faq for details.advanced search: by author, subreddit...this post was submitted on 12 Sep 2013494 points (97% upvoted)shortlink: remember mereset passwordloginSubmit a new linkSubmit a new text postdogswearinghatssubscribeunsubscribe13,106 readers~10 users here nowThe worlds #1 destination for dogs wearing hats.
created by TheT0KERa community for 2 yearsmessage the moderatorsMODERATORSTheT0KERbigbreathein24teflonsteveGobble45The_Real_Artabout moderation team »493494495My model greyhound LOVES when the hats come out! He'll do anything for extra attn. & cookies. (i.imgur.com)submitted 1 year ago by 42Deadsquirrels23 commentssharecancelsorry, this has been archived and can no longer be voted onloading...all 23 commentssorted by: besttopnewhotcontroversialoldrandomq & a[–]fan_fanington 13 points14 points15 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onWhen I"m stressed out, I come to /r/dogswearinghats or (/r/aww) and it always makes me feel better. This dog is hilarious and makes me feel really happy right now. Thanks for posting!
permalink[–][deleted] 7 points8 points9 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onThat has to be the happiest dog I have ever seen!
permalink[–]behindthespine 6 points7 points8 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onI love this dog. I want to squeeze him. permalink[–]lilgillie 4 points5 points6 points 1 year ago (3 children)sorry, this has been archived and can no longer be voted onthat look on his face is supremely adorable, post more pics if you can!
permalink[–]42Deadsquirrels[S] 2 points3 points4 points 1 year ago (2 children)sorry, this has been archived and can no longer be voted onHEHEHE! Oh MAN, can I do that! Xander is seriously the happiest dog I've ever met. You can't help but smile when he's wagging that tail. :)
permalinkparent[–]lilgillie 1 point2 points3 points 1 year ago (1 child)sorry, this has been archived and can no longer be voted onYou're so lucky to have him! What a sweetie! I can't imagine ever being in a bad mood around him :)
permalinkparent[–]42Deadsquirrels[S] 3 points4 points5 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onUntil he eats tampons out of the trash, or needs his stomach pumped because he's eaten yet another toy, WHOLE. Dude keeps us on our toes. Best way to keep a marriage strong is to be 'teamed up' on a common goal, right? permalinkparent[–]okdanasrsly 3 points4 points5 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onhe is absolutely precious! usually i don't find greyhounds to be the most super adorable of breeds, but this guy totally takes the cake, cuteness-wise! i want to snuggle him big time!
permalink[–]neurosciencey 2 points3 points4 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onAw, reminds me of my greyhound...especially the little crooked bottom teeth. ;)
permalink[–]Pibbles 2 points3 points4 points 1 year ago (1 child)sorry, this has been archived and can no longer be voted onNo one believes that my dog is part greyhound, but I see pics like this and I see obvious similarities in their faces.
permalink[–]42Deadsquirrels[S] 3 points4 points5 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onAt Petco-Smart, they have little kits so you can DNA test your dog. It should shut up any 'haters'. I think I saw it for 40 bucks. :)
permalinkparent[–]pursemeatballs 2 points3 points4 points 1 year ago (1 child)sorry, this has been archived and can no longer be voted onI love him. His smile is contagious. What's his name?
permalink[–]42Deadsquirrels[S] 2 points3 points4 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onHis name is Xander. He's a huge goof!
permalinkparent[–]pixelinaa 2 points3 points4 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onAdorable.
permalink[–]bortnib 2 points3 points4 points 1 year ago (2 children)sorry, this has been archived and can no longer be voted onhis face just makes me want to hug him. what a great dog letting you put all these hats on him
permalink[–]42Deadsquirrels[S] 1 point2 points3 points 1 year ago (1 child)sorry, this has been archived and can no longer be voted onXander Doobers is a rescued AKC grey. He's one of those rare non-racers. Poor fella came to us as a 'last chance' home. He was to be destroyed for being aggressive. (IDIOTS! Zero aggression, just needed training & a schedule) He just turned 2 years old and will lick you to death before he'd EVER harm you. He'll do ANYTHING to make you smile on a bad day. He's my best buddy & we feel blessed to have him in our pack. (2 retired racing ladies keep him in line)
permalinkparent[–]bortnib 1 point2 points3 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onoh wow! even just by the pics he looks like the least aggressive dog ever. hes so lucky to have found you and im sure you are so grateful to have found him too. Its amazing how much an animal can thrive being in the right environment! permalinkparent[–]JokeTwoSmoints 2 points3 points4 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onhe's just so adorably silly haha
permalink[–]Hello_Again 2 points3 points4 points 1 year ago (2 children)sorry, this has been archived and can no longer be voted onHe is too cute. I have to ask, where did you get that hat on the right? I'd love to get that for my Airedale/Schnauzer.
permalink[–]42Deadsquirrels[S] 2 points3 points4 points 1 year ago (1 child)sorry, this has been archived and can no longer be voted onI made it! I sell 'snoods' on Etsy. Our store name is Greycraftcorner. Every dime goes directly to Greyhound Options to help with the costs of getting retired racers off the tracks, transported, vetted, & fostered until we find their perfect forever home. A greyhound literally saved my life, so now I do this to help the new guys as they transition to retired life. permalinkparent[–]Hello_Again 1 point2 points3 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onThat's absolutely amazing! I know what I'm getting my pup for Christmas. They're really cute, but the adorable model sold it to me. permalinkparent[–]UltimateXavior 1 point2 points3 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted on"snoop dogg putting some hats on" permalink[–][deleted] 1 point2 points3 points 1 year ago (0 children)sorry, this has been archived and can no longer be voted onThis is the hardest I have ever laughed.
