This invention is directed to methods of treating conditions which present with low bone mass, particularly osteoporosis, frailty, an osteoporotic fracture, a bone defect, childhood idiopathic bone loss, alveolar bone loss, mandibular bone loss, bone fracture, osteotomy, bone loss associated with periodontitis, or prosthetic ingrowth comprising administering prostaglandin agonists which are EP4 receptor selective prostaglandin agonists. This invention is especially directed to those methods wherein the EP4 receptor selective agonist is a compound of Formula I:

This invention relates to methods and pharmaceutical compositions comprising prostaglandin agonists which are useful to prevent bone loss, restore or augment bone mass and to enhance bone healing including the treatment of conditions which present with low bone mass and/or bone defects in vertebrates, and particularly mammals, including humans. This invention specifically relates to methods and pharmaceutical compositions comprising EP4 receptor selective prostaglandin agonists.
Osteoporosis is a systemic skeletal disease, characterized by low bone mass and deterioration of bone tissue, with a consequent increase in bone fragility and susceptibility to fracture. In the U.S., the condition affects more than 25 million people and causes more than 1.3 million fractures each year, including 500,000 spine, 250,000 hip and 240,000 wrist fractures annually. Hip fractures are the most serious consequence of osteoporosis, with 5-20% of patients dying within one year, and over 50% of survivors being incapacitated.
The elderly are at greatest risk of osteoporosis, and the problem is therefore predicted to increase significantly with the aging of the population. Worldwide fracture incidence is forecasted to increase three-fold over the next 60 years, and one study has estimated that there will be 4.5 million hip fractures worldwide in 2050.
Women are at greater risk of osteoporosis than men. Women experience a sharp acceleration of bone loss during the five years following menopause. Other factors that increase the risk include smoking, alcohol abuse, a sedentary lifestyle and low calcium intake.
There are currently two main types of pharmaceutical therapy for the treatment of osteoporosis. The first is the use of anti-resorptive compounds to reduce the resorption of bone tissue.
Estrogen is an example of an anti-resorptive agent. It is known that estrogen reduces fractures. In addition, Black, et al. in EP0605193A1 report that estrogen, particularly when taken orally, lowers plasma levels of LDL and raises those of the beneficial high density lipoproteins (HDL""s). However, estrogen fails to restore bone back to young adult levels in the established osteoporotic skeleton. Furthermore long-term estrogen therapy has been implicated in a variety of disorders, including an increase in the risk of uterine cancer, endometrial cancer and possibly breast cancer, causing many women to avoid this treatment. The significant undesirable effects associated with estrogen therapy support the need to develop alternative therapies for osteoporosis that have the desirable effect on serum LDL but do not cause undesirable effects.
A second type of pharmaceutical therapy for the treatment of osteoporosis is the use of anabolic agents to promote bone formation and increase bone mass. This class of agents is expected to restore bone to the established osteoporotic skeleton.
Certain prostagladin agonists are disclosed in GB 1478281, GB1479156 and U.S. Pat. Nos. 4,175,203, 4,055,596, 4,175,203, 3,987,091 and 3,991,106 as being useful as, for example, renal vasodilators.
U.S. Pat. No. 4,033,996 discloses certain 8-aza-9-oxo(and dioxo)-thia-11,12-secoprostaglandins which are useful as renal vasodilators, for the prevention of thrombus formation, to induce growth hormone release, and as regulators of the immune response.
French patent no. 897,566 discloses certain amino acid derivatives for the treatment of neurological, mental or cardiovascular disease.
J. Org. Chem. 26; 1961; 1437 discloses N-acetyl-N-benzyl-p-aminophenylmercaptoacetic acid.
U.S. Pat. No. 4,761,430 discloses certain arylbenzenesulfonamide compounds as lipid-lowering agents.
U.S. Pat. No. 4,443,477 discloses certain sulphonamidophenylcarboxylic acids as lipid lowering agents.
U.S. Pat. No. 3,528,961 discloses certain xcex5-caprolactam derivatives as dyes.
U.S. Pat. No. 3,780,095 discloses certain acylated anilinocarboxylic acids as choleretics.
U.S. Pat. No. 4,243,678 discloses certain acylhydrocarbylaminoalkanoic acids as having utility in the treatment of gastric ulcers, as sebaceous gland excretion inhibitors and for combatting skin inflammation.
U.S. Pat. No. 4,386,031 discloses certain N-benzoyl-xcfx89-anilinoalkanecarboxylic acids as antiallergic agents, thrombotic aggregation inhibitors, antiinflammatory agents and lipid-lowering agents.
In addition to osteoporosis, approximately, 20-25 million women and an increasing number of men have detectable vertebral fractures as a consequence of reduced bone mass, with an additional 250,000 hip fractures reported yearly in America alone. The latter case is associated with a 12% mortality rate within the first two years and with a 30% rate of patients requiring nursing home care after the fracture. While this is already significant, the economic and medical consequences of convalescence due to slow or imperfect healing of these bone fractures is expected to increase, due to the aging of the general population.
Estrogens have been shown (Bolander et al., 38th Annual Meeting Orthopedic Research Society, 1992) to improve the quality of the healing of appendicular fractures. Therefore, estrogen replacement therapy should be effective as a method for the treatment of fracture repair. However, patient compliance with estrogen therapy is relatively poor due to its side effects, including the resumption of menses, mastodynia, an increased risk of uterine cancer, an increased perceived risk of breast cancer, and the concomitant use of progestins. In addition, men are likely to object to the use of estrogen treatment. The need exists for a therapy which would be beneficial to patients who have suffered debilitating bone fractures and which would increase patient compliance.
It has been demonstrated that prostaglandin E2 (PGE2) can restore lost bone in an ovariectomized (OVX) rat model, a model for postmenopausal osteoporosis. Ke, H. Z., et al., Bone, 23:249-255, 1998. However there are severe side effects associated with PGE2. Jee, W. S. S. and Ma, Y. F., Bone, 21:297-304, 1997.
Although there are a variety of osteoporosis therapies, there is a continuing need and a continuing search in this field of art for alternative osteoporosis therapies. In addition, there is a need for bone fracture healing therapies. Also, there is a need for therapy which can promote bone re-growth into skeletal areas where defects exist such as defects caused or produced by, for example, tumors in bone. Further, there is a need for therapy which can promote bone re-growth into skeletal areas where bone grafts are indicated.
This invention is directed to methods of treating a condition which presents with low bone mass in a mammal comprising administering to said mammal an EP4 receptor selective agonist, a prodrug thereof or a pharmaceutically acceptable salt of said EP4 receptor selective agonist or of said prodrug.
This invention is particularly directed to such methods wherein said condition is osteoporosis, frailty, an osteoporotic fracture, a bone defect, childhood idiopathic bone loss, alveolar bone loss, mandibular bone loss, bone fracture, osteotomy, bone loss associated with periodontitis, or prosthetic ingrowth. In preferred methods of this invention, the EP4 receptor selective agonist is administered systemically, e.g., orally, subcutaneously, intramuscularly or via aerosol. In other preferred methods of this invention, the EP4 agonist is administered locally.
The methods of this invention are especially useful wherein said condition is frailty.
The methods of this invention are also especially useful wherein said condition is osteoporosis.
The methods of this invention are also especially useful wherein said condition is bone fracture or osteoporotic fracture.
Preferred EP4 selective agonists for use in the methods of this invention include compounds of Formula I:
prodrugs thereof and pharmaceutically acceptable salts of said compounds or said prodrugs, wherein:
Q is COOR3, CONHR4 or tetrazol-5-yl;
A is a single or cis double bond;
B is a single or trans double bond;
U is
R2 is xcex1-thienyl, phenyl, phenoxy, monosubstituted phenyl and monosubstituted phenoxy, said substituents being chloro, fluoro, phenyl, methoxy, trifluoromethyl or (C1-C3)alkyl;
R3 is hydrogen, (C1-C5)alkyl, phenyl or p-biphenyl;
R4 is COR5 or SO2R5; and
R5 is phenyl or (C1-C5)alkyl.
A preferred group of EP4 receptor selective agonists of Formula I for use in the methods of this invention are those compounds of Formula I wherein Q is 5-tetrazolyl and U is
forming compounds having the formula IA,
Particularly preferred compounds within this group include 5S-(4-(3-chloro-phenyl)-3R-hydroxy-butyl)-1-(6-(2H-tetrazol-5-yl)-hexyl-pyrrolidin-2-one; 5S-(3R-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl)-1-(6-(2H-tetrazol-5-yl)-hexyl)-pyrrolidin-2-one; 5R-(3S-hydroxy-4-phenyl-but-1-enyl)-1-[6-(1H-tetrazol-5-yl)-hexyl]-pyrrolidin-2-one; and 5S-(3R-hydroxy-4-phenyl-butyl)-1-[6-(1H-tetrazol-5-yl)-hexyl]-pyrrolidin-2-one.
Another preferred group of EP4 receptor selective agonists of Formula I for use in the methods of this invention are those compound of Formula I wherein Q is COOH. Particularly preferred compounds within this group include 7-{2S-[3R-hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-heptanoic acid; 7-(2S-(3R-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid; 7-{2S-[4-(3-chloro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-heptanoic acid; 7-(2S-(3R-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid; and 7-[2R-(3S-hydroxy-4-phenyl-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-heptanoic acid.
Preferably post-menopausal women and men over the age of 60 are treated. Also preferred are individuals regardless of age who have significantly reduced bone mass, i.e., greater than or equal to 1.5 standard deviations below young normal levels.
In the methods of this invention, conditions which present with low bone mass include such conditions as, for example, osteoporosis, childhood idiopathic bone loss, alveolar bone loss, mandibular bone loss, bone fracture, osteotomy, bone loss associated with periodontitis and prosthetic ingrowth.
Methods for treating xe2x80x9csecondary osteoporosisxe2x80x9d are also included within the methods of this invention. xe2x80x9cSecondary osteoporosisxe2x80x9d includes glucocorticoid-induced osteoporosis, hyperthyroidism-induced osteoporosis, immobilization-induced osteoporosis, heparin-induced osteoporosis and immunosuppressive-induced osteoporosis in a vertebrate, e.g., a mammal (including a human being). These methods are carried out by administering to said vertebrate, e.g., mammal, a xe2x80x9csecondary osteoporosisxe2x80x9d treating amount of an EP4 receptor selective prostaglandin agonist, a prodrug thereof or a pharmaceutically acceptable salt of said EP4 receptor selective prostaglandin agonist or of said prodrug.
Yet another aspect of this invention is directed to methods for strengthening a bone graft, inducing vertebral synostosis, enhancing long bone extension, enhancing bone healing following facial reconstruction, maxillary reconstruction and/or mandibular reconstruction in a vertebrate, e.g., a mammal (including a human being), comprising administering to said vertebrate, e.g., a mammal which has undergone facial reconstruction, maxillary reconstruction or mandibular reconstruction, a bone enhancing amount of an EP4 receptor selective prostaglandin agonist, a prodrug thereof or a pharmaceutically acceptable salt of said EP4 receptor selective prostaglandin agonist or of said prodrug. The active EP4 receptor selective prostaglandin agonists of this invention may be applied locally to the site of bone reconstruction or may be administered systemically.
