A method of increasing the growth of stem cells by mixing the stem cells with a growth medium that has been conditioned by an incubation with placental tissue. The method increases the expansion of the stem cell population.

Stem cells have the ability to divide for indefinite periods in culture and to give rise to specialized cells. Typically, stem cells are divided into two main groups: adult stem cells and embryonic stem cells. Stem cells may also be generated through artificial means such as nuclear transfer, cytoplasmic transfer, cell fusion, parthenogenesis and reprogramming. Isolated stem cells can give rise to many types of differentiated cells, and can be used to treat many types of diseases.
Adult stem cells are undifferentiated but are present in differentiated tissues, and are capable of differentiation into the cell types from the tissue that the adult stem cell originated. Adult stem cells have been derived from various sources, such as the nervous system (McKay, 1997, Science 276:66-71; Shihabuddin, et al., 1999, Mol. Med Today 5:474-480); bone marrow (Pittenger, et al., 1999, Science 284:143-147; Pittenger, M. F. and Marshak, D. R. (2001) In: Mesenchymal stem cells of human adult bone marrow. Marshak, D. R., Gardner, D. K., and Gottlieb, D. eds. (Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press) 349-374); adipose tissue (Gronthos, et al., 2001, J Cell. Physiol. 189:54-63), dermis (Toma, et al., 2001, Nature Cell Biol. 3:778-784); pancreas and liver (Deutsch, el al., 2001, Development 128:871-881). Stem cells have also been isolated from umbilical cord (Rogers, et al., 2004, Best Pract Res Clin Obstet Gynaecol. 18(6):893-908; Wang et al., 2004, Stem Cells 22(7):1330-1337; Surbek, et al, 2002, Ther Umsch. 59(11):577-582; and placenta (Yen et al., 2005, Stem Cells 23(1):3-9), each of which is incorporated by reference herein in its entirety. It is believed that stem cells of the adult type are also found in smooth muscle tissue, striated muscle tissue, cardiac muscle tissue, bone tissue, bone spongy tissue, cartilage tissue, pancreatic ductal tissue, spleen tissue, thymus tissue, tonsil tissue, Peyer's patch tissue, lymph nodes tissue, thyroid tissue, epidermis tissue, dermis tissue, subcutaneous tissue, heart tissue, lung tissue, vascular tissue, endothelial tissue, blood cells, bladder tissue, kidney tissue, digestive tract tissue, esophagus tissue, stomach tissue, small intestine tissue, large intestine tissue, adipose tissue, uterus tissue, eye tissue, lung tissue, testicular tissue, ovarian tissue, prostate tissue, connective tissue, endocrine tissue, and mesentery tissue.
Several patents disclose various aspects of adult stem cells. For example, U.S. Pat. No. 5,556,783 discloses methods of culturing hair follicle stem cells, while U.S. Pat. No. 5,486,359 discloses methods of isolating human mesenchymal stem cells. U.S. Pat. Nos. 4,714,680, 5,061,620, and 5,087,570 provide examples of hematopoietic stem cells. Each of these patents is incorporated by reference herein in its entirety.
Embryonic stem cells are undifferentiated cells derived from the embryo. Typically these cells are extracted from the inner cell mass of a blastocyte and when cultured under the unique conditions, either alone or in combination with a variety of feeder cells, the embryonic stem cells maintain euploid karyotype, do not undergo senescence, and retain the ability to differentiated into cells of the endodermal, ectodermal, and mesodermal lineages.
These cells have the potential to become a wide variety of specialized cell and tissue types, which can then be used for basic research, drug discovery, and treatment (or prevention) of many types of diseases. Patents describing aspects of embryonic stem cells include U.S. Pat. No. 6,506,574 to Rambhatla, U.S. Pat. No. 6,200,806 to Thomson, U.S. Pat. No. 6,432,711 to Dinsmore, and U.S. Pat. No. 5,670,372 to Hogan, each of which is incorporated by reference herein in its entirety. Importantly, murine embryonic stem cells can be cultured indefinitely under the presence of leukemia inhibitory factor (LIF), which maintains their undifferentiated state. In contrast, human embryonic stem cells are not responsive in the same manner to LIF, thus stimulating the invention of numerous methodologies to expand them. Unfortunately, many such methodologies involve the use of either murine feeder cells or other animal components, hence limiting the therapeutic potential of these cells. Furthermore, even when established cell lines, such as the federally approved embryonic stem cells, are cultured in murine-free conditions, contamination is still present as recently reported (Martin, et al., 2005, Nat Med 11:228-232, which is incorporated by reference herein in its entirety). Accordingly, one object of the invention disclosed is to provide novel methods of expanding stem cells in absence of animal components, said invention being applicable to a variety of stem cells, including embryonic stem cells.
The importance of technologies associated with expansion of stem cells, both of adult and/or embryonic derivation is illustrated by the numerous preclinical and clinical uses of these cells in treatment of a wide range of diseases.
One of the earliest clinical uses of stem cells was for performing bone marrow transplants in patients with hematological malignancies in which hematopoietic stem cells derived from the donor bone marrow were administered into the recipient subsequent to providing said recipient with a sufficient dose of radiation and/or chemotherapy in order to ablate not only the hematological malignancy but also non-malignant hematopoiesis. The administration of, non-malignant hematopoietic stem cells resulted in donor-specific hematopoiesis and in some patients, cure of the malignancy. This was first described by Thomas et al in 1957, who reported that large volumes of donor bone marrow could be safely infused in patients with acute leukemia following myeloablation and that donor-specific hematopoiesis was established (Thomas, et al., 1957, N Engl J Med 257:491-496, which is incorporated by reference herein in its entirety). The identification of similar hematopoietic stem cell activity in the peripheral blood led to development of techniques used to mobilize and harvest peripheral blood hematopoietic stem cells for use in transplantation settings. For example, the use of GM-CSF and G-CSF in enhancing the number of peripheral blood hematopoietic stem cells was reported in the clinical situation of autologous transplantation subsequent to high dose chemotherapy (Peters, et al., 1993, Blood 81:1709-1719; Sheridan, et al., 1992, Lancet 339:640-644, each of which is incorporated by reference herein in its entirety).
In addition to treatment of hematological malignancies, stem cells have been utilized in the context of therapy for solid tumors. The dose limiting variable in cancer chemotherapy is bone marrow toxicity. Accordingly, in 1958, Kurnick et al performed an autologous bone marrow transplant to demonstrate ability of infused bone marrow to allow use of very high doses of chemotherapy and/or radiation therapy (Kurnick, et al., 1958, Ann Intern Med 49:973-986, which is incorporated by reference herein in its entirety). The use of autologous hematopoietic cell transplants combined with high dose chemo/radiotherapy for solid tumors has been extensively investigated for breast (Peppercorn, et al., 2005, Cancer 104:1580-1589; Dillman, et al., 2005, Am J Clin Oncol 28:281-288), colon (Leff, et al., 1986, J Clin Oncol 4:1586-1591; Franchi, et al., 1994, Eur J Cancer 30A:1420-1423), lung (Ziske, et al., 2002, Anticancer Res 22:3723-3726), nasopharyngeal cancer (Chen, et al., 2003, Jpn J Clin Oncol 33:331-335), and other types of cancers (Gratwohl, et al., 2004, Ann Oncol 15:653-660), each of which is incorporated by reference herein in its entirety.
The identification of the type 1 transmembrane protein/adhesion molecule, the sialomucin CD34 as a marker of hematopoietic stem cells led to the use of CD34+ cell selection as a means of concentrating hematopoietic stem cell activity (Civin, et al., 1984, J Immunol 133:157-165, which is incorporated by reference herein in its entirety). Specifically, it was demonstrated that although bone marrow mononuclear cells contain approximately 1-4% CD34+ cells, the administration of these cells, but not bone marrow depleted of CD34+ cells, into lethally irradiated baboons led to hematopoietic reconstitution (Berenson, et al., 1988, J Clin Invest 81:951-955, which is incorporated by reference herein in its entirety). Clinical development of purified CD34+ cells as a source of stem cells was originally sought as a method of performing bone marrow transplant without contamination of donor T cells. This would in theory stop development of graft versus host disease, one of the main causes of allogeneic transplant associated morbidity and mortality (Ferrara, et al., 2005, Clin Adv Hematol Oncol 3:415-419, 428, which is incorporated by reference herein in its entirety). Unfortunately, clinical evidence demonstrated that patients receiving purified CD34+ stem cell grafts, although having a lower incidence of graft versus host disease, also had a higher incidence in leukemic relapse due to an immunologically mediated graft versus leukemia effect that is absent when donor bone marrow grafts are depleted of T cells (Martino, et al., 2000, Haematologica 85:1165-1171; Butt, et al., 2003, Leuk Lymphoma 44:1509-1513, each of which is incorporated by reference herein in its entirety). The critical importance of bone marrow derived T cells in the induction and upkeep of graft versus leukemia effects was illustrated in studies of leukemic patients who have relapsed and were subsequently treated by infusion of donor T cells. This induced a long-term remission in the patients that had major relapse (Kolb, H. J., 1998, Vox Sang 74 Suppl 2:321-329; Guglielmi, et al., 2002, Blood 100:397-405, each of which is incorporated by reference herein in its entirety). Furthermore, it was also observed that under some conditions, bone marrow derived non-CD34 cells of the osteoblast lineage have a role in facilitating engraftment in allogeneic settings (Good, R. A., 2000, World J Surg 24:797-810, which is incorporated by reference herein in its entirety). Despite these potential drawbacks, clinical use of CD34+ cells both from mobilized peripheral blood, as well as bone marrow, during autologous transplantation for high dose chemo/radiation therapy was considered to be a useful approach (Korbling, et al., 2001, Blood 98:2900-2908; Pecora, et al., 2001, Bone Marrow Transplant 27:1245-1253; Pecora, A. L., 1999, Bone Marrow Transplant 23 Suppl 2:S7-12, each of which is incorporated by reference herein in its entirety). This is due to the fact that in this setting, neither facilitator cells are needed, since the graft is autologous, and the CD34+ selection substantially clears the marrow of contaminating tumor cells, so that the risk of tumor relapse is lessened as opposed to using non-purified bone marrow (Preti, et al., 2001, Cytotherapy 3:85-95; Siena, et al., 2000, J Clin Oncol 18:1360-1377; Vannucchi, et al., 1998, Br J Haematol 103:610-617, each of which is incorporated by reference herein in its entirety).
The use of hematopoietic stem cells has also been described for "reprogramming" the immune system to induce an antigen-specific state of non-responsiveness called tolerance. Specifically, this use can be divided into two main areas: the use of stem cells to induce donor-specific tolerance during allogeneic or xenogeneic transplantation, and the use of stem cells to induce tolerance in situations of autoimmunity. Although common mechanisms of tolerance maintenance such as generation of T regulatory cells, effector T cell depletion, and effector T cell anergy have been described in both types of tolerance, the mechanism of induction seems to be different; therefore we will describe them individually.