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Movie Night: "The Great Gatsby" - The NoVA Banned Book Club (Fairfax, VA)
Bow Tie Cinemas - Reston Town Center 11
We recently read Fitzgerald's "The Great Gatsby" and the movie has been readapted to the big screen. Check out the movie's trailer here: https://www.youtube.com/watch?v=rARN6agiW7o
The more the merrier so invite your family & friends (just indicate so when your RSVP). On Tuesday evening I'll send out an e-mail on where to meet exactly, my cell phone #, etc. The movie starts at 7:30pm so we should plan on meeting there by 7:15pm. Adult tickets are $12. Hope to see you guys there!
PS I'll probably get there a little earlier and there are plenty of good restaurants/bars nearby. If you're up for grabbing a bite and/or drink with me then let me know and we can plan something!
Laur Enjoyed the movie, but would have preferred a different soundtrack. I left feeling inspired to buy a pink suit. And I certainly wouldn't mind meeting up at RTC again! 1 · May 16 Lorenzo Great group as usual and enjoyed the discussions! The movie was..interesting. I thought the acting was great. 1 · May 16 Melissa Long I had a really good time. 1 · May 15 Rhett Great group! Enjoyed many parts of the film. 1 · May 15 A former member just me now 0 · May 15 Lorenzo I sent a message with details to those who have RSVP'ed. Most importantly, we'll meet in the main lobby of the cinema at 7:15pm. 0 · May 14 A former member might bring a friend along 1 · May 12 Laur I'm game for meeting up early for food/drinks. And there are usually some good wine specials at RTC on Wednesdays. 0 · May 12 View all 6 replies
Lorenzo Sounds great! Deah's suggestion works for me. I am a bit of a wino too so if you know of any place that has wine specials then let me know. 0 · May 12 Laur Uno's always has good specials. You can also check out the happy hour menus for Mon Ami Gabi if you're a wino. :) 0 · May 12 Laur http://www.monamigabi... This place is definitely a bit posh, which could set the mood for watching Gatsby. But I'm up for anywhere! 0 · May 12 A former member i love wine. lols any place with the aestheics of a speak easy? 1 · May 12 Lorenzo Mon Ami Gabi is such a great place!! Are others interested in meeting there? I tried their escargots a while back...pretty tasty ;-) 0 · May 12 Rhett Think it is still a good happy hour. Lagunitas Pilsner on draft. Reasonable sliders. 0 · May 12 Rhett !!! 0 · May 12 Deah Hester I will try to bring my husband. Lorenzo, there is an Uno's right next door and I think they have good Happy Hour special from 5-7 maybe. 2 · May 12 10 attended
Resident Bukowski Scholar, Co-Organizer
$2.00 At each event
Deah Hester, Rhett
This covers: Dues cover the cost and maintenance of the website.
- “$2.00 cash paid at each event or online with PayPal (preferred method).”

value: This website is a service used to organize online groups that host in-person and virtual events for people with similar interests. It was founded in 2002 by Chairman Scott Heiferman and four co-founders.


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Select item 240456911.Trans-illuminated laser speckle imaging of collateral artery blood flow in ischemic mouse hindlimb.Meisner JK, Niu J, Sumer S, Price RJ.J Biomed Opt. 2013 Sep;18(9):096011. doi: 10.1117/1.JBO.18.9.096011.PMID: 24045691 Free PMC ArticleSimilar articles Select item 263382972.Mechanisms of Amplified Arteriogenesis in Collateral Artery Segments Exposed to Reversed Flow Direction.Heuslein JL, Meisner JK, Li X, Song J, Vincentelli H, Leiphart RJ, Ames EG, Blackman BR, Blackman BR, Price RJ.Arterioscler Thromb Vasc Biol. 2015 Nov;35(11):2354-65. doi: 10.1161/ATVBAHA.115.305775. Erratum in: Arterioscler Thromb Vasc Biol. 2015 Nov;35(11):e58. Blackman, Brett R [added]. 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Schaper W, Schmitz-Rixen T.Arterioscler Thromb Vasc Biol. 2004 Sep;24(9):1664-8.PMID: 15242864 Free ArticleSimilar articles Select item 251944707.Impaired arteriogenesis in syndecan-1(-/-) mice.Tang GL, Weitz K.J Surg Res. 2015 Jan;193(1):22-7. doi: 10.1016/j.jss.2014.08.002.PMID: 25194470 Similar articles Select item 149630078.Collateral artery growth (arteriogenesis) after experimental arterial occlusion is impaired in mice lacking CC-chemokine receptor-2.Heil M, Ziegelhoeffer T, Wagner S, Fernández B, Helisch A, Martin S, Tribulova S, Kuziel WA, Bachmann G, Schaper W.Circ Res. 2004 Mar 19;94(5):671-7.PMID: 14963007 Free ArticleSimilar articles Select item 209301479.Role of PECAM-1 in arteriogenesis and specification of preexisting collaterals.Chen Z, Rubin J, Tzima E.Circ Res. 2010 Nov 26;107(11):1355-63. doi: 10.1161/CIRCRESAHA.110.229955.PMID: 20930147 Free PMC ArticleSimilar articles Select item 1116627510.Revascularization in the rabbit hindlimb: dissociation between capillary sprouting and arteriogenesis.Hershey JC, Baskin EP, Glass JD, Hartman HA, Gilberto DB, Rogers IT, Cook JJ.Cardiovasc Res. 2001 Feb 16;49(3):618-25.PMID: 11166275 Similar articles Select item 1576801511.The effect of gradual or acute arterial occlusion on skeletal muscle blood flow, arteriogenesis, and inflammation in rat hindlimb ischemia.Tang GL, Chang DS, Sarkar R, Wang R, Messina LM.J Vasc Surg. 2005 Feb;41(2):312-20.PMID: 15768015 Free ArticleSimilar articles Select item 1902805312.Limb ischemia after iliac ligation in aged mice stimulates angiogenesis without arteriogenesis.Westvik TS, Fitzgerald TN, Muto A, Maloney SP, Pimiento JM, Fancher TT, Magri D, Westvik HH, Nishibe T, Velazquez OC, Dardik A.J Vasc Surg. 2009 Feb;49(2):464-73. doi: 10.1016/j.jvs.2008.08.077.PMID: 19028053 Free PMC ArticleSimilar articles Select item 775818713.Recovery of disturbed endothelium-dependent flow in the collateral-perfused rabbit ischemic hindlimb after administration of vascular endothelial growth factor.Bauters C, Asahara T, Zheng LP, Takeshita S, Bunting S, Ferrara N, Symes JF, Isner JM.Circulation. 