This invention relates to immunological reagents and methods specific for a mammalian, transmembrane protein termed Pgp, having a non-specific efflux pump activity established in the art as being a component of clinically-important multidrug resistance in cancer patients undergoing chemotherapy. The invention provides methods for developing and using immunological reagents specific for certain mutant forms of Pgp and for wild-type Pgp in a conformation associated with substrate binding or in the presence of ATP depleting agents. The invention also provides improved methods for purifying hematopoietic stems cells expressing Pgp and diagnostic and therapeutic methods for cancer cells expressing Pgp.

1. Field of the Invention
The present invention relates to the use of immunological reagents specific for a human transmembrane efflux pump protein (P-glycoprotein) in a biochemical conformation adopted in the presence of certain cytotoxic, lipophilic drugs that are substrates for P-glycoprotein, in the presence of cellular ATP depleting agents, and by certain mutant embodiments of Pgp. The invention provides such immunological reagents for immunodiagnostic and therapeutic uses, for isolating lymphocytes and hematopoietic stem cells, and for anticancer drug development.
2. Background of the Invention
Many human cancers express intrinsically or develop spontaneously resistance to several classes of anticancer drugs, each with a different structure and different mechanism of action. This phenomenon, which can be mimicked in cultured mammalian cells selected for resistance to certain plant alkaloids or antitumor antibiotics such as colchicine, vinblastine and doxorubicin (former known as Adriamycin), is generally referred to as multidrug resistance ("MDR"; see Roninson (ed)., 1991, Molecular and Cellular Biology of Multidrug Resistance in Tumor Cells, Plenum Press, N.Y., 1991; Gottesman et al., 1991, in Biochemical Bases for Multidrui Resistance in Cancer, Academic Press, N.Y., Chapter 11 for reviews). The MDR phenotype presents a major obstacle to successful cancer chemotherapy in human patients.
MDR frequently appears to result from decreased intracellular accumulation of drug as a consequence of increased drug efflux related to alterations at the cellular plasma membrane. When mutant cell lines having the MDR phenotype are isolated, they are found to express an ATP-dependent non-specific molecular "pump" protein (generally known as P-glycoprotein) that is located in the plasma membrane and keeps the intracellular accumulation of an anti-cancer drug low enough to evoke the drug-resistance phenotype. This protein (which has been determined to be the gene product of the MDR 1 gene in humans) facilitates active (i.e., energy-dependent) drug efflux from the cell, against a concentration gradient of (generally) lipophilic compounds, including many cytotoxic drugs.
The gene encoding P-glycoprotein (which is also known as gp 170-180 and the multidrug transporter) has been cloned from cultured human cells by Roninson et al. (see co-owned U.S. Pat. No. 5,206,352, issued Apr. 27, 1993, having an effective filing date of Mar. 28, 1986), and is generally referred to as MDR 1. The protein product of the MDR 1 gene, most generally known as P-glycoprotein ("Pgp"), is a 170-180 kilodalton (kDa) transmembrane protein having the aforementioned energy-dependent efflux pump activity.
Molecular analysis of the MDR1 gene indicates that Pgp consists of 1280 amino acids distributed between two homologous halves (having 43% sequence identity of amino acid residues), each half of the molecule comprising six hydrophobic transmembrane domains and an ATP binding site within a cytoplasmic loop. Only about 8% of the molecule is extracellular, and carbohydrate moieties (approximately 30 kDa) are bound to sites in this region (Chen et al., 1986, Cell 47: 381-387).
Expression of Pgp on the cell surface is sufficient to render cells resistant to many (but not all) cytotoxic drugs, including many anti-cancer agents. Pgp-mediated MDR appears to be an important clinical component of tumor resistance in tumors of different types, and MDR1 gene expression correlates with resistance to chemotherapy in different types of cancer.
Because Pgp is involved in the resistance of different types of human malignancies to conventional chemotherapy, the expression of Pgp is an important diagnostic and prognostic factor which in many cases helps the physician to choose the most effective combination of chemotherapeutic drugs and to monitor the efficacy of treatment. One way Pgp expression has been evaluated is by detecting the binding of specific immunological reagents (antibodies) to tumor samples. However, frequently the expression level of Pgp in tumor cells is low and cannot be reproducibly detected by routine immunological methods. In addition, there are few immunological or other reagents specific for functionally-active Pgp (which are the only forms of Pgp that are clinically relevant). Thus, there is a need in the art to increase the sensitivity and specificity of immunological and immunohistochemical methods for detecting functional Pgp expression.
Pgp is also constitutively expressed in many normal cells and tissues (see Cordon-Cardo et al., 1990, J. Histochem. Cytochem. 38: 1277; and Thiebaut et al., 1987, Proc. Natl. Acad. Sci. USA 84: 7735 for reviews). In hematopoietic cells, Neyfakh et al. (1989, Exp. Cancer Res. 185: 496) have shown that certain subsets of human and murine lymphocytes efflux Rh123, a fluorescent dye that is a Pgp substrate, and this process can be blocked by small molecule inhibitors of Pgp. It has been demonstrated more recently that Pgp is expressed on the cell-surface membranes of pluripotent stem cells, NK cells, CD4- and CD8-positive T lymphocytes, and B lymphocytes (Chaudhary et al., 1992, Blood 80: 2735; Drach et al., 1992, Blood 80: 2729; Kimecki et al., 1994, Blood 83: 2451; Chaudhary et al., 1991, Cell 66: 85). Pgp expression on the cell surface membranes of different subsets of human lymphocytes has been extensively documented (Coon et al., 1991, Human Immunol. 32: 134; Tiirikainen et al., 1992, Ann. Hematol. 65: 124; Schluesener et al., 1992, Immunophannacology 23: 37; Gupta et al., 1993, J. Clin. Immunol. 13: 289). Although recent studies suggest that Pgp plays a role in normal physiological functions of immune cells (Witkowski et al., 1994, J. Immunol. 153: 658; Kobayashi et al., 1994, Biochem. Pharmacol. 48: 1641; Raghu et al., 1996, Exp. Hernatol. 24:1030-1036, as disclosed more fully in co-pending U.S. patent application, Ser. No. 08/658,583, filed Jun. 7, 1996, incorporated by reference herein in its entirety), the physiological role of Pgp in normal immune cells has remained unclear to date.
Expression of Pgp in hematopoietic cells provides an effective means for identifying and punfyng lymphocytes and hematopoietic stem cells. As described more completely in co-owned and/or co-pending U.S. Pat. No.5,434,075, issued Jul. 18, 1995 and U.S. Ser. No.08/032,056, filed Mar. 16, 1993, functional Pgp assays (such as fluorescent dye efflux) and immunochemical methods (such as fluorescence activated cell sorting (FACS) analysis) can in theory be used to purify lymphocytes and hematopoietic stem cells.
However, the levels of expression of Pgp on stem cells are low, and consequently the amount of an immunological reagent such as a monoclonal antibody (mAb) bound to a hematopoietic stem cell membrane using conventional procedures is generally not high enough to efficiently separate Pgp-positive cells by any conventional immunological technique (such as FACS, immunomagnetic particle separation, cell panning, or other methods known in the art). Thus, there remains a need in this art to improve the efficiency of methods for using Pgp expression to specifically purify lymphocytes and hematopoietic stem cells from biological sources.
Once the central role in MDR played by Pgp was uncovered, agents with a potential for reversing MDR phenotypes were developed that target Pgp. Several classes of drugs, including calcium channel blockers (e.g., verapamil), immunosuppresants (such as cyclosporines and steroid hormones), calnodulin inhibitors, and other compounds, were found to enhance the intracellular accumulation and cytotoxic action of Pgp-transported drugs (Ford et al., 1990, Pharm. Rev. 42: 155). Many of these agents were found to inhibit either drug binding or drug transport by Pgp (Akiyama et al., 1988, Molec. Pharm. 33: 144; Horio et al., 1988, Proc. Natl. Acad. Sci. USA 84: 3580). Some of these agents themselves were found to bind to and be effluxed by Pgp, suggesting that their enhancing effects on the cytotoxicity of Pgp substrates are due, at least in part, to competition for drug binding sites on this protein (Cornwell et al., 1986, J Bio.. Chem. 261: 792 1; Tamai, 1990, J. Biochem. Molec. Biol. 265: 16509).
Many of these agents, however, also have strong, deleterious side effects at physiologically-achievable concentrations. These systemic side effects severely limit the clinical use of these agents as specific inhibitors of Pgp or for negative selection against Pgp-expressing tumor cells. Most of the known MDR-reversing drugs used in clinical trials have major side effects unrelated to inhibition of Pgp, such as calcium channel blockage (verapamil) or immunosuppression (cyclosporines and steroids). Similarly, targeting of cytotoxic drugs to Pgp-expressing cells is capable of compromising normal tissue function in normal cells (such as kidney, liver, colonic epithelium, etc.) that normally express Pgp. These drawbacks restrict the clinically-achievable dose of such agents and ultimately, their usefulness.
Immunological reagents, specifically such agents linked to cytotoxic molecules or detectably labeled reporter molecules, provide an alternate and specific way for identifying cells expressing Pgp at the cell surface and specifically delivering cytotoxic substances directly to such cells. Immunological reagents specific for extracellular epitopes of Pgp, such as anti-Pgp antibodies, offered the prospect of specificity, since antibodies should target only Pgp. However, it has also been recognized that only antibodies which react with an extracellular epitope of Pgp are expected to react with the protein in the plasma membrane of intact cells and thereby inhibit the MDR phenotype in such cells. Antibodies directed to the cytoplasmic portion of Pgp, on the other hand, are unlikely to be useful for reversal of MDR.
In addition, antibody binding to Pgp was expected to have a more-prolonged inhibitory effect than that caused by transient binding of a competitive inhibitor. Such reagents may also provide a means for delivering cytotoxic agents specifically to Pgp-expressing tumor cells in regimens aimed to selective killing of such cells.
Monoclonal antibodies specific for Pgp are known in the art.
Hamada et al., 1986, Proc. Natl. Acad. Sci. USA 83: 7785 disclose the mAbs MRK-16 and MRK-17, produced by immunizing mice with doxorubicin-resistant K-562 human leukemia cells. MRK- 16 mAb was also reported to modulate vincristine and actinomycin D transport in resistant cells, and MRK-17 was shown to specifically inhibit growth of resistant cells with these drugs.
Meyers et al., 1987, Cancer Res. 49: 3209 disclose mAbs HYB-241 and HYB-612, which recognize an external epitope of Pgp.
O'Brien et al., 1989, Proc. Amer. Assoc. Cancer Res. 30:Abs 2114 disclose that mAbs HYB-241 and HYB-612 increased the accumulation of vincristine and actinomycin D in tumor cells and increased the cytotoxicity of combinations of these drugs with verapamil.
Tsuruo et al., 1989, Jpn. J Cancer Res. 80: 627 reported that treatment of athyrnic mice that had been previously inoculated with drug resistant human ovarian cancer cells with the mAb MRK 16 caused regression of established subcutaneous tumors.
Hamada et al., 1990, Cancer Res. 50: 3167 disclosed a recombinant chimeric antibody that combines the variable region of MRK-16 with the F.sub.c portion of a human antibody, and showed this chimeric antibody to be more effective than MRK-16 mAb in increasing cytotoxicity in vitro.
Pearson et al., 1991, J. Natl. Cancer Inst. 88: 1386 disclosed that MRK-16 mAb increased the in vivo toxicity of vincristine to a human MDR colon cancer cell line grown as a xenograft in nude mice. The in vitro potentiation of drug cytotoxicity by MRK-16 mAb was, however, weak relative to known chemical inhibitors of Pgp action, and was apparently limited to only two Pgp substrates (vincristine and actinomycin D), having no effect on cytotoxicity by doxorubicin.
Cinciarelli et al., 1991, Int. J. Cancer 47: 533 disclosed a mouse IgG.sub.2a mAb, termed MAb657, having cross reactivity to Pgp-expressing human MDR cells. This mAb was shown to increase the susceptibility of MDR cells to human peripheral blood lymphocyte-mediated cytotoxicity, but was not shown to have an inhibitory effect on the drug efflux activity of Pgp.
Arcesi et al., 1993, Cancer Res. 53: 310-317 disclosed mnAb 4E3 that binds to extracellular epitopes of Pgp but does not disrupt drug efflux or potentiate MDR drug-induced cytotoxicity.
Mechetner and Roninson, in co-owned and/or co-pending U.S. Pat. No. 5,434,075, issued Jul. 18, 1995, and in U.S. Ser. No. 08/032,056, filed Mar. 16, 1993, disclosed mAb UIC2, having specificity for extracellular Pgp epitopes. This antibody was also shown to effectively inhibit Pgp-mediated drug efflux in MDR cells, and to reverse the MDR phenotype in vitro thereby, for a number of structurally and functional different cytotoxic compounds and all tested chemotherapeutic drugs known to be substrates for Pgp-mediated drug efflux.
The production of UIC2 mAb demonstrated the usefulness of the development of mAbs specific for extracellular epitopes of Pgp that were capable of inhibiting drug efflux activity. As evidenced by the mAbs developed in the prior art, production of extracellular epitope-specific mAbs does not necessarily result in mAbs that can affect drug efflux. There thus remains in the art a need for developing methods for producing mAbs that are capable of inhibiting drug efflux activity in Pgp. There also remains a need in the art for methods for developing more sensitive mAbs and methods to improve the sensitivity of currently available mAbs for the detection of Pgp expression in cancer cells in vivo, for improved cancer diagnostics and therapeutic applications with both normal and tumor cells expressing Pgp. There also remains a need in the art to develop more specific and efficient tools for the isolation of lymphocytes and hematopoietic stem cells, especially pluripotent and totipotent stem cells.