The possibility of bone marrow hematopoietic stem cells having the utility of inducing tolerance to a grafted organ was first elaborated on by Owens in the 1940s. In studies demonstrating that in utero mixing of blood in the context of shared circulation between two genetically different cows, he observed bilateral transplantation tolerance in adulthood. Accordingly, he postulated that the original sharing of circulation may have contributed to the state of tolerance which theoretically should not have existed due to the genetic disparity between the siblings. Furthermore, definitive roles for using stem cells to induce tolerance came from Billingham and Medawar in the 1950s in experiments showing injection of donor bone marrow cells into neonates allowed for tolerance to the donor antigen when the animal reached adulthood (Slavin, S., 2002, Int J Hematol 76 Suppl 1:172-175, which is incorporated by reference herein in its entirety). In animal models it has been demonstrated that bone marrow cells contribute to generation of a donor-specific tolerogenic state which is associated with chimeric hematopoiesis. The combination of donor-specific bone marrow transplant, with solid organs, has been used in some clinical situations to induce complete tolerance to the grafted organ without the need for chronic, continuous immune suppression (George, et al., 2002, Immunol Res 26:119-129, which is incorporated by reference herein in its entirety). Unfortunately, wide spread use of bone marrow induced tolerance is limited by the fact that bone marrow transplantation is associated with a high degree of morbidity and mortality during the myeloablative phase. In addition, the possibility of graft versus host disease is another pitfall to the full-scale implementation. In order to overcome this, several methods of inducing partial chimerism, or mini-chimerism are being investigated through the use of non-myeloid ablative techniques such as donor-specific transfusions combined with anti CD154 antibodies (Seung, et al., 2003, J Clin Invest 112:795-808, which is incorporated by reference herein in its entirety). Induction of organ tolerance by hematopoietic stem cells is believed to occur through both thymic dependent (Noris, et al., 2001, J Am Soc Nephrol 12:2815-2826, which is incorporated by reference herein in its entirety), and independent (van Pel, et al., 2003, Transpl Immunol 11:375-384, which is incorporated by reference herein in its entirety) mechanisms. Specifically, donor hematopoietic cells generate a variety of both lymphoid and non-lymphoid cells that express the same antigens found in the donor organ, but somehow redirect the immune system not to attack these specific antigens, while maintaining responses against other antigens not related to the graft. One mechanism that is postulated to occur is the thymic stromal tissue in the recipient becomes populated with donor-derived cells. These cells then act at the level of negative selection in order to induce apoptosis of T cells reactive to the donor antigen in a similar way to which the immune system deletes autoreactive T cells during thymic selection (Shizuru, et al., 2000, Proc Natl Acad Sci USA 97:9555-9560, which is incorporated by reference herein in its entirety). Another mechanism of tolerance involves the persistent presentation of donor antigen in absence of costimulatory molecules. This was demonstrated in one situation by the fact that persistence of T cells from the donor bone marrow is essential in maintaining tolerance (Xu, et al., 2004, J Immunol 172:1463-1471, which is incorporated by reference herein in its entirety). The mesenchymal component of the bone marrow produces a cell population that consitutively secretes immune inhibitory factors such as IL-10 and TGF-b while presenting antigens (Liu, et al., 2004, Transplant Proc 36:3272-3275; Togel, et al., 2005, Am J Physiol Renal Physiol 289:F31-42, each of which is incorporated by reference herein in its entirety). This is believed to further inhibit immunity in an antigen specific manner. During T cell activation, two general signals are required for the T cell in order to mount a productive immune response, the first signal is recognition of antigen, and the second is recognition of costimulatory or coinhibitory signals. Mesenchymal cells present antigens to T cells but provide a coinhibitory signal, thus specifically inhibiting T cells that recognize them, and other cells expressing similar antigens. Finally, the fact that CD34+ cells express the T cell killing molecule FasL has been postulated as another mechanism of tolerogenesis. Indeed transplantation of bone marrow from mice with a mutated FasL did not induce tolerogenesis in recipients (George, et al., 1998, Nat Med 4:333-335, which is incorporated by reference herein in its entirety).
The potential of using hematopoietic cell transplantation for autoimmunity derives from the belief that the immune system can be deleted and recapitulated, but in such a manner to "reset the clock" so that autoreactive T cells will not re-appear (Muraro, et al, Renewing the T cell repertoire to arrest autoimmune aggression. Trends Immunol., e-published on Jan. 4, 2006, which is incorporated by reference herein in its entirety). Specifically, it is known that the process of autoimmunity requires the failure of several self-tolerance mechanisms before clinical presentation appears. These include: a) self-reactive T cell deletion in the thymus; b) anergy/deletion of self reactive T cells in the periphery; c) failure of the regulatory T cell activity; and d) the presence of inflammation or antigen release in order to allow expansion of the autoreactive T cell clone. During autoimmunity the failure of all of these systems is usually a culmination of environmental and genetic factors occurring over a protracted period of time. Accordingly if the immune system could be made to "start anew" the normal tolerogenic processes would again be reactivated and the disease would be cured, at least temporarily. To date clinical use of autologous stem cells has been performed for a variety of autoimmune indications, including rheumatoid arthritis (Jantunen, et al., 1999, Scand J Rheumatol 28:69-74, which is incorporated by reference herein in its entirety), multiple sclerosis (Karussis, et al., 2004, J Neurol Sci 223:59-64; Brodsky, et al., 1999, Curr Opin Oncol 11:83-86, each of which is incorporated by reference herein in its entirety), systemic lupus erythromatosis (Brunner, et al., 2002, Arthritis Rheum 46:1580-1584; Burt, et al., 2006, Jama 295:527-535, each of which is incorporated by reference herein in its entirety), and systemic sclerosis (Viganego, et al., 2000, Curr Rheumatol Rep 2:492-500, which is incorporated by reference herein in its entirety). According to a report in 2005, approximately 700 patients in total have received an autologous stem cells for autoimmune diseases with a positive benefit/risk ratio that has led to initiation of phase III prospective randomized controlled trials (Tyndall, et al., 2005, Clin Exp Immunol 141:1-9, which is incorporated by reference herein in its entirety).
Induction of tolerance through hematopoietic stem cell transplantation, either from bone marrow or peripheral blood sources possesses the intrinsic danger of bone marrow failure during ablation of the recipient immune system. Although non-myeloablative protocols are under development, even these carry the risk of immune suppression due to the lymphoablation. Accordingly there is a need in the art to develop novel methods of either expanding hematopoietic stem cells ex vivo in large enough quantities to guarantee graft take, as well as methods of in vivo expanding the stem cells and their progeny so that the period under which the transplant recipient is immunosuppressed is minimized.
Stem cell therapy has also been performed in the context of administration of mesenchymal stem cells, without the hematopoietic component, for induction of tolerance. It was demonstrated in a murine model that flk-1+Sca-1-mesenchymal cell transplantation leads to permanent donor-specific immunotolerance in allogeneic host and results in long-term allogeneic skin graft acceptance (Deng, et al., 2004, Exp Hematol 32:861-867, which is incorporated by reference herein in its entirety). Other studies have shown that mesenchymal stem cells are inherently immunosuppressive through production of PGE-2, interleukin-10 and expression of the tryptophan catabolizing enzyme indoleamine 2,3,-dioxygenase as well as Galectin-1 (Kadri, et al., 2005, Stem Cells Dev 14:204-212; Ryan, et al., 2005, J Inflamm (Lond) 2:8, each of which is incorporated by reference herein in its entirety). These stem cells also have the ability to non-specifically modulate the immune response through the suppression of dendritic cell maturation and antigen presenting abilities (Beyth, et al., 2005, Blood 105:2214-2219; Aggarwal, et al., 2005, Blood 105:1815-1822, each of which is incorporated by reference herein in its entirety). Functional induction of allogeneic T cell apoptosis was also demonstrated using freshly isolated, irradiated, or long-term cultured mesenchymal stem cells (Plumas, et al., 2005, Leukemia 19:1597-1604, which is incorporated by reference herein in its entirety). Others have also demonstrated that mesenchymal stem cells have the ability to preferentially induce expansion of antigen specific T regulatory cells with the CD4+CD25+ phenotype (Maccario, et al., 2005, Haematologica 90:516-525, which is incorporated by reference herein in its entirety). Supporting the potential clinical utility of such cells, it was previously demonstrated that administration of mesenchymal stem cells inhibits antigen specific T cell responses in the murine model of multiple sclerosis, experimental autoimmune encephalomyelitis, leading to prevention and/or regression of pathology (Zappia, et al., 2005, Blood 106:1755-1761, which is incorporated by reference herein in its entirety). Safety of infusing mesenchymal stem cells was illustrated in studies administering 1-2.2×106 cells/kg in order to enhance engraftment of autologous bone marrow cell. No adverse events were associated with infusion, although level of engraftment remained to be analyzed in randomized trials (Koc, et al., 2000, J Clin Oncol 18:307-316, which is incorporated by reference herein in its entirety). In a matched pair analysis study, it was demonstrated that in vitro expanded mesenchymal stem cells reduced both acute and chronic graft versus host disease in the allogeneic bone marrow transplant setting. Clinical administration of mesenchymal stem cells was reported in a patient suffering severe, grade IV graft versus host disease in the liver and gut subsequent to bone marrow transplant. Administration of 2×106 cells/kg on day 73 after bone marrow transplant lead to a long term remission of graft versus host disease, which was maintained at the time of publication, 1 year subsequent to administration of the mesenchymal stem cells (Le Blanc, et al., 2004, Lancet 363:1439-1441, which is incorporated by reference herein in its entirety). A feasibility study in 46 patients receiving mesenchymal cells prior to transplant revealed a favorable safety profile and is encouraging further dose finding studies (Lazarus, et al., 2005, Biol Blood Marrow Transplant 11:389-398, which is incorporated by reference herein in its entirety). Unfortunately, mesenchymal cell expansion is relatively slow and in many situations is not practical for widespread clinical use. The development of novel methods of expanding stem cell populations, as for example the methods thought in the present invention, are likely to increase use of this therapeutically promising cell population.