1995 Jun 1;91(11):2802-9.PMID: 7758187 Free ArticleSimilar articles Select item 2070549314.Variations in surgical procedures for hind limb ischaemia mouse models result in differences in collateral formation.Hellingman AA, Bastiaansen AJ, de Vries MR, Seghers L, Lijkwan MA, Löwik CW, Hamming JF, Quax PH.Eur J Vasc Endovasc Surg. 2010 Dec;40(6):796-803. doi: 10.1016/j.ejvs.2010.07.009.PMID: 20705493 Free ArticleSimilar articles Select item 1238825815.Blood monocyte concentration is critical for enhancement of collateral artery growth.Heil M, Ziegelhoeffer T, Pipp F, Kostin S, Martin S, Clauss M, Schaper W.Am J Physiol Heart Circ Physiol. 2002 Dec;283(6):H2411-9.PMID: 12388258 Free ArticleSimilar articles Select item 1880202316.Vascular endothelial growth factor-A specifies formation of native collaterals and regulates collateral growth in ischemia.Clayton JA, Chalothorn D, Faber JE.Circ Res. 2008 Oct 24;103(9):1027-36. doi: 10.1161/CIRCRESAHA.108.181115.PMID: 18802023 Free PMC ArticleSimilar articles Select item 1466008617.Therapeutic angiogenesis/arteriogenesis in the chronic ischemic rabbit hindlimb: effect of venous basic fibroblast growth factor retroinfusion.Lebherz C, von Degenfeld G, Karl A, Pfosser A, Raake P, Pachmayr F, Scholz D, Kupatt C, Boekstegers P.Endothelium. 2003;10(4-5):257-65.PMID: 14660086 Similar articles Select item 1455384018.Enhanced hindlimb collateralization induced by acidic fibroblast growth factor is dependent upon femoral artery extraction.Hershey JC, Corcoran HA, Baskin EP, Gilberto DB, Mao X, Thomas KA, Cook JJ.Cardiovasc Res. 2003 Oct 1;59(4):997-1005.PMID: 14553840 Similar articles Select item 2043106119.Endothelial nitric oxide synthase deficiency causes collateral vessel rarefaction and impairs activation of a cell cycle gene network during arteriogenesis.Dai X, Faber JE.Circ Res. 2010 Jun 25;106(12):1870-81. doi: 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Patent US5048374 - Vehicle speed control system for motor vehicle having an automatic ... - Google PatentsSearch Images Maps Play YouTube News Gmail Drive More »Sign inAdvanced Patent SearchPatentsA vehicle speed control system comprising an automatic speed control system for controlling the vehicle velocity to a set velocity selected by an operator, and automatic transmission control system for controlling the speed stage of the automatic transmission of the vehicle in response to the deviation...http://www.google.com/patents/US5048374?utm_source=gb-gplus-sharePatent US5048374 - Vehicle speed control system for motor vehicle having an automatic transmission control systemAdvanced Patent SearchPublication numberUS5048374 APublication typeGrantApplication numberUS 07/413,765Publication dateSep 17, 1991Filing dateSep 28, 1989Priority dateNov 29, 1986Fee statusLapsedAlso published asDE3786894D1, DE3786894T2, DE3789513D1, DE3789513T2, EP0270036A2, EP0270036A3, EP0270036B1, EP0469645A2, EP0469645A3, EP0469645B1, EP0473204A2, EP0473204A3, US5088351Publication number07413765, 413765, US 5048374 A, US 5048374A, US-A-5048374, US5048374 A, US5048374AInventorsOsamu Miyake, Nobuyasu Suzumura, Shoji KawataOriginal AssigneeAisin Seiki Kabushiki KaishaExport CitationBiBTeX, EndNote, RefManPatent Citations (15), Referenced by (7), Classifications (25), Legal Events (4) External Links: USPTO, USPTO Assignment, EspacenetVehicle speed control system for motor vehicle having an automatic transmission control systemUS 5048374 AAbstract A vehicle speed control system comprising an automatic speed control system for controlling the vehicle velocity to a set velocity selected by an operator, and automatic transmission control system for controlling the speed stage of the automatic transmission of the vehicle in response to the deviation between the actual vehicle velocity and the set vehicle velocity of the automatic speed control system. The automatic transmission control system comprises two operating modes including a first mode when the automatic speed control system is controlling the vehicle velocity, and a second mode when the automatic speed control system is not controlling the vehicle velocity, and the automatic transmission control system prohibits the shift of the speed stage to a fixed speed stage when the automatic speed control system is controlling the vehicle velocity. The automatic speed control system does not detect a shift of the speed stage when the automatic speed control system is controlling the throttle valve during shifting of the speed stage. The automatic speed control system controls a throttle valve in response to an abnormal state of the transmission.
