The invention provides recombinant antibody molecules comprising antigen binding regions derived from the heavy and/or light chain variable regions of a donor anti-CD3 antibody, e.g. OKT3, and which have anti-CD3 binding specificity, preferably of affinity similar to that of OKT3. The recombinant antibody is preferably a humanized antibody and may be a chimeric or CDR-grafted antibody. A method is disclosed for preparing CDR-grafted humanized antibodies in which, in addition to the CDR""s non-human antibody residues are preferably used at positions 23, 24, 49, 71, 73 and 78 of the heavy chain variable region and at positions 46, 48, 58, and 71 of the light chain variable region. The recombinant, especially the humanized, anti-CD3 antibodies may be used for in vivo therapy or diagnosis.

The present invention relates to a recombinant antibody molecule (RAM), and especially a humanised antibody molecule (HAM), having specificity for an antigen present in the T-cell receptor-CD3 complex of most T-cells, to a process for its production using recombinant DNA technology and to its therapeutic use.
In the present application, the term xe2x80x9crecombinant antibody moleculexe2x80x9d (RAM) is used to describe an antibody produced by an process involving the use of recombinant DNA technology, including any analogues of natural immunoglobulins or their fragments. The term xe2x80x9chumanised antibody moleculexe2x80x9d (HAM) is used to describe a molecule having an antigen binding site derived from an immunoglobulin from a non-human species, remaining immunoglobulin-derived parts of the molecule being derived from a human immunoglobulin. The antigen binding site may comprise either complete variable domains fused onto constant domains or one or more complementarity determining regions grafted onto appropriate framework regions in the variable domains. The abbreviation xe2x80x9cMAbxe2x80x9d is used to indicate a monoclonal antibody.
In the description, reference is made to a number of publications by number. The publications are listed in numerical order at the end of the description.
Natural immunoglobulins have been known for many years, as have the various fragments thereof, such as the Fab, (Fabxe2x80x2)2 and Fc fragments, which can be derived by enzymatic cleavage. Natural immunoglobulins comprise a generally Y-shaped molecule having an antigen-binding site towards the end of each upper arm. The remainder of the structure, and particularly the stem of the Y, mediates the effector functions associated with immunoglobulins.
Natural immunoglobulins have been used in assay, diagnosis and, to a more limited extent, therapy. However, such uses, especially in therapy, have been hindered by the polyclonal nature of natural immunoglobulins. A significant step towards the realisation of the potential of immunoglobulins as therapeutic agents was the discovery of techniques for the preparation of monoclonal antibodies of defined specificity (ref. 1). However, most MAbs are produced by fusions of rodent spleen cells with rodent myeloma cells. They are therefore essentially rodent proteins. There are very few reports of the production of human MAbs.
Since most available MAbs are of rodent origin, they are naturally antigenic in humans and thus can give rise to an undesirable immune response termed the HAMA (Human Anti-Mouse Antibody) response. Therefore, the use of rodent MAbs as therapeutic agents in humans is inherently limited by the fact that the human subject will mount an immunological response to the MAb and will either remove it entirely or at least reduce its effectiveness. Thus, in practice, MAbs of rodent origin may not be used in patients for more than one or a few treatments as a HAMA response soon develops rendering the MAb ineffective as well as giving rise to undesirable reactions.
Proposals have therefore been made for making non-human MAbs less antigenic in humans. Such techniques can be generically termed xe2x80x9chumanisationxe2x80x9d techniques. These techniques generally involve the use of recombinant DNA technology to manipulate DNA sequences encoding the polypeptide chains of the antibody molecule.
Early methods for humanising MAbs involved production of chimeric antibodies in which an antigen binding site comprising the complete variable domains of one antibody is linked to constant domains derived from another antibody. Methods for carrying out such chimerisation procedures are described in EPO120694 (Celltech Limited), EP0125023 (Genetech Inc. and City of Hope), EP-A-0 171496 (Res. Dev. Corp. Japan), EP-A-0 173 494 (Stanford University), and WO 86/01533 (Celltech Limited). This latter Celltech application (WO 86/01533) discloses a process for preparing an antibody molecule having the variable domains from a mouse MAb and the constant domains from a human immunoglobulin. Such humanised chimeric antibodies, however, still contain a significant proportion of non-human amino acid sequence, i.e. the complete non-human variable domains, and thus may still elicit non-human variable domains, and thus may still elicit some HAMA response, particularly if administered over a prolonged period [Begent al al Br. J.Cancer, 62: 487 (1990)].
WO 86/01533 also describes the production of an antibody molecule comprising the variable domains of a mouse MAb, the CHl and CL domains of a human immunoglobulin, and a non-immunoglobulin-derived protein in place of the Fc portion of the human immunoglobulin.
In an alternative approach, described in EP-A-0239400 (Winter), the complementarity determining regions (CDRs) of a mouse MAb have been grafted onto the framework regions of the variable domains of a human immunoglobulin by site directed mutagenesis using long oligonucleotides. There are 3 CDRs (CDR1, CDR2 and CDR3) in each of the heavy and light chain variable regions. Such CDR-grafted humanised antibodies are much less likely to give rise to a HAMA response than humanised chimeric antibodies in view of the much lower proportion of non-human amino acid sequence which they contain.
The earliest work on humanising MAbs by CDR-grafting was carried out on MAbs recognising synthetic antigens, such as the NP or NIP antigens. However, examples in which a mouse MAb recognising lysozyme and a rat MAb recognising an antigen on human T-cells respectively were humanised by CDR-grafting are shown by Verhoeyen et al (ref. 2) and Riechmann et al (ref. 3). The preparation of CDR-grafted antibody to the antigen on human T cells is also described in WO 89/07452 (Medical Research Council).
In Riechmann et al it was found that transfer of the CDR regions alone (as defined by Kabat refs. 4 and 5) was not sufficient to provide satisfactory antigen binding activity in the CDR-grafted product. Riechmann et al found that it was necessary to convert a serine residue at position 27 of the human sequence to the corresponding rat phenylalanine residue to obtain a CDR-grafted product having satisfactory antigen binding activity. This residue at position 27 of the heavy chain is within the structural loop adjacent to CDR1. A further construct which additionally contained a human serine to rat tyrosine change at position 30 of the heavy chain did not have a significantly altered binding activity over the humanised antibody with the serine to phenylalanine change at position 27 alone. These results indicate that changes to residues of the human sequence outside the CDR regions, in particular in the loop adjacent to CFR1, may be necessary to obtain effective antigen binding activity for CDR-grafted antibodies which recognise more complex antigens. Even so the binding affinity of the best CDR-grafted antibodies obtained was still significantly less than the original MAb.