A preferred dosage is about 0.001 to about 100 mg/kg/day of an EP4 receptor selective agonits, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug. An especially preferred dosage is about 0.01 to about 10 mg/kg/day of an EP4 receptor selective agonist, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.
The phrase xe2x80x9ccondition(s) which presents with low bone massxe2x80x9d refers to a condition where the level of bone mass is below the age specific normal as defined in standards by the World Health Organization xe2x80x9cAssessment of Fracture Risk and its Application to Screening for Postmenopausal Osteoporosis (1994). Report of a World Health Organization Study Group. World Health Organization Technical Series 843xe2x80x9d. Included in xe2x80x9ccondition(s) which presents with low bone massxe2x80x9d are primary and secondary osteoporosis, as described above. Also included is periodontal disease, alveolar bone loss, post-osteotomy and childhood idiopathic bone loss. The phrase xe2x80x9ccondition(s) which presents with low bone massxe2x80x9d also includes long term complications of osteoporosis such as curvature of the spine, loss of height and prosthetic surgery.
The phrase xe2x80x9ccondition(s) which presents with low bone massxe2x80x9d also refers to a vertebrate, e.g., a mammal known to have a significantly higher than average chance of developing such diseases as are described above including osteoporosis (e.g., post-menopausal women, men over the age of 50). Other bone mass augmenting or enhancing uses include bone restoration, increasing the bone fracture healing rate, replacing bone graft surgery entirely, enhancing the rate of successful bone grafts, bone healing following facial reconstruction or maxillary reconstruction or mandibular reconstruction, prosthetic ingrowth, vertebral synostosis or long bone extension.
The methods of this invention may also be used in conjunction with orthopedic devices such as spinal fusion cages, spinal fusion hardware, internal and external bone fixation devices, screws and pins.
Those skilled in the art will recognize that the term bone mass actually refers to bone mass per unit area which is sometimes (although not strictly correctly) referred to as bone mineral density.
The term xe2x80x9ctreatingxe2x80x9d, xe2x80x9ctreatxe2x80x9d or xe2x80x9ctreatmentxe2x80x9d as used herein includes preventative (e.g., prophylactic), palliative and curative treatment.
By xe2x80x9cpharmaceutically acceptablexe2x80x9d it is meant the carrier, vehicle, diluent, excipients, and/or salt must be compatible with the other ingredients of the formulation, and not deleterious to the recipient thereof.
The expression xe2x80x9cprodrugxe2x80x9d refers to compounds that are drug precursors which, following administration, release the drug in vivo via some chemical or physiological process (e.g., a prodrug on being brought to the physiological pH or through enzyme action is converted to the desired drug form). Exemplary prodrugs upon cleavage release the corresponding drug compound.
The expression xe2x80x9cpharmaceutically acceptable saltxe2x80x9d refers to nontoxic anionic salts containing anions such as (but not limited to) chloride, bromide, iodide, sulfate, bisulfate, phosphate, acetate, maleate, fumarate, oxalate, lactate, tartrate, citrate, gluconate, methanesulfonate and 4-toluene-sulfonate. The expression also refers to nontoxic cationic salts such as (but not limited to) sodium, potassium, calcium, magnesium, ammonium or protonated benzathine (N,Nxe2x80x2-dibenzylethylenediamine), choline, ethanolamine, diethanolamine, ethylenediamine, meglamine (N-methyl-glucamine), benethamine (N-benzylphenethylamine), piperazine or tromethamine (2-amino-2-hydroxymethyl-1,3-propanediol).
The methods of this invention result in bone formation resulting in decreased fracture rates. This invention makes a significant contribution to the art by providing methods that increase bone formation resulting in prevention, retardation, and/or regression of osteoporosis and related bone disorders.
This invention is also directed to compounds selected from 7-{2S-[3R-hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-heptanoic acid; 5S-(4-(3-chloro-phenyl)-3R-hydroxy-butyl)-1-(6-(2H-tetrazol-5-yl)-hexyl-pyrrolidin-2-one; 7-(2S-(3R-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid; 7-{2S-[4-(3-chloro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-heptanoic acid; and 5S-(3R-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl)-1-(6-(2H-tetrazol-5-yl)-hexyl)-pyrrolidin-2-one.
This invention is also particularly directed to 7-{2S-[3R-hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl)-heptanoic acid.
This invention is also particularly directed to 5S-(4-(3-chloro-phenyl)-3R-hydroxy-butyl)-1-(6-(2H-tetrazol-5-yl)-hexyl-pyrrolidin-2-one.
This invention is also particularly directed to 7-(2S-(3R-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid.
This invention is also particularly directed to 7-{2S-[4-(3-chloro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-heptanoic acid.
This invention is also particularly directed to 5S-(3R-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl)-1-(6-(2H-tetrazol-5-yl)-hexyl)-pyrrolidin-2-one.
Other features and advantages will be apparent from the specification and claims which describe the invention.
Any EP4 receptor selective agonist may be used as the EP4 receptor selective agonist of this invention. EP4 selective agonists are compounds which have an IC50 at the EP1, EP2 and EP3 receptor which is at least 10-fold greater than the IC50 at the EP4 receptor subtype. For example, 7-(2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid is an EP4 receptor selective PGE2 agonist with an EP4 receptor binding IC50 of 16 nM. At all other EP receptor subtypes, including the EP1, EP2 and EP3 receptor subtypes, the IC50 for 7-(2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid is greater than 3200 nM.
The compounds of Formula I may be prepared as disclosed in commonly assigned U.S. Pat. No. 4,177,346, which is incorporated herein by reference.
The EP4 receptor selective agonists used in the methods of this invention are all adapted to therapeutic use as agents that stimulate bone formation and increase bone mass in a vertebrates, e.g.; mammals, and particularly humans. Since bone formation is closely related to the development of osteoporosis and bone related disorders, the agonists used in the methods of this invention, by virtue of their action on bone, prevent, arrest and/or regress osteoporosis.
The utility of the EP4 selective agonists used in the methods of the present invention as medical agents in the treatment of conditions which present with low bone mass (e.g., osteoporosis) in vertebrates, e.g., mammals (especially humans and particularly female humans) is demonstrated by the activity of those agonists in conventional assays, including a receptor binding assay, a cyclic AMP assay, an in vivo assay and a fracture healing assay, all of which are described below. Such assays also provide a means whereby the activities of the EP4 selective agonists can be compared to each other and with the activities of other known compounds and compositions. The results of these comparisons are useful for determining dosage levels in a vertebrates, e.g., mammals, including humans, for the treatment of such diseases.
The activity of anabolic bone agents in stimulating bone formation and increasing bone mass can be tested in intact male or female rats, sex hormone deficient male (orchidectomy) or female (ovariectomy) rats.
Male or female rats at different ages (such as 3 months of age) can be used in the study. The rats are either intact or castrated (ovariectomized or orchidectomized), and subcutaneously injected or gavaged with prostaglandin agonists at different doses (such as 1, 3, or 10 mg/kg/day) for 30 days. In the castrated rats, treatment is started at the next day after surgery (for the purpose of preventing bone loss) or at the time bone loss has already occured (for the purpose of restoring bone mass). During the study, all rats are allowed free access to water and a pelleted commercial diet (Teklad Rodent Diet #8064, Harlan Teklad, Madison, Wis.) containing 1.46% calcium, 0.99% phosphorus and 4.96 IU/g of Vitamin D3. All rats are given subcutaneous injections of 10 mg/kg calcein on days 12 and 2 before sacrifice. The rats are sacrificed. The following endpoints are determined:
Femoral Bone Mineral Measurements
The right femur from each rat is removed at autopsy and scanned using dual energy X-ray absorptiometry (DXA, QDR 1000/W, Hologic Inc., Waltham, Mass.) equipped with xe2x80x9cRegional High Resolution Scanxe2x80x9d software (Hologic Inc., Waltham, Mass.). The scan field size is 5.08xc3x971.902 cm, resolution is 0.0254xc3x970.0127 cm and scan speed is 7.25 mm/second. The femoral scan images are analyzed and bone area, bone mineral content (BMC), and bone mineral density (BMD) of whole femora (WF), distal femoral metaphyses (DFM), femoral shaft (FS), and proximal femora (PF) are determined.
Tibial Bone Histomorphometric Analyses
The right tibia is removed at autopsy, dissected free of muscle, and cut into three parts. The proximal tibia and the tibial shaft are fixed in 70% ethanol, dehydrated in graded concentrations of ethanol, defatted in acetone, then embedded in methyl methacrylate (Eastman Organic Chemicals, Rochester, N.Y.).
Frontal sections of proximal tibial metaphyses at 4 and 10 xcexcm thickness are cut using a Reichert-Jung Polycut S microtome. The 4 xcexcm sections are stained with modified Masson""s Trichrome stain while the 10 xcexcm sections remain unstained. One 4 xcexcm and one 10 xcexcm sections from each rat are used for cancellous bone histomorphometry.
Cross sections of tibial shaft at 10 xcexcm thickness are cut using a Reichert-Jung Polycut S microtome. These sections are used for cortical bone histomorphometric analysis.
Cancellous bone histomorphometry: A Bioquant OS/2 histomorphometry system (RandM Biometrics, Inc., Nashville, Tenn.) is used for the static and dynamic histomorphometric measurements of the secondary spongiosa of the proximal tibial metaphyses between 1.2 and 3.6 mm distal to the growth plate-epiphyseal junction. The first 1.2 mm of the tibial metaphyseal region needs to be omitted in order to restrict measurements to the secondary spongiosa. The 4 xcexcm sections are used to determine indices related to bone volume, bone structure, and bone resorption, while the 10 xcexcm sections are used to determine indices related to bone formation and bone turnover.
I) Measurements and calculations related to trabecular bone volume and structure: (1) Total metaphyseal area (TV, mm2): metaphyseal area between 1.2 and 3.6 mm distal to the growth plate-epiphyseal junction. (2) Trabecular bone area (BV, mm2): total area of trabeculae within TV. (3) Trabecular bone perimeter (BS, mm): the length of total perimeter of trabeculae. (4) Trabecular bone volume (BV/TV, %): BV/TVxc3x97100. (5) Trabecular bone number (TBN, #/mm): 1.199/2xc3x97BS/TV. (6) Trabecular bone thickness (TBT, xcexcm): (2000/1.199)xc3x97(BV/BS). (7) Trsabecular bone separation (TBS, xcexcm): (2000xc3x971.199)xc3x97(TV-BV).
II) Measurements and calculations related to bone resorption: (1) Osteoclast number (OCN, #): total number of osteoclast within total metaphyseal area. (2) Osteoclast perimeter (OCP, mm): length of trabecular perimeter covered by osteoclast. (3) Osteoclast number/mm (OCN/mm, #/mm): OCN/BS. (4) Percent osteoclast perimeter (%OCP, %): OCP/BSxc3x97100.
III) Measurements and calculations related to bone formation and turnover: (1) Single-calcein labeled perimeter (SLS, mm): total length of trabecular perimeter labeled with one calcein label. (2) Double-calcein labeled perimeter (DLS, mm): total length of trabecular perimeter labeled with two calcein labels. (3) Inter-labeled width (ILW, xcexcm): average distance between two calcein labels. (4) Percent mineralizing perimeter (PMS, %): (SLS/2+DLS)/BSxc3x97100. (5) Mineral apposition rate (MAR, xcexcm/day): ILW/label interval. (6) Bone formation rate/surface ref. (BFR/BS, xcexcm2/d/xcexcm): (SLS/2+DLS)xc3x97MAR/BS. (7) Bone turnover rate (BTR, %/y): (SLS/2+DLS)xc3x97MAR /BVxc3x97100.