What is claimed is: 1. A speed control system for a vehicle having an automatic transmission with a plurality of shiftable speed stages, the automatic transmission being capable of shifting from an operating speed stage to another speed stage and capable of maintaining an operating speed stage when in a normal condition, as required by a deviation between the actual vehicle velocity and throttle opening, the speed control system, comprising:velocity setting means operative when activated for controlling the throttle to maintain a set vehicle velocity and deactivated for discontinuing the set vehicle velocity; transmission control means governed by the set vehicle velocity for either maintaining the operating speed stage or alternatively shifting from the operating speed stage to another required speed stage in accordance with the actual vehicle velocity and the operating speed stage of the automatic transmission; failure detecting means for detecting the inability of the automatic transmission either to maintain the operating seed stage or alternatively the inability to shift to the required speed stage; and deactivating means responsive to the failure detecting means for deactivating the velocity setting means in response to the detection of the inability to maintain the operating speed stage or alternatively detection of the inability to shift from the operating speed stage to another required speed stage. 2. The system of claim 1, further comprising means for rendering the deactivating means inoperative to deactivate the velocity setting means at times when the transmission control means requires shifting from the operating speed stage to another speed stage higher than the operating speed stage.
3. The system of claim 2, wherein the automatic transmission control means includes means for inhibiting a required down shifting of the speed stage to a lower speed stage when the actual vehicle velocity and the throttle opening are both greater than preset levels.
4. A vehicle speed control system according to claim 1, further comprising means for deactivating the velocity setting means at times when said actual vehicle velocity slows to a predetermined value.
5. The system of claim 1, wherein the automatic transmission includes a plurality of solenoids, selectively energized for determining the operating speed stage, and said failure detecting means includes means for detecting both an electrical short and open condition of each of said plurality of solenoids.
6. The system of claim 5, wherein said means for detecting the electrical short and open condition includes means for sampling output data of the automatic transmission, comparing input data to each solenoid with output data from each solenoid, and detecting a failure of a respective solenoid when the corresponding output data is equal to the corresponding input data.
7. The system of claim 1 wherein the automatic transmission control means includes means for requiring the shifting of the speed stage of the automatic transmission of the vehicle in response to both the deviation between the actual vehicle velocity and the set vehicle velocity, and the actual vehicle velocity.
8. The system of claim 7 wherein the transmission control means includes delay means for delaying the required shifting of the speed stage of the automatic transmission of the vehicle.
9. The system of 8 wherein the delay means includes means for shortening the delay period of the required shifting when the deviation between the actual vehicle velocity and the set vehicle velocity is greater than a predetermined value.
10. A vehicle speed control system according to claim 1 wherein the automatic transmission control means includes means for suppressing a required shift of the speed stage to the next speed stage in response to at least one of the actual vehicle velocity and the throttle opening.
11. The system of claim 10 wherein the automatic transmission control means includes means for prohibiting a required shift from the operating speed stage to a higher speed stage at times when the vehicle velocity and the throttle opening are below predetermined levels.
12. A method of operating an automatic speed control systems at times when an automatic transmission, having a plurality of solenoid operated shiftable speed stages, is abnormally caused to shift to another speed stage or abnormally prevented from shifting from an operating speed stage to another speed stage and the speed control system is activated for a selected vehicle speed setting, the method comprising:(a) sampling solenoid output data of the automatic transmission; (b) comparing the sampled solenoid output data with solenoid input data; (c) detecting a solenoid failure when solenoid output data is equal to solenoid input data; and (d) deactivating the speed control system at times when a solenoid failure for a speed stage is detected, and the automatic transmission requires shifting from the operating speed stage to a speed stage lower than the operating speed stage. Description
This is a division of application Ser. No. 07/126,901, filed Nov. 30, 1987 now abandoned.