Very recently Queen et al (ref. 6) have described the preparation of a humanised antibody that binds to the interleukin 2 receptor, by combining the CDRs of a murine MAb (anti-Tac) with human immunoglobulin framework and constant regions. The human framework regions were chosen to maximise homology with the anti-Tac MAb sequence. In addition computer modelling was used to identify framework amino acid residues which were likely to interact with the CDRs or antigen, and mouse amino acids were used at these positions in the humanised antibody.
In WO 90/07861 Queen et al propose four criteria for designing humanised immunoglobulins. The first criterion is to use as the human acceptor the framework from a particular human immunoglobulin that is unusually homologous to the non-human donor immunoglobulin to be humanised, or to use a consensus framework from many human antibodies. The second criterion is to use the donor amino acid rather than the acceptor if the human acceptor residue is unusual and the donor residue is typical for human sequences at a specific residue of the framework. The third criterion is to use the donor framework amino acid residue rather than the acceptor at positions immediately adjacent to the CDRs. The fourth criterion is to use the donor amino acid residue at framework positions at which the amino acid is predicted to have a side chain atom within about 3 xc3x85 of the CDRs in a three-dimensional immunoglobulin model and to be capable of interacting with the antigen or with the CDRs of the humanised immunoglobulin. It is proposed that criteria two, three of four may be applied in addition or alternatively to criterion one, and may be applied singly or in any combination.
WO 90/07861 described in detail the preparation of a single CDR-grafted humanised antibody, a humanised antibody having specificity for the p55 Tac protein of the IL-2 receptor. The combination of all four criteria, as above, were employed in designing this humanised antibody, the variable region frameworks of the human antibody Eu (refs. 4 and 5) being used as acceptor. In the resultant humanised antibody the donor CDRs were as defined by Kabat et al (refs. 4 and 5) and in addition the mouse donor residues were used in place of the human acceptor residues, at positions 27, 30, 48, 66, 67, 89, 91, 94, 103, 104, 105 and 107 in the heavy chain and at positions 48, 60 and 63 in the light chain, of the variable region frameworks. The humanised anti-Tac antibody obtained is reported to have an affinity for p55 of 3xc3x97109 Mxe2x88x921, about one-third of that of the murine MAb.
OKT3 is a mouse IgG2a/k MAb which recognises an antigen in the T-cell receptor-CD3 complex and has been approved for use in many countries throughout the world as an immunosuppressant in the treatment of acute allograft rejection [Chatenoud et al (ref. 7), and Jeffers et al (ref. 8) However, in view of the murine nature of this MAb, a significant HAMA response, with a major anti-idiotype component, may build up on use. Clearly, it would be highly desirable to diminish or abolish this undesirable HAMA response by suitable humanisation or other recombinant DNA manipulation of this very useful antibody and thus enlarge its area of use. It would also be desirable to apply the techniques of recombinant DNA technology more generally to this useful antibody to prepare RAM products.
Moreover, we have further investigated the preparation of CDR-grafted humanised antibody molecules and have identified a hierarchy of positions within the framework of the variable regions (i.e. outside both the Kabat CDRs and structural loops of the variable regions) at which the amino acid identities of the residues are important for obtaining CDR-grafted products with satisfactory binding affinity. This has enabled us to establish a protocol for obtaining satisfactory CDR-grafted products which may be applied very widely irrespective of the level of homology between the donor immunoglobulin and acceptor framework. The set of residues which we have identified as being of critical importance does not coincide with the residues identified by Queen et al (ref. 6).
Accordingly the present invention provides an RAM comprising antigen binding regions derived from the heavy and/or light chain variable regions of a donor anti-CD3 antibody and having anti-CD3 binding specificity, and preferably having an anti-CD3 binding affinity similar to that of OKT3.
Typically the donor anti-CD3 antibody is a rodent MAb.
The RAM of the invention may comprise antigen binding regions from any suitable anti-CD3 antibody, typically a rodent anti-CD3 MAb, e.g. a mouse or rat anti-CD3 MAb. The RAM may comprise a recombinant version of whole or a major part of the amino acid sequence of such a MAb. Also the RAM may comprise only the variable region (VH and/or VL) or one or more CDRs of such a MAb. Especially the RAM may comprise amino acid sequences, whether variable region, CDR or other, derived from the specific anti-CD3 MAb (OKT3) hereinafter specifically described with reference to FIGS. 1 and 2.
Preferably the RAM of the present invention is a humanised antibody molecule (HAM) having specificity for CD3 and having an antigen binding site wherein at least one of the complementarity determining regions (CDRs) of the variable domain, usually at least two and preferably all of the CDRs, are derived from a non-human anti-CD3 antibody, e.g. a rodent anti-CD3 MAb.
The RAM may be a chimeric antibody or a CDR-grafted antibody.
Accordingly, in preferred embodiments the invention provides an anti-CD3 CDR-grafted antibody heavy chain having a variable region domain comprising acceptor framework and donor CD3 binding regions wherein the framework comprises donor residues at at least one of positions 6, 23 and/or 24, 48 and/or 49, 71 and/or 73, 75 and/or 76 and/or 78 and 88 and/or 91.
More preferably, the heavy chain framework of the preferred embodiment comprises donor residues at positions 23, 24, 49, 71, 73 and 78 or at positions 23, 24 and 49. The residues at positions 71, 73 and 78 of the heavy chain framework are preferably either all acceptor or all donor residues.
In particularly preferred embodiments the heavy chain framework additionally comprises donor residues at one, some or all of positions 6, 37, 48 and 94. Also it is particularly preferred that residues at positions of the heavy chain framework which are commonly conserved across species, i.e. positions 2, 4, 25, 36, 39, 47, 93, 103, 104, 106 and 107, if not conserved between donor and acceptor, additionally comprise donor residues. Most preferably the heavy chain framework additionally comprises donor residues at positions 2, 4, 6, 25, 36, 37, 39, 47, 48, 93, 94, 103, 104, 106 and 107.
In addition the heavy chain framework optionally comprises donor residues at one, some or all of positions:
1 and 3,
72 and 76,
69 (if 48 is different between donor and acceptor),
38 and 46 (if 48 is the donor residue),
80 and 20 (if 69 is the donor residue),
67,
82 and 18 (if 67 is the donor residue),
91,
88, and
any one or more of 9, 11, 41, 87, 108, 110 and 112.