Cortical bone histomorphometry: A Bioquant OS/2 histomorphometry system (RandM Biometrics, Inc., Nashville, Tenn.) is used for the static and dynamic histomorphometric measurements of tibial shaft cortical bone. Total tissue area, marrow cavity area, periosteal perimeter, endocortical perimeter, single labeled perimeter, double labeled perimeter, and interlabeled width on both periosteal and endocortical surface are measured, and cortical bone area (total tissue areaxe2x80x94marrow cavity area), percent cortical bone area (cortical area/total tissue areaxc3x97100), percent marrow area (marrow cavity area/total tissue areaxc3x97100), periosteal and endocortical percent labeled perimeter [(single labeled perimeter/2+double labeled perimeter)/total perimeterxc3x97100], mineral apposition rate (interlabeled width/intervals), and bone formation rate [mineral apposition ratexc3x97[(single labeled perimeter/2+double labeled perimeter)/total perimeter] are calculated.
Statistics
Statistics can be calculated using StatView 4.0 packages (Abacus Concepts, Inc., Berkeley, Calif.). The analysis of variance (ANOVA) test followed by Fisher""s PLSD (Stat View, Abacus Concepts Inc., 1918 Bonita Ave, Berkeley, Calif. 94704-1014) are used to compare the differences between groups.
cDNAs representing the complete open reading frames of the human EP4 receptors are generated by reverse transcriptase polymerase chain reaction using oligonucleotide primers based on published sequences (1) and RNA from primary human lung cells (EP4) as templates. cDNAs are cloned into the multiple cloning site of pcDNA3 (Invitrogen Corporation, 3985B Sorrento Valley Blvd., San Diego, Calif. 92121) and used to transfect 293-S human embryonic kidney cells via calcium phosphate co-precipitation. G418-resistant colonies are expanded and tested for specific [3H]PGE2 binding. Transfectants demonstrating high levels of specific [3H]PGE2 binding are further characterized by Scatchard analysis to determine Bmax and Kds for PGE2. The lines selected for compound screening have approximately 256,400 receptors per cell and a Kd=2.9 nM for PGE2 (EP4). Constitutive expression of the receptor in parental 293-S cells is negligible. Cells are maintained in RPMI supplemented with fetal bovine serum (10% final) and G418 (700 ug/ml final).
cAMP responses in the 293-S/EP4 lines are determined by detaching cells from culture flasks in 1 ml of Ca++ and Mg++ deficient PBS via vigorous pounding, adding serum-free RPMI to a final concentration of 1xc3x97106 cells/ml, and adding 3-isobutyl-1-methylxanthine (IBMX) to a final concentration of 1 mM. One milliliter of cell suspension is immediately aliquoted into individual 2 ml screwcap microcentrifuge and incubated for 10 minutes, uncovered, at 37xc2x0 C., 5% CO2, 95% relative humdity. The compound to be tested is then added to cells at 1:100 dilutions such that final DMSO or ethanol concentrations is 1%. Immediately after adding compound, the tubes are covered, mixed by inverting two times, and incubated at 37xc2x0 C. for 12 minutes. Samples are then lysed by incubation at 100xc2x0 C. for 10 minutes and immediately cooled on ice for 5 minutes. Cellular debris is pelleted by centrifugation at 1000xc3x97g for 5 minutes, and cleared lysates are transferred to fresh tubes. cAMP concentrations are determined using a commercially available cAMP radioimmunoassay kit RIA (NEK-033, DuPont/NEN Research Products, 549 Albany St., Boston, Mass. 02118) after diluting cleared lysates 1:10 in cAMP RIA assay buffer (included in kit). Typically, the cells are treated with 6-8 concentrations of the compound to be tested in 1 log increments. EC50 calculations are performed on a calculator using linear regression analysis on the linear portion of the dose response curves.
References
1. Regan, J. W. Bailey, T. J. Pepperl, D. J. Pierce, K. L. Bogardus, A. M. Donello, J. E. Fairbairn, C. E. Kedzie, K. M. Woodward, D. F. and Gil, D. W. 1994 Cloning of a Novel Human Prostaglandin Receptor with Characteristics of the Pharmaclogically Defined EP2 Subtype. Mol. Pharmacology 46:213-220.
Membrane Preparation: All operations are performed at 4xc2x0 C. Transfected cells expressing prostaglandin E2 type 1 receptors (EP1), type 2 (EP2), type 3 (EP3) or type 4 (EP4) receptors are harvested and suspended to 2 million cells per ml in Buffer A [50 mM Tris-HCl (pH 7.4), 10 mM MgCl2, 1 mM EDTA, 1 mM Pefabloc peptide, (Boehringer Mannheim Corp., Indianapolis, Ind.), 10 uM Phosporamidon peptide, (Sigma, St. Louis, Mo.), 1 uM pepstatin A peptide, (Sigma, St. Louis, Mo.), 10 uM elastatinal peptide, (Sigma, St. Louis, Mo.), 100 uM antipain peptide, (Sigma, St. Louis, Mo.)]. The cells are lysed by sonification with a Branson Sonifier (Model #250, Branson Ultrasonics Corporation, Danbury, Conn.) in 2 fifteen second bursts. Unlysed cells and debris are removed by centrifugation at 100xc3x97g for 10 min. Membranes are then harvested by centrifugation at 45,000xc3x97g for 30 minutes. Pelleted membranes are resuspended to 3-10 mg of protein per ml of Buffer A, the protein concentration being determined by the method of Bradford [Bradford, M., Anal. Biochem., 72, 248 (1976)]. Resuspended membranes are then stored frozen at xe2x88x9280xc2x0 C. until use.
Binding Assay: Frozen membranes prepared as above are thawed and diluted to 0.5 mg/ml (for rat EP2) or 0.3 mg/ml (for rat EP4) in Buffer A above. Frozen membranes prepared as above are thawed and diluted to 0.5 mg/ml (for rat EP2) or 0.3 mg/ml (for rat EP4) in Buffer A above. One volume of membrane preparation is combined with 0.05 volume test compound or buffer and one volume of 3 nM 3H-prostaglandin E2 (#TRK 431, Amersham, Arlington Heights, Ill.) in Buffer A. The mixture (205 xcexcL total volume) is incubated for 1 hour at 25xc2x0 C. The membranes are then recovered by filtration through type GF/C glass fiber filters (#1205-401, Wallac, Gaithersburg, Md.) using a Tomtec harvester (Model Mach II/96, Tomtec, Orange, Conn.). The membranes with bound 3H-prostaglandin E2 are trapped by the filter, while the buffer and unbound 3H-prostaglandin E2 pass through the filter into waste. Each sample is then washed 3 times with 3 ml of 50 mM Tris-HCl (pH 7.4), 10 mM MgCl2, 1 mM EDTA. The filters are then dried by heating in a microwave oven. To determine the amount of 3H-prostaglandin bound to the membranes, the dried filters are placed into plastic bags with scintillation fluid and counted in a LKB 1205 Betaplate reader (Wallac, Gaithersburg, Md.). IC50s are determined from the concentration of test compound required to displace 50% of the specifically bound 3H-prostaglandin E2.
293S cells expressing either the rat EP2 or EP4 prostaglandin E2 receptors are generated according to methods known to those skilled in the art. Typically, PCR (polymerase chain reaction) primers corresponding to the 5xe2x80x2 and 3xe2x80x2 ends of the published full length receptor are made according to well known methods disclosed above and are used in an RT-PCR reaction using total RNA from rat kidney for both EP2 and EP4 receptors.
The full length coding sequence for the rat EP2 receptor is made as disclosed in Nemoto et al., Prostaglandins, 1997, 54, 713-725. The full length coding sequence for the rat EP4 receptor is made as disclosed in Sando et al., Biochem. Biophys. Res. Comm., 1994, 200, 1329-1333. These full length receptors are used to prepare 293S cells expressing the EP2 and EP4 receptors.
The full length coding sequence for the EP1 receptor is made as disclosed in Funk et al., Journal of Biological Chemistry, 1993, 268, 26767-26772. The full length coding sequence for the EP2 receptor is made as disclosed in Regan et al., Molecular Pharmacology, 1994, 46, 213-220. The full length coding sequence for the EP3 receptor is made as disclosed in Regan et al., British Journal of Pharmacology, 1994, 112, 377-385. The full length coding sequence for the EP4 receptor is made as disclosed in Bastien, Journal of Biological Chemistry, 1994, 269,11873-11877. These full length receptors are used to prepare 293S cells expressing the EP1, EP2, EP3 and EP4 receptors.
293S cells expressing either the human EP1, EP2, EP3 or EP4 prostaglandin E2 receptors are generated according to methods known to those skilled in the art. Typically, PCR (polymerase chain reaction) primers corresponding to the 5xe2x80x2 and 3xe2x80x2 ends of the published full length receptor are made according to the well known methods disclosed above and are used in an RT-PCR reaction using the total RNA from human kidney (for EP1), human lung (for EP2), human lung (for EP3) or human lymphocytes (for EP4) as a source. PCR products are cloned by the TA overhang method into pCR2.1 (Invitrogen, Carlsbad, Calif.) and identity of the cloned receptor is confirmed by DNA sequencing.
293S cells (Mayo, Dept. of Biochemistry, Northwestern Univ.) are transfected with the cloned receptor in pcDNA3 by electroporation. Stable cell lines expressing the receptor are established following selection of transfected cells with G418.
Clonal cell lines expressing the maximal number of receptors are chosen following a whole cell 3H-PGE2 binding assay using unlabeled PGE2 as a competitor.
Fracture Technique: Sprage-Dawley rats at 3 months of age are anesthetized with Ketamine. A 1 cm incision is made on the anteromedial aspect of the proximal part of the right tibia or femur. The following describes the tibial surgical technique. The incision is carried through to the bone, and a 1 mm hole is drilled 4 mm proximal to the distal aspect of the tibial tuberosity 2 mm medial to the anterior ridge. Intramedullary nailing is performed with a 0.8 mm stainless steel tube (maximum load 36.3 N, maximum stiffness 61.8 N/mm, tested under the same conditions as the bones). No reaming of the medullary canal is performed. A standardized closed fracture is produced 2 mm above the tibiofibular junction by three-point bending using specially designed adjustable forceps with blunt jaws. To minimize soft tissue damage, care is taken not to displace the fracture. The skin is closed with monofilament nylon sutures. The operation is performed under sterile conditions. Radiographs of all fractures are taken immediately after nailing, and rats with fractures outside the specified diaphyseal area or with displaced nails are excluded. The remaining animals are divided randomly into the following groups with 10-12 animals per each subgroup per time point for testing the fracture healing. The first group receives daily gavage of vehicle (water: 100% Ethanol=95:5) at 1 ml/rat, while the others receive daily gavage from 0.01 to 100 mg/kg/day of the compound to be tested (1 ml/rat) for 10, 20, 40 and 80 days.
At 10, 20, 40 and 80 days, 10-12 rats from each group are anesthetized with Ketamine and sacrificed by exsanguination. Both tibiofibular bones are removed by dissection and all soft tissue is stripped. Bones from 5-6 rats for each group are stored in 70% ethanol for histological analysis, and bones from another 5-6 rats for each group are stored in a buffered Ringer""s solution (+4xc2x0 C., pH 7.4) for radiographs and biomechanical testing which is performed.
Histological Analysis: The methods for histologic analysis of fractured bone have been previously published by Mosekilde and Bak (The Effects of Growth Hormone on Fracture Healing in Rats: A Histological Description. Bone, 14:19-27, 1993). Briefly, the fracture site is sawed 8 mm to each side of the fracture line, embedded undecalcified in methymethacrylate, and cut frontals sections on a Reichert-Jung Polycut microtome in 8 xcexcm thick. Masson-Trichrome stained mid-frontal sections (including both tibia and fibula) are used for visualization of the cellullar and tissue response to fracture healing with and without treatment. Sirius red stained sections are used to demonstrate the characterisitics of the callus structure and to differentiate between woven bone and lamellar bone at the fracture site. The following measurements are performed: (1) fracture gapxe2x80x94measured as the shortest distance between the cortical bone ends in the fracture, (2) callus length and callus diameter, (3) total bone volume area of callus, (4) bony tissue per tissue area inside the callus area, (5) fibrous tissue in the callus, and (6) cartilage area in the callus.