SUMMARY OF THE INVENTION It is an object of the present invention to maintain a comfortable ride for a vehicle having both an automatic transmission control system and an automatic speed control system.
BRIEF DESCRIPTION OF THE INVENTION FIG. 1 is an circuit diagram showing the electric control system for controlling operation of an automatic speed control system and an automatic transmission control system according to one embodiment of the present invention;
FIG. 5 is a flow chart showing a sub routine SHIFT JUDGMENT for the CPU;
DESCRIPTION OF PREFERRED EMBODIMENT Referring to FIG. 1, there is shown an electric circuit of a vehicle speed control system for a motor vehicle having an automatic transmission control system (an automatic transmission control means) and an automatic speed control system (an automatic speed control means or a throttle opening control means). In this embodiment, the electrical control device is mainly comprised of a microcomputer CPU. The CPU has a microprocessor unit MPU (not shown), a programmable timer PTM (not shown), input ports P1 through P18, output ports P21 through P30, a random access memory RAM (not shown) and a read only memory ROM (not shown). The MPU has two accumulators A and B (not shown) and an index register X (not shown). The MPU executes a program stored in the ROM.
A battery BE is connected to a regulated voltage supply CON via an ignition switch IG. The battery BE is a battery established in the motor vehicle generally. The regulated voltage supply CON regulates the voltage of the battery BE to 5 volts. The microcomputer CPU, an input interface circuit lP and an output interface circuit OP are supplied with the 5 volts from the voltage supply CON.
A speed sensor SP2 comprises a read switch for generating a frequency signal proportional to the rotational speed of an output shaft (not shown) of the automatic transmission by a permanent magnet (not shown) connected to the output shaft. The speed sensor SP2 is connected to the input port P2 of the CPU via an input interface comprising resistors R4, R5, R6 and R7, and a transistor Q2. When the speed sensor SP2 is closed, the input port P2 changes to the "H" level. When the speed sensor SP2 is opened, the input port P2 changes to the "L" level.
A mode switch MS for setting the mode in the vehicle speed control to an economy mode, a power mode or an auto drive mode is comprised of an economy mode switch MS-E, a power mode switch MS-P and an auto drive mode switch MS-A. In the economy mode, the economy mode switch MS-E is turned ON, and the power mode switch MS-P and the auto drive mode switch MS-A are turned OFF. In the power mode, the power mode switch MS-P is turned ON, and the economy mode switch MS-E and the auto drive mode switch MS-A are turned OFF. In the auto drive mode, the auto drive mode switch MS-A is turned ON, and the economy mode switch MS-E and the power mode swtich MS-P are turned OFF. The power mode switch MS-P and the auto drive mode switch MS-A are connected to the input ports P6 and P7 of the CPU, respectively, via input interfaces comprising resistors R11, R12, R13 and R14, and buffers DR4 and DR5. When the power mode switch MS-P is turned ON or OFF, the input port P6 is changed to the "H" level or the "L" level, respectively. When the auto drive mode switch MS-A is turned ON or OFF, the input port P7 is changed to the "H" lever or the "L" level, respectively.
A throttle opening sensor SS is for detecting the opening degree of the throttle valve (not shown) and comprises three contacts L1, L2 and L3 for dividing the throttle opening in to eight stages TH0 through TH7 and a contact IDL for detecting the engine idling condition. The relationship between the throttle opening THO through TH7 and and the states of contacts L1, L2 and L3 is as follows:
______________________________________THROTTLE    STAGES OF CONTACTSOPENING     L1           L2     L3______________________________________TH0         OFF          OFF    OFFTH1         OFF          OFF    ONTH2         OFF          ON     ONTH3         OFF          ON     OFFTH4         ON           ON     OFFTH5         ON           ON     ONTH6         ON           OFF    ONTH7         ON           OFF    OFF______________________________________
A parking brake switch RK is turned ON when a parking brake (not shown) is operated. A set switch SP and resume switch RS for the vehicle speed control system are each comprised of a push button type switch wherein the switch is turned ON when the switch is pushed and is turned OFF when the switch is not pushed. A vacuum switch VS is for detecting vacuum in a surge tank (not shown). Such vacuum is required to operate the vehicle speed control system. The vacuum switch VS is turned ON when the vacuum pressure is higher than a preset value (In this case the vacuum pressure approaches atmospheric pressure). The parking brake switch PK, the set switch SP, the resume switch RS and the vacuum switch VS are connected to the input ports P14, P15, P16 and P17 of the CPU, respectively, via input interfaces comprising diodes D3, D4, D5 and D6, resistors R31, R34, R37 and R40, and transistors Q6, Q7, Q8 and Q9. When each of switches PK, SP, RS and VS is tuned ON, the input ports P14, P15, P16 and P17 are changed to the "H" level, respectively. When each of switches PK, SP, RS and VS is turned OFF, the input ports P14, P15, P16 and P17 are changed to the "L" level.