There is evidence that embryonic stem cells are also capable of inducing immunological tolerance. Indeed, coculture of alloreactive T cells with embryonic T cells demonstrated an antigen-specific inhibitory effect (Li, et al., 2004, Stem Cells 22:448-456, which is incorporated by reference herein in its entirety). Data is still preliminary in this area, and the problem of embryonic stem cells inducing teratomas currently precludes their use for this indication. An alternative method of immune modulation using embryonic stem cells is the generation of defined immunological cells that can be used directly, or tailored to possess specific desired properties through modification of culture conditions or gene manipulation. For example, it was demonstrated that the murine model of multiple sclerosis, experimental autoimmune encephalomyelitis can be successfully treated with dendritic cells generated from embryonic stem cell cultures that have been manipulated to present the MOG autoantigen in the presence of TRAIL, a molecule known to induce T cell apoptosis (Hirata, et al., 2005, J Immunol 174:1888-1897, which is incorporated by reference herein in its entirety). Generation of such tailor-made immunological cells would greatly expand the clinical armamentarium of immunotherapy, however, this is limited by the currently lack of methodologies for expanding stem cells in a GMP/GTP compliant and feasible manner.
One of the main therapeutic uses for stem cells is in the area of regenerative medicine. The concept of regenerative medicine is to restore or enhance the ability of tissues to self-organize and heal themselves following endogenous or exogenous injury. Although examples of the use of stem cells for tissue regeneration are almost limitless, several are overviewed below. This should not be taken as an exhaustive literature review, but rather a general discussion for example purposes in order to stimulate one skilled in the art to further investigate this field.
Bone marrow stem cells have been extensively investigated for repair of myocardial tissue subsequent to infarction. Early studies by Orlic demonstrated that administration of GFP c-kit+, lineage−, bone marrow into ligation induced myocardial infarct area resulted in regeneration of myocardial and endothelial tissue by the donor cells (Orlic, et al., 2001, Nature 410:701-705, which is incorporated by reference herein in its entirety). Subsequent studies have used mesenchymal bone marrow cells treated with the DNA methyltransferase inhibitor 5-aza-cytidine to not only transdifferentiate into myocardial tissue, but also to improve left ventricular ejection fraction and inhibit cardiac remodeling (Tomita, et al., 1999. Circulation 100:II247-256, which is incorporated by reference herein in its entirety). Importantly, similar experiments were performed in porcine models of infarction, also indicating improvement in cardiac function (Tomita, et al., 2002, J Thorac Cardiovasc Surg 123:1132-1140, which is incorporated by reference herein in its entirety). Accordingly, clinical experiments were performed administering autologous bone marrow cells directly into the myocardium during coronary bypass grafting. In a series of experiments initiated in 1999, 5 patients treated had no adverse effects, with objective vascularization enhancement in the area of stem cell administration as detected by nuclear imaging (Hamano, et al., 2001, Jpn Circ J 65:845-847, which is incorporated by reference herein in its entirety). A subsequent study administering AC133 purified bone marrow stem cells into the infarct area in 12 patient during bypass grafting demonstrated a marked improvement in left ventricular ejection fraction, a decreased rate of remodeling, and improved perfusion (Stamm, et al., 2004, Thorac Cardiovasc Surg 52:152-158, which is incorporated by reference herein in its entirety). Administration of stem cells into coronary circulation or directly into the myocardium has also been performed both in the angina setting, as well as subsequent to cardiac infarct in order to enhance angiogenesis, and prevent remodeling, respectively. In patients with end stage angina, administration of autologous bone marrow cells using the NOGA catheter system in 14 patients resulted in improved ejection fraction from a baseline of 20% to 29% (P=0.003) and a reduction in end-systolic volume (P=0.03) in the treated patients. Furthermore, electromechanical mapping revealed significant mechanical improvement of the injected segments (P<0.0005) at 4 months after treatment (Perin, et al., 2003, Circulation 107:2294-2302, which is incorporated by reference herein in its entirety). Improvements were also notably maintained in the same patient population at 1-year follow-up (Perin, E., 2004, Int J Cardiol 95 Suppl 1:S45-46, which is incorporated by reference herein in its entirety). Transcoronary administration of bone marrow cells in patients post-myocardial infarction induced an improvement at 6 months in regional and global LV function, increased thickness of the infarcted wall, and showed a reduction in myocardial remodeling as determined by a decrease in the end-systolic volume (Fernandez-Aviles, et al., 2004, Circ Res 95:742-748, which is incorporated by reference herein in its entirety). In another study, patients post myocardial infarction were transplanted with autologous bone marrow cells via a balloon catheter placed into the infarct-related artery during balloon dilatation (percutaneous transluminal coronary angioplasty), resulting in decreased infarct size, improved wall motion score, and a decrease in ventricular remodeling (Strauer, et al., 2002, Circulation 106:1913-1918, which is incorporated by reference herein in its entirety). Randomized trials are currently underway using autologous bone marrow stem cells for increasing cardiac function post myocardial infarction although results are still controversial and inconclusive (Assmus, et al., 2002, Circulation 106:3009-3017; Cleland, et al., 2006, Eur J Heart Fail 8:105-110, each of which is incorporated by reference herein in its entirety). In addition to bone marrow hematopoietic cells, other types of stem cells have been utilized for improvement in myocardial activity, perfusion, and decreasing ventricular remodeling. These include mesenchymal stem cells (Chen, et al., 2004, Chin Med J (Engl) 117:1443-1448, which is incorporated by reference herein in its entirety), endothelial stem cells (Aoki, et al., 2005, J Am Coll Cardiol 45:1574-1579, which is incorporated by reference herein in its entirety), and skeletal myoblasts (Ye, et al., 2006. Exp Biol Med (Maywood) 231:8-19, which is incorporated by reference herein in its entirety). A limiting factor in presently used cellular therapies for myocardial dysfunction is the lack of ability to induce transdifferentiation of the stem cells into the desired cardiac tissue in a directed manner. Additionally, methods do not exist for expanding sufficient numbers of semi-differentiated progenitor stem cells that possess a high proclivity for repairing the heart. This drawback is in part due to lack of proper culture mediums for expansion of such unique cell populations. The current invention addresses this issue.
The importance of stem cells inducing regeneration of other organ systems has been shown in a variety of settings. In a pathological setting, it was reported that bone marrow derived stem cells are the precursors of stomach epithelial tissue in Helicobacter pylori infected mice that progresses to the develop stomach cancer (Houghton, et al., 2004, Science 306:1568-1571, which is incorporated by reference herein in its entirety). In a therapeutic setting, administration of Green Fluorescent Protein (GFP) bone marrow stem cells into rats with ethanol-induced ulcers resulted in generation of GFP expressing, cytokeratin-positive epithelial cells and vimentin-positive interstitial cells, contributing to a decreased pathology in the stem cell recipients (Komori, et al., 2005, J Gastroenterol 40:591-599, which is incorporated by reference herein in its entirety). The human bone marrow derived Flk1(+)/CD31(−)/CD34(−) cell population was reported to transdifferentiated into a variety of tissues, including stomach epithelium when injected into non-obese diabetic, severe combined immunodeficient (NOD-SCID) mice, thus suggesting human stem cells also possess such transdifferentiation ability (Fang, et al., 2003, J Hematother Stem Cell Res 12:603-613, which is incorporated by reference herein in its entirety). Stomach-homing capacity to injured tissue of human stem cells was demonstrated human mesenchymal stem cells infused systemically in NOD-SCID mice that received radiation to the abdominal area. This resulted in a specific rise in stem cell engraftment exclusively to the irradiated areas (Francois, et al., Local irradiation induces not only homing of human Mesenchymal Stem Cells (hMSC) at exposed sites but promotes their widespread engraftment to multiple organs: A study of their quantitative distribution following irradiation damages. Stem Cells, e-published on Dec. 8, 2005, which is incorporated by reference herein in its entirety) It is anticipated that since stem cells can selectively home to the injured stomach area, addition of factors to allow expansion once already homed into the injured tissue will increase therapeutic efficacy of stem cell therapies. The invention teaches methods of expanding cells that have already homed to an injured tissue.
The use of stem cells has also been applied to liver disease. It is known that partial hepatectomy leads to mobilization of an AC133+ stem cell population in clinical situations (Gehling, et al., 2005, J Hepatol 43:845-853, which is incorporated by reference herein in its entirety). Furthermore, studies using carbon tetrachloride induced liver injury have demonstrated a therapeutic effect of bone marrow flk-1+ cell infusion (Fang, et al., 2004, Transplantation 78:83-88, which is incorporated by reference herein in its entirety). It is believed that liver damage induces expression of several chemokines, including stromal derived factor-1 (SDF-1) which attracts stem cells into the damaged areas (Hatch, et al., 2002, Cloning Stem Cells 4:339-351, which is incorporated by reference herein in its entirety). Therapeutic mobilization of endogenous stem cells using granulocyte colony stimulating factor (G-CSF) has also demonstrated protective effects in liver injury models (Quintana-Bustamante, et al., 2006, Hepatology 43:108-116, which is incorporated by reference herein in its entirety). It is anticipated that since stem cells can selectively home to the injured hepatic area, addition of factors to allow expansion once already homed into the injured tissue will increase therapeutic efficacy of stem cell therapies. The invention teaches methods of expanding cells that have already homed to an injured tissue.
Stem cells have also been useful for treatment of neurological deficiencies in a variety of situations. Administration of fetal stem cells in the form of mesenchphalic tissue into the striatal area of Parkinson's disease (PD) patients have demonstrated that grafted dopaminergic neurons can reinnervate the striatum, restore regulated dopamine release and movement-related frontal cortical activation, and result in observable clinical benefit (Lindvall, et al., 2004, NeuroRx 1:382-393, which is incorporated by reference herein in its entirety). Patients suffering from stroke have also been treated by implantation of autologous mesenchymal stem cells into the middle cerebral arterial territory. Improvements were seen in some functional indexes such as the Barth's score (Bang, et al., 2005, Ann Neurol 57:874-882; Rabinovich, et al., 2005, Bull Exp Biol Med 139:126-128, each of which is incorporated by reference herein in its entirety). A wide variety of neurological indications are currently under investigation for amenability to stem cell therapy (Kulbatski, et al., 2005, Curr Drug Targets 6:111-126; Zhu, et al., 2005, Curr Drug Targets 6:97-110, each of which is incorporated by reference herein in its entirety). Unfortunately, ethical issues associated with the use of fetal tissue, as well as inability to define the activities and functions of neurally injected stem cells hampers progress in the field. Development of novel culture and expansion methodologies for stem cell applications is therefore an important area of issue.