In the preferred embodiments of the present invention described above and hereinafter, reference is made to CDR-grafted antibody products comprising acceptor framework and donor antigen binding regions. It will be appreciated that the invention is widely applicable to the CDR-grafting of anti-CD3 antibodies in general. Thus, the donor and acceptor antibodies may be anti-CD3 antibodies derived from animals of the same species and even same antibody class or sub-class. More usually, however, the donor and acceptor antibodies are derived from animals of different species. Typically, the donor anti-CD3 antibody is a non-human antibody, such as a rodent MAb, and the acceptor antibody is a human antibody.
In the CDR-grafted antibody products of the present invention, the donor CD3 binding region typically comprises at least one CDR from the donor antibody. Usually the donor antigen binding region comprises at least two and preferably all three CDRs of each of the heavy chain and/or light chain variable regions. The CDRs may comprise the Kabat CDRs, the structural loop CDRs or a composite of the Kabat and structural loop CDRs and any combination of any of these. Preferably, the antigen binding regions of the CDR-grafted heavy chain variable domain comprise CDRs corresponding to the Kabat CDRs at CDR2 (residues 50-65) and CDR3 (residues 95-100) and a composite of the Kabat and structural loop CDRs at CDR1 (residues 26-35).
The residue designations given above and elsewhere in the present application are numbered according to the Kabat numbering (refs. 4 and 5). Thus the residue designations do not always correspond directly with the linear numbering of the amino acid residues. The actual linear amino acid sequence may contain fewer or additional amino acids than in the strict Kabat numbering corresponding to a shortening of, or insertion into, a structural component, whether framework or CDR, of the basic variable domain structure. For example, the heavy chain variable region of the anti-Tac antibody described by Queen et al (ref. 6) contains a single amino acid insert (residue 52a) after reside 52 of CDR2 and a three amino acid insert (residues 82a, 82b and 82c) after framework residue 82, in the Kabat numbering. The correct Kabat numbering of residues may be determined for a given antibody by alignment at regions of homology of the sequence of the antibody with a xe2x80x9cstandardxe2x80x9d Kabat numbered sequence.
The invention also provides in a further preferred embodiment a CDR-grafted antibody light chain having a variable region domain comprising acceptor framework and donor CD3 binding regions wherein the framework comprises donor residues at at least one of positions 1 and/or 3 and 46 and/or 47. Preferably the CDR grafted light chain of this preferred embodiment comprises donor residues at positions 46 and/or 47.
The invention also provides in a yet further preferred embodiment a CDR-grafted antibody light chain having a variable region domain comprising acceptor framework and donor CD3 binding regions wherein the framework comprises donor residues at at least one of positions 46, 48, 58 and 71.
More preferably in this latter embodiment, the framework comprises donor residues at all of positions 46, 48, 58 and 71.
In particularly preferred embodiments of the above preferred embodiments, the light chain framework additionally comprises donor residues at positions 36, 44, 47, 85 and 87. Similarly positions of the light chain framework which are commonly conserved across species, i.e. positions 2, 4, 6, 35, 49, 62, 64-69, 98, 99, 101 and 102, if not conserved between donor and acceptor, additionally comprise donor residues. Most preferably the light chain framework additionally comprises donor residues at positions 2, 4, 6, 35, 36, 38, 44, 47, 49, 62, 64-69, 85, 87, 98, 99, 101 and 102.
In additions the light chain framework of the above preferred embodiments optionally comprises donor residues at one, some or all of positions:
1 and 3,
63,
60 (if 60 and 54 are able to form at potential saltbridge),
70 (if 70 and 24 are able to form a potential saltbridge),
73 and 21 (if 47 is different between donor and acceptor),
37 and 45 (if 47 is different between donor and acceptor), and
any one or more of 10, 12, 40, 80, 103 and 105.
Preferably, the antigen binding regions of the CDR-grafted light chain variable domain comprise CDRs corresponding to the Kabat CDRs at CDR1 (residue 24-34), CDR2 (residues 50-56) and CDR3 (residues 89-97).
The invention further provides in a fourth aspect a CDR-grafted antibody molecule comprising at least one CDR-grafted heavy chain and at least one CDR-grafted light chain as defined above.
The CDR-grafted and humanised antibody molecules and chains of the present invention may comprise: a complete antibody molecule, having full length heavy and light chains; a fragment thereof, such as a Fab, (Fabxe2x80x2)2 or FV fragment; a light chain or heavy chain monomer or dimer; or a single chain antibody, e.g. a single chain FV in which heavy and light chain variable regions are joined by a peptide linker; or any other CDR-grafted or humanised antibody product with anti-CD3 binding specificity. Similarly the CDR-grafted heavy and light chain variable region may be combined with other antibody domains as appropriate.
Also the CDR-grafted or humanised heavy or light chains or antibody molecules of the present invention may have attached to them an effector or reporter molecule. For instance, it may have a macrocycle, for chelating a heavy metal atom, or a toxin, such as ricin, attached to it by a covalent bridging structure. Alternatively, the procedures of recombinant DNA technology may be used to produce an immunoglobulin molecule in which the Fc fragment of CH3 domain of a complete immunoglobulin molecule has been replaced by, or has attached thereto by peptide linkage, a functional non-immunoglobulin protein, such as an enzyme or toxin molecule.
For CDR-grafted antibody products, any appropriate acceptor variable region framework sequences may be used having regard to class/type of the donor antibody from which the antigen binding regions are derived. Preferably, the type of acceptor framework used is of the same/similar class/type as the donor antibody. Conveniently, the framework may be chosen to maximise/optimise homology with the donor antibody sequence particularly at positions close or adjacent to the CDRs. However, a high level of homology between donor and acceptor sequences is not important for application of the present invention. The present invention identifies a hierarchy of framework residue positions at which donor residues may be important or desirable for obtaining a CDR-grafted antibody product having satisfactory binding properties. The present invention advantageously enables the preparation of CDR-grafted antibody products having binding affinities similar to, and even in some cases better than the corresponding donor antibody product, e.g. OKT3 product. Preferably, the CDR-grafted antibody products of the invention have binding affinities of at least about 105 Mxe2x88x921, preferably at least about 108 Mxe2x88x921 and especially within the range 108xe2x88x921012 Mxe2x88x921. In principle, the present invention is applicable to any combination of donor and acceptor antibodies irrespective of the level of homology between their sequences. A protocol for applying the invention to any particular donor-acceptor antibody pair is given hereinafter. Examples of human frameworks which may be used are KOL, NEWM, REI, EU, LAY and POM (refs. 4 and 5); for instance KOL and NEWM for the heavy chain and RE1 for the light chain and EU, LAY and POM for both the heavy chain and the light chain.