Biomechanical Analysis: The methods for biomechanical analysis have been previously published by Bak and Andreassen (The Effects of Aging on Fracture Healing in Rats. Calcif Tissue Int 45:292-297, 1989). Briefly, radiographs of all fractures are taken prior to the biomechanical test. The mechanical properties of the healing fractures are analyzed by a destructive three- or four-point bending procedure. Maximum load, stiffness, energy at maximum load, deflection at maximum load, and maximum stress are determined.
Fracture Technique: Female or male beagle dogs at approximately 2 years of age are used under anesthesia in the study. Transverse radial fractures are produced by slow continuous loading in three-point bending as described by Lenehan et al. (Lenehan, T. M.; Balligand, M.; Nunamaker, D. M.; Wood, F. E.: Effects of EHDP on Fracture Healing in Dogs. J Orthop Res 3:499-507; 1985). A wire is pulled through the fracture site to ensure complete anatomical disruption of the bone. Thereafter, local delivery of prostaglandin agonists to the fracture site is achieved by slow release of compound delivered by slow release pellets or by administration of the compounds in a suitable formulation such as a paste gel solution or suspension for 10, 15, or 20 weeks.
Histological Analysis: The methods for histologic analysis of fractured bone have been previously published by Peter et al. (Peter, C. P.; Cook, W. O.; Nunamaker, D. M.; Provost, M. T.; Seedor, J. G.; Rodan, G. A. Effects of alendronate on fracture healing and bone remodeling in dogs. J. Orthop. Res. 14:74-70, 1996) and Mosekilde and Bak (The Effects of Growth Hormone on Fracture Healing in Rats: A Histological Description. Bone, 14:19-27,1993). Briefly, after sacrifice, the fracture site is sawed 3 cm to each side of the fracture line, embedded undecalcified in methymethacrylate, and cut on a Reichert-Jung Polycut microtome in 8 xcexcm thick of frontal sections. Masson-Trichrome stained mid-frontal sections (including both tibia and fibula) are used for visualization of the cellullar and tissue response to fracture healing with and without treatment. Sirius red stained sections are used to demonstrate the characteristics of the callus structure and to differentiate between woven bone and lamellar bone at the fracture site. The following measurements are performed: (1) fracture gapxe2x80x94measured as the shortest distance between the cortical bone ends in the fracture, (2) callus length and callus diameter, (3) total bone volume area of callus, (4) bony tissue per tissue area inside the callus area, (5) fibrous tissue in the callus, (6) cartilage area in the callus.
Biomechanical Analysis: The methods for biomechanical analysis have been previously published by Bak and Andreassen (The Effects of Aging on Fracture Healing in Rats. Calcif Tissue Int 45:292-297, 1989) and Peter et al. (Peter, C. P.; Cook, W. O.; Nunamaker, D. M.; Provost, M. T.; Seedor, J. G.; Rodan, G. A. Effects of Alendronate On Fracture Healing And Bone Remodeling In Dogs. J. Orthop. Res. 14:74-70, 1996). Briefly, radiographs of all fractures are taken prior to the biomechanical test. The mechanical properties of the healing fractures are analyzed by a destructive three- or four-point bending procedures. Maximum load, stiffness, energy at maximum load, deflection at maximum load, and maximum stress are determined.
Administration of the EP4 receptor selective agonists according to the methods of this invention can be via any mode which delivers the EP4 receptor selective agonist systemically and/or locally (e.g., at the site of the bone fracture, osteotomy, or orthopedic surgery). These methods include oral routes, parenteral, intraduodenal routes, etc. Generally, the compounds of this invention are administered orally, but parenteral administration (e.g., intravenous, intramuscular, transdermal, subcutaneous, rectal or intramedullary) may be utilized, for example, where oral administration is inappropriate for the target or where the patient is unable to ingest the drug.
The methods of this invention are used for the treatment and promotion of healing of bone fractures and osteotomies by the local application (e.g., to the sites of bone fractures of osteotomies) of EP4 receptor selective agonists. The EP4 receptor selective agonists of this invention are applied to the sites of bone fractures or osteotomies, for example, either by injection of the compound in a suitable solvent (e.g., an oily solvent such as arachis oil) to the cartilage growth plate or, in cases of open surgery, by local application thereto of the compound in a suitable vehicle, carrier or diluent such as bone-wax, demineralized bone powder, polymeric bone cements, bone sealants, etc. Alternatively, local application can be achieved by applying a solution or dispersion of the compound in a suitable carrier or diluent onto the surface of, or incorporating it into solid or semi-solid implants conventionally used in orthopedic surgery, such as dacron-mesh, gel-foam and kiel bone, or prostheses.
In any event, the amount and timing of compounds administered will, of course, be dependent on the subject being treated, on the severity of the affliction, on the manner of administration and on the judgment of the prescribing physician. Thus, because of patient to patient variability, the dosages given herein are a guideline and the physician may titrate doses of the drug compound to achieve the treatment (e.g., bone mass augmentation) that the physician considers appropriate for the patient. In considering the degree of treatment desired, the physician must balance a variety of factors such as bone mass starting level, age of the patient, presence of preexisting disease, as well as presence of other diseases (e.g., cardiovascular disease).
In general an amount of an EP4 receptor selective agonist is used that is sufficient to augment bone mass to a level which is above the bone fracture threshold (as detailed in the World Health Organization Study previously cited herein).
The EP4 receptor selective agonist compounds used in the methods of this invention are generally administered in the form of a pharmaceutical composition comprising at least one of the compounds of this invention together with a pharmaceutically acceptable vehicle or diluent. Thus, the EP4 receptor selective agonist can be administered individually in any conventional oral, intranasal, parenteral, rectal or transdermal dosage form.
For oral administration the pharmaceutical composition can take the form of solutions, suspensions, tablets, pills, capsules, powders, and the like. Tablets containing various excipients such as sodium citrate, calcium carbonate and calcium phosphate are employed along with various disintegrants such as starch and preferably potato or tapioca starch and certain complex silicates, together with binding agents such as polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often very useful for tabletting purposes. Solid compositions of a similar type are also employed as fillers in soft and hard-filled gelatin capsules; preferred materials in this connection also include lactose or milk sugar as well as high molecular weight polyethylene glycols. When aqueous suspensions and/or elixirs are desired for oral administration, the compositions of this invention can be combined with various sweetening agents, flavoring agents, coloring agents, emulsifying agents and/or suspending agents, as well as such diluents as water, ethanol, propylene glycol, glycerin and various like combinations thereof.
For purposes of parenteral administration, solutions in sesame or peanut oil or in aqueous propylene glycol can be employed, as well as sterile aqueous solutions of the corresponding water-soluble salts. Such aqueous solutions may be suitably buffered, if necessary, and the liquid diluent first rendered isotonic with sufficient saline or glucose. These aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal injection purposes. In this connection, the sterile aqueous media employed are all readily obtainable by standard techniques well-known to those skilled in the art.
For purposes of transdermal (e.g.,topical) administration, dilute sterile, aqueous or partially aqueous solutions (usually in about 0.1% to 5% concentration), otherwise similar to the above parenteral solutions, are prepared.
Methods of preparing various pharmaceutical compositions with a certain amount of active ingredient are known, or will be apparent in light of this disclosure, to those skilled in the art. For examples of methods of preparing pharmaceutical compositions, see Remington""s Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 19th Edition (1995).
The purpose of this study is to determine whether an EP4 receptor selective agonist, and specifically 7-(2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid, can restore bone mass in OVX rat model.
Sprague-Dawley male rats were ovariectomized (OVX) at 3.5 months of age. Ten months after OVX, the rats were treated by subcutaneous injection with vehicle or 7-(2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid at 30 mg/kg/day for 28 days. The rats were autopsied. The right femur from each rat was removed at autopsy and scanned using dual energy x-ray absorptiometry (DEXA, QDR 1000/W, Hologic Inc., Waltham, Mass.) equipped with xe2x80x9cRegional High Resolution Scanxe2x80x9d software (Hologic Inc., Waltham, Mass.). The scan field size was 5.08xc3x971.902 cm, resolution is 0.0254xc3x970.0127 cm and the scan speed was 7.25 mm/second. The femoral scan images were analyzed. Bone mineral content (BMC) and bone mineral density (BMD) of whole femora (WF), distal femoral metaphysis (DFM), and femoral shaft (FS) were determined according to the method described in H. Z. Ke et al., Droloxifene, a New Estrogen Antagonist/Agonist, Prevents Bone Loss in Ovariectomized Rats. ENDOCRINOLOGY 136;2435-2441, 1995.
Compared with OVX rats treated with vehicle, OVX rats treated with 7-(2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid showed total femoral BMC increased by 17%, total femoral BMD increased by 13%, distal femoral BMC increased by 8%, distal femoral BMD increased by 8%, femoral shaft BMC increased by 20%, and femoral shaft BMD increased by 18%. No PGE2-like side effects were seen in rats treated with 7-(2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid.
This data shows that 7-(2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl)-heptanoic acid, an EP4 receptor selective PGE2 agonist, stimulates bone formation and restores bone in established osteopenic OVX rats.
NMR spectra were recorded on a Varian Unity 400 spectrometer (Varian Co., Palo Alto, Calif.) at about 23xc2x0 C. at 400 MHz for proton nuclei. Chemical shifts are expressed in parts per million. The peak shapes are denoted as follows: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; bs=broad singlet. Atmospheric pressure chemical ionization (APCI) mass spectra were obtained on a Fisons Platform II Spectrometer. Where the intensity of chlorine or bromine-containing ions are described the expected intensity ratio was observed (approximately 3:1 for 35Cl/37Cl-containing ions) and 1:1 for 79Br/81Br-containing ions) and the intensity of only the lower mass ion is given.
Medium pressure chromatography was performed using a Biotage purification system (Biotage, Dyax Corporation, Charlottesville, Va.) under nitrogen pressure. Flash chromatography was performed with either Baker Silica Gel (40 xe2x96xa1m) (J. T. Baker, Phillipsburg, N.J.) or Silica Gel 60 (EM Sciences, Gibbstown, N.J.) in glass columns under low nitrogen pressure. Radial Chromatography was performed using a Chromatotron (model 7924T, Harrison Research). Preparative Chromatography was performed using Analtech Uniplates Silica Gel GF (20xc3x9720 cm) (Analtech, Inc. Newark, Del.). Dimethylformamide (DMF), tetrahydrofuran (THF), and dichloromethane (CH2Cl2) used as reaction solvents were the anhydrous grade supplied by Aldrich Chemical Company (Milwaukee, Wis.). The term xe2x80x98concentratedxe2x80x99 refers to removal of solvent at water aspirator pressure on a rotary evaporator. The abbreviation xe2x80x98hxe2x80x99 stands for hours. The term xe2x80x9cTBAFxe2x80x9d refers to tetrabutylammonium fluoride. The term xe2x80x9cDMAPxe2x80x9d refers to dimethylaminopyridine. The terms xe2x80x9cdichloromethanexe2x80x9d and xe2x80x9cmethylene chloridexe2x80x9d are synonymous and are used interchangeably throughout this specification and in the Examples and Preparations.