The olefin-hydrogen effluent vapor stream from a dehydrogenation process is separated by a cryogenic separation method utilizing a cryogenic separation system. The method does not require external refrigeration and reheats and portions an expander feed stream to extract energy and controls the warm end and cold end temperature differences in the primary heat exchanger to provide energy savings and economical operation and material use.

The function of a dehydrogenation process, when utilized in hydrocarbon processing, is to convert paraffinic hydrocarbons to their corresponding olefins.
Thus, propane is converted to propylene: EQU C.sub.3 H.sub.8 =C.sub.3 H.sub.6 +H.sub.2
or isobutane is converted to isobutylene: EQU iC.sub.4 =iC.sub.4 H.sub.8 +H.sub.2
These are two examples of the use of the process.
The reactor effluent stream from such dehydrogenation processes contains these components in the vapor phase. A cryogenic process is employed to condense the C.sub.3 or C.sub.4 components as a liquid phase product. The residual vapor stream is required free of the C.sub.3 or C.sub.4 components to minimize their losses.
A method and apparatus for the separation of C.sub.4 hydrocarbons from a gaseous mixture containing the same are described in U.S. Pat. Nos. 5,414,188, issued May 9, 1995, and 5,505,048, issued Apr. 9, 1996, hereby incorporated by reference.
These patents are directed to the separation of the C.sub.4 hydrocarbons in a series of cooling for condensation, phase separation, and expansion process steps with refrigeration by indirect heat exchange between expansion products to save energy and power consumption.
It is desirable to provide a new and improved cryogenic separation process and apparatus system for the
It is desirable to provide a new and improved cryogenic separation process and apparatus system for the separation of C.sub.3 or C.sub.4 gaseous mixtures with hydrogen, such as produced in dehydrogenation processes to permit the use of lower Cost and more economical materials of construction in the process, to increase the extraction of energy in the process, to reduce the size of heat exchangers employed in the process and apparatus, and to otherwise improve the efficiency and economy of the separation process and apparatus system.

Cryogenic separation process for the recovery of components from the products of a dehydrogenation reactor


A method for reducing a size of an image of a class file is provided. In this method, a first stack map that is associated with a first bytecode of a method and a second stack map that is associated with a second bytecode of the method are provided. Next, an operation of the second bytecode is applied to determine an effect on the first stack map. The effected first stack map defines a resulting stack map. The resulting stack map is then compared with the second stack map. If the resulting stack map matches the second stack map, the second stack map is removed from the class file. The removal of the second stack map operates to reduce the size of the image of the class file.