Also the constant region domains of the products of the invention may be selected having regard to the proposed function of the antibody in particular the effector functions which may be required. For example, the constant region domains may be human IgA, IgE, IgG or IgM domains. In particular, IgG human constant region domains may be used, especially of the IgG1 and IgG3 isotypes, when the humanised antibody molecule is intended for therapeutic uses, and antibody effector functions are required. Alternatively, IgG2 and IgG4 isotypes may be used when the humanised antibody molecule is intended for therapeutic purposes and antibody effector functions are not required, e.g. for simple blocking of the T-cell receptor-CD3 complex.
However, the remainder of the antibody molecules need not comprise only protein sequences from immunoglobulins. For instance, a gene may be constructed in which a DNA sequence encoding part of a human immunoglobulin chain is fused to a DNA sequence encoding the amino acid sequence of a polypeptide effector or reporter molecule.
Preferably the CDR-grafted heavy and light chain and antibody molecule products are produced by recombinant DNA technology.
Thus in further aspects the invention also includes DNA sequences coding for the RAMs, HAMs and CDR-grafted heavy and light chains, cloning and expression vectors containing the DNA sequences, host cells transformed with the DNA sequences and processes for producing the CDR-grafted antibody molecules comprising expressing the DNA sequences in the transformed host cells.
The general methods by which the vectors may be constructed, transfection methods and culture methods are well known per se and form no part of the invention. Such methods are shown, for instance, in references 9 and 10.
The DNA sequences which encode the anti-CD3 donor amino acid sequences may be obtained by methods well known in the art. For example the anti-CD3 coding sequences may be obtained by genomic cloning, or cDNA cloning from suitable hybridoma cell lines, e.g. the OKT3 cell line hereinafter specifically described. Positive clones may be screened using appropriate probes for the heavy and light chain genes in question. Also PCR cloning may be used.
DNA coding for acceptor, e.g. human acceptor, sequences may be obtained in any appropriate way. For example DNA sequences coding for preferred human acceptor frameworks such as KOL, REI, EU and NEWM, are widely available to workers in the art.
The standard techniques of molecular biology may be used to prepare DNA sequences coding for the chimeric and CDR-grafted products. Desired DNA sequences may be synthesised completely or in part using oligonucleotide synthesis techniques. Site-directed mutagenesis and polymerase chain reaction (PCR) techniques may be used as appropriate. For example oligonucleotide directed synthesis as described by Jones et al (ref. 17) may be used. Also oligonucleotide directed mutagenesis of a pre-exising variable region as, for example, described by Verhoeyen et al (ref. 2) or Riechmann et al (ref. 3) may be used. Also enzymatic filling in of gapped oligonucleotides using T4 DNA polymerase as, for example, described by Queen et al (ref. 6) may be used.
Any suitable host cell/vector system may be used for expression of the DNA sequences coding for the CDR-grafted heavy and light chains. Bacterial e.g. E. coli, and other microbial systems may be used, in particular for expression of antibody fragments such as FAb and (Fabxe2x80x2)2 fragments, and especially FV fragments and single chain antibody fragments e.g. single chain FVs. Eucaryotic, e.g. mammalian, host cell expression systems may be used, in particular, for production of larger CDR-grafted antibody products, including complete antibody molecules. Suitable mammalian host cells include CHO cells and myeloma or hybridoma cell lines.
Thus, according to a further aspect the present invention provides a process for producing an anti-CD3 RAM which process comprises:
(a) producing in an expression vector an operon having a DNA sequence which encodes an antibody heavy chain wherein at least one CDR of the variable domain is derived from a donor anti-CD3 antibody and remaining immunglobulin-derived parts of the antibody chain are derived from an acceptor immunoglobulin;
and/or
(b) producing in an expression vector an operon having a DNA sequence which encodes a complementary antibody light chain wherein at least one CDR of the variable domain is derived from a donor anti-CD3 antibody and the remaining immunoglobulin-derived parts of the antibody chain are derived from an acceptor immunoglobulin;
(c) transfecting a host cell with the or each vector;
and
(d) culturing the transfected cell line to produce the RAM.
The RAM may comprise only heavy or light chain-derived polypeptide, in which case only a heavy chain or light chain polypeptide coding sequence is used to transfect the host cells.
For production of RAMs comprising both heavy and light chains, the cell line may be transfected with two vectors. The first vector may contain an operon encoding a light chain-derived polypeptide and the second vector may contain an operon encoding a heavy chain-derived polypeptide. Preferably, the vectors are identical except in so far as the coding sequences and selectable markers are concerned so as to ensure as far as possible that each polypeptide chain is equally expressed. Alternatively, a single vector may be used, the vector including the sequences encoding both light chain- and heavy chain-derived polypeptides.
The DNA in the coding sequences for the light and heavy chains may comprise cDNA or genomic DNA or both. However, it is preferred that the DNA sequence encoding the heavy or light chain comprises, at least partially, genomic DNA. Most preferably, the heavy or light chain encoding sequence comprises a fusion of cDNA and genomic DNA.
The present invention also includes therapeutic and diagnostic compositions comprising the RAMs, HAMs and CDR-grafted light and heavy chains and molecules of the invention and uses of such compositions in therapy and diagnosis.
Accordingly in a further aspect the invention provides a therapeutic or diagnostic composition comprising a RAM, HAM or CDR-grafted antibody heavy or light chain or molecule according to previous aspects of the invention in combination with a pharmaceutically acceptable carrier, diluent or excipient.
Accordingly also the invention provides a method of therapy or diagnosis comprising administering an effective amount of a RAM, HAM or CDR-grafted antibody heavy or light chain or molecule according to previous aspects of the invention to a human or animal subject.
The RAM, HAM and CDR-grafted products of the present invention may be used for any of the therapeutic uses for which anti CD3 antibodies, e.g. OKT3, have been used or may be used in the future. For example, the products may be used as ummunosuppressants, e.g. in the treatment of acute allograft rejection.
A preferred protocol for obtaining CDR-grafted antibody heavy and light chains in accordance with the present invention is set out below together with the rationale by which we have derived this protocol. This protocol and rationale are given without prejudice to the generality of the invention as hereinbefore described and defined.
Protocol
It is first of all necessary to sequence the DNA coding for the heavy and light chain variable regions of the donor antibody, to determine their amino acid sequences. It is also necessary to choose appropriate acceptor heavy and light chain variable regions, of known amino acid sequence. The CDR-grafted chain is then designed starting from the basis of the acceptor sequence. It will be appreciated that in some cases the donor and acceptor amino acid residues may be identical at a particular position and thus no change of acceptor framework residue is required.