Methods and reagents for preparing and using immunological agents specific for P-glycoprotein

Board of Trustees of the University of Illinois


A low-pressure feedwater preheater or heater in power station plants, heated with bleed steam and having a two-pass tube bundle of the tube sheet type of construction, has a carrier design for the tube bundle having side plates (9) running parallel to the tube bundle and one or more supporting plates (8) disposed perpendicularly to the tube bundle. In the center of the bundle between the cold and the hot leg, a partition (11) extends along the entire length of the bundle leg. Arranged on either side of the partition (11) are suction tubes (12) through which non-condensable gases are drawn off in the zones of lowest pressure. According to the invention, the supporting plates (8) are each made of a single piece and are thus continuous, and the partition (11) consists of individual sheet-metal parts which are connected to the supporting plates (8). In one embodiment, the sheet-metal parts of the partition (11) and the supporting plate (8) are connected to one another by indentations (16) on each side of the sheet-metal part (11) which faces the supporting plate (8) and by a corresponding opening (17) in the supporting plate (8).

1. Field of the Invention
The invention relates to a low-pressure feedwater preheater or district heater, which is heated by bleed steam and has two-pass tube bundles of the tube-sheet type of construction, for power station plants and relates in particular to a design for the tube-bundle carrier.
2. Discussion of Background
Low-pressure feedwater preheaters are generally known. They are arranged in a power station plant between the condenser and the deaerator/feedwatertank and serve to preheat the feedwater in a steam/water circuit by heat exchange between steam and feedwater flowing in tubes. To this end, the steam is bled from a low-pressure turbine and directed into the preheater, where it condenses on the tubes and heats the feedwater. In a district-heating power station, the heater serves to reheat heating water which flows through a district-heating circuit.
Air (non-condensable gases) passes with the bled steam as well as from the atmosphere, in particular in the case of preheaters operated in the vacuum range, into the preheater and hinders the heat transfer. This hindrance is removed by deaearation of the apparatus by drawing off these non-condensable gases, for which reason various measures have to be taken in order to carry away the gases as completely as possible.
The feedwater flows via a two-pass system from a water box which is subdivided into two parts and is arranged at one end of the preheater. This two-pass system consists of a bundle of tubes, which are either designed in a straight line together with a reverse flow water box or are of U-shaped design. The water flows from the first part of the water box through the tubes via the revese flow water box or via the U back into the second part of the water box, the cold half of the tubes being designated as cold leg and the hot half being designated as hot leg. In feedwater preheaters of the tube-sheet type of construction, the tube bundles are each firmly anchored in a plate which is arranged in a position adjacent to the water box. Furthermore, the tubes are held and supported by tube carriers, which in each case consist of supporting plates and longitudinal support bars.
The steam bled from the turbine passes through the steam inlet opening into the low-pressure feedwater preheater and is distributed by a baffle plate, so that it first of all flows into an annular steam space between the shell and the tube bundle and then penetrates radially into the tube bundle along the entire length of the latter and condenses on the tube surfaces. There is a free space, the so-called bundle lane, in the center of the tube bundle. Direct inflow of the steam into the bundle lane is prevented by side plates arranged on the outside. Arranged in the bundle lane are one or more deaeration tubes. The latter have a plurality of suction openings through which the non-condensable gases are drawn off. The gases collect in the zone in which the lowest pressure prevails. It has been found that the zone of lowest pressure does not run parallel to the tubes and along the center of the bundle lane but shifts gradually from the center of the bundle to the cold leg of the tube bundle. It therefore runs obliquely from the center of the deflection side of the tubes to the cold inlet side of the tubes at the water box. It is not possible to place a suction tube ideally in this zone of lowest pressure, since such a suction tube would have to be placed at the tube-sheet side on or in the cold leg.
Described in DE 28 20 736 is a feedwater preheater which, in the bundle lane between the cold and the hot leg of the tube bundle, has a partition which extends along the entire length of the tube bundle. Due to the partition, a zone of lowest pressure which does not extend into the tube bundle is obtained in the bundle lane on either side of and in proximity to the partition. Here, suction tubes having deaeration openings through which the non-condensable gases are drawn off are arranged on each side of the partition. This ensures that there are deaeration tubes at the points of lowest pressure in both the hot and the cold bundle half.
The partition, which is continuous over the bundle length, is welded to the side plates of the bundle carriers. The perforated supporting plate consisting of two halves is likewise welded to the side plates and the partition. From the technical point of view, the welded joints are readily suitable for this type of preheater; however, the fabrication of the bundle carriers with this welded construction is very time-consuming, involves a great deal of work and is very costly.


Devices, systems, and methods for printing indicia on latex are presented. A system for printing includes a latex surface supported by a mold on a rotary mount, a printer, a base plate supporting print heads, a rotational drive system for selectively rotating the mold during printing, and a printer engine programmed to print indicia onto a substantially contiguous area of the latex surface, from an intermediate border to and including a distal end. In a system with multiple molds moving along a path, the system may include a chassis for supporting the base plate, printer, and/or rotational drive system, and a motion controller for moving the chassis near the molds. Latex articles may be imprinted with indicia that is decorative and/or informative.

Described herein is a system and method for printing indicia on a latex surface; for example, on a multi-layer latex condom.
Plain condoms are boring. Variations in condom shape, texture, and color have not sufficiently increased consumer interest in the use of condoms, especially among young people. Thus, there is a need in the art for manufacturing a more appealing and interesting latex condom.
There is also a related need in the art for safely printing any of a variety of indicia onto the latex surface of any of a variety of products, including those intended for medical uses.

This patent is subject to a terminal disclaimer.

Systems and methods for printing indicia on latex surfaces in motion


A first computer sends a list of possible security associations to a second computer in a message according to a protocol of an application layer, a security parameter index being contained in the message for each security association. The second computer selects a security association and transmits it or an indication of the security association selected by it to the first computer.