An actuator ACT1 for the automatic transmission comprises shift solenoids SL1 and SL2 and lockup solonoid SL3. The shift solenoids are for shifting of automatic transmission gearing. The relationship between the shift solenoids SL1 and SL2 with respect to the times they are electrically energized and electrically deenergized and the condition of the automatic transmission gearing are as follows:
______________________________________SHIFTSOLE-    SPEED STAGESNOIDS    1st speed 2nd speed 3rd speed                                O/D speed______________________________________SL1      Energized Energized De-     De-                        energized                                energizedSL2      De-       Energized Energized                                De-    energized                   energized______________________________________
The lockup solenoid SL3 is for controlling a lockup clutch (not shown) for accomplishing direct engagement an output shaft of an engine (not shown) with an input shaft of a torque converter (not shown). When the lockup solenoid SL3 is energized, the lockup clutch is engaged and therefore the lockup operation is accomplished. When the lockup solenoid SL3 is deenergized, the lockup clutch is disengaged and therefore the lockup operation is released. The solenoids SL1, SL2 and SL3 are operated in accordance with the condition of the output ports P21, P23 and P25 of the CPU respectively. A buffer DR11, a transistor Q21 and a diode D11 comprise an output interface for the shift solenoid SL1. When the output port P21 is changed to the "H" level, the solenoid SL1 is deenergized. When the output port P21 is changed to the "L" level, the solenoid SL1 is energized. A buffer DR12 and resistors R51 and R54 comprise a fail detecting circuit for the solenoid SL1. The solenoid SL1 is connected to the input port P22 via the fail detecting circuit. When the wire of the solenoid or the wire between the output interface and the solenoid is cut or broken, the input port P22 is changed to the "H" level. When the wire between the output interface and the solenoid SL1 is short-circuited to ground, the port P22 is changed to the "L" level. A buffer DR13, a transistor Q22 and a diode D12 comprise an output interface of the solenoid SL2. A buffer DR13 and resistors R52 and R55 comprise a fail detecting circuit for the solenoid SL2. A buffer DR15, a transistor Q23 and a diode D13 comprise an output interface for the solenoid SL3. A buffer DR16 and resistors R53 and R56 comprise a fail detecting circuit for the solenoid SL3. When the output port P23 is changed to the "H" level, the solenoid SL2 is deenergized. When the output port P23 is changed to the "L" level, the solenoid SL2 is energized. When the wire of the solenoid SL2 or the wire between the output interface of the solenoid SL2 and the solenoid SL2 is cut or broken, the input port P24 is changed to the "H" level. When the wire between the output interface of the solenoid SL2 and the solenoid SL2 is short-circuited to ground, the port P24 is changed to the "L" level. When the output port P25 is changed to the "H" level, the solenoid SL3 is deenergized. When the output port P25 is changed to the "L" level, the solenoid SL3 is energized. When the wire of the solenoid SL3 or the wire between the output interface of the solenoid SL3 and the solenoid SL3 is cut or broken, the input port P26 is changed to the "H" level. When the wire between the output interface of the solenoid SL3 and the solenoid SL3 is short-circuited to ground, the port P26 is changed to the "L" level.
After step 50, a sub routine MODE CHANGE (step 60), a sub routine MAP CHANGE (step 80), a sub routine SHIFT JUDGMENT (step 100), a sub routine LOCKUP DATA SET (step 110), a sub routine SHIFT CONTROL (step 120) and a sub routine AUTOMATIC SPEED CONTROL (step 180) are executed continuously, and step 50 is executed again after these are completed.
The sub routine MAP CHANGE as shown in FIG. 4 is executed after sub routine MODE CHANGE. When flag F-ECT-A/D is "1" (step 81), or when flag F-ECT-A/D is "0" and flag F-ACC is "1" and variable Vdev is not lower than 0.5 kilometer per hour (step 81 and step 84, 85 and 86), a constant ECT-SMAP is set to an address pointer Ps of the shift data and a constant ECT-LMAP is set to an address pointer P1 of the lockup data (step 82 and 83). When flag F-ECT-A/D is "0" and flag F-ACC is "0" and variable TH is equal to constant THmax (step 81, 84 and 90), or when flag F-ECT-A/D is "0" and flag F-ACC is "0" and variable TH is equal to constant THmax and flag F-ACC-SEN is "1" (step 81, 84, 90 and 91), a constant ECT-SMAP is set to an address pointer Ps of the shift data and a constant ECT-LMAP is set to an address pointer P1 of the lockup data (step 82 and 83) after flag F-ACC is set to "1" and timer T-DEL-UP is set to "0" (step 92 and 93). Variable Vdev is calculated in step 85 as the absolute value of the deviation between a present vehicle velocity Vc and the setting vehicle velocity Vset. The present vehicle velocity Vc is calculated by signals from the speed sensor SP1 and the speed sensor SP2. The setting vehicle velocity Vset is the vehicle velocity at the time which the set switch SP is turned OFF. Variable TH is the present throttle opening. Constant THmax is the maximum throttle opening, and the value of the THmax is "TH7". Thus, step 90 is a judgment step to determine whether the present throttle opening is "TH7". Flag F-ACC-SEN is set to "1" at timer interrupt, which is described below, when the amount of the throttle opening is large. Flag F-ACC is set to "1" if the present throttle opening is "TH7" or if the throttle opening is rapidly increased when the acceleration is not stepped at ECT-A/D mode. Flag F-ACC is set to "0" if the deviation between the present vehicle velocity Vc and the setting vehicle velocity Vset is smaller than 0.5 kilometer per hour when the acceleration is stepped at ECT-A/D mode. In this way, the value of the head address of ECT shift data (ECT-SMAP) is set to the address pointer Ps, and the value of the head address of ECT lockup data (ECT-LMAP) is set to the address pointer P1 at the ECT mode, at the time that the deviation Vdev is large but the accelerator is stepped at ECT-A/D mode, and at the time that the throttle opening is a maximum or moves quickly when the accelerator is not stepped at ECT-A/D mode. Therefore ECT shift and lockup data are used for shift and lockup judgment in this case.