EP4 receptor selective agonists in the treatment of osteoporosis

Abstract: wherein the variables are as defined in the specification.


A semiconductor device may include a substrate including a first surface and a second surface, a through electrode penetrating the substrate to include a protrusion that protrudes from the second surface of the substrate, and a front side bump electrically coupled to the through electrode and disposed on the first surface of the substrate. The semiconductor device may include a first passivation pattern disposed on the first surface of the substrate to substantially surround a sidewall of the front side bump and may be formed to include an uneven surface, and a second passivation pattern disposed on the second surface of the substrate to include an uneven surface. The protrusion of the through electrode may penetrate the second passivation pattern to protrude from the uneven surface of the second passivation pattern.

1. Technical Field
Various embodiments of the present disclosure generally relate to semiconductor devices including through electrodes, semiconductor packages including the same, electronic systems including the same, memory cards including the same, and methods of manufacturing the same.
2. Related Art
Ultra small-sized semiconductor devices with large capacities are increasingly in demand with the development of smaller and higher performance electronic products. A plurality of semiconductor chips may be assembled in a single semiconductor package to increase a data storage capacity of the semiconductor device. That is, the data storage capacity of the semiconductor device may be readily increased using a multi-chip package technique.
However, even though the multi-chip package technique is used to increase the data storage capacity of the semiconductor device, there may be limitations associated with obtaining a sufficient space needed for electrical connections between the plurality of semiconductor chips in the multi-chip package as the number of the semiconductor chips increases. Recently, through silicon vias (TSVs) have been proposed to resolve the limitations associated with the multi-chip package techniques. The TSVs may be formed to penetrate a plurality of chips at a wafer level, and the chips stacked in the package may be electrically and physically connected to each other by the TSVs. Accordingly, if the TSVs are employed in the packages, the performance and the storage capacity of the packages may be improved.

Semiconductor devices having through electrodes, semiconductor packages including the same, electronic systems including the same, and methods of manufacturing the same


A self-contained, transportable system for conducting any of hydrologic tests or water sampling techniques without the need for retooling or readjusting the system. The system includes a borehole unit for conducting the tests in a borehole, and a data acquisition unit for monitoring and recording the results of the tests. The borehole unit includes, connected in serial relation, a pair of borehole packers for forcing the unit in place in the borehole, several pressure transducers for obtaining hydraulic measurements, a pump located between the borehole packers for withdrawing fluid and a fluid injector for injecting fluid. The data-acquisition system includes flow meters for monitoring fluid injection and fluid withdrawals, a data-acquisition module for monitoring a fluid pressure from the pressure transducers, power source and a computer interface. The overall system includes a pressure manifold for supplying gas to the packers, transducer and injectors, a hand-held electric water-level sounder, and a pressure regulator. The system can be used to measure ambient hydraulic pressure, fluid injection or fluid withdrawal tests for inferring formation properties and tracer tests for inferring properties of the formation controlling chemical migration. Additionally, the system can be used in contaminated groundwater environments because it is easy to decontaminate.