1. As a first step donor residues are substituted for acceptor residues in the CDRs. For this purpose the CDRs are preferably defined as follows:
The positions at which donor residues are to be substituted for acceptor in the framework are then chosen as follows, first of all with respect to the heavy chain and subsequently with respect to the light chain.
2. Heavy Chain
2.1 Choose donor residues at all of positions 23, 24, 49, 71, 73 and 78 of the heavy chain or all of positions 23, 24 and 49 (71, 73 and 78 are either all donor or all acceptor).
2.2 Check that the following have the same amino acid in donor and acceptor sequences, and if not preferably choose the donor: 2, 4, 6, 25, 36, 37, 39, 47, 48, 93, 94, 103, 104, 106 and 107.
2.3 To further optimise affinity consider choosing donor residues at one, some or any of:
i. 1, 3
ii. 72, 76
iii. If 48 is different between donor and acceptor sequences, consider 69
iv. If at 48 the donor residue is chosen, consider 38 and 46
v. If at 69 the donor residue is chosen, consider 80 and then 20
vi. 67
vii. If at 67 the donor residue is chosen, consider 82 and then 18
viii. 91
ix. 88
x. 9, 11, 41, 87, 108, 110, 112
3. Light Chain
3.1 Choose donor at 46, 48, 58 and 71
3.2 Check that the following have the same amino acid in donor and acceptor sequences, if not preferably choose donor: 2, 4, 6, 35, 38, 44, 47, 49, 62, 64-69 inclusive, 85, 87, 98, 99, 101 and 102
3.3 To further optimise affinity consider choosing donor residues at one, some or any of:
i. 1, 3
ii. 63
iii. 60, if 60 and 54 are able to form a potential saltbridge
iv. 70, if 70 and 24 are able to form a potential saltbridge
v. 73 and 21, if 47 is different between donor and acceptor
vi. 37 and 45, if 47 is different between donor and acceptor
vii. 10, 12, 40, 80, 103, 105
Rationale
In order to transfer the binding site of an antibody into a different acceptor framework, a number of factors need to be considered.
1. The extent of the CDRs
The CDRs (Complementary Determining Regions) were defined by Wu and Kabat (refs. 4 and 5) on the basis of an analysis of the variability of different regions of antibody variable regions. Three regions per domain were recognised. In the light chain the sequences are 24-34, 50-56, 89-97 (numbering according to Kabat (ref. 4), Eu Index) inclusive and in the heavy chain the sequences are 31-35, 50-65 and 95-102 inclusive.
When antibody structures became available it became apparent that these CDR regions corresponded in the main to loop regions which extended from the xcex2 barrel framework of the light and heavy variable domains. For H1 there was a discrepancy in that the loop was from 26 to 32 inclusive and for H2 to loop was 52-56 and for L2 from 50 to 53. However, with the exception of H1 the CDR regions encompassed the loop regions and extended into the xcex2 strand frameworks. In H1 residue 26 tends to be a serine and 27 a phenylalanine or tyrosine, residue 29 is a phenylalanine in most cases. Residues 28 and 30 which are surface residues exposed to solvent might be involved in antigen-binding. A prudent definition of the H1 CDR therefore would include residues 26-35 to include both the loop region and the hypervariable residues 33-35.
It is of interest to note the example of Riechmann et al (ref. 3), who used the residue 31-35 choice for CDR-H1. In order to produce efficient antigen binding residue 27 also needed to be recruited from the donor (rat) antibody.
2. Non-CDR residues which contribute to antigen binding
By examination of available X-ray structures we have identified a number of residues which may have an effect on net antigen binding and which can be demonstrated by experiment. These residues can be sub-divided into a number of groups.
2.1 Surface residues near CDR [all numbering as in Kabat et al (ref. 4)].
2.1.1. Heavy Chainxe2x80x94Key residues are 23, 71 and 73. Other residues which may contribute to a lesser extent are 1, 3 and 76. Finally 25 is usually conserved but the murine residue should be used if there is a difference.
2.1.2 Light Chainxe2x80x94Many residues close to the CDRs, e.g. 63, 65, 67 and 69 are conserved. If conserved none of the surface residues in the light chain are likely to have a major effect. However, if the murine residue at these positions is unusual, then it would be of benefit to analyse the likely contribution more closely. Other residues which may also contribute to binding are 1 and 3, and also 60 and 70 if the residues at these positions and at 54 and 24 respectively are potentially able to form a salt bridge i.e. 60+54; 70+24.
2.2 Packing residues near the CDRs.
2.2.1. Heavy Chainxe2x80x94Key residues are 24, 49 and 78. Other key residues would be 36 if not a tryptophan, 94 if not an arginine, 104 and 106 if not glycines and 107 if not a threonine. Residues which may make a further contribution to stable packing of the heavy chain and hence improved affinity are 2, 4, 6, 38, 46, 67 and 69. 67 packs against the CDR residue 63 and this pair could be either both mouse or both human. Finally, residues which contribute to packing in this region but from a longer range are 18, 20, 80, 82 and 86. 82 packs against 67 and in turn 18 packs against 82. 80 packs against 69 and in turn 20 packs against 80. 86 forms an H bond network with 38 and 46. Many of the mouse-human differences appear minor e.g. Leu-Ile, but could have an minor impact on correct pacing which could translate into altered positioning of the CDRs.
2.2.2 Light Chainxe2x80x94Key residues are 48, 58 and 71. Other key residues would be 6 if not glutamine, 35 if not tryptophan, 62 if not phenylalanine or tryosine, 64, 66, 68, 99 and 101 if not glycines and 102 if not a threonine. Residues which make a further contribution are 2, 4, 37, 45 and 47. Finally residues 73 and 21 and 19 may make long distance packing contributions of a minor nature.
2.3. Residues at the variable domain interface between heavy and light chainsxe2x80x94In both the light and heavy chains most of the non-CDR interface residues are conserved. If a conserved residue is replaced by a residue of different character, e.g. size or charge, it should be considered for retention as the murine residue.
2.3.1 Heavy Chainxe2x80x94Residues which need to be considered are 37 if the residue is not a valine but is of larger side chain volume or has a charge or polarity. Other residues are 39 if not a glutamine, 45 if not a leucine, 47 if not a tryptophan, 91 if not a phenylalanine or tyrosine, 93 if not an alanine and 103 if not a tryptophan. Residue 89 is also at the interface but is not in a position where the side chain could be of great impact.
2.3.2. Light Chainxe2x80x94Residues which need to be considered are 36, if not a tyrosine, 38 if not a glutamine, 44 if not a proline, 46, 49 if not a tyrosine, residue 85, residue 87 if not a tyrosine and 98 if not a phenylalanine.