The sub routine LOCKUP DATA SET, as shown in FIG. 6, is executed after sub routine SHIFT JUDGMENT. The deviation Vdev is set to the absolute value of the difference between the present vehicle velocity Vc and the setting vehicle velocity (step 112). When the deviation Vdev is larger than 2 kilometers per hour (step 113), flag F-EN-L/U is set to "0" (step 114), timer T-DIS-L/U is set to 5 seconds (step 115) and timer T-DIS-L/U is started (step 116). The timer T-DIS-L/U is decreased 1 bit according to a predetermined time, and becomes "0" after 5 seconds. When the deviation Vdev is smaller than 0.5 kilometers per hour (steps 113 and 117), flag F-EN-L/U is set to "1" (step 118). Flag F-EN-L/U is a lockup allowance flag. If the flag F-EN-L/U is "1", then the engaging of the lockup clutch is enablod, and if the flag F-EN-L/U is "0" than the engaging of the lockup clutch is not enabled during sub routine SHIFT CHANGE, described later. These operations are not executed if flag F-ECT-A/D is "0" (ECT mode).
Referring to FIGS. 7 through 9 again, when the mode is ECT-A/D mode (flag F-ECT-A/D) is "1") and the acceleration is not operated (flag F-ACC is "0"), steps 132 through 143 are executed except for the case when the present vehicle velocity is lower than 50 kilometers and the setting speed stage in step 123 is 3rd speed. When the present vehicle velocity is lower then 50 kilometers and the setting speed stage in step 123 is 3rd speed, steps 132 through 160 are skipped. In this way, the shift for 3rd speed is not done, and the time of the shift of the speed stage is decreased, so the feeling to the vehicle occupants is more comfortable. The step 131 is able to change to the step in FIG. 36. In this routine, if each of the vehicle velocity Vc and the throttle opening TH is small, then the shift for O/D speed is not done. Also, if the throttle opening TH is large, then the shift for 1st and 2nd stage is not done. In step 132 through 143, if the shift of the speed stage is to a higher stage, the index register X is set to the value of constant ECT-A/D-UT, and sub routine TEMPORARY TIMER SET is executed. Further, if flag F-DEL-UP is "0", then timer T-DEL-UP is set to 4 seconds and the timer is started. In step 138, the sub routine UP SHIFT PROHIBITION is executed. Timer T-DEL-UP is an up shift delay timer, and delays the change of the speed stage until the timer become "0". In steps 132 through 143, if the shift of the speed stage is a down shift, the index register X is set to a constant ECT-A/D-DT, and sub routine TEMPORARY TIMER SET is executed. Further, timer T-DIS-UP is set to 5 seconds and the timer is started. Also, flag F-DEL-UP is set to "0".
The sub routine GET MAX TRQ is shown in FIG. 18. The index register X has been set to the value calculated as the variable Vc in the sub routine GET TRQ. In step 291, the value in register A is replaced by a value calculated by subtracting 2 from the last value in the register A. The value in the index register X is replaced by a value calculated by multiplying the last value in the index register X by 3 and adding the value in the register A. Further, the value in the index register X is replaced by a value calculated by adding the constant TBL-MT to the last value in the index register X. In step 294, the register A is set to the contents of the address indicated by the index register X. The constant TBL-MT indicates the head address of the maximum torque data, and the register A contains a value which indicates the maximum torque of the vehicle, corresponding to the variable TN and the maximum torque data shown in FIG. 26. The variable TN shows the present torque, and the variable Vc shows the present vehicle velocity.
Referring to the sub routine SHIFT CHANGE (FIG. 7, 8 and 9) again, in step 144 (FIG. 8), if timer T-DEL-UP is "0" then flag F-DEL-UP is set to "0" and the shift of the speed stage is allowed. After that, if the timers T-T1, T-T2, T-T3, T-T4 and T-T5 are all "0" (the shift of speed stage is not done at the present time), then the variables T1-TEM through T5-TEM are set to the timers T-T1 through T-T5, and timers T-T1, T-T2 and T-T4 are started. If one of the timers T-T1 through T-T5 is not "0", the deviation Vdev is set to the absolute value of the difference between the present vehicle velocity and the setting vehicle velocity. When the deviation is larger than 3 kilometers per hour, the values of the timers T-T1 through T-T5 are changed by subtracting the value of the passage time of the timers from the value of the temporary timers. The value of the passage time of the timers is calculated by subtracting the value of the present time of the timers form the value of the last temporary timers. If the changed value is less than zero, the changed value is replaced with zero. In this way, if the deviation Vdev is large, the value of the timers is decreased, and the shift of the speed stage occurs earlier. As shown in FIG. 19, when the deviation Vdev is small, the speed stage is not shifted for small changes of the throttle opening. As shown in FIG. 18, when the deviation is large, the shift of the speed stage is done for small changes of the throttle opening. In this way, when the deviation Vdev is large, the sensitivity is increased.