The invention relates to subsurface hydrology and engineering geology and, in general, the invention concerns a borehole testing apparatus for conducting hydrologic tests and researching the mechanisms controlling fluid movement and chemical transport in fractured rock aquifers.
Study of the subsurface beneath the ground to identify the quantity and quality of ground water and characterize the movement of contaminants in the subsurface is a concern of many Federal and state regulatory agencies which are responsible for ground-water resources and for the oversight of ground-water remediation. Such study is needed to ensure that technically feasible regulatory decisions can be made. It is also of interest to government agencies responsible for remediation of property. For example, the U.S. Geological Survey routinely conducts cooperative investigations with other federal agencies and state and local governments involving the characterization of ground-water quantity and quality and the migration of contaminants in the subsurface, requiring the hydrologic testing and the collection of water samples from subsurface boreholes for geochemical analysis. Furthermore, many private engineering companies are engaged in similar types of site characterization for corporate clients engaged in the remediation of contaminated ground water.
Hydrologic testing and the collection of water samples for geochemical analysis have often been conducted using downhole equipment to isolate and test water in a discrete interval in a borehole. Individual components used in borehole testing and geochemical analysis, such as pumps, borehole packers, pressure transducers and various types of data acquisition equipment are available from a number of manufacturers.
Commonly, a borehole is drilled into the ground to allow access to the subsurface and testing equipment is lowered into the borehole. In most cases, the borehole equipment is constructed for a specific type of test or geochemical sampling and is typically fixed in place. However, a site may require more than a single type of test to adequately study the site. Further, the physical dimensions of the borehole equipment and the need for various peripheral components at land surface for data collection make such borehole testing equipment cumbersome and not readily transportable from site to site. This is disadvantageous because testing at more than a single borehole, sometimes a series of boreholes, is frequently necessary to characterize a ground-water site.
Therefore, there is a serious need in the art for an integrated borehole testing equipment design which is capable of isolating and testing a discrete interval of a borehole, and which is transportable, easily assembled, and capable of conducting multiple types of hydrologic tests as well as collecting water samples for geochemical analysis. Moreover, with the increased need in recent years for remediation of contaminated ground water, there is an increased interest in equipment which can conduct testing of contaminated ground water. However, the application of downhole equipment capable at isolating a discrete interval of a borehole for hydrologic testing or geochemical sampling at sites having contaminated ground water has not been actively considered because of problems, such as the contamination of the downhole equipment. Therefore, there is a further need in the art for testing equipment that can operate in contaminated ground-water sites.
Prior art of interest includes U.S. Pat. Nos. 5,934,375 (Peterson); 4,435,843 (Jageler); and 3,799,733 (Ringgenberg). The Peterson patent discloses a deep well system for obtaining artesian samples. The system comprises a borehole packer, a pump for withdrawing fluids, and a sample chamber for holding the fluids. A portion of the sample is discharged to flush the system, but the system does not discharge tracer fluid into the borehole or extract tracer fluid to estimate properties of the subsurface At affecting chemical migration therein. The Jageler patent discloses an apparatus for testing borehole fluids. The apparatus comprises a packer, an electrical section, a hydraulic section, and a mechanical section. The Ringgenberg patent discloses a system for servicing a well, taking fluid measurements, and taking fluid samples. The system comprises a borehole packer, a pump for withdrawing fluids, a sampling chamber for holding fluids, and pressure and temperature recording devices.
The borehole and data acquisition equipment disclosed in these patents do not address the needs for multiple testing at multiple sites, some of which may be contaminated. The equipment is not transportable, easily assembled, capable of conducting multiple types of hydrologic tests or performing geochemical sampling, nor is the equipment suitable for use at sites having contaminated ground water.
In accordance with the invention, a self-contained, transportable borehole system for conducting hydrologic tests or water sampling is provided which meets the needs described above and overcomes other problems associated with the prior art.
The borehole system has a borehole unit for conducting the hydrologic tests in a borehole, and a data acquisition unit for monitoring and recording the results of the tests. The borehole unit comprises, housed in protective shrouds and connected together by union connectors in serial relation, first and second borehole packers, a plurality of pressure transducers for obtaining pressure measurements, a pump unit, located between the borehole packers, for withdrawing fluid from a test interval, and an injection unit, located between the borehole packers, for injecting fluid into a test interval. The data-acquisition unit is comprised of at least one flow meter for monitoring fluid injection by the injection unit, at least one flow meter for monitoring fluid withdrawal by the pump unit, and a data-acquisition module for providing a digital record of flow meter readings and pressure measurements.
Advantageously, the data acquisition unit further comprises rechargeable batteries for providing power for the unit and for the flow meters.
Preferably, the borehole system has a single, portable pressure source and a pressure manifold for supplying gas to operate the injector unit, to operate the transducer, and to inflate the borehole packers. Advantageously, a fluid reservoir is connected to both the fluid injection unit and to a pressure regulator which regulates the air pressure applied to the fluid reservoir during fluid injection.
The borehole system may be used for conducting measurements of ambient hydraulic pressure, fluid injection or fluid withdrawal tests for inferring formation properties, or tracer tests for inferring properties of the formation controlling chemical migration.
Preferably, the pressure transducers include an upper transducer that measures hydraulic pressure above a test interval, an intermediate transducer that measures hydraulic pressures within the test interval, and a lower transducer for measuring the hydraulic pressure below the test interval. Further, the intermediate and lower transducers are preferably located above the borehole packers.
Advantageously, the data-acquisition unit electronically stores digital records of collected data and displays the data as it is collected.
Preferably, transducer cables and pump cables are prewired in the borehole unit, and the union connectors between segments comprise screw-disconnect union ball joints.
The borehole system is very simple to decontaminate after use in contaminated ground-water or other sources of contamination, and to this end, there are provided connecting tubes made of stainless steel, polytetrafluoroethylene tubing for transporting water within the system, stainless-steel fittings, a stainless-steel pump and a polytetrafluoroethylene coated pump cable.

U.S. Geological Survey, Water Resources Division, Branch of Regional Research Eastern Region, 12201 Sunrise Valley Dr., MS:


Techniques for implementing a streaming transaction in a transaction based data storage system is disclosed. In an embodiment, a server computer system stores a dataset comprising a plurality of files where the dataset comprises a corresponding version number. The server computer system receives a command to modify the dataset and, in response, starts a streaming transaction for the dataset. During the streaming transaction, the server computer system receives a plurality of updates to the dataset, wherein each of the plurality of updates modifies one or more files of the plurality of files. The server computer system executes the plurality of updates in the dataset without modifying the corresponding version number of the dataset. When the server computer system commits the streaming transaction to the dataset, the server computer system increments the corresponding version number of the dataset.

The approaches described in this section are approaches that could be pursued, but not necessarily approaches that have been previously conceived or pursued. Therefore, unless otherwise indicated, it should not be assumed that any of the approaches described in this section qualify as prior art merely by virtue of their inclusion in this section.
A data pipeline system is a distributed data storage system that may include a plurality of datasets and that is programmed or capable of executing specified transformations of raw datasets or derived datasets into other derived datasets. A dataset is a digital representation of a set of files and metadata regarding the set of files. A dataset may store multiple versions of a file as the file is updated.
To provide version control and efficient access, datasets are updated using a transaction based system. Transactions allow for discrete updates to a dataset. When a transaction is opened, the server computer system grants permission to one or more computing devices and/or programs to update a dataset. While the transaction is open, updates may be received for the dataset, but are not viewable until the transaction is committed. When the transaction is committed, the state of the dataset is finalized and the dataset is viewable to other users and programs. A version number of the dataset is incremented to indicate that a new version of the dataset has been stored. Two-phase commit is a common technique in relational database systems for ensuring that only committed changes become usable.
Datasets may be modified by different types of transactions. A snapshot transaction replaces all files in the dataset with a new version or stores a snapshot of all the current files in the dataset. An append transaction adds a new version to an existing file or existing files in a dataset. An update transaction adds a new file to a dataset. Each transaction is identified by the version number and/or a transaction identifier.
When a query is received for data from the dataset, the server computer system uses the transactional nature of the system to build a dataset. First, the server computer system identifies a snapshot of the dataset. Second, the server computer system identifies each transaction that occurred after the snapshot of the dataset. Finally, the server computer system builds the dataset by applying the identified transactions to the identified snapshot of the dataset. By storing updates as completed transactions, the server computer system can effectively roll back the dataset to any transaction identifier.
While transaction based updates are useful for efficiently responding to queries and performance of version control, the transaction based nature is less effective for low-latency data requirements. Some types of datasets, such as financial market pricing datasets, are most effective when they are updated frequently as prices may change rapidly. Ideally, an application accessing financial market pricing datasets would receive real-time values, thereby effectively matching the values queried from the dataset to the current financial market prices.
While a transaction is open and not fully committed, the updates in the transaction are not visible to other applications or computing devices. Thus, to use a transaction based system to provide low-latency data, a server computer system must open transactions, update data, and commit the transactions at the speed at which the data changes, thereby requiring the opening and committing of a relatively large number of transactions in a short period. The opening and closing of many transactions to deliver real-time data is extremely computationally expensive and inefficient.
Thus, there is a need for a technique for handling streaming updates while maintaining the benefits of a transaction based system.

Processing streaming data in a transaction-based distributed database system


A bi-directional stepping motor in which a rotor is rotatable in steps of 180 degrees each, the stepping motor comprising a rotor comprising a permanent magnet and rotatably mounted about an axis and providing a permanent magnetic field; a first electrical coil and a second electrical coil; a stator on which the first electrical coil is mounted; and a control circuit, coupled to the first and second electrical coils, for applying electrical pulses independently to each coil and for controlling the polarity thereof, the coils producing magnetic fields in response to the pulses and wherein the rotor is rotatable in response to the magnetic fields; wherein each step of 180 degrees is effectuated by providing to the first coil, a first pulse of a first polarity and a second pulse of a second polarity; and to said second coil, a pulse of the second polarity simultaneously with the providing of the second pulse to the first coil; wherein during the providing of the first pulse to the first coil, there is no pulse being provided to the second coil.