Present-day communication systems are usually based on what is known as the OSI layer model (Open System Interconnection layer model), which shares different communication tasks between computers on different communication layers.
The hierarchically structured OSI layer model includes what is known as the application layer, on which customary application protocols, such as for example for communication between electronic mail programs, Internet browser programs or else other application programs, are executed.
Arranged hierarchically under it are what are known as network layers, which are responsible for transparent communication between computers communicating with one another and possibly switching processors lying in between in a communication link. Examples of network layers are layers which provide a communication between computers according to the Internet Protocol (IP) or according to the Transport Control Protocol (TCP) or else according to the User Datagram Protocol (UDP).
Known application protocols of layer 7, using the Internet Protocol (IP) on OSI layer 3, provide not only their core functionality but usually also a more or less extensive set of security mechanisms for the protection of the communication conducted by the application protocol (protocol on the level of the application layer in the OSI layer model). Among the security objectives to be achieved in this process, the authentication of the messages according to the application protocol and also their integrity and confidentiality are of very great importance.
Until now, two possibilities have been known for achieving these security objectives. On the one hand, the application protocol itself may be protected, that is to say a cryptographic protection may take place on the level of the application layer. Since such an application protocol on the basis of IP runs above the network layer, considered hierarchically, reference is also made in this case to an assured security on the level of the application layer. For this purpose, modifications of the application protocol itself are usually required, or the security mechanisms are already contained in the specification of the respective application protocol.
On the other hand, it is known to protect the application protocols in one of the network layers lying under the application layer, for example according to the IP or the TCP. As a solution for security on the level of the network layer, in other words on the network level, the architectures according to IPsec (see, "Understanding the IPsec Protocol Suite, Secure Virtual Private Network Solutions," Timestep Corporation, December 1998) or TLS (Transport Layer Security) are known.
IPsec represents an expansion of the Internet Protocol (IP) communication protocol of OSI layer 3, by which security services are provided on the level of the network layer. According to the IPsec protocol, cryptographic protection is provided in a transparent way for every type of application which uses the protocol according to IPsec of the network layer for communication on the level of the application layer.
As described in "Understanding the IPsec Protocol Suite, Secure Virtual Private Network Solutions," the Encapsulating Security Payload protocol (ESP) is used for IPsec to protect the user data transmitted, in particular for authentication and for encryption, and the Authentication Header (AH) protocol is used for pure authentication of the communication partners.
Within the security protocol IPsec, what are known as security associations (SA) are used between the computers communicating with one another. Such a security association is to be understood hereafter as meaning a set of security parameters for a communication link to be set up later, which include in addition to the key material to be used for a cryptographic method further parameters and security policy information, such as for example the cryptographic algorithms to be used, a maximum service life of keys or a maximum service life of the security association itself, or else identification data for identifying the communication partners.
Furthermore, the security association may also include in particular the cryptographic parameters defined in IETF RFC 2407: IP Security Domain of Interpretation, a selection of which is presented below. The security association may, for example, include:                a security parameter index, with which the security association between two entities communicating with one another, preferably computers communicating with one another, is unequivocally identified in each case,        a security association life type, with which the unit in which the service life of the security association is measured, optionally seconds or kilobytes, is indicated,        a security association lifetime, in other words the indication of a point in time or a time duration until which or during which the indicated security association is valid for a communication between the two associated entities, and consequently the indicated security parameters can be used for a communication,        an indication of an encryption algorithm to be used in a later communication link using this security association, which algorithm is used according to the ESP,        an indication of an authentication algorithm to be used within a later communication link,        an indication of the key length,        an indication of possibly existing key rings to be used.
Protocols for negotiating security associations are described in IETF RFC 2409: The Internet Key Exchange (IKE) and Internet Engineering Task Force: Draft-IETF-KINK-KINK-00.txt. These independent protocols, which run independently of an application protocol, are very complex and computationally intensive and are consequently not suitable for implementation in a communication device having only a low computing capacity, in particular in a mobile communication terminal. Furthermore, according to the methods described in IETF RFC 2409 and Draft-IETF-KINK-KINK-00.txt, an independent communication is required for negotiating the security associations. To illustrate the disadvantages described above of the methods described in IETF RFC 2409 and Draft-IETF-KINK-KINK-00.txt, these methods are briefly outlined in the text which follows.
The protocol according to IETF RFC 2409 has two phases. In a first phase, what is known as an IKE security association, in other words a cryptographically protected communication channel, is produced between two computers communicating with one another. On the basis of the IKE security association, a second phase can be subsequently executed, during which the actual IPsec security associations are generated. According to IETF RFC 2409, use of what is known as the "Diffie-Hellman Key Exchange" protocol, which necessitates complex asymmetrical cryptographic computations in the computers, is required in the first phase.
In particular, the current capability of processor units in mobile communication terminals, such as mobile radio telephones, stands in the way of efficient use of this method. It should further be noted that the IKE protocol is based on the comprehensive ISAKMP standard. Altogether, the implementation of IKE is very complex and prone to errors, making the protocol only very poorly suited for communication terminals with only low computing capacity.
The protocol described in Draft-IETF-KINK-KINK-00.txt, referred to as the KINK protocol, prescribes the use of what is known as the Kerberos standard. This is likewise very extensive and complex and rather inefficient in its implementation on a mobile communication terminal with low computing capacity.
Furthermore, what is known as the Session Initiation Protocol (SIP), as described in IETF RFC 2543: Session Initiation Protocol, is an example of an application protocol. The SIP is a protocol which is defined on the level of the application layer and specifies the signaling for controlling communication sessions. Communication sessions controlled by SIP are, for example, telephone conversations or multimedia conferences over the Internet. The SIP uses either "request" messages from a client to a server, or "response" messages from a server to the client.
A further protocol on the application layer is the Hypertext Transfer Protocol (HTTP).
In Ericsson, Nokia and Nortel, "Security Mode Setup for the IMS Registration," 3GPP TSG SA WG 3 Security-S3##19, S3-010326, Jul. 4-6, 2001, it is described for a future UMTS protocol that SIP is used between a mobile communication terminal and a mobile radio network computer for negotiating which algorithm is to be used for protecting integrity. This procedure is only very poorly suited for practical implementation, since the restriction to the negotiation of an algorithm which is to be used in a future communication link for protecting integrity is not adequate to satisfy present-day requirements within the scope of mobile multimedia communication.

System for negotiating security association on application layer


This hydraulic system is provided with: a hydraulic pump; a pilot-type control valve; an electromagnetic proportional valve; a controller; and a pilot pressure switching unit which is capable of switching the electromagnetic proportional valve supply pressure to a first pressure during a normal operation, or to a second pressure lower than the first pressure. The control valve is provided with a bleed-off passage, and is capable of controlling the operating oil pressure supplied to the actuator, according to the opening area thereof. During an emergency operation, the electromagnetic proportional valve supply pressure is switched from the first pressure to the second pressure, the electromagnetic proportional valve is brought into a fully opened state, and the operating oil discharge amount from the hydraulic pump increases and decreases, and the operating oil pressure increases and decreases, and thus the operating speed of the actuator is controlled.

In recent years, an electric operation system which electrically controls a control valve of a hydraulic system is mounted on an operating system of a hydraulic working machine. In the electric operation system, an operation signal from an operation lever is input to the controller, and an electromagnetic proportional valve is operated according to a drive signal from the controller. By the operation of the electromagnetic proportional valve, a pilot pressure of the control valve of the hydraulic system is controlled.
The electric operation system can perform advanced control by executing a control logic in the controller and is an important technique for realizing energy saving, low noise, optimum control, and the like which are required for hydraulic working machine in recent years.
In the electric operation system, when an electric circuit unit breaks down, the controller cannot control the electromagnetic proportional valve. Therefore, it is preferable that the electric operation system has an emergency operation device for responding during a failure of the electric operation system (for example, Patent Literature 1). An example of the electric operation system including the emergency operation device is illustrated in FIG. 7.
In the electric operation system illustrated in FIG. 7, during a normal operation, when an operation lever 9 of an operation box 20 is operated, a drive signal based on the operation thereof is output from a controller 2 and input to an electromagnetic proportional valve 4 via an amplifier 3. When the electromagnetic proportional valve 4 operates and a pilot pressure is supplied to a control valve 27, a spool of the control valve 27 moves, and an operating oil pressure is supplied to an actuator 5. Thereby, a drive direction and an operation speed of the actuator 5 are controlled.
When a failure such as disconnection occurs in an electric circuit unit of the electric operation system, a power supply switching switch 22 is switched to an emergency operation side. An emergency operation switch 21 incorporated in the operation box 20 is switched in conjunction with an operation of the operation lever 9 and when a side of the electromagnetic proportional valve 4 is energized, a pilot pressure is supplied to the control valve 27, and the actuator 5 is driven.

Hydraulic system and emergency operation method


A conjugated diolefin copolymer rubber is produced by copolymerization of a conjugated diolefin and an aromatic vinyl compound and has a primary amino group and an alkoxysilyl group bonded to the copolymer chain. The conjugated diolefin copolymer rubber contains the aromatic vinyl compound in an amount of 5 to 60 wt %. The aromatic vinyl compound is densely distributed in one terminal of the copolymer chain, and is scarcely distributed in the other terminal. The conjugated diolefin copolymer rubber has a temperature difference (ΔTg) of 25° C. or more. The conjugated diolefin copolymer rubber has low hysteresis loss properties as well as improved abrasion resistance, rupture characteristics, and road grip characteristics without impairing wet skid characteristics. A method for producing the conjugated diolefin copolymer rubber is also disclosed.