2.4. Variable-Constant region interfacexe2x80x94The elbow angle between variable and constant regions may be affected by alterations in packing of key residues in the variable region against the constant region which may effect the position of VL and VH with respect to one another. Therefore it is worth noting the residues likely to be in contact with the constant region. In the heavy chain the surface residues potentially in contact with the variable region are conserved between mouse and human antibodies therefore the variable region contact residues may influence the V-C interaction. In the light chain the amino acids found at a number of the constant region contact points vary, and the V and C regions are not in such close proximity as the heavy chain. Therefore the influences of the light chain V-C interface may be minor.
2.4.1. Heavy Chainxe2x80x94Contact residues are 7, 11, 41, 87, 108, 110, 112.
2.4.2. Light Chainxe2x80x94In the light chain potentially contacting residues are 10, 12, 40, 80, 83, 103 and 105.
The above analysis coupled with our considerable practical experimental experience in the CDR-grafting of a number of different antibodies have lead us to the protocol given above.
The present invention is now described, by way of example only, with reference to the accompanying FIGS. 1-13.


A display device includes a substrate including a display region arranged with pixels, and a terminal part on the outside of the display region, a polarizer overlapping the display region and the terminal part, the polarizer arranged with first wirings, a retardation plate overlapping the display region and the terminal part, the retardation plate between the pixels and the polarizer, and second wirings arranged in a direction intersecting the direction the first wirings is arranged, wherein an interval the first wirings is arranged is narrower than an interval the second wirings is arranged, the first wirings and the second wirings are arranged sandwiching a dielectric body, the first wirings and the second wirings are connected to a lead-out line, and an end of the retardation plate at the terminal part is closer to the display region than an end of the polarizer.

A liquid crystal display device utilizing the electro-optical effects of liquid crystals and organic EL display devices using organic electroluminescence (organic EL) devices have been developed practically used as display devices used in electric appliances and electronic devices. In addition, a touch panel, which is a display device in which a touch sensor is mounted on a display region, has been rapidly spreading. Touch panels are especially required for mobile information terminals including smartphones.
As a method of arranging a touch sensor electrode (transmission electrode, reception electrode) in an organic EL display device, there is a method of attaching a separately manufactured touch sensor or a method of forming a touch sensor electrode on a display region. For example, Japanese Patent Laid Open Patent Publication No. 2008-197757 discloses the use of a wire grid arranged on a polarizer as a touch sensor electrode.


Various aspects of one or more methods, apparatuses and data processor program products capable of enabling administration of an athleticism development program are disclosed herein. These various aspects include facilitating an athleticism training routine by an athleticism development program subscriber and facilitating a present athleticism level assessment by the athleticism development program subscriber after facilitating the athleticism training routine. After facilitating the present athleticism level assessment, an attained standardized athleticism level corresponding to the present athleticism level evaluation is determined. The implementation of standardized athleticism levels is advantageous as it supports a measurable plan of progress for motivating a subscriber and trainer to meet their individual and mutual goals.

The disclosures herein relate generally to athletic and fitness development programs and more particularly to one or more methods, apparatuses and data processor program products capable of enabling administration of a levels-based athleticism development program.
Many people engage in fitness training routines with the intent and expectation of achieving and/or maintaining a desired level of athleticism. In some cases, such people are athletes who are no longer actively involved in a previous sport or sports. In other cases, such people are not athletes per se, but do have a competitive spirit generally associated with athleticism and have a desire to improve their level of fitness.
In each case, such people generally have a desire to both measure their progress against some defined standard and to assess the development of their fitness and athleticism among their peers in a competitive environment such that the results of their hard work are recognized. The assessment of their fitness and athletic development is a key motivating factor that positively reinforces their quest towards achieving and/or maintaining their desired level of athleticism and fitness.
Self-directed general fitness activities facilitated via a health club membership or home gym are a common approach for engaging in a fitness training routine. A facility such as a local health club or a home gym generally provides the equipment necessary to improve a person""s level of fitness. However, persons motivated by factors such as recognition of their hard work, competition among peers, assessment of their progress and often loose motivation as a result of the seemingly static measures and limited feedback offered by activities and routines associated with traditional fitness development programs and facilities.
It is not that the health clubs and home gyms do not provide the equipment or tools needed to develop an athletic level of fitness. But, the individual motivation and knowledge needed to develop and/or measure balance from an athleticism perspective is often lacking. Consequently, at least a portion of health club members engage personal trainers. However, even when the training equipment is available and the knowledge from personal trainers is available, limitations associated with quantitatively assessing their athletic development and physical fitness against a set of athletic fitness standards or against peers in a self-challenging and competitive environment still exists.
Research by American Sports Data, Inc. and The International Health, Racquet and Sportsclub Association (IHRSA) verifies that traditional fitness development programs suffer from several limitations with respect to developing a desired athleticism level, quantifying a desired level of fitness and/or athleticism, and motivating one towards their desired athleticism level. One limitation is that traditional fitness development programs are not based on standardized development levels for allowing comparative assessments between program subscribers or club members. Another limitation is that there is no standard manner for determining an attained level of fitness and/or athleticism. Yet another limitation is that there is no targeted and/or quantified feedback for leveraging a fitness routine in a manner that contributes to improving a level of athleticism.
Accordingly, an athleticism development program capable of being administered in a manner that overcomes the limitations associated with a traditional fitness and/or athleticism development program is useful.

Method, apparatus and data processor program product capable of enabling administration of a levels-based athleticism development program


Peptides and mimetics of selected domains of mammalian serum amyloid A isoform 2.1 (SAA2.1) and compounds and compositions thereof are provided that enhance the effect on macrophage cholesterol ester hydrolase activity and/or inhibit acyl CoA:cholesterol acyl transferase activity. Methods of using these compositions in the treatment and/or prevention of atherosclerosis as well as coronary heart disease and cardiovascular disease are also provided.

Cardiovascular disease, including coronary heart disease caused by atherosclerosis, is the single largest killer of adults in North America (2002 Heart and Stroke Statistical Update). The development and progression of atherosclerosis in coronary arteries can lead to heart attacks and angina. In 1999 it was estimated that 12.6 million Americans had coronary heart disease. Approximately 1 in 5 deaths in 1999 were due to coronary heart disease, with a total US and Canadian mortality of over 500,000 and 42,000 individuals, respectively. It is estimated that over 102 million American adults have blood cholesterol levels that are either border-line high risk, or high risk of developing coronary heart disease. In addition to the immediate social and economic burden that heart attacks have on our health care system, there also is the considerable cost associated with the aftermath of a coronary heart disease event. About 25% of males and 38% of females will die one year after a heart attack, and death by coronary heart disease tends to occur during a person's peak productive years (BRFSS [1997], MMWR vol. 49, No. SS-2, Mar. 24, 2000, CDC/NCHS). There is also a further economic burden of coronary heart disease associated with premature and permanent disability of the labor force. In 1998, over $10 billion was paid to Medicare beneficiaries for coronary heart disease (Health Care Financing Review, Statistical Supplement [2000], HFCA).