The present invention is directed to stepping motors, and in particular, to an improved 2-step bi-directional stepping motor that is can reliably run in either direction merely dependent upon a specific pulse-current scheme used therewith. The present invention, among other applications, is particularly suited for use in connection with electronic quartz wristwatches or other micro-mechanical devices.
xe2x80x9cMulti-Stepxe2x80x9d stepping motors, having multiple stator and/or rotor poles, or those of the so-called xe2x80x9cLavetxe2x80x9d type, are well known in the art. For example, while the xe2x80x9cmultiple-stepxe2x80x9d motors run in both directions depending on the direction of the xe2x80x9crotor-drivingxe2x80x9d electromagnetic field, the xe2x80x9cLavetxe2x80x9d type motor may have one preferred rotational direction achievable through the rotor""s xe2x80x9crest position.xe2x80x9d The prior art provides a plurality of examples illustrating the foregoing constructions.
Alternating rectangular pulse-current schemes to drive rotors in 180 degree steps are also known. For example, in one such known configuration, an alternating rectangular current-pulse is applied to the motor coil, and the rotor magnet follows the changed magnetic flux in the stator, being pushed from its rest position to its opposite stable rest position, 180 degrees rotated. To continue the rotation of the rotor, the current-pulse is reversed and the rotor is pushed in the same rotational direction back to its starting-position, thus completing one revolution in two steps. FIG. 6 exemplifies the relevant portion of a prior art stepping motor that is operable in the foregoing manner. As can be readily seen, typical of such a stepping motor is the specific positioning of the xe2x80x9crest position,xe2x80x9d which may be chosen as a function of the magnetic vector of the electromagnetic flux generated by the motor coil. Geometries of the rotor or stator cutouts and/or the use of small permanent magnets can also play a significant part in the operation of such a stepping motor.
The foregoing exemplary stepping motor is widely used in electronic timepieces, notwithstanding its perceived deficiency in being unable to run in both a clockwise and counter-clockwise direction. Being able to run in both directions would be significantly advantageous, for example, so as to be able to perform a quick xe2x80x9ctime-settingxe2x80x9d function in radio controlled watches.
Bipolar current-pulse sequences have been developed to make the foregoing type of stepping motor run in a reverse direction. For example, the rotor may be accelerated for a short distance (i.e. 45 degrees) from its rest position in the reverse direction by a first initial pulse so that when it passes its stator flux axis, a reversed pulse pushes it to the opposite stable position. Thereafter, it may again drop 45 degrees backwards, against the rotational direction, into its opposite stable Rest Position when the coil current is off. Unfortunately however, this particular stepping motor design provides for rough running, and additionally, due to the additionally required pulse per step, leads to higher power consumption while running in reverse. Furthermore, if the reverse pulse is not accurately applied in intensity and time, the rotor may undesirably pass its alternating rest positions thus leading to an undesirable xe2x80x9cone step per revolutionxe2x80x9d result.
It can thus be seen that it is difficult to ensure a reliable 2-Step reverse run in known xe2x80x9cLavetxe2x80x9d type stepping motors. However, the prior art is not without its attempts to do so.
For example, U.S. Pat. No. 4,361,790 attempts to overcome the foregoing deficiencies by the implementation of alternating the current direction within two respective coils. In particular, the two coils always operate simultaneously by a certain current-pulse scheme, which alleges, among other things, the elimination of the need to double the coil volume. U.S. Pat. No. 4,361,790 proposes a current-pulse scheme in which four steps per revolution, each of which is 90 degrees, are made. Such is the case in both the clockwise and counter-clockwise direction.
However, the present inventors still believe further improvements over the state of the art are necessary, both to overcome the foregoing explicitly perceived deficiencies as well as to overcome deficiencies that would be apparent to those skilled in the art. For example, one desirous feature is to construct a smooth running xe2x80x9cLavetxe2x80x9d type stepping motor that can make two steps per revolution, in both the clockwise or counter-clockwise direction. Secondly, it is desirous to provide such a stepping motor that does not have a preferred rotational direction. In particular, it is desired that the shape of the current-pulses, applied to the motor coils, determine the direction of rotation of the rotor. The present invention achieves the foregoing and below mentioned objectives, as well as overcomes the perceived deficiencies in the prior art.
It is therefore an object of the present invention to provide an improved stepping motor that overcomes the foregoing perceived deficiencies.
It is another object of the present invention to provide an improved stepping motor that more smoothly runs in both the forward and reverse direction.
It is yet another object of the present invention to provide an improved stepping motor that does not increase the power consumption from that seen in prior art bi-directional stepping motors, namely, it is an object of the present invention to provide an improved current-pulse scheme that has the same power consumption regardless of the rotational direction of the rotor.
It is yet another object of the present invention to provide an improved stepping motor that merely needs to rely on the pulse-current scheme for its directional rotation.
Still other objects and advantages of the invention will in part be obvious and will in part be apparent from the specification.
The invention accordingly comprises the features of construction, combination of elements and arrangement of parts and sequence of steps which will be exemplified in the construction, illustration and description hereinafter set forth, and the scope of the invention will be indicated in the claims.
Generally speaking, in accordance with the present invention, a stepping motor in which a rotor is rotatable in steps of 180 degrees each, is provided. In one embodiment, the stepping motor comprises a rotor comprising a permanent magnet and rotatably mounted about an axis and providing a permanent magnetic field; a first electrical coil and a second electrical coil; a stator comprising three pole faces arranged around the rotor and arms on which the electrical coils are mounted; and a control circuit, coupled to the first and second electrical coils, for applying electrical pulses independently to each coil and for controlling the polarity thereof, the coils producing magnetic fields in response to the pulses and wherein the rotor is rotatable in response to the magnetic fields; wherein each step of 180 degrees is effectuated by providing to the first coil, a first pulse of a first polarity and a second pulse of a second polarity; and to said second coil, a pulse of the second polarity simultaneously with the providing of the second pulse to the first coil; wherein during the providing of the first pulse to the first coil, there is no pulse being provided to the second coil.
Furthermore, the rotation of the second 180 degree step is preferably achieved by providing to the first coil, a third pulse of the second polarity and a fourth pulse of the first polarity; and to said second coil, a pulse of the first polarity simultaneously with the providing of the fourth pulse to the first coil; wherein during the providing the third pulse to the first coil, there is no pulse being provided to the second coil.
In a particular arrangement, each step of 180 degrees is effectuated by a rotation of the rotor a first X degrees and then by a further rotation of the rotor the remaining 180-X degrees, wherein X is a predetermined number greater than zero. In a specific configuration, the value of X is approximately 30 degrees and the three pole faces are arranged at an angular spacing of approximately 120 degrees with respect to each other. In this manner, the rotation of the rotor the approximately X degrees is achieved by providing to the first coil, a first pulse of a first polarity; and wherein the rotation of the rotor the approximately 180-X degrees to complete a step is achieved by providing to the first coil a second pulse of a second polarity simultaneously with the providing of a pulse of the second polarity to the second coil. The second step of 180 degrees is effectuated by the additional rotation of the rotor X degrees and then by a further rotation of the rotor the remaining 180-X degrees; wherein the additional rotation of the rotor X degrees is achieved by providing to the first coil, a third pulse of the second polarity; and wherein the rotation of the rotor the approximately 180-X degrees to complete the second step is achieved by providing to the first coil a fourth pulse of the first polarity simultaneously with the providing of a pulse of the first polarity to the second coil; whereby during the providing of the third pulse to the first coil, there is no pulse being provided to the second coil.
In a preferred embodiment, the stepping motor constructed in accordance with the present invention is incorporated into a timepiece, such as a wristwatch.


A mounting structure is provided for an NOx reduction system for a hydraulic excavator. The NOx reduction system includes a PM collection filter to be arranged in an exhaust passage of an engine to collect suspended particulate matter in exhaust gas from the engine, a urea SCR catalyst for reducing and purifying nitrogen oxides in the exhaust gas from the engine, a urea water spay device for spraying a urea water into the exhaust passage of the engine, a urea water feeder for feeding the urea water to the urea water spray device, and a urea water storage tank for storing therein the urea water to be guided to the urea water feeder. In the mounting structure, the counterweight is provided with a recessed section formed therein, the urea SCR catalyst is arranged in the recessed section, and the PM collection filter is arranged in the engine compartment. The urea SCR catalyst and PM collection filter are arranged to form an “L” pattern.

As a conventional technology of this sort, there is one disclosed in Patent Document 1. Disclosed in this Patent Document 1 are an NOx reduction system, which includes an upstream-side, first aftertreatment device for collecting particulates contained as suspended particulate matter in exhaust gas from an engine, that is, a PM collection filter and also a downstream-side, second aftertreatment device for purifying and detoxifying nitrogen oxide (NOx), that is, a reduction catalyst, and also a construction that the PM collection filter and the reduction catalyst are arranged in a turned square "U" pattern via a piping which forms an exhaust passage of the engine.
Mounting of such an NOx reduction system is also desired in a construction machine. The above-mentioned conventional technology disclosed in the Patent Document 1 will hereinafter be described with respect to a mounting structure for the NOx reduction system, which can be contemplated upon mounting the NOx reduction system on a construction machine, for example, a hydraulic excavator.
FIG. 5 is a plan view showing certain essential parts of the conventionally-contemplated mounting structure for the NOx reduction system for the construction machine, and FIG. 6 is a view as seen in the direction of arrow B in FIG. 5.
As shown in these FIGS. 5 and 6, the hydraulic excavator is provided with a revolving upperstructure 40 mounted on a travel base. This revolving upperstructure 40 is provided with an engine compartment 41 and a counterweight 42 arranged adjacent the engine compartment 41 at a rear position of a body to maintain stability of the body. An engine 43 and a hydraulic pump 44 drivable by the engine 43 are arranged in the engine compartment 41. Upon application of the above-mentioned conventional technology disclosed in the Patent Document 1 to such a hydraulic excavator, it may be possible to contemplate a construction that as shown in FIG. 5, a PM collection filter 46 and a reduction catalyst 48 are arranged side by side in a lateral direction of the construction machine such that the reduction catalyst 48 is located on an outboard side, the engine 43 is communicated to the PM collection filter 46 via a piping 45, the PM collection filter 46 is communicated to the reduction catalyst 48 via a piping 47, and a muffler 49 is arranged at a part of the reduction catalyst 48 to release exhaust gas into the atmosphere. Specifically, it may be possible to contemplate a construction that the PM collection filter 46 and the reduction catalyst 48 are arranged via the piping 47 in a turned square "U" pattern as seen in plan.
Patent Document 1: JP-A-2005-155404

Mounting structure for NOx reduction device for construction machine

Hitachi Construction Machinery Co., Ltd.


A medical device includes a clevis, a first jaw, and a second jaw. The first jaw includes a first proximal gripping portion and a first distal gripping portion, and the first proximal gripping portion includes a first needle alignment portion. The second jaw is coupled to the clevis and includes a second proximal gripping portion and a second distal gripping portion, and the second proximal gripping portion includes a second needle alignment portion. The first and second needle alignment portions are located opposite each other when in the closed orientation. The first and second needle alignment portions define a clamp path when in the closed orientation within which the curved portion of the needle is received when the second jaw is in the open orientation. The first and second needle alignment portions are configured to rotate the needle when moving to the closed orientation.