A conjugated diolefin rubber exhibiting excellent abrasion resistance, rupture characteristics, and grip characteristics as well as superior operational stability (e.g., wet skid resistance) has been desired as a tire material in order to deal with a recent demand for safety and operational stability of vehicles.
As a rubber material with a low hysteresis loss, natural rubber, polyisoprene rubber, polybutadiene rubber, and the like have been known. However, these rubber materials exhibit poor wet skid resistance.
As a method for decreasing hysteresis loss without impairing wet skid resistance, a method of introducing functional groups into the terminals of styrene-butadiene copolymers having various structures and polymerized in a hydrocarbon solvent using an organolithium initiator has been proposed. A styrene-butadiene copolymer obtained by modifying or coupling the polymer terminal with a tin compound (see Patent Document 1) and a styrene-butadiene copolymer obtained by modifying the polymer terminal with an isocyanate compound (see Patent Document 2) have been known. These modified polymers exhibit a reduced hysteresis loss and excellent abrasion resistance and rupture characteristics without showing reduced wet skid resistance particularly when using a composition containing carbon black as a reinforcing agent.
On the other hand, a method of using a rubber composition containing silica or a mixture of silica and carbon black as a reinforcing agent as a rubber material for tires has been proposed recently. A tire tread formed using silica or a mixture of silica and carbon black exhibits low rolling resistance and provides excellent operational stability represented by wet skid resistance. However, a vulcanizate thereof exhibits a low tensile strength and abrasion resistance. The above modified styrene-butadiene copolymer can produce an excellent rubber material for tires exhibiting superior abrasion resistance and rupture characteristics if carbon black is used as a reinforcing agent, but the effect is small when silica is used as a reinforcing agent.
In order to improve the tensile strength and the abrasion resistance of a vulcanizate containing silica or a mixture of silica and carbon black, a rubber composition containing a polymer into which a functional group having affinity with silica is introduced has been proposed. A method for producing a polymer by reacting silicon tetrahalide, trihalosilane, or the like has been proposed in Patent Document 3. Patent Document 4 discloses a method of producing a polymer modified by a halogenated silane compound. Patent Document 5 discloses a diene rubber with an alkyl silyl group introduced therein and Patent Document 6 discloses a diene rubber with a halogenated silyl group introduced therein. Patent Document 7 discloses a diene rubber into which a tertiary amino group and an alkoxysilyl group are introduced.
Although some improvement in properties can be seen by using these modified polymers in the composition comprising silica or a mixture of silica and carbon black, improvement in tensile strength and abrasion resistance of the vulcanizate is not sufficient. In addition, the effect of lowering the hysteresis loss is not necessarily satisfactory in the composition using a mixture of silica and carbon black, particularly when the proportion of carbon black is increased. Moreover, a silica-containing composition exhibits inferior processability as compared with a carbon black-containing composition, resulting in an increased processing cost. Processability tends to become worse when a polymer in which the above-described functional groups having affinity with silica are introduced.
Generally known modified polymers are classified into those suitable for use mainly with carbon black and those suitable for use with silica. When altering the type of a reinforcing agent used in manufacturing tires, rubbers to be used must be reselected. In addition, when a mixture of silica and carbon black is added, the effect obtained has a correlation with the ratio of silica and carbon black irrespective of the type of the modified polymer used.
Polymers modified by introducing an amino group are thought to be effective in either a composition to which carbon black is added or a composition to which silica is added. Examples of such polymers for use with carbon black that have been proposed include (1) polymers obtained by introducing an amino group into the polymer terminal using a lithium amide initiator (see Patent Documents 8 to 12) and (2) styrene-butadiene copolymers having different structures polymerized using an organolithium initiator, of which the terminals are modified with a nitrogen-containing compound such as a urea compound (see Patent Document 13), a dialkylamino benzophenone compound (see Patent Documents 14 and 15), or a lactam compound (see Patent Document 16). Diene rubbers with an amino group introduced therein have been disclosed as polymers for use with silica (see Patent Documents 17 to 19).
A certain degree of improvement in the properties has been achieved by the modified polymers obtained using these methods in both the compositions containing silica and the compositions containing carbon black. Although the above prior art contain detailed descriptions of the method of introducing an amino group, the prior art only disclose general information concerning the relationship between the structure and performance of the polymers.
As a novel conjugated diolefin copolymer rubber having good processability and well-balanced low hysteresis loss, abrasion resistance, and rupture characteristics without impairing wet skid properties which is used together with either carbon black or silica, the inventors of the present invention have previously proposed a copolymer rubber of a conjugated diolefin and an aromatic vinyl compound having a primary amino group and an alkoxysilyl group which are bonded to the copolymer chain (see Patent Documents 20 and 21). Rubber compositions having various improved rubber characteristics mentioned above can be obtained by using the proposed technology. However, in order to further improve operational stability and safety of vehicles, a copolymer rubber having excellent wet skid properties, abrasion resistance, a low rebounding coefficient, and road surface gripping characteristics in both dry and rainy conditions (dry grip and wet grip) has been desired.    Patent Document 1: JP-A-57-55912    Patent Document 2: JP-A-61-141741    Patent Document 3: JP-B-49-36957    Patent Document 4: JP-B-52-5071    Patent Document 5: JP-A-1-188501    Patent Document 6: JP-A-5-230286    Patent Document 7: JP-A-7-233217    Patent Document 8: JP-A-59-38209    Patent Document 9: JP-B-5-1298    Patent Document 10: JP-A-6-279515    Patent Document 11: JP-A-6-199923    Patent Document 12: JP-A-7-53616    Patent Document 13: JP-A-61-27338    Patent Document 14: JP-A-58-162604    Patent Document 15: JP-A-58-189203    Patent Document 16: JP-A-61-43402    Patent Document 17: JP-A-1-101344    Patent Document 18: JP-A-64-22940    Patent Document 19: JP-A-9-71687    Patent Document 20: JP-A-2003-171418    Patent Document 21: JP-A-2004-67987

Conjugated diolefin copolymer rubber, method for producing the same, rubber composition and tire


A motor vehicle including an exhaust system downstream of an internal combustion engine and which comprises several exhaust gas aftertreatment devices for cleaning the exhaust gas from the internal combustion engine, wherein the exhaust system includes an SCR catalytic converter and at least one additional catalytic converter, wherein the additional catalytic converter is designed as an NOx storage catalytic converter or an oxidation catalytic converter, wherein a control device is provided for controlling at least one operating component of the motor vehicle influencing the temperature of the exhaust gas, wherein the control device is designed to control the operating component for setting the temperature of the exhaust gas depending on at least route information dependent on an expected travel distance and conversion information relating to a temperature-dependent pollutant conversion rate of at least one exhaust gas aftertreatment device of the exhaust system.

When operating internal combustion engines, in particular diesel engines, environmentally harmful and/or health-damaging substances, such as nitrogen oxides (NOx), carbon monoxide (CO), unburned hydrocarbons (CmHn) and soot particles are sometimes emitted. This creates the need for exhaust gases to be cleaned or undergo aftertreatment in the first place, before being emitted into the atmosphere. In some cases, legal limits have to be considered. Such aftertreatment or purification of the exhaust gas is generally done by exhaust gas aftertreatment devices, such as catalytic converters or particulate filters.
Oxidation catalytic converters are often used for this purpose, whereby carbon monoxide and unburned hydrocarbons are removed by the residual oxygen present in the exhaust gas. However, NOx storage catalytic converters are also frequently used to remove nitrogen oxides. These are operated in an initial phase (storage phase), wherein the nitrogen oxides are stored in storage components of the NOx storage catalytic converter. This can continue, until saturation of the memory components. Regeneration or discharge of the storage components then follows in a second phase (regeneration phase or rich operation). This means that the exhaust gas is "enriched," i.e., more fuel is injected than is burned in the combustion chamber of the internal combustion engine, such that the exhaust gas has an increased proportion of unburned hydrocarbons. Due to chemical reactions of the unburned hydrocarbons with the nitrogen oxides stored in the NOx storage catalytic converter, the storage components of the NOx storage catalytic converter are discharged.
What's more, SCR catalytic converters (SCR: selective catalytic reduction) may be used to remove nitrogen oxides. A urea solution (e.g., AdBlue®) is dosed through an injection unit and injected into the exhaust gas, before or as it enters the SCR catalytic converter. Chemical reactions then convert the nitrogen oxides in the SCR catalytic converter into carbon dioxide, elemental nitrogen, and water.
DE 10 2014 216 217 A1 discloses a method for reducing NOx emissions in the exhaust gases from an internal combustion engine of a motor vehicle. Here, an exhaust system downstream of the internal combustion engine comprises an NOx storage catalytic converter, an oxidation catalytic converter, and an SCR catalytic converter. Here, data relating to an expected travel distance are evaluated, whereby the regeneration and storage phases of the NOx storage catalytic converter and the dosing of the urea solution are controlled as a function thereof.
DE 10 2016 224 430 A1 discloses a method for the regeneration of an exhaust gas aftertreatment device. In particular, distance data is used for optimizing the regeneration phases using, e.g., an exhaust-gas heating device.
The object of the invention is to indicate a motor vehicle, in which particularly efficient cleaning of the exhaust gas takes place, in particular in terms of the overall pollutant conversion rate.

Motor vehicle and a method for operating a motor vehicle


A sheet conveying device including a conveying section configured to convey a sheet in a predetermined conveying direction, a discharge section configured to discharge the sheet onto a stacking surface and configured to support the sheet, which is discharged by the discharge section, from below, a first contact portion provided on the stacking surface at a middle portion of the stacking surface in a width direction orthogonal to the conveying direction; a second contact portion provided on the stacking surface at one side with respect to the first contact portion in the width direction; and a third contact portion provided on the stacking surface at an other side with respect to the first contact portion in the width direction.

JP-U-H06-23955 discloses an example of a sheet conveying device in a related art. The sheet conveying device includes a conveying section and a discharge section. The conveying section conveys a sheet in a predetermined conveying direction. The discharge section forms a part of the conveying section. The discharge section discharges the sheet onto a stacking surface positioned on a downstream side of the discharge section in the conveying direction. The stacking surface extends in the conveying direction and a width direction orthogonal to the conveying direction. The stacking surface is configured to support the sheet, which is discharged by the discharge section, from below.
A first contact portion is provided on the stacking surface at a middle portion of the stacking surface in the width direction. An upper end of the first contact portion is positioned above the stacking surface and extends in the conveying direction. The upper end of the first contact portion is configured to come into contact with the sheet, which is discharged by the discharge section, from below.
In the sheet conveying device, when a sheet is discharged onto the stacking surface, a middle portion of the sheet in the width direction is raised by the upper end of the first contact portion that extends in the conveying direction. Accordingly, since the sheet is curved in an inverted U-shape in the cross-section thereof perpendicular to the conveying direction, the downward bending of the middle portion of the sheet in the width direction, which is caused by gravity, may be suppressed. That is, it is possible to make the sheet, which is placed on the stacking surface, to have stiffness by the first contact portion. In the sheet conveying device, a plurality of sheets are stably discharged and stacked onto the stacking surface in this way.