1. Field of the Invention
The present invention relates to computer programming languages and, more particularly, to methods and systems of reducing a program size.
2. Description of the Related Art
A stack based computer language, such as JAVA™ (hereinafter referred to as "Java"), basically puts and gets variables to and from a stack. As a result, the contents of the stack need to be identified at any given point of a program. To identify the contents of the stack, classes contain stack maps, which describe the state of the stack being executed at various points of the program. The stack maps constitute a significant amount of program size. For example, in Java, stack maps typically constitute 10% of a read-only memory (ROM) image. Having a small ROM image is particularly important for resource-constrained consumer devices such as cellular telephones, two-way pagers, and personal organizers. In view of the foregoing, there is a need to provide methods and systems for reducing the size of a program.

Methods and systems for reducing a program size


According to various aspects of the present disclosure, there are provided various exemplary embodiments of covers, systems and methods for covering boards and deck components. In one exemplary embodiment, a cover generally includes an upper portion and generally downwardly extending side portions. The cover's upper and side portions are configured to generally cover at least the top and sides of the outdoor deck component. The cover's side portions may include through-holes to allow venting of moisture from an area between the cover and the outdoor deck component when the cover is positioned on the outdoor deck component.

The statements in this section merely provide background information related to the present disclosure and may not constitute prior art.
Decks are typically constructed of wood. These decks are often constructed using standard nails, screws, and other conventional construction techniques to secure the various component parts of the deck together. For example, nails are often used to secure the floorboards to support members and to construct other features of the deck. By way of further example, a deck may include side rails for enclosing the deck structure and/or integrated benches for providing seating.
Decks are often constructed of wood that has been chemically impregnated to retard decomposition. The treated wood typically includes various additives, some of which may be particularly harmful to children and pets. For example, treated wood commonly used in the past for constructing decks included arsenic, which is potentially dangerous to users of the deck.
In addition, extensive maintenance of wood components is normally required, particularly in geographic areas subject to extreme weather changes or harsh weather conditions (e.g., excessive rain or sunlight, etc.). For example, periodic treatment of the deck surface with a water sealer or similar protecting agent is normally required to slow damage to the deck (e.g., splitting of the wood, etc.) caused by weather conditions. Further, a stain or similar type of material for maintaining a desired color of the deck is often used, and typically requires periodic application. These protecting agents and stains add costs to the maintenance of the deck. Additionally, despite efforts to maintain the condition of the deck (e.g., sealing the deck yearly to inhibit damage, etc.), floorboards, railing boards, and other deck parts often must be replaced as a result of continuous exposure to outdoor elements.
Plus, the standard wood nails that are typically used with deck construction often loosen from the wood and become raised. In addition to their unsightliness, these raised nails can result in a potentially dangerous condition that may cause injury to users of the deck (e.g., injury to a user's foot while walking on the deck, etc.).

This patent is subject to a terminal disclaimer.

Covers, systems, and methods for covering outdoor deck components


A power starting circuit used in a DC power supply circuit has a double-pole, double-throw switch electrically connected to a positive pole of a DC power supply and controls a conducting state of the circuit. A first capacitor provides a voltage signal for a first switch circuit and controls the conducting state of the first switch circuit in a continuous charging and discharging process. The first switch circuit controls the conducting state of the DC power supply circuit. A maintaining signal input end provides a maintaining signal for the first capacitor. The power starting circuit can achieve an over discharge protection function for the control circuit. Moreover, when the switch is turned on, it can automatically switch the circuit off so as to save power if the electrical product does not need to continue to work.

The following generally relates to a power starting circuit and, more particularly, to a power starting circuit for controlling a DC power supply.
As shown in FIG. 1, a currently known power starting circuit used in a hand-held power tool comprises: a switch SW11 having a first position end electrically connected to the positive pole of a DC power DC10 (the DC power DC10 is a battery pack) and a second position end electrically connected to a diode D11, a resistor R13 and a transistor Q11 respectively. The diode D11 and the resistor R13 are electrically connected to a resistor R11, a resistor R12, a transistor Q12 and a resistor R14 to form a switching circuit 100 for controlling the conducting state of an output end 11. When the switch SW1 is turned off, if the voltage of the battery pack 10 is higher than the break-over voltage of the diode D11, the transistor Q12, Q11 are on, and the circuit is started; if the voltage of the battery pack 10 is lower than the break-over voltage of the diode D11, the transistor Q12, Q11 are off, and the power supply is stopped to prevent the over discharge of the battery, wherein the break-over voltage of the diode D11 is close to the over discharge protection voltage of the battery pack 10. In the operation process, the currently known starting circuit can achieve an over discharge protection function of the battery, but it can not achieve a power saving function by automatically turned-off when the power of the battery is sufficient but not be used.


A media drive is disclosed. In one embodiment, a media drive can comprise: a disk holder, a drive laser, a disk reader, and a luminescence detector. The disk holder can be capable of applying a rotational force to a media disk. The drive laser is disposed so that light from the laser will be incident on the media disk when it is disposed in the disk holder. The disk reader is disposed to receive data from the media disk. A method for authenticating a media disk is also disclosed. The method can comprises: disposing the article in a reader, illuminating at least a portion of the article with the information reader, monitoring a luminescence emission of the illuminated article, and determining if the luminescence emission is consistent with an authentic article. If a luminescent material is present, the information reader excites the luminescent material to create a luminescence emission.

In the field of data storage media, digital content owners, such as music companies, movie studios, video game manufacturers, computer software manufacturers, and the like, desire increased flexibility in the distribution of their digital content onto various forms of data storage media. A major problem confronting the various makers and users of data storage media such as compact discs (CD), digital versatile discs (DVD), enhanced video discs (EVD), recordable compact discs (CD-R) and recordable digital versatile discs (DVD-R) is the unauthorized reproduction or copying of information by unauthorized manufacturers, sellers, and/or users. Such unauthorized reproduction or duplication of data storage media is often referred to as piracy. Piracy may occur in a variety of ways, including consumer level piracy at the point of end use as well as wholesale duplication of data, substrate and anti-piracy information at the commercial level. Regardless of the manner, piracy of data storage media deprives legitimate digital content providers and manufacturers of significant revenue and profit.
Attempts to stop piracy at the consumer level have included the placement of electronic anti-piracy signals on information carrying substrates along with the information sought to be protected. The machine readers and players of such data storage media are configured to require the identification of such anti-piracy signals prior to allowing access to the desired information. Theoretically, consumer level duplications are unable to reproduce these electronic anti-piracy signals on unauthorized copies and hence result in duplicates and copies that are unusable.
However, numerous technologies to thwart such consumer level anti-piracy technologies have been and continue to be developed. Moreover, commercial level duplications have evolved to the point that unauthorized duplicates may now contain the original electronic anti-piracy circuit, code, etc. For example, commercial level duplication methods include pit copying, radio frequency (RF) copying, "bit to bit" copying and other mirror image copying techniques which result in the placement of the anti-piracy signal on the information carrying substrate of the duplicate along with the information sought to be protected. Other technologies commonly used by hackers include the modification of the computer code in order to remove anti-piracy (also referred to as copy-protection or copy-proofing) features and enable unlimited access to the data.
It would be desirable to have a data storage media that can be easily identified as to being authentic or pirated. One anti-piracy technology aimed at combating these more sophisticated consumer and commercial level reproduction and copying practices involves the placement of 'tags' or authentication markers in substrates used in the construction of data storage media. Such tags or authentication markers can be detected at one or more points along the data storage media manufacturing or distribution chain or by the end use reader or player used to access the data on a particular data storage media.
The automated identification of plastic compositions used in data storage media is very desirable for a variety of applications, such as recycling, tracking the manufacturing source, antipiracy protection, and others. Furthermore, it may be desirable for automated authentication of the data storage media.
Despite the foregoing, there still remains a desire for an authentication detector and a method for using the same.

Media drive with a luminescence detector and methods of detecting an authentic article


An improved carding process utilizing a brush attached to a cylinder carding machine for inhibiting the buildup of napped fibers. Mounted just downstream of the transfer region where carding takes place, the brush is stationary with respect to the revolving cylinders. The brush is disposed across the face of the carding drum with the nylon bristles of the brush engaged in the card-cloth wires of the drum. The bristles continuously brush, straighten and force napped fibers down into the wires of the carding cylinder, which results in a thicker, heavier web of carded fibers and virtually eliminates buildup of fluffy fibers on the carding drum as well as the input drum.