Patients currently have a choice of a number of different drugs to treat cardiovascular disease/coronary heart disease. These drugs fall into various classes, including antihypertensives and antihyperlipidemics. Although these products have been shown to be beneficial in reducing the progression of coronary heart disease and preventing heart attacks, they can be limited in their effectiveness in some individuals because of low tolerability and, in some cases, mitigation of drug efficacy by the compensatory effects of the liver (Turley, S. D. (2002) Am. J. Managed Care 8 (2 Suppl):S29-32).
The accumulation of lipids, especially cholesterol, in several aortic and arterial cell-types, such as macrophages and smooth muscle cells, is the defining pathologic feature of atherosclerosis (Gotlieb et al. (1999) Blood Vessels. In Pathology. Rubin, E. and Farber, J. L., editors. Lippincott-Raven, Philadelphia, New York. 481-530). Major investigative efforts are being expended to understand two central issues related to this problem. The first relates to the mechanism by which cholesterol is delivered to, and taken up by, these cells. The second relates to the process by which these cells export and rid themselves of excessive cholesterol. In the treatment and prevention of atherosclerosis, one of the aims is to limit the intracellular accumulation of large quantities of cholesterol that adversely influence the viability of these cells, thereby eventually altering the structural integrity of the blood vessels.
An analogous set of events occurs in acute tissue injury. Such injuries result in local cell death and set in motion local inflammation and the systemic acute phase response (Fantone, J. C. and Ward, P. A. (1994) Inflammation. In Pathology. Rubin, E. and Farber, J. editors. Lippincott, Philadephia. 32-6). Alterations in local cholesterol processing are important components of this process. At sites of acute tissue injury, dying cells release large quantities of cell debris that includes cell membrane fragments rich in cholesterol (Fantone, J. C. and Ward, P. A. (1994) Inflammation. In Pathology. Rubin, E. and Farber, J. editors. Lippincott, Philadephia. 32-6). As part of acute inflammation, macrophages arriving at sites of injury ingest these fragments for further processing and thereby acquire a considerable cholesterol load, becoming foam cells, analogous to those seen in atherosclerosis. During acute tissue injury and the consequent acute inflammatory process, a cholesterol removal mechanism is required to mobilize the cholesterol either for excretion or re-use.
The physiological role of one of the major acute phase (AP) proteins synthesized by the liver in response to tissue injury, serum amyloid A (SAA), is directly related to these events and processes. SAA represents a group of four polymorphic proteins, encoded by a multigene family, that have been conserved for over 600 million years (Jensen et al. (1997) J. Immunol. 158:384-392; Santiago et al. (2000) J. Exp. Zool. 288: 3335-344). Isoforms SAA1.1 and SAA2.1 are present in plasma in acute phase tissue injury and are the most thoroughly investigated.
The nomenclature for serum amyloid A was revised in 1999, as there was a recognized need by researchers for a systematic nomenclature of the multiple SAA genes in human and animal models and for their allelic variants (Amyloid: Int. J. Exp. Clin. Invest. 1999 6:67-70). The major revision was the re-designation of the mouse Saa1 and Saa2 genes. Based upon chromosomal mapping, it appears that the mouse Saa2 locus corresponds to human SAA1. Therefore, the mouse nomenclature was changed to be fully compatible with the human nomenclature.
The following Tables set forth the revised nomenclature for SAA mouse and human proteins as well as their corresponding sequences. These tables are based upon the disclosure in 1999 in Amyloid: Int. J. Exp. Clin. Invest. 6:67-70. The tables presented herein have been modified, however, to clarify alignment and provide numbering for residue (−1) of mouse isoform SAA3 comprising an additional amino acid.
TABLE IMouse SAA proteinsNewOld−11234567891011121314151617181920212223242526SAA1.1SAA2GFFSFIGEAFQGAGDMWRAYTDMKEASAA1.2SJL/JSAA1.3mc1SRSAA1.4mc2SSAA1.5mm1VHSAA1.6mm2SAA2.1SAA1VHSAA2.2CE/JVHLSAA3QRWVQMKGSRSKSAA4SAA5DWYFRTWLRNL272829303132333435363738394041424344454647484950515253SAA1.1SAA2GWKDGDKYFHARGNYDAAQRGPGGVWASAA1.2SJL/JSAA1.3mc1RSAA1.4mc2SAA1.5mm1SAA1.6mm2SAA2.1SAA1NNSSAA2.2CE/JSAA3SRASAA4SAA5NYQNAQYEQSI545556575859606162636465666768697071727374757677787980SAA1.1SAA2AEKISDARESFQEFFGRGHEDTMADQESAA1.2SJL/JSAA1.3mc1SAA1.4mc2GSAA1.5mm1SAA1.6mm2SAA2.1SAA1GAISAA2.2CE/JASAA3KVVKTHASRFSAA4SAA5KITSKYGLLNHLTLQTK↑[NRYYFGIR]81828384858687888990919293949596979899100101102103SAA1.1SAA2ANRHGRSGKDPNYYRPPGLPAKYSAA1.2SJL/JDSAA1.3mc1SAA1.4mc2SAA1.5mm1SAA1.6mm2DSAA2.1SAA1DSAA2.2CE/JDSAA3EWHFAKRSAA4SAA5EEWNHFEEF
TABLE IIHuman SAA proteinsNewOld123456789101112131415161718192021222324252627SAA1.1SAA1αRSFFSFLGEAFDGARDMWRAYSDMREASAA1.2SAA1βSAA1.3SAA1γSAA1.4SAA1δSAA1.5SAA1βSAA2.1SAA2αSAA2.2SAA2βSAA4ESWRSFFKEALQGVGDMGRAYMDIMIS282930313233343536373839404142434445464748495051525354SAA1.1SAA1αNYIGSDKYFHARGNYDAAKRGPGGVWASAA1.2SAA1βASAA1.3SAA1γASAA1.4SAA1δASAA1.5SAA1βASAA2.1SAA2αASAA2.2SAA2βASAA4MHQNSNRYLYARGNYDAAQRGPGGVWA555657585960616263646566676869707172737475767778798081SAA1.1SAA1αAEAISDARENIQRFFGHGAEDSLADQASAA1.2SAA1βVDSAA1.3SAA1γSAA1.4SAA1δVNSAA1.5SAA1βVSAA2.1SAA2αVNLTSAA2.2SAA2βVNLTRSAA4AKLISRSRVYLQGLISTVLEDSKSNEK↑[DYYLFGNS]828384858687888990919293949596979899100101102103104SAA1.1SAA1αANEWGRSGKDPNHFRPAGLPEKYSAA1.2SAA1βSAA1.3SAA1γSAA1.4SAA1δSAA1.5SAA1βSAA2.1SAA2αKRSAA2.2SAA2βKRSAA4AEEWGRSGKDPDRFRPDGLPKKYThe nomenclature for SAA proteins employed in this patent application corresponds to the revised nomenclature as set forth in the above Tables. However, it must be appreciated that journal references published prior to this 1999 revision and patent applications filed prior to this 1999 revision may use the old nomenclature, thus, for example referring to mouse Saa1 as mouse Saa2 and vice versa.