A Bluetooth based security system utilized to provide ad-hoc security services to secured assets. Such a Bluetooth based security system comprises a secured device (SD) equipped with Bluetooth (BT) technology; a plurality of Bluetooth Access Points (BTAPs) located at designated points to establish a BT link with the secured device (SD); and a security server (SS) connected to all BTAPs and arranged to provide access control and security services for the secured device (SD), wherein the security server (SS) obtains attribute information of the secured device (SD), including an unique device identification (ID) and a last known location of the secured device (SD), activates a lock with the secured device (SD), and sends location information of a designated BTAP and an unlock code to the secured device (SD), via the designated BTAP.

Electronics devices such as notebook and laptop computers have become increasingly compact and portable and, as a result, increasingly vulnerable to unauthorized use, theft or loss. This is because these portable computers are small, expensive and may contain very valuable information.
Many computers, especially portable computers, have been secured from unauthorized use, theft or loss by mechanisms based on principles of prevention, deterrence or recovery. Prevention mechanisms may include physical locking devices which lock portable computers to docking stations. Deterrence mechanisms may include myriad alarm systems which employ various deterrence methods, including sound and visual alarms to deter an unauthorized person or a thief from stealing the portable computers. Recovery mechanisms may include various systems for providing automatic disabling of portable computers and/or for locating and tracking stolen portable computers for recovery via existing radio communication infrastructures or existing cellular network infrastructures.
One typical example of computer tracking systems for locating stolen computers is the use of a software (location tracking program) installed to instruct the computer to call a third party monitoring service at regular intervals. When the computer calls the monitoring service, the computer establishes a data link and transmits data to the monitoring service that identifies the computer. When the monitoring service receives a call from the user's computer, the monitoring service is able to determine the location of the computer by utilizing Caller ID. The location of the computer may then be forwarded to a law enforcement agency so that the lost or stolen computer can be retrieved by the law enforcement agency.
Alternatively, the location tracking program may also be installed to identify if an e-mail is being sent from the lost or stolen computer and compare a sender address to a predetermined owner address. If the sender address matches the owner address, the e-mail is sent unimpeded. However, if the sender address does not match with the sender address, then the e-mail is redirected to a third party such as a law enforcement agency to notify that the computer may have been stolen. However, such location tracking systems are not optimal because a third party monitoring service is required.
Another example location tracking systems are known as Radio Frequency Identification (RFID) systems which are available to uniquely identify and track devices equipped with RFID tags as disclosed, for example, in U.S. Pat. No. 6,232,870 for Applications For Radio Frequency Identification Systems issued to Garber et al., U.S. Pat. No. 6,100,804 for Radio Frequency Identification System issued to Brady et al., U.S. Pat. No. 5,963,134 for Inventory System Using Articles With RFID Tags issued to Bowers et al., and U.S. Pat. No. 5,838,253 for Radio Frequency Identification Label issued to Wurz et al. A typical RFID tag (also known as transponder) consists of a semiconductor chip having RF circuits, control logic, memory and an antenna (and a battery in the case of active tags) mounted to a substrate for providing remote identification. However, such RFID systems require dedicated wireless communications, and contain no general wireless data communications capabilities. Another drawback is that the user has purchase the RFID tags, the tag reader, and setup the environment specifically for the RFID service. RFID tags can also be cost prohibitive as each RFID tag can vary from 50 cents to $150 based on the desired capabilities.
Accordingly, there is a need for a new type of asset security and wireless tracking system for electronic devices such as portable computers that can be easily and effectively implemented with an industry standard communications wireless technology such as Bluetooth™ to provide access control, tracking and security services of varying complexity without any additional hardware overheads.


In an outer ring, a first ball filling slot which extends from a first raceway groove to a first end face of one side of an axial direction is formed, and in an inner ring, a second raceway groove and a second ball filling slot which extends from the second raceway groove to a second end face of one side of the axial direction are formed. Circumferential extension grooves which extend from one of the first ball filling slot and the second ball filling slot by a predetermined length in a circumferential direction at the opposite sides of the circumferential direction of the ball filling slot and extend from the first raceway groove to the first end face in the axial direction are formed.

1. Field of the Invention
The present invention relates to a ball bearing, and more particularly, to a full ball type deep groove ball bearing which is suitably used as a touchdown bearing in a turbo-molecular pump.
2. Related Art
In general, ball filling slots for mounting balls are formed in full ball type deep groove ball bearings as shown in Japanese patent publication No. JP 11-325085A. The ball filling slot extends over inner and outer rings so as to be formed substantially in a circular-shape. The ball filling slot is designed to have a size slightly smaller than the size of the balls such that the ball smoothly rotates and does not escape therefrom. Namely, the ball filling slot is designed to have a size so as to form a small interference to the balls.
In such a full ball type deep groove ball bearing, for mounting the balls, either of the following methods is used. One is a method of directly pressing a ball placed in the vicinity of the ball filling slot into the bearing. The other is a method of inserting a jig (a wedge) into the ball filling slot or a space between the inner ring and the outer ring in the vicinity of the ball filling slot to widen a gap between the inner and outer rings first, and then inserting the balls into the bearing in a state that the interference is hardly remained.
However, in the method of pressing the ball placed in the vicinity of the ball filling slot into the bearing, if the rigidity of the bearing rings is high, a large force for pressing the ball is required so that the balls and the periphery of the ball filling slot of the bearing ring can be easily damaged. The periphery of the ball filling slot can be easily deformed, when a ball is pressed into the slot. On the other hand, as for the method of inserting the wedge into the ball filling slot or the space between the inner ring and the outer ring in the vicinity of the ball filling slot first and then inserting the balls in a state that the interference is hardly remained, this method cannot be used if the gap between the outer diameter of the outer circumferential surface of the inner ring and the inner diameter of the inner circumferential surface of the outer ring is very small, and the wedge cannot be inserted into the space between the inner and outer rings.
On the other hand, conventionally, there is a turbo-molecular pump in which a rotary shaft is magnetically supported with respect to a housing a magnetic bearing with a non-contact state at the time of normal operation and a ball bearing is used as a touchdown bearing to mechanically support the rotary shaft by contacting with the rotary shaft when a vacuum environment is destroyed due to an operation error (artificial error) of a vacuum apparatus decompressed by the turbo-molecular pump such as a switching error of a valve of the vacuum apparatus for example, or due to an emergency state such as the stoppage of power supply. (See, for example, JP 11-190348A.)
Among turbo-molecular pumps having such a construction, there is a turbo-molecular in which a strong 2-pole permanent magnet (functioning as a rotor of a motor) is fixed at one of the rotary shaft and the housing in order to rotate the rotary shaft in high speed with respect to the housing. In this turbo-molecular pump, overcurrent according to the law of electromagnetic induction occurs in the other of the rotary shaft and the housing in which the 2-pole permanent magnet is not mounted due to a magnetic field generated by the 2-pole permanent magnet, and heat is generated in the other of the rotary shaft and the housing in which the 2-pole permanent magnet is not mounted.
It has been known that the heat is not radiated well to the outside of the pump and remains in the pump when the 2-pole permanent magnet is fixed at the housing so that the heat is generated in the rotary shaft. Therefore, this configuration is not suitable. Accordingly, in the turbo-molecular pump, the 2-pole permanent magnet is usually fixed at the rotary shaft so that the heat due to the mount of the 2-pole permanent magnet efficiently radiates to the outside of the pump.
For manufacturing such a turbo-molecular pump, the manufacturing is generally conducted by mounting a ball bearing in a housing first and then passing a 2-pole permanent magnet fixed to a rotary shaft through an inner circumferential surface of an inner ring. Accordingly, the assembling of the ball bearing and the rotary shaft is simplified.
Under such circumferences, the following problem has been found. If bearing rings of the ball bearing are formed of a metal material having high hardness such as a stainless steel or bearing steel to which a hardening treatment is performed in order to increase an impact strength, the inner ring is magnetized to become a magnet when the 2-pole permanent magnet fixed to the rotary shaft passes the inside of the inner circumferential surface of the inner ring. Accordingly, overcurrent may flow in the rotary shaft by a magnetic field generated by the magnetized inner ring, so that the heat is generated in the rotary shaft. In other words, upon normal operation, when the rotary shaft rotates relative to the housing and the ball bearing as the touchdown bearing is in such a condition that the outer ring is fixed to the housing and that the inner ring is not brought into contact with the rotary shaft and does not rotate relative to the outer ring, the heat is generated in the rotary shaft due to the magnetization of the inner ring.
Meanwhile, if the inner ring is formed of a metal material having a weak magnetization property in order to avoid the problem that the heat is generated in the rotary shaft due to the magnetization of the inner ring, the hardness of the inner ring becomes lower so that the inner ring is weak against impact or the cost of the inner ring significantly becomes higher.


A process and catalyst blend for selectively producing mercaptans and sulfides from alcohols. The alcohol is reacted with hydrogen sulfide, in the presence of a catalyst blend containing a hydrotreating catalyst and a dehydration catalyst to convert the alcohol to mercaptan or sulfide in one-pass. The alcohols can include primary and secondary alcohols. The mercaptan or sulfide having less than about 30% unreacted alcohol contained therein.