This invention relates to the manufacture of textiles, and more particularly to the carding of fibers, including an improved method of carding wool and fine-fiber hairs, and apparatus for carrying out the method.
In the manufacture of textiles, an implement or machine called a card or carder is utilized to process fiber stock such as grease wool or picked fleece, to disentangle, arrange and collect together the individual fibers in an arrangement suitable for spinning. The carding process, historically a handicraft, was mechanized in the mid to late eighteenth century.
A card employs "card cloth" or wire "clothing" which comprises a matrix of wire tines or teeth, set closely spaced apart in rows, in a suitable foundation such as leather, rubber or synthetic material. The wire tines are usually bent to a uniform angle, all pitched in the same general direction, the points providing in concert a surface for the action of carding fiber stock such as wool. In the carding process, two opposed wired surfaces are moved relatively in opposite directions, the wired surfaces of the adjacent cards being close to each other, but not touching or meshing. The surfaces are disposed such that the points of the wire clothing of the respective surfaces pitch in opposite directions, working point against point, and this opposition provides, in part, the tension applied to fibers disposed between the card surfaces.
Carding subjects fiber stock, a tight mass of tangled, matted fibers having nonuniform and erratic distribution throughout the mass, to forces set up between two card-cloth surfaces. Fiber stock is applied to a first one of the surfaces, where it is held, entangled among the wires. Carding takes place when there is forcible transfer of individual fibers from the first card-cloth surface to the other surface. Fibers of the stock are held by friction on the card-cloth wires of the first card. Wires of the empty card, passing near the wires of the card charged with fibers, snag and hold individual fibers of the mass, causing tension in the fibers, and it is the tension built up in the fibers in overcoming the frictional forces holding the fibers which effectuates carding. When the tension is sufficient to overcome the frictional force holding a fiber to the wire(s) of the first surface, the fiber is released, forcibly extracted and transferred to the surface of the second card with a whipping action induced by spring temper of the wires. The transferred fibers are thus collected and laid down in a coalescent web or sheet of intermeshed fibers termed a bart or sliver. The purpose of carding is to achieve a uniform distribution of density of fibers in a bart necessary to allow maximum control by the hand spinner in drafting, or by machines in drawing and spinning yarn. Depending on the type of fiber carded and the kind of yarn desired, one carding may suffice, while other fibers and blends of fibers may require additional cardings to achieve the degree of evenness and uniformity of distribution of the fibers desired for spinning. The carding process can be repeated by stripping the bart of partially carded fibers from the second surface, reapplying the batt to the first surface and carding the fibers again to further open, straighten and redistribute the fibers. Whether performed manually as a handicraft, or effected on a large scale by industrial machinery, the carding process is essentially the same.
Artisans who spin yarn by hand often prepare fiber stock for spinning by utilizing hand-held cards, which look like flat, rectangular paddles with handles; the paddles have card cloth affixed to one side. Other artisans and hand spinners prepare fibers for spinning on small, two-cylinder carding machines, called "drum carders," which have card-cloth wires disposed on the surfaces of two cylinders or drums of different size, rotating with their surfaces tangentially adjacent and parallel to each other. Large industrial carding machines employ multiple stages of carders, each stage having small worker and stripper drums set successively closer to a large main cylinder or swift, thus subjecting the fiber stock to repeated carding, and a thorough opening, cleaning and straightening of the fiber. The carded fibers are removed from the main cylinder by a dolling cylinder, as described below.
Small, drum carders utilized by artisans employ two rotating cylinders journaled on a wooden frame: a small diameter input drum or "licker-in," and a larger diameter carding drum or swift, the swift rotating considerably faster than the licker-in. Although some of these machines are powered by small electric motors, the process is essentially a handicraft, the machines being fed and stripped by hand, and commonly operated manually by a hand crank. Fiber stock loaded onto a feed pan adjacent the licker-in is impaled by the wire teeth of the smaller cylinder, pulled in and held on the surface of the cylinder as it rotates. The carding takes place along a narrow gap between the surfaces of the licker-in and the swift, called the transfer region, where the surfaces approach closely but do not touch or intermesh. Some of the fibers projecting above the teeth of the licker-in are readily caught by the wire clothing on the surface of the swift, which is moving rapidly in the opposite direction relative to the licker-in, the wires working point against point, pulling the fibers away against resistance from the smaller cylinder, thus disentangling, straightening and opening the mat of fibers, the extracted fibers cohering in the wires of the swift. After a web or batt of carded fiber of sufficient thickness is collected on the swift, the machine is stopped and the web manually stripped or doffed, and further processed by additional carding, or by spinning.
The carding process is inherently complex for the artisan because of the wide range of properties of fibers and various blends of fibers, and handicraft carding can be extremely demanding of the time and patience of the practitioner, often requiring that the fibers be passed through the drum carder several times, using different settings of drum separation, various densities of card-cloth wires, and/or different relative speeds of the drum surfaces. The spacing and parallelism of the cylinders, the pitch and density of the teeth, the relative speed of the drum surfaces, the manner and rate at which the fiber stock is input into the feed tray, and the properties (and mixes) of the fibers, are all critical factors which affect the efficacy of the carding process. Considerable experience and skill are required of the artisan to successfully card fiber with a drum carder.
The carding of fine-fiber hair such as alpaca, angora rabbit, cashmere, mohair, etc., is more difficult than processing ordinary coarse wool, because such fine fibers generally are shorter and smoother, having fewer convolutions and epidermal scales than most wool, and therefore less disposed to be held by friction on the card-cloth wires and grasped by the moving card-cloth wires of the opposed surface. Consequently, when the shorter, smoother fibers are caught by the moving wire clothing of the carding drum, many of the fibers are pulled up but not extracted from the fiber stock disposed on the input drum, while other fibers are easily released from the wire clothing of the input drum; therefore, little tension is developed in the fibers, resulting in ineffectual carding. Fibers are left protruding above the surface of both the carding and input drums, where they interfere with the carding process and build up rapidly above the wire tines to form a villous or fluffy nap. The buildup of napped fibers is believed to be caused, also, in part, by static electricity, wherein static charges induced in the smoother fibers by the motion of the drums causes the loosened fibers to extend outward from the surface of the drum. The napped fibers clog the carder, resulting in an excessively fluffy bart of partially and incompletely carded fibers. The problem of excessive fluffing is seen also in the carding of shorter, fine-fiber wools such as those produced, for example, by merino and rambouillet breeds of sheep, The problem is conventionally solved by one or more of the following: providing a more dense card clothing for processing the smooth, short fibers; adjusting the gap between the drums of the carder; or additional passes of the fluffy batts through the carding machine. However, these conventional solutions are expensive, labor intensive, time consuming, and not always effective.
Brushes are known in certain industrial carding machines; however, such brushes are not employed intrinsically in the carding process, but perform some other function apart from carding, as to extract material from rotating cylinders, or to clean waste material from particular elements of the machine. For example, in the latter instance, a rotating drum having bristles disposed in a spiral configuration cleans residual lint from the flats of cotton carders. In the former instance, a "fancy roller," utilizing long, brush-like wire clothing, working back against back in relation to the card-cloth wires of the swift, brushes up or lifts a web of carded fibers above the surface of the swift, so that the web is in a position favorable for transfer to a dolling cylinder, the element which removes the carded web from the swift.
It is therefore a principle object of my invention to provide an improved carding machine.
Another object of the invention is to provide an improved process for carding wool and fine fibers, and to provide apparatus for carrying out the process.
It is another object of my invention to provide an improved drum carder for artisans who prepare fiber for spinning.
Another object of the invention is to provide apparatus in a carding machine which inhibits the buildup of fluffy nap in a web of carded fibers.
Another object of the invention is to provide apparatus in a cylinder carding machine which prevents clogging of the transfer region of the carding mechanism with napped fibers.
Another object of the invention is to provide apparatus in a carding machine which inhibits the buildup of napped fibers on the input cylinder of the machine.
Yet another object of the invention is to provide apparatus in a cylinder carding machine which facilitates carding a wide range of stock fibers and fiber blends of different staple length, smoothness and cohesiveness, without changing the configuration of the carding machine cylinders and the card-cloth wires.
Still another object of my invention is to provide an improved cylinder carding machine having means for forcing napped fibers on the carding cylinder into the card-cloth wires of the carding machine.
Another object of the invention is to provide an improved cylinder carding machine having a brush with bristles engageable with the card clothing of the carding cylinder to brush napped fibers into the card-cloth wires positioned on an outer portion of the cylinder.

Carding machine having a fine-fiber brush


Embodiments of the invention provide systems and methods for locking a seat belt with respect to a fixed base frame to prevent movement of a movable seat portion in the event of a crash or crash-like condition on an aircraft.