When timer T-T1 is "0" or the timer T-T1 ends, timer T-T3 is started (step 163). The end of timer T-T1 is at the time that the timer T-T1 is nearly "0". After that, when timer T-DIS-UP is "0" and flag F-DWN is "1" and when the timer T-DIS-UP is "0", Flag F-DWN is "0" and flag F-DIS-UP is "0", the data of the speed stage is sent to the output ports P21 and P23 (step 167). In short, when the up shift is not allowable, the shift of the speed stage of the transmission is not executed.
After the timer operation, sub routine SOLENOID ABNORMAL DETECTING is executed (step 175). This routine is an abnormal detecting means. Referring to FIG. 1, when the output port P21 is high level, if the solenoid SL1 is normal, the input port P22 is turned to low level, and if the solenoid SL1 is opened, the input port P22 is turned to high level. When the output port P21 is low level, if the solenoid SL1 is normal, the input port P22 is turned to high level, and if each side of the solenoid SL1 is short circuited, then the input port P22 is turned to low level. In this way, if the solenoid SL1 is normal, when the output of the output port P21 is "1" (high), the input of the input port P22 is changed to "0" (low), and when the output is "0" (low), the input is "1" (high). The relation of the other output ports P23 and P25 and input ports P24 and P26 are same. Accordingly, if the value sent to the output port of a solenoid is equal to the value received at the input port of the solenoid, the failure of the solenoid is detected. Referring to FIG. 15, if the output data for solenoids is equal to the input data, then flag F-FAIL is set to "1" and the failure of the solenoid is able to be detected. When the solenoid is normal, flag F-FAIL is set to "0".
Referring to FIG. 10 again, if the timer T-T4 is not "0", then step 197 is executed. If the timer T-T4 is "0" and the timer T-T5 is "0", then flag F-TH-HLD is set to "0" and step 197 is executed. If the timer T-T4 is "0" and the timer T-T5 is not "0", then the fixed duty rate control is executed in steps 208 and 209 in FIG. 11. Referring to FIG. 11, the variable THOUT is set to the value of register B that is stored with the value of the throttle opening after the shift of the speed stage. In step 209, the output port P29 is turned to low level, and the output ports P27 and P28 are controlled to keep the duty rate corresponding to the value of the variable THOUT (that is the fixed duty rate control).
In step 197, if flag SW-A/D-MAIN is "0", then flag F-A/D-CAN is set to "1", flag F-A/D-SET is set to "0" and the step 216 is executed. If flag SW-A/D-MAIN is "1" and flag SW-A/D-SET is "1", then the flag F-A/D-CAN is set to "0"and flag F-A/D-SET is set to "1". Further, timer T-DEL-MODE is set to one second and the timer is started. Thus, the timer T-DEL-MODE is started at ECT-A/D mode when the set switch SP is turned ON and delays the change of modes. For example, this relationship is shown in FIG. 21. When the mode is changed, the-speed stage is apt to shift, a shift of the speed stage does not occur at the same time the automatic speed control is set. Referring to FIG. 11 again, when flag F-ECT-A/D is "1", and flag F-FAIL is "0", the SPEED CONTROL routine (step 205) is executed. In this step, the vehicle velocity is held by controlling the duty rate of the output for the release valve RV and the control valve CV. Then flag F-TH-HLD is set to "0" (step 207), and the fixed duty rate control is cancelled. If the flag F-FAIL is "1", then flag F-A/D-CAN is set to "1", the automatic control is cancelled, and flag F-TH-HLD is set to "0". If the flag F-ECT-A/D is "0", then the sequence of steps 204 through 207 is not executed. In this routine, if the automatic transmission control system has a failure, the automatic speed control is cancelled. When a shift of the speed stage occurs by control of the throttle valve by the automatic speed control, if the solenoid of the transmission has failed, then the shift may be an unexpected shift. For example, when the solenoid SL2 fails to hold the ON state, the speed stage of the transmission can be only 1st speed or O/D speed. At this time, if the automatic speed control changes the stage very often between 1st speed and O/D speed due to changes in the grade of the road, the shock on the vehicle is very large and dangerous. But, in the present embodiment, this danger does not occur. Here, when the solenoid fails, the danger of a shift to the upper side of the speed stage is smaller than for the lower side. Thus, when a shift to the upper side of the speed stage occurs, the automatic speed control may be continued. This case is shown in FIG. 35. In step 204-a, when the flag F-FAIL is "1", if a shift to 3rd stage or O/D stage occurs, then the speed control is executed.
Referring to FIG. 12 again, in steps 210 through 215, the cancelling of the automatic speed control is executed. If flag SW-A/D-RES is "1", then flag F-A/D-CAN is set to "0", and the automatic speed control is not cancelled. When flag SW-A/D-RES is "0", if flag SW-PKB is "1", or if flag SW-BK is "1", or if flag SW-D is not "1", then flag F-A/D-CAN is set to "1", and the automatic speed control is cancelled.
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