Mercaptans, which are also known as thiols, are organic compounds that are used in diverse applications. Some mercaptans are used as pipeline gas odorants, agricultural supplements, or as polymerization process modifiers. When mercaptans are present in large quantities, they are generally considered to be malodorous and highly undesirable. But in low concentrations these same compounds are used as fragrances and flavorings. When used in this manner they can be an effective and economical replacement of more expensive natural products in a wide variety of perfumed articles such as soaps, detergents, powders, perfume and cologne, and even as flavoring agents. For example, mercaptans are commonly used in perfumes and cleaning supplies to impart a lavender scent. In foods and other products, mercaptan compounds are widely used to reproduce the taste and fragrance associated with black currant or grapefruit.
The variety of tastes and flavorings available can be attributed to the numerous mercaptan compounds that can be synthesized. Yet, while numerous compounds can be made, economical processes for the production of even relatively simple mercaptans are not available in some cases. In other cases, the purity of known processes require difficult separation steps to obtain mercaptans of a desired purity.
A common process to produce mercaptans is by catalytically reacting an olefin with hydrogen sulfide in the presence of a solid acid catalyst. One drawback to this typical type of reaction is that the raw material costs associated with olefins can be high. In order to reduce raw material costs, prior attempts have been made to produce mercaptans from cheaper raw materials, such as alcohols. Water is a product of reaction in the synthesis of mercaptans from alcohols. Generally, one mole of water is produced for each mole of mercaptan produced. Because the solid acid catalysts that are used for converting olefins to mercaptans can be poisoned by moisture, they are not suitable for use in synthesis of mercaptans from alcohols.
A need exists for a process for selective production of mercaptans from alcohols that results in essentially complete conversion of the alcohol to the desired mercaptan. A further need exists for a process that achieves essentially complete conversion of the alcohol to the mercaptan in one process step.

Process and catalyst for synthesis of mercaptans and sulfides from alcohols


A cost effective, pressure-sealed, multi-page paper assembly is configured as a mailer-type of assembly for use as a business form, direct mail piece or other document to distribute information to end-users. The multi-page paper assembly is formed from a single ply or sheet of paper that permits use of such cost-saving technologies as laser printing and imaging of the single ply or sheet. The multi-page paper assembly includes a central transverse line of cross fold perforations and a plurality of lines of fold assist perforations that permit the single ply or sheet to be folded into a desired Z-fold, C-fold, eccentric C-fold, V-fold, double parallel-fold or other folded configuration having a number of inboard panels to serve as pages of the assembly. Deposits of pressure-activated cohesive along one or both surfaces of the single ply or sheet are placed to define those panels to serve as inboard boards and to adhere portions of the ply or sheet when folded. Cohesive deposits can form seals and thereby a secure seal multi-page paper assembly when pressure-sealed to protect contents. Use of low-tack deposits can form unsecured seals for use with direct mail pieces. Removal of stub portions of the paper assembly permits the sealed assembly to be opened in a book-like manner along the sealed cross fold perforations and access provided to the multiple of pages contained therein.

Multi-page paper assemblies, such as mailer-type assemblies, used for business forms, direct mail pieces, information and promotional booklets, and other multi-page volumes are formed to distribute and supply information and/or instruments, e.g., having a certain dollar value, to end users. Such multi-page paper assemblies are typically constructed of a multiple of plies or sheets of suitable paper. The multiple plies or sheets of paper are typically printed, imaged or otherwise processed to dispose data and information on one or more surfaces of each ply or sheet. Thereafter, the multiple of plies or sheets are typically collated, stacked and joined, e.g., adhered or bound by adhesive or cohesive, along at least one longitudinal or one transverse edge to form a binder. The binder permits the multiple plies or sheets to be held together as an assembly and to configure the assembly in a book like manner. The plies or sheets of paper serve as pages of the assembly and can be readily accessible to an end user. Such an assembly can be further configured to provide a sealed assembly whereby adhesive or cohesive can be deposited on each ply or sheet at certain locations to join or seal adjacent pages, and in some cases to seal the assembly around its perimeter. Such sealed, multi-page sealed assemblies can be distributed to end users by hand and/or by mailing through the U.S. Postal Service.
Printing and binding multiple plies or sheets of paper suitable for forming a multi-page assembly includes a number of steps such as printing each surface of each ply or sheet, depositing sufficient adhesive or cohesive along certain portions of the plies or sheets and collating and binding the multiple of plies and sheets such that the completed assembly is bound and opens in a book like manner. Multiple process steps and plies or sheets of paper required to form a sealed, multi-page assembly, as described, contribute to the overall manufacturing costs and time to produce such an assembly and, hence, can increase the expense of producing this type of multi-page document. Thus, it is desirable to provide a sealed, multi-page paper assembly configured as a mailer type of assembly that is produced with less materials and by a reduced number of process steps, e.g., by permitting the use of laser printing or imaging techniques, such that manufacturing time and costs are reduced and a cost effective sealed, multi-page document is provided.

Cross folded, pressure sealed multi-page paper assembly and methods of making same


A mechanism is provided for achieving autonomic computing self-healing utilizing meta level reflection. In a base level, a monitor detects an error in a production environment. The base level provides a reification message comprising data about the error to a meta level. A reasoning system in the meta level receives the reification message and analyzes the data using knowledge of computational components in the base level. The reasoning system identifies a self-healing action for the error and returns a reversion message comprising a signal to implement the self-healing action. Responsive to receiving the signal, the base level implements the self-healing action.

Various approaches have been proposed for handling errors or failures in computers. In some examples, error-handling is not separated from hardware. In other examples, the complexity of automated problem-solving is not addressed.
It is very difficult, with the current state of the art in computing, for a system or application experiencing an error to automatically examine possible recovery approaches, and select a recovery approach. It is very complex for the programmer to deal with an error in the same space utilized by the application experiencing the error.
Thus there is a need for methods and systems that reduce to a manageable level the complexity of automated error handling, recovery, problem-solving.

System and method for achieving autonomic computing self-healing, utilizing meta level reflection and reasoning

International Business Machines Corporation


A needle device having a plurality of needles with applied tips arranged so as to define a plurality of contact edges. In one embodiment one or more oblique edges is provided relative to an intermediate horizontal edge or a plurality of oblique edges with adjacent edges sharing a common needle point vertex. The number and needle grading (e.g., fine/sharper tips; medium and heavy grades) are represented by the different contact edges of the needle device. The number of each needle sub-sets is also preferably varied to provide enhanced flexibility as the application as in cosmetic tattooing. A tool for supporting is also preferably provided as in a reciprocating needle device support is provided. The needle device is particularly well suited for providing permanent eye liner makeup in view of its adaptability to accommodate different skin textures and skin surface presentations.

For centuries efforts have been made to achieve enhanced coloration of skin for a myriad of reasons. Historically, a variety of implements have been used to indelibly color the skin, ranging from sharply pointed bones, teeth, thorns, guitar strings, safety pins, wood, plastic or any metallic object sufficient to penetrate the skin with color so as to leave a visible mark. These colorful or black and white marks, also known as tattoos, have been applied for adornment, symbolic, patriotic, ethnic or religious identification and artistic expression on a human canvas of skin. Interest also exists in the area of cosmetic, corrective or camouflage tattooing (medical micropigmentation) as a method to provide, for example, permanent makeup, or restore color and/or symmetry to the eyebrows, lips, eyelids, breasts, scars and skin conditions from losses suffered by aging, birth defects (cleft lip), cancer treatment (alopecia) or skin changes (hypopigmentation-vitiligo).
Tools to implement a desired intradermal, coloration of the skin with a penetration implement have also evolved over the years. These include early hand tools such as bamboo sticks with strapped needles (a technique still common today in parts of Asia) and motor driven intradermal injection devices. These devices are relied on to inject ink, dye or other marking material (referred to hereafter collectively as "ink" for brevity) just under the skin, so that the ink is retained within the skin and the color of the ink injection pattern is visible. The motorized devices normally comprise a skin-penetrating needle which has the capacity to retain some quantity of ink, a mechanism to reciprocate the needle for repeated punctures of the skin to implant the ink under the skin in the desired pattern, and a housing for the device which the operator holds and often uses to guide the device. There is also typically an off-on switch and power source for the reciprocating drive mechanism. With some devices the operator repeatedly dips the needle into an ink pool to coat the needle, while other devices have built-in reservoirs for the ink from which the ink is fed continuously to the needle.
A number of different devices, particularly with different types of reciprocating needle drives, have been disclosed over the years. Typical of such devices are those described in U.S. Pat. No. 2,840,076 (Robbins: 1976); U.S. Pat. No. 4,508,106 (Angres: 1985); U.S. Pat. No. 4,644,952 (Patips et al.: 1987); U.S. Pat. No. 4,798,582 (Sarath et al.: 1989); and U.S. Pat. No. 5,279,552 (Magnet: 1984).
Examples of pen like (non-motorized) skin marking devices can be seen in U.S. Pat. No. 4,655,912 (Burton: 1987) and U.S. Pat. No. 5,810,862 (Pilamanis: 1998).
Intradermal pigment injection instruments or implements include the use of singular needles. Multi-tip array needles are also featured in the above noted Angres, Pilmanis, and Sarath patents as well as in U.S. Pat. No. 6,030,404 (Lawson et al.: 2000). Reference is also made to the technical report "The Manufacture of Needle Specifically For Tattooing"© Eikon Device Inc., October 1995 with June 1996 revisions, which report is incorporated herein by reference. The needles are designed for penetrating the skin to deliver the ink with the depth being based on insertion depth which, in turn, is dependent upon a variety of factors including the taper and/or diameter of the penetrating needle, the resistance level of the material being penetrated (e.g., the toughness of the skin) as well as environmental factors such as the dryness of the needle and/or skin being penetrated and whether there is wetting or lubrication material on the needle or on the skin being penetrated (or other body material). The invention can also be used for other medical, non-skin use such as medical pigment application to non-skin areas like the cornea of an eye, although the preferred usage and many of the parameters set forth herein are directed at skin penetration.
The age of the person can have an influence on the resistance level to penetration as aging skin tends to lose in collagen level and turgor so as to become more resistant to needle penetration and is also more susceptible to tissue damage from tattoo needles. One's skin also typically becomes thinner than in one's youth, thus playing a role relative to ink penetration levels.
"Single point" needles are typically relatively larger needles that are designed and used alone relative to the holder for line generation (e.g., single line or areas following extensive multiple line repetition). In view of their size, these single point needles typically are more traumatic on the skin leading to greater puffing etc., which can make ink application more difficult and less error free (e.g., if puffing and distortion initiates while the ink application is ongoing in the same area). Single point needles do allow, however, for high definition location application particularly in difficult to reach areas or when attempting to set initial external boundary regions.
Multiple needle configurations such as that described in Lawson are used to penetrate the skin over large areas. They are, however, not always well suited for areas such as those described above where a certain skin topography, desired ink configuration and/or a body arrangement makes the particular array arrangement of the multi-array needles ill suited for a desired use. This entails having to switch out multiple needle types to accommodate the application requirement or the use of a plurality of different holder/needle set combinations.
Furthermore, microscopic examination of tattooed skin to determine the depth and location of ink or colorant means (e.g., particles (pigments)) reveals pigment lying as deep as 2.6 mm in some applications. This depth may be acceptable on some parts of the body, but is excessive on others (e.g., consider the thickness of eyelid skin at 0.5-0.75 mm). Placing colorants too deeply results in skin changes and scarring as well as impaired visibility of the pigment. Considering that, unlike many body tattoos, in excess of 90% of cosmetic tattoos are placed on the face (e.g., eyeliner application) where they are constantly visible without the benefit of a covering (clothes), there exists a particularly high level of concern with respect to the final look and quality of the cosmetic tattoo application. That is, there is little room for visible errors when dealing with cosmetic surgery on the face such as the lips, eyelids, brow, etc. Also, from the foregoing, it is also apparent the putting colorants into the skin is quite variable from a variety of perspectives. This variation is particularly prevalent when dealing with cosmetic surgery on the face where the topography changes and skin texture changes can be more dramatic even in small areas of application. Because of this, there often exists in the prior art techniques a requirement to repeat an application (e.g., eyeliner application repeats) due to poor or improper penetration and/or the providing of insufficient pigment. This is especially undesirable and inefficient in that the initial procedure (which requires a healing process as well) is at least in part a wasted effort.