Aircraft and other passenger transport vehicle seats are often designed to allow passengers to move from a fully seated upright position to a partially or fully reclined position. These seats may allow only a slight backward motion, or they may be seats that are convertible to a bed, i.e., to a substantially flat bed position so that the passenger may lie down. Premium seats for civil aircrafts typically offer a number of various seat positions, from the most upright position to a full flat or a lie flat bed position. In some instances, these seats may also be surrounded by a privacy shell or partition, which allows seat motion and other passenger movement without interrupting the space of the passenger behind. In that case, the seats are usually composed of a fixed base frame, a movable part used as a seat pan, a back rest, and one or more electrical actuators for seat position adjustment. From upright to the bed-like position, the movable part of the seat will move forward or backward, allowing or causing the backrest to recline or raise without interfering with the privacy shell.
The passenger seat belt is typically attached to the movable part of the seat, in order to retain the passenger in any position. It is important for the seat belt to function to both hold the passenger in place during normal activities, such as take-off and landing and normal air turbulence, as well as to safely restrain the passenger in place during extreme turbulence or crash/crash-like conditions.
Airline guidelines generally require that passenger seats must sustain crash conditions when in upright position. The main issue to solve is thus how to transfer the load due to the passenger mass up to the seat attachment point, knowing that the seat deflection should be limited. This has been done previously either using electromagnetic locker pins to secure the movable part of the seat to the base frame in its upright position or using the brake of the actuator to sustain the load. Both solutions add weight and cost to the design. Accordingly, improved seat belts and securing functions are needed.


Polycyclic aromatic hydrocarbon (PAH) functionalized isobutylene copolymers, methods for making a PAH functionalized isobutylene copolymer comprising combining a halogenated copolymer with a PAH in a solvent under basic conditions at a temperature ranging from 30° C. to 150° C., the use of these PAH functionalized copolymers as a dispersant in elastomeric nanocomposite compositions comprising a halobutyl rubber matrix and nanoparticles of graphite or graphene, and a tire innerliner or innertube produced from these elastomeric nanocomposite compositions is disclosed.

Halobutyl rubbers, which are halogenated isobutylene/isoprene copolymers, are the polymers of choice for best air retention in tires for passenger, truck, bus, and aircraft vehicles. Bromobutyl rubber, chlorobutyl rubber, and halogenated star-branched butyl rubbers can be formulated for specific tire applications, such as tubes or innerliners. The selection of ingredients and additives for the final commercial formulation depends upon the balance of the properties desired, namely, processability and tack of the green (uncured) compound in the tire plant versus the in-service performance of the cured tire composite. Examples of halobutyl rubbers are bromobutyl (brominated isobutylene-isoprene rubber or BIIR), chlorobutyl (chlorinated isobutylene-isoprene rubber or CIIR), star-branched butyl (SBB), EXXPRO™ elastomers (brominated isobutylene-co-p-methyl-styrene) copolymer (otherwise known as BIMSM), etc.
For rubber compounding applications, traditional small sub-micron fillers such as carbon black and silica are added to halobutyl rubbers to improve fatigue resistance, fracture toughness and tensile strength. More recently, methods to alter product properties and improve air barrier properties in halobutyl rubbers have been developed that comprise adding nanofillers apart from these traditional fillers to the elastomer to form a "nanocomposite." Nanocomposites are polymer systems containing inorganic particles with at least one dimension in the nanometer range (see, for example, WO 2008/042025).
Common types of inorganic particles used in nanocomposites are phyllosilicates, an inorganic substance from the general class of so called "nanoclays" or "clays" generally provided in an intercalated form wherein platelets or leaves of the clay are arranged in a stack in the individual clay articles with interleaf spacing usually maintained by the insertion of another compound or chemical species between the adjacent lamellae. Ideally, intercalation inserts into the space or gallery between the clay surfaces. Ultimately, it is desirable to have exfoliation, wherein the polymer is fully dispersed with the individual nanometer-size clay platelets.
The extents of dispersion, exfoliation, and orientation of platy nanofillers such as organosilicates, mica, hydrotalcite, graphitic carbon, etc., strongly influence the permeability of the resulting polymer nanocomposites. The barrier property of a polymer in theory is significantly improved, by an order of magnitude, with the dispersion of just a few volume percent of exfoliated high aspect-ratio platy fillers, due simply to the increased diffusion path lengths resulting from long detours around the platelets. Nielsen, J. Macromol. Sci. (Chem.), vol. A1, p. 929 (1967), discloses a simple model to determine the reduction in permeability in a polymer by accounting for the increase in tortuosity from impenetrable, planarly oriented platy fillers. Gusev et al., Adv. Mater., vol. 13, p. 1641 (2001), discloses a simple stretched exponential function relating the reduction of permeability to aspect ratio times volume fraction of the platy filler that correlates well with permeability values numerically simulated by direct three-dimensional finite element permeability calculations.
To maximize the effect of aspect ratio on permeability reduction, it is therefore useful to maximize the degree of exfoliation and dispersion of the platelets, which are generally supplied in the form of an intercalated stack of the platelets. However, in isobutylene polymers, dispersion and exfoliation of platy nanofillers requires sufficient favorable enthalpic contributions to overcome entropic penalties. As a practical matter, it has thus proven to be very difficult to disperse ionic nanofillers such as clay into generally inert, nonpolar, hydrocarbon elastomers. The prior art has, with limited success, attempted to improve dispersion by modification of the clay particles, by modification of the rubbery polymers, by the use of dispersion aids, and by the use of various blending processes.
Due to the difficulties encountered in dispersing ionic nanoclays in nonpolar elastomers, graphitic carbon has been explored as an alternative platy nanofiller. For example, elastomeric compositions comprising graphite nanoparticles are described in U.S. Pat. No. 7,923,491.
US 2006/0229404 discloses a method for making compositions of an elastomer with exfoliated graphite in which the diene monomers are polymerized in the presence of 10 phr or more exfoliated graphite so that the graphite is intercalated with the elastomer.
U.S. Pat. No. 8,110,026 describes a process for producing a functional graphene sheet (FGS) based on exfoliation of oxidized graphite suitable for a high degree of dispersion in a polymer matrix for use in a nanocomposite.
Nano graphene platelets (NGPs) obtained through rapid expansion of graphite have become commercially available as of late. These NGPs have graphitic surfaces, as opposed to graphene oxide platelets of oxidized graphitic surfaces, and are quite compatible with hydrocarbon based non-polar butyl halobutyl rubbers. However, a high degree of exfoliation and dispersion of NGPs without agglomerations and aggregations cannot be achieved by solid compounding or solution mixing of these nanoparticles into halobutyl rubbers.
There is a need, therefore, for improving the dispersion of graphite and graphene nanofillers in elastomeric nanocomposite compositions comprising halobutyl rubbers useful for tires, air barriers, and other things requiring air retention, in order to improve the air impermeability of those compositions. The present invention fulfills this need by providing a novel graphite and graphene nanofiller dispersant useful in isobutylene-based elastomer/nanofiller nanocomposite compositions that results in these nanocomposite compositions having improved air barrier properties and that are suitable for use as a tire innerliner or innertube.

Polycyclic aromatic hydrocarbon functionalized isobutylene copolymers for dispersing graphene and graphite


A metallized thermoplastic label includes a first skin layer containing a thermoplastic and a first cavitating agent, wherein the first skin layer has a first side and a second side, the first skin layer is cavitated, and the first side of the first skin layer is adapted to be used in contact with a cold glue adhesive. The label further includes a cavitated core layer and a second skin layer that is metallized. The particle size of the cavitating agent in the core layer is selected to achieve a desired appearance, especially luster, of the metal layer.