The embodiments described herein relate to grasping tools, more specifically to medical devices, and still more specifically to endoscopic tools. More particularly, the embodiments described herein relate to medical devices that include jaws having a self-righting needle alignment portion that can be used, for example, in surgical applications.
Known techniques for Minimally Invasive Surgery (MIS) employ instruments to manipulate tissue that can be either manually controlled or controlled via computer-assisted teleoperation. Many known MIS instruments include a therapeutic or diagnostic end effector (e.g., forceps, a cutting tool, or a cauterizing tool) mounted on a wrist mechanism at the distal end of an extension (also referred to herein as the main tube or shaft). During an MIS procedure, the end effector, wrist mechanism, and the distal end of the main tube can be inserted into a small incision or a natural orifice of a patient to position the end effector at a work site within the patient's body. The optional wrist mechanism can be used to change the end effector's orientation with respect to the main tube to perform the desired procedure at the work site. Known wrist mechanisms generally provide the desired degrees of freedom (DOFs) for movement of the end effector. For example, for forceps or other grasping tools, known wrist mechanisms are often able to change the pitch and yaw of the end effector with reference to the main tube. A wrist may optionally provide a roll DOF for the end effector, or the roll DOF may be implemented by rolling the main tube. An end effector may optionally have additional mechanical DOFs, such as grip or knife blade motion. In some instances, wrist and end effector mechanical DOFs may be combined. For example, U.S. Pat. No. 5,792,135 (filed May 16, 1997) discloses a mechanism in which wrist and end effector grip DOFs are combined.
To enable the desired movement of the wrist mechanism and end effector, known instruments include tension members (e.g., cables, cable/hypotube combinations, tension bands) that extend through the main tube of the instrument and that connect the wrist mechanism to a transmission or actuator (also referred to herein as a backend mechanism). The backend mechanism moves the cables to operate the wrist mechanism. For computer-assisted systems, the backend mechanism is motor driven and can be operably coupled to a processing system to provide a user interface for a doctor to control the instrument.
Patients benefit from continual efforts to improve the effectiveness of MIS methods and tools. For example, reducing the size and/or the operating footprint of the main tube and wrist mechanism can allow for smaller entry incisions, thereby reducing the negative effects of surgery, such as pain, scarring, and undesirable healing time. But, producing small diameter medical instruments that implement the clinically desired functions for minimally invasive procedures can be challenging. Specifically, simply reducing the size of known wrist mechanisms by "scaling down" the components will not result in an effective solution because required component and material properties do not scale.
Another way to reduce the negative effects of surgery is to minimize the number of times that the wrist mechanism or end effector is moved into and out of the operation area of the patient. For example, some medical instruments have end effectors with gripping jaws coupled to the wrist mechanism, which can be used for performing clinical functions including cutting, dissection, incising, cauterizing, destroying tissue, and suturing. Such known medical instruments have an opposing set of jaws that can be operated to contact tissue and interact with other medical instruments. Some instruments include jaws that can grip a surgical needle therebetween and manipulate the needle to perform suturing functions. However, it can be difficult to orient the needle between the jaws of these instruments at a desired orientation for suturing. Further, it can be challenging to maintain the orientation of the needle when gripped between the jaws and while maneuvering the needle under surgical conditions, such as while maneuvering the needle within a small operating area and within a lubricious environment. Moreover, these conventional instruments are typically withdrawn from the clinical operations area during use by the operator to switch instruments (e.g., between a tissue gripping instrument and a suturing instrument) or modify the needle orientation according to surgical needs and conditions, so as to avoid the high risk of the needle slipping during such adjustments. Such manipulations can slow the suturing process and increase related challenges for maintaining a sterile clinical environment.
Thus, a need exists for improved endoscopic tools including tools for performing suturing clinical functions. Improvements may include wrist mechanisms having jaws with one or more gripping portions providing enhanced retention of surgical needles for suturing, and improved flexibility for quickly changing or modifying the needle orientation during use.

Medical devices having tissue grasping surfaces and features for manipulating surgical needles


A dehider includes an elongated handle having side-by-side circular disks, one of which is stationary and the other is rotary. A pinion gear is power-driven to rotate the rotary disk.

Power-operated handheld dehiders are well known in the meat processing industry. Generally speaking, these tools have oppositely reciprocating cutting blades, usually disk-shaped with serrated outer edges. The adjacent cutting disks are driven in opposite cutting oscillations, typically by a pair of oscillating pushrods connected to an eccentric drive mechanism driven by an air motor carried on the tool. As the pushrods oscillate, the teeth on one rotary disk move past the teeth on the oppositely-moving rotary disk. This produces a shearing and cutting action that separates the hide from the carcass.
Although this dehider design has proved effective in the industry for many years, various improvements have been developed over time to address certain problems that have arisen from using these dehiders. Among these problems are the noise and vibration caused by using the tool over prolonged periods of time on a production line. The oscillating nature of the eccentric-driven pushrods and the cutting blades produces substantial vibration. And excessive vibration of these handheld dehiders as well as other handheld power tools can lead to workplace hazards such as hand-arm vibration syndrome, such as carpal tunnel syndrome. These problems have been recently addressed by OSHA standards to protect workers from the ill effects of constantly using handheld vibrating power tools such as eccentric-driven dehiders.
To reduce vibration experienced with oscillating dehiders, recent design approaches have been taken. These include reducing the weight of the tool and/or counterbalancing the moving mass of the pushrods, as examples. As for dehiders in particular, the oscillating blades are driven at high speeds during use, typically at speeds in excess of 6,500 strokes per minute ("spm"). The tool tends to slow down when pressure is applied and can return to a much higher no-load speed when the cutting load is released. The no-load speed can momentarily return to at least 8,000 spm, for example. This causes worker fatigue as well as excessive vibration forces being transmitted from the handheld device. Attempts to reduce vibration have also included motor speed controls and speed governors to smooth out the accelerations that cause undue vibrations which otherwise occur during normal changes in load applied by the cutting blades during use. Examples of such speed controls are disclosed in U.S. Pat. No. 7,722,448 to Gwyther and U.S. Pat. No. 4,901,400 to Karubian.
Thus, there is an ongoing need to produce a handheld dehider that is light in weight and is essentially devoid of vibration problems caused during normal use over extended periods of time. The tool should also maintain standard cutting efficiencies and be easy to repair and operate at a reasonable cost.

This patent is subject to a terminal disclaimer.


A method of performing an analysis on internal hard drive(s), which includes an analysis of the file management system, individual files that exist on the hard drive, developing a Unique ID for each program (i.e., executable) file and continuously analyzing (i.e., scanning) the hard drive(s) to detect physical changes in the previously analyzed program files. The method may be implemented on a computer unit, which can be a 32/64-bit Microsoft PC O/S, or a 32/64-bit Microsoft Server O/S.

As electronic connectivity, networking and automation expands in business and organizations, as data (information) and file transfers become faster, and while the number of unauthorized programs, viruses, malicious mobile code, spyware, etc., grows to new levels on a daily basis, there is an increasing need for a new technology that organizations can utilize to address these threats in a manner that is not limited by traditional file signature methods of protecting the internal file contents of 32/64-bit Microsoft computer hard drive(s) and devices that connect to 32/64-bit Microsoft computers.
As an example, a Microsoft 32-bit computer, or a 64-bit Microsoft computer, may be connected to a network, which is connected to the Internet, which is further connected to several other networks via gateways that are referred to in this document as "connection points", which may allow data to be automatically transferred to/from any "connection point" to that particular computer. As the internal threat of corporate espionage grows greater and greater on a daily basis, the installation of unauthorized software from those individuals who have the authority to manage and control networks is also growing on a daily basis. The terms Microsoft PC, Microsoft Server, Microsoft computer, Microsoft 32-bit computer, and/or any other similar variations and combinations using Microsoft to describe a specific computer, device and/or server may be used interchangeably to mean a computer, device and/or server on which a Microsoft operating system (O/S) is implemented.
The overall problem that exists in most of the technology utilized in today's modern computing environment, is that the number of viruses, malicious mobile code, spyware, unauthorized programs has grown to such a large number, that it requires, at a minimum, daily electronic updates to maintain a file signature database on known viruses, etc., which could require hours to receive a daily update and scan an individual computer for the possible new viruses. Furthermore, the computer must not only be scanned for new viruses, but also for previous known viruses that are still being electronically transmitted across the Internet. The requirement to receive an update and scan an individual computer requires such a significant amount of network and computer resources that it significantly affects a users ability to utilize a networked computer during a normal daily scan for possible viruses, etc.
Because of the problems described above, a new technology (i.e., a utility) is needed that can take advantage of an existing operating system (O/S) file management system to perform an analysis of any internal hard drive(s) in a computer (e.g., as used herein a "Microsoft computer" is defined to include any computer running a computer and/or server O/S from Microsoft Corporation of Redmond Wash.). The utility may determine all executable program files, regardless of the file naming convention used to name the files and use an algorithm to establish a unique ID for each executable program file located on the hard disk. When the analysis is complete, the utility may continue to cycle (i.e., scan) all of the hard drive(s) to insure (i.e., maintain) the integrity of each specific program file, detect any unauthorized change to any specific program file, and detect any new unauthorized program file that might be installed anywhere within the computers internal hard drive(s).
Information system industry experts, who understand the latest features and capabilities about operating system (O/S) design and development and that are familiar with kernel operations, are aware that the ability to intercept kernel events is a major key to providing adequate security to an O/S. While the 32/64-bit Microsoft O/S kernel provides a mechanism to intercept many events, there is currently no known mechanism available that would allow a security solution to receive a kernel event or notification that a file transfer protocol (i.e., a file download/upload) is about to start, for 100% of the programs that have the ability to transfer files in and out of a networked computer. Because there is no such kernel mechanism currently available, and no known development plans at this time in future O/S's (e.g., Vista, etc.), to intercept and control 100% of all program file transfer activity, a solution is needed that provides the ability to detect physical changes in existing program files and detect the installation of new program files. Preferably, the solution may co-exist as an O/S utility with the O/S file management system and have the ability to independently perform an analysis (i.e., scan) and maintain the authorized integrity on every program file that exists within the Microsoft computer.

Analyzing internal hard drives and continuously authenticating program files within a 32/64 bit operating system


A user interface automation framework is described. A system records multiple user interface screenshots during a session of a user interacting with a user interface application executing on a host computer. The system records metadata associated with the host computer during the session. The system executes a test of the user interface application based on the multiple user interface screenshots and the metadata.

The subject matter discussed in the background section should not be assumed to be prior art merely as a result of its mention in the background section. Similarly, a problem mentioned in the background section or associated with the subject matter of the background section should not be assumed to have been previously recognized in the prior art. The subject matter in the background section merely represents different approaches, which in and of themselves may also be inventions.
Test automation is the use of software to control the execution of tests, the comparison of actual outcomes to predicted outcomes, the setting up of test preconditions, and other test control and test reporting functions. A test automation framework is not a single tool to perform some specific testing task, but a set of tools that provide support for automated software testing in a unified manner, thereby providing a common platform for an automation engineer to do a job. A test automation framework integrates function libraries, test data sources, object details and various reusable modules. A test automation framework is responsible for defining the format in which to express expectations, creating a mechanism to drive an application being tested, executing the tests, and reporting the results. If there is change to any test case for an application that is being tested, only the test case file needs to be updated, such that the driver script and startup script remain the same.

Automation framework for testing user interface applications