Intradermal color introducing needle device, and apparatus and method involving the same


A linearized ferroelectric capacitor includes a ferroelectric dielectric medium; a doped region in the ferroelectric dielectric medium, the doped region having a charge of a first polarity which divides the medium into two sections of opposing polarized domains; and a pair of electrodes on the ferroelectric medium having a contact potential of a second, opposite polarity to that of the doped region for reinforcing and stabilizing the polarized domains in each section.

Many materials are used as dielectrics in capacitors. These include glasses, ceramics, non-conductive metal oxides, plastics and polymers, and minerals of various types. The disadvantage of these non-ferroelectric dielectrics are the low dielectric constant of most of these materials. Most of these materials have relative permittivity values in the range of 1 to 100. A low dielectric constant requires that a capacitor be physically large to obtain large capacitance values, so the primary disadvantage is the size and weight of capacitors made with these materials. High dielectric ceramics gain their properties from an instability of the crystalline lattice which causes the ceramic to change phases with temperature. In certain phases the instability results in large values of permittivity, but the permittivity changes markedly with temperature.
Ferroelectric ceramics have also been used as capacitor dielectrics. The disadvantage of ferroelectric capacitors for use in most circuit applications is hysteresis. The hysteresis creates two disadvantages: it lowers the effective permittivity, and it creates switching effects which are unwanted in most circuits. When a voltage is first applied to a ferroelectric capacitor a large amount of charge is absorbed. This makes it appear as if the capacitor has a large dielectric constant. However, when the applied voltage is refumed to zero, only a fraction of the charge that went into the capacitor flows back out. The bulk of the charge is stored more or less permanently as remanent polarization in the capacitor. The only way to remove the remanent polarization electrically is to apply a reverse bias of the same magnitude as applied originally, which causes a reverse switching of the remanent polarization. This hysteresis means that the true dielectric constant of the capacitor as used in the circuit is actually much lower than the value computed when the remanent charge is included.


A method and apparatus for imparting a positive phase slope (i.e., a negative group delay) to a narrowband signals is disclosed which adjusts the phases of the various frequency components of a signal in a manner opposite to that of a delay line. The invention also permits the amount of phase slope to be adjusted, electronically, without the need for electro-mechanical apparatus or the interchange of cables. Furthermore, embodiments of the present invention are advantageous in that at the center of the operating band they maintain the phase of the delayed signal. These results are obtained in an illustrative embodiment of the present invention which divides the signal to be delayed into two signals which traverse different signal paths and are recombined in a signal combiner. The first signal path advantageously comprises a gain control block. The second signal path advantageously comprises a second gain control block and a fixed line delay which is not found in the first signal path. The amount of phase slope imparted to the signal can be adjusted by varying the gain (or attenuation) of the respective gain control blocks.

In electronic circuits it is often advantageous to temporally delay a narrowband electric signal. Such a delay can be realized by increasing the physical path traversed by the signal, thereby altering the phases of the various frequency components in an amount proportional to their frequency. In some applications, however, it is advantageous to expedite a narrowband signal (i.e., make the signal appear to have propagated a shorter distance than it actually did).

Method and apparatus for imparting positive phase slope to a narrowband signal


A novel polymer containing silphenylene and polyether structures in the backbone is used to formulate a photosensitive composition having improved reliability.

In the prior art, photosensitive protective films for semiconductor devices and photosensitive dielectric films for multilayer printed circuit boards are formed of photosensitive polyimide, epoxy resin, and silicone compositions. While many photosensitive materials are known for the protection of such substrates and circuit boards, Patent Document 1 discloses a photosensitive silicone composition having improved flexibility.
Patent Document 2 discloses a photosensitive silicone composition based on a silphenylene structure-containing silicone polymer. Although this photosensitive silicone composition has improved chemical resistance to liquid photoresist strippers, the cured product can spall off from the substrate or crack during a thermal cycle test (repetitive 1,000 cycles of −25° C./10 min holding and 125° C./10 min holding). A further improvement in reliability is desired.

Polymer containing silphenylene and polyether structures


The present invention provides a method for preparing HSV vectors. The method comprises co-transfecting a source vector and a mutating cassette together into a population of appropriate host cells, such that homologous recombination occurs between the mutating cassette and the source vector whereby the mutating cassette replaces a region of the HSV genome. The mutating cassette has a unique restriction site not present in the sequence of the vector. The method further comprises plaquing the co-transfected host cells, selecting plaques in which recombination has occurred between the source vector and the mutating cassette, and isolating the viral DNA from the plaques. The isolated viral DNA is digested with a restriction endonuclease appropriate for cleaving the viral DNA at the unique restriction site within the mutating cassette to produce two viral polynucleotides. Following purification, the two viral polynucleotides can be ligated to form an HSV vector comprising the two viral polynucleotides. Alternatively, the two isolated viral polynucleotides can be recombined with an insertion cassette to form an HSV vector comprising the insertion cassette at the former locus of the unique restriction site. The present invention further provides a mutant vector, particularly an HSV vector constructed in accordance with the method for the present invention. The present invention further provides a multigene HSV vector, particularly a multigene HSV vector for cancer therapy.