This invention relates to metallized labels generally, and more specifically, but without limitation, to die cut or patch metallized labels for use with cold glue, and to films for making such labels. In one aspect, the invention relates to the use of such labels on substrates, where the label forms a destruct bond with the substrate such that the label delaminates. In another aspect, the invention relates to labels which possess the beneficial properties of the known thermoplastic labels, but which exhibit improved initial adhesion and destruct bonding, such as delamination.
In many label applications, it is desirable that the label stock from which the labels are cut be a film of polymeric material rather than paper. Polymeric film can provide one or more of the following properties lacking in paper: durability, strength, water resistance, curl resistance, abrasion resistance, gloss, transparency and other properties. Obviously, the polymeric material must meet the practical requirements of commercial manufacture and use. Material costs must be competitive. The film must be formable by a suitable commercial process, such as cast film extrusion or blown film extrusion, requiring that the molten film material be flowable to the correct degree to accomplish proper film formation. The formed film must be capable of hot-stretching without deleterious effect, because it is generally advantageous to hot-stretch and anneal the formed film, so as to orient the film and impart a stiffness to the film that is different in machine and cross directions. The film must have a printable face and be die-cuttable and matrix-strippable when used in a pressure-sensitive label construction.
In the die cut, cold glue label applications, there are certain other desired characteristics. Cold glues are widely used as an economical alternative to wrap-around or pressure-sensitive labels. Cold glue labels must be capable of providing a means for drying the adhesive and providing good initial adhesion, while not exhibiting visual defects.
Olefin blends have been found in many respects to be a preferred material to meet the demands of polymeric, die-cut label manufacture. The relatively low cost of the olefinic resins, and their high strength allowing for low caliper film, tend to minimize overall material cost. Hot-stretched polypropylene and/or polyethylene provides stiffness in the machine direction even at relatively low-caliper thicknesses for adequate print registration and dispensing while providing sufficiently low tensile modulus and, in particular, sufficiently high elongation in the cross direction to allow good conformability.
Olefin blends have been found useful to produce, on an economical basis, printable 3.5 mil (caliper measured) coextruded labels die-cut from biaxially hot-stretched film that are durable, strong and water-resistant, and that generally have good structural and surface properties and performance characteristics. Biaxial stretching also makes an important contribution to performance by creating substantial differentials between machine direction (MD) and cross direction (CD) stiffnesses, tensile moduli, elongations, and other properties.
Paper labels have been used in the art in conjunction with a cold glue adhesive for application to glass and plastic containers. The paper labels exhibit good initial adhesion, adhesive drying, and destruct bonds. A metallized layer may be applied to paper labels to achieve a metallized graphic appearance. However, the inherent surface roughness of some papers does not encourage a highly reflective mirror-like surface.
U.S. Pat. No. 4,060,168 discloses a label assembly that includes a strip or web of backing material with a series of printed labels arranged in serial order. The backing material includes a die cut portion opposed to the label, which remains adhered to the label upon application of the label to a container. The cut portion may include printing and serves as a promotional item with improved pilfer resistance. U.S. Pat. No. 4,060,168 is incorporated herein by reference.
U.S. Pat. No. 5,061,334 discloses a high speed labeling machine and method for transferring labels to articles, such as cans. The labeling machine includes a device for supplying articles to be labeled, a label transfer wheel provided with individual groups of closely-spaced vacuum holddown ports to enable a succession of closely-spaced labels to be supported by the transfer wheel and carried by the transfer wheel to the articles, and a pressure-severing device for releasing the label from a carrier stock for subsequent transfer from a transfer wheel to the article to be labeled. Sprocket holes are utilized to precisely align a label. The labels may be die cut on the printing press and retained on the carrier by pinpoint attachments or they may be die cut as the label is advanced. In one particular embodiment, single layer label tapes with release agent on one surface and a pressure sensitive adhesive on the other surface provide added advantages of smaller spool sizes and faster speeds. Single layer label tapes also make tamper-evident packaging very versatile and advantageous. Another aspect disclosed in this patent is the significantly improved economics by reducing the number of layers to one instead of two for normal pressure sensitive labels as well as by eliminating the cost and problems associated with using glue applicators with many types of labeling products. U.S. Pat. No. 5,061,334 is incorporated herein by reference.
U.S. Pat. No. 5,174,852 discloses a method for attaching labels to containers where the labels are removed from a stack by rolling contact with an adhesive surface coated with a cold glue. Additionally, prior to termination of the removing step, localized hot glue coating is applied to the foremost label in the stack. After the label has thus been coated with cold glue and hot glue, it is peeled off the adhesive surface and pressed into contact of its glue-coated backside with a container. The provision that the hot glue coating is applied to the label while it is still held in the magazine ensures that the shape and position of the hot glue coating is fully independent of the speed of the labeling operation. U.S. Pat. No. 5,174,852 is incorporated herein by reference.
U.S. Pat. No. 5,288,548 discloses a multilayer label stock structure comprising a label stock of polypropylene film having on one side thereof a highly printable blend of (A) alkaline acrylate copolymer and (B) an interpolymer of an alkyl acrylate, an alkyl methacrylate and an alkyl acrylate acid. U.S. Pat. No. 5,288,548 is incorporated herein by reference.
U.S. Pat. No. 5,486,253 discloses a method of labeling containers utilizing a labeler having a multiple port vacuum drum. The multiple port vacuum drum has a first cavity which is supplied with one level of vacuum suitable for picking up label segments from a cutter with limited tension. The vacuum drum has a second cavity, which is supplied with another, higher level of vacuum suitable for firmly griping the label segments as an adhesive or the like is applied to the label segment. The second cavity may be further divided into a label application segment with lower vacuum pressure. The cutter drum has an arcuate cavity to which vacuum is supplied at the end first contacting the labeling material web with reduced vacuum being provided at the label cut off point and label release point. U.S. Pat. No. 5,486,253 is incorporated herein by reference.
U.S. Pat. No. 5,897,722 discloses a process for applying labels to articles by first adhering a limited portion of the label to the article and subsequently making additional portions of the label adhesive so that they will adhere to the article after the label is affixed to the article. The leading and trailing edges of the label may be bonded to the article or each other to affix the label to a container or other such article. The portions of the label between the leading and trailing edges may be either provided with an adhesive agent that is in a non-adhesive state or be formed from a label material which is not normally adhesive but can be rendered adhesive through an additional process step. For example, coated polyethylene film can be applied to a container by means of a hot melt adhesive and then subsequently heated to a temperature at which the polyethylene material becomes adhesive causing it to stick to the container. Other labeling materials can be printed with an adhesive that remains substantially non-adhesive until heated or activated by means of radiation, application of a chemical activating agent or by application of mechanical force. U.S. Pat. No. 5,897,722 is incorporated herein by reference.
U.S. Pat. No. 6,083,342 discloses a system and method for rapidly labeling a blow-molded hollow container made of a plastic composition subject to out-gassing after de-molding. Otherwise, conventional high speed silk screen labeling technology and processing is employed shortly after container fabrication de-molding in order to accurately deposit a continuous liquid layer of conventional silk screen ink over an external container surface area designated for application thereon of a self-adhering pre-printed label. The label is rapidly and accurately transfer applied over the silk screen coated area immediately after the same has dried, and even while out-gassing is still occurring from the de-molded to container. However, the protective ink barrier coating prevents formation of bubbles beneath the label. Those gases in the container wall resident beneath the label-affixed area can then nevertheless out-gas by migrating out of the container internal wall surface. Also, labeling may be silk screen imprinted on the container surface in a designated label-receiving area, and/or on non-designated surface area concurrently with barrier coating of the designated area. A gas-blocking clear ink barrier layer may be silk screened over the silk screen imprinting, and then the label applied over the gas-blocking layer of clear ink when dry. The label may be "peelable" for customer rebate return without thereby altering the labeling information remaining on the container. The pre-printed label may be a replicate of the label-covered imprinting so that the "trade dress" need not be altered by the peelable label, and so that the container surface area available for permanent labeling is maximized. U.S. Pat. No. 6,083,342 is incorporated herein by reference.
U.S. Pat. No. 6,139,935 discloses an oxygen-absorbing label consisting of a base sheet, adhesive on the base sheet for securing the base sheet to an internal surface of a container, an oxygen-absorbing composition on the base sheet, and a cover sheet secured to the base sheet to confine the oxygen-absorbing composition there between. The cover sheet is fabricated of material which will permit oxygen to pass therethrough but will not stain due to the oxidation of the oxygen-absorbing composition or due to contact with materials in the container. A plurality of the foregoing labels are mounted on a web to be used with conventional labeling equipment. U.S. Pat. No. 6,139,935 is incorporated herein by reference.


A sensor includes a body member, a volume change body, and a detection member. The body member has a flat plate-like shape, a first end in a first direction being supported, and a storage space opening at at least one of both end faces in a thickness direction. The volume change body, whose volume changes depending on an amount of a target, is supported by the body member so that at least a part of the volume change body is stored in the storage space. The detection member is in contact with a second end in the first direction of the body member, and detects stress caused by the change in the volume of the volume change body.

One of the known sensors has a volume change body, whose volume changes depending on the amount of a target (for example, molecules that constitute a gas or a liquid), and detects stress caused by the change in the volume of the volume change body. For example, the sensor described in Patent Literature 1 has a body member, the volume change body, and a detection member. The body member has a flat plate-like shape. The volume change body changes in the volume by receiving the target and covers both end faces in the thickness direction of the body member. The detection member, whose tip is supported, extends from the end of the body member. The detection member includes a piezoresistive element and detects the stress caused by the change in the volume of the volume change body.

Sensor, detection method, and sensor manufacturing method