Gene transfer technology has wide-ranging utility in a number of applications relating to biological research and the treatment of disease. Central to this technology is a vector for introducing expression cassettes into target cells such that the cassettes can be expressed in the target cells. Examples of such vectors include naked DNA vectors (such as plasmids), viral vectors (such as adeno-associated viral vectors) (Berns et al., Annals of the New York Academy of Sciences, 772, 95-104 (1995)), adenoviral vectors (Bain et al., Gene Therapy, 1, S68 (1994)), herpesvirus vectors (Fink et al., Ann. Rev. Neurosci., 19, 265-87 (1996)), packaged amplicons (Federoff et al., Proc. Nat. Acad. Sci. USA, 89, 1636-40 (1992)), pappiloma virus vectors, picornavirus vectors, polyoma virus vectors, retroviral vectors, SV40 viral vectors, vaccinia virus vectors, and other vectors. Once a given type of vector is selected, its genome must be manipulated for use as a background vector, after which it must be engineered to incorporate exogenous polynucleotides.
A particularly attractive vector system employs recombinant herpes simplex virus (HSV) vectors. HSV attacks the human nervous system after a primary infection of the dermal or mucosal tissues. HSV naturally enters axonal portions of sensory ganglia and is transported to the somas of the cells, whereupon the viral DNA is released to the nucleus (Stevens, Microbiol. Rev., 53, 318-32 (1989); Roizman & Sears, in Field's Virology, 2d ed. Raven Press, Fields et al., eds. 1795-1841 (1990)). At that point, the wild-type virus either initiates a lytic cycle characterized by viral replication, or enters a latent phase. During the lytic phase, HSV gene expression proceeds through a well-characterized cascade typified by three discrete phases: Immediate Early (.alpha. or IE), Early (.beta. or E), and Late (.gamma. or L). Recombinant HSV viruses can be induced to enter latency by inactivating immediate early genes required for replication (e.g., d120, DeLuca et al., J. Virol., 56, 558 (1985)). During the latent phase, the virus persists in an episomal form for the life of its host (Mellerick and Fraser, Virology, 158, 265-75, (1987); Rock and Fraser, J. Virol., 55, 849-52, (1985)), neither interfering with neuronal function nor inducing any autoimmune response (Ramakrishnan et al., J. Virol., 68, 1864-70 (1994); Fruh et al., Nature, 375, 415 (1995)).
The HSV viral genome is well characterized, as is its life cycle, and the functions of more than 80 native coding polynucleotides are largely defined. HSV coding polynucleotides are generally contiguous linear sequences, thus facilitating genetic engineering of mutant vectors. Furthermore, as roughly half of the viral genes are dispensable for growth in cell culture, the possibility exists of deleting large segments of the HSV genome to accommodate transgenic material (Roizman & Sears, supra; Glorioso et al., in Viral Vectors, Academic Press, New York (Kaplitt & Loewy, eds.) 1-23 (1995)). Theoretically, up to 30 KB of the HSV genome can thus be replaced with exogenous material without requiring complementary host cells for propagation of the virus.
As the HSV genome is so well characterized, its genome is readily manipulated for use as a background vector. For example, it is possible to produce vectors deficient for essential loci as long as the missing translation product is otherwise provided, such as via a helper virus or complementary cell line. In fact, of the IE loci (ICP0, ICP4, ICP22, ICP27, and ICP47), only two (ICP4 and ICP27) are required for viral replication. In the absence of these loci, only other immediate early loci are expressed, and the virus is rendered replication-incompetent in non-complementing cell lines. A replication-deficient HSV virus can be induced to exist in host neural tissue in its persistent latent phase without any apparent pathological effect on the host (Ramakrishnan et al., supra), and defective HSV vectors can persist in a similar state in non-neuronal cells as well.
HSV vectors deficient for only one essential IE locus remain highly cytotoxic to infected cells, largely due to expression of the remaining IE gene products (DeLuca et al., J. Virol., 56, 558-70 (1985); Johnson et al., J. Virol., 66, 2952-65 (1992)). However, further research has indicated that mutants deleted for ICP4, ICP22, and ICP27 demonstrate reduced cytotoxicity over singly deficient vectors (see, e.g., Wu et al., J. Virol., 70(9), 6358-69 (1996)). Furthermore, the deletion of other loci reduces the cytotoxicity of HSV viruses (e.g., ICP47 (including U.sub.s 10-11), UL41, VP16, UL24; see, e.g., Jacobson et al., J. Virol., 63(4), 1839-43 (1989); Johnson, J. Virol., 68(10), 6347-62 (1994)). As such, multideficient HSV vectors represent a desirable choice for gene transfer technology.
Of course, production of multideficient viruses is one step in developing vectors for expression cassette transfer. The second step is engineering a vector containing exogenous expression cassettes. In many applications of gene transfer technology, the expression of multiple transgenes within the target cells is desirable. For example, coordinate expression of multiple cytokine sequences together with a sequence encoding an activator for an antitumor pro-drug is a potentially effective cancer therapy. Preferably, a vector is engineered such that the multiple transgenes are independently controlled (e.g., each is under regulatory control of a separate promoter) to optimize the expression kinetics. Because replication-deficient HSV vectors can transiently or constitutively express exogenous expression cassettes (Fink et al., Ann Rev. Neurosci, 19, 265-287 (1996)), HSV vectors containing multiple exogenous cassettes represent powerful agents for gene transfer applications.
Although attractive choices for gene transfer technology, HSV vectors have not been widely utilized for these therapies as of yet, primarily due to the difficulty in engineering vectors using standard procedures. The standard method for engineering mutant HSV viruses is to cotransfect host cells with the source virus and a polynucleotide comprising the desired mutation flanked by regions homologous to the target site within the HSV genome. Within the host cell, homologous recombination produces desired mutant HSV viruses less than 5% of the time, with the efficiency generally proportionate to the size of the flanking regions. Aside from inherently low efficiency, recombinant viruses often grow at markedly reduced rates vis-a-vis unmodified parental viruses within host cells, and so are easily overgrown. Thus, where the source vector is already deficient for native loci (particularly essential loci) viral growth can be substantially compromised, and the efficiency of recombination reduced accordingly. Therefore, screening plaques for desired recombinants is a laborious process, and the process becomes incrementally more tedious with multideficient HSV vectors, especially so where the desired mutation is not readily selectable.
Methods for increasing the efficiency of recombination have been attempted, but each presents significant drawbacks for quickly developing novel transfer vectors. Site-specific recombinases such as the cre-lox recombination system (reviewed in Kilby et al. Trends in Genetics, 9, 413-21 (1993)), have been employed to facilitate introduction of exogenous material into viral genomes (see, e.g., Gage et al., J. Virol., 66, 6509-15 (1992)). By these methods, exogenous polynucleotides are introduced into a recombinase recognition site previously engineered into the HSV genome. Recombinants can be selected by assaying for a reporter construct also present within the cassette. Furthermore, source viruses in which the recombinase recognition site is in a locus conferring growth benefits (e.g., the tk locus) do not enjoy the growth advantages over recombinants as seen in traditional methods. Thus, recombinase-mediated production of vectors is a more efficient method for producing HSV vectors than the co-precipitation method, routinely producing desired recombinants roughly 10% of the time, and in some instances site-specific recombination can be significantly more efficient (Rasty et al., Meth. Mol. Genet., 7, 114-30 (1995)). Despite the gain in efficiency, recombinase-mediated production of vectors presents two significant drawbacks. The first is that the method necessarily incorporates the entirety of any plasmid containing the desired insertion sequence, generally a bacterial plasmid for cloning a desired exogenous cassette. Of course, this requirement partially obviates the advantage inherent in the HSV genome's potentially large capacity to accommodate foreign DNA. Secondly, and more significantly, because the recombinase recognition site is retained (in duplicate) within a recombinant vector, subsequent rounds of site-specific recombination greatly disrupt the vectors and can result in randomization of the genome. Thus, site-specific recombination is not a preferred method for generating HSV vectors comprising multiple transgenes.
In order to enhance the efficiency of vector production, some attempts have been made to introduce unique restriction sites into the HSV genome. For example, in one system, all of the XbaI sites within the HSV genome are removed, and the RSAI site in the unique short region of the genome is changed to an XbaI site (Rixon & McLauchlan, J. Gen. Virol., 71, 2931-39 (1990)). Similarly, another system involves the introduction of a unique PacI site within the LAT region of an HSV mutant lacking 4.1 KB in one LAT copy (i.e., the HFEM mutant) (Huang et al., Gene Therapy, 1, 300-06 (1994)). With either of these systems, double-digestion of source vector DNA with the appropriate restriction enzyme permits efficient insertion of cassettes. Furthermore, screening of recombinants is achieved using a selectable marker (i.e., .beta.-galactosidase), which increases overall efficiency. However, these methods are limited to introducing exogenous DNA only at these loci, and generally only permit introduction of single cassettes.
Engineering HSV vectors to include multiple cassettes has proven difficult. Not only are the aforementioned methods inefficient for developing multigene HSV vectors, but certain properties of the HSV genome have thwarted attempts to create multiple insertions using standard means. For example, during replication, the HSV virus reacts to foreign DNA inserted at or near repeat elements. In some cases, a single expression cassette can be effectively introduced, but when the insertion of a second cassette is attempted, the virus reacts by recombining and shuffling its genome, substantially destabilizing the vector. In other instances, insertion of exogenous DNA disrupts viral function as well. Where multiple cassettes contain homologous regions to each other (i.e., a similar promoter, similar polyadenylation sequences, etc.), intragenomic recombination also can occur to destabilize the vector.
In view of the foregoing, there exists a need for a method for efficiently producing multideficient HSV vectors, especially multideficient HSV vectors that can be effectively employed in several successive rounds of mutagenesis. There also exists a need for a method for efficiently inserting exogenous polynucleotides into the HSV genome, and especially a means of creating multigene HSV vectors. There exists a further need for vectors having a multiplicity of transgenes for expression in a host cell (i.e., multigene vectors), and particularly for multigene HSV vectors having independently regulated transgenes.
The present invention provides an efficient method for creating mutant HSV vectors. The present invention further provides a multigene HSV vector having independently regulated transgenes. In particular, the present invention provides an HSV vector for cancer therapy deficient for native HSV polynucleotides and containing multiple pharmacologically active therapeutic transgenes. These and other advantages of the present invention, as well as additional inventive features, will be apparent from the description of the invention provided herein.

University of Pittsburgh of the Commonwealth System of Higher Education


A rotary Wankel type engine system is disclosed, wherein the crankshaft is supported by a center main roller bearing. The rotor apexes are sealed by two piece apex seals and by side seals. The engine system further includes an oiling system which permits oil flow through the center bearing and through lubrication passages in the engine rotor to provide a means for cooling the engine rotor under high loading conditions. Riblets are defined in the combustion face of the rotor to promote aerodynamic lateral stratified charge control to minimize atomized fuel charge loss to surface wetting before ignition.

The present invention relates to rotary Wankel type engines, and more particularly to such an engine which provides improved operation in the form of improved fuel charging.
Exemplary rotary type engines are shown in U.S. Pat. Nos. 3,996,898; 3,694,113; 3,228,183; and 3,259,115, and in German (Democratic Republic) patent document no. 26759.
Rotary Wankel type engine systems known in the art suffer from various drawbacks which have prevented such systems from achieving maximum engine performance and reliability. Such problems include high leakage of the rotor apex and side seals, causing overheating and lubrication contamination, and crankshaft bending for lack of a sufficient center main bearing support, causing side housing gouging and limiting maximum engine RPM under high compression firing loads.
The German patent document no. 26759 (Straube) illustrates a sealing arrangement apparently used in a rotary engine. This arrangement does not appear to provide an effective end seal in contact with the side seals. It also appears that the side seals restrict movement of the apex seals; the side seals will tend to jamb against the apex seals from the effects of rotation of the rotor.
U.S. Pat. No. 3,228,183 discloses a rotary internal combustion engine which is apparently a two stage diesel type engine. While the reference discloses two spark plugs, fuel is not injected laterally between the sparkplugs for stratified area charge. Moreover, the spark plugs are not equally spaced from the injectors, and therefore the system does not provide maximum stratification.
U.S. Pat. Nos. 3,259,115 and 3,694,113 describe rotary engine designs which employ a crankshaft supported by split journal type bearing assemblies. A disadvantage of such bearing assemblies is that they fail to provide adequate lubrication flow to the crankshaft and engine components, due to the relatively tight tolerance fit of the bearing journals around the crankshaft.
U.S. Pat. No. 3,996,898 describes an apex seal arrangement for a rotary engine. The dual apex seals do not seal at the side walls laterally. Separate corner seal elements are employed.
It would therefore represent an advance in the art to provide an improved rotary Wankel type engine system with improved apex and side sealing, and with improved center support bearing means for proper support of the crankshaft during high RPM loading.