SAA isoforms SAA1.1 and SAA2.1 are produced primarily by hepatocytes in response to various causes of tissue injury and inflammation (Morrow et al. (1981) Proc. Natl. Acad. Sci. USA 78:4718-4722). Synthesis of SAA1.1 and 2.1 by the liver is induced by cytokines such as interleukin-1, interleukin-6, and tumor necrosis factor, which are released by activated macrophages, and which act through a set of downstream effectors in the hepatocyte cytoplasm and nucleus (Edbrooke et al. (1991) Cytokine 3:380-388; Betts et al. (1993) J. Biol. Chem. 268:25624-25631; Ray et al. (1999) J. Biol. Chem. 274:4300-430810; and Sipe et al. (1987) Lymphokine Res. 6:93-101). Maximum transcription rates for the SAA1.1 and 2.1 genes are seen 3-4 hours following tissue injury, and within 18-24 hours of injury the plasma concentration of these two proteins rises from 1-5 μg/mL to 500-1000 μg/mL (500-1000-fold increase) (McAdam et al. (1978) J. Clin. Invest. 61:390-394; McAdam, K. P., Sipe, J. D. (1976) J. Exp. Med. 144:1121-1127). Once secreted from hepatocytes, SAA1.1 and 2.1 are found predominantly in the high density lipoprotein (HDL) fraction and form 30-80% of the HDL apolipoproteins, resulting in a major reorganization of the apolipoprotein composition of the HDL fraction (Benditt et al. (1979) Proc. Natl. Acad. Sci. USA 76: 4092-4096; Hoffman, J. S. and Benditt, E. P. (1982) J. Biol. Chem. 257:10518-10522).
At present there is debate whether the observed increase in SAA expression during tissue injury is associated with a beneficial role against atherosclerotic lesions, or whether increased SAA levels are in fact associated with a role in developing atherosclerosis.
Elevated levels of SAA isoforms are observed during the early pathological vascular events leading to atherosclerosis before clinical symptoms are evident. (reviewed in Kisilevsky, R. and Tam, S.-P. (2002) Pediatric Pathol. and Mol. Med. 21: 291-303). This elevation has led some researchers to suggest that SAA levels may play a causative, contributing role in atherogenesis (Jousilahti et al. (2001) Atherosclerosis 156:451-456; Kumon et al. (1998) Scand. J. Immunol. 48:419-424; Liuzzo et al. (1994) N. Engl. J Med. 331:417-424; Ridker et al. (1998) Circulation 98:839-844; Rosenthal, C. J. and Franklin, E. C. (1975) J. Clin, Invest. 55:746-753; Steinmetz et al. (1989) Biochim Biophys. Acta. 1006:173-178; Van Lenten et al. (1995) J. Clin. Invest. 96:2758-2767).
However, there have also been reports of SAA and isoforms thereof promoting the efflux of cholesterol from macrophages.
For example, high density lipoprotein-serum amyloid A (HDL-SAA) has been shown to have reduced ability to accept cholesterol from low density lipoprotein/very low density lipoprotein (LDL/VLDL), ensuring that HDL, in its afferent route, arrives at macrophages carrying as little cholesterol as possible (Kisilevsky et al. (1996) Amyloid 3: 252-260). Thus, this form of HDL has a greater capacity to accept cholesterol from cholesterol-laden macrophages. HDL-SAA has also been demonstrated to have a 3 to 4-fold higher affinity for macrophages when compared to HDL alone. Further, an increase was observed in the number of HDL-SAA binding sites on macrophages obtained from animals with an AP inflammatory reaction. Competition studies with macrophages (Kisilevsky, R. and Subrahmanyan, L. (1992) Lab. Invest. 66: 778-785) showed that unlabelled HDL-SAA, but not HDL alone, effectively displaced radiolabeled HDL-SAA. This preferential displacement by HDL-SAA is likely indicative of the presence of SAA receptors on the macrophages. Such SAA receptors are separate and additional to the binding sites for apoA-1 on the macrophages (Kisilevsky, R. and Subrahmanyan, L. (1992) Lab. Invest. 66: 778-785; U.S. Pat. No. 6,004,936). The presence of SAA receptors is further supported by the demonstration that HDL-SAA was in clathrin coated pits shortly after binding to macrophages. These pits and the resulting endosomes are consistent with the concept of receptor-mediated endocytosis, a process that is dependent on cell surface heparin sulphate, to which SAA binds effectively (Ancsin, J. and Kisilevsky, R. (1999) J. Biol. Chem. 274:7172-7181; Rocken, C. and Kisilevsky, R. (1997) Amyloid 4: 259-273).
More recent studies have demonstrated that SAA enhances HDL uptake by macrophages (Banka et al. (1995) J. Lipid Res. 36:1058-10865) and has an affinity for cholesterol (Liang, J. S. and Sipe, J. D. (1995) J. Lipid Res. 36:37-46). Using synthetic peptides corresponding to residues 1-18 and 40-63 of human apoSAA1 (now referred to as SAA1.1) and residues 1-18 of human apoSAA4 (now referred to as SAA4) it was shown that apoSAA1 but not apoSAA4 binds cholesterol at the amino terminal region (Liang et al. (1996) J. Lipid Res. 37:2109-2116).
Furthermore murine SAA2.1, but not murine SAA1.1, was shown to inhibit macrophage acyl CoA:cholesterol acyl transferase (ACAT) activity in culture in intact murine macrophages and in their post-nuclear homogenates in a dose-dependent manner (Ely et al. (2001) Amyloid 8:169-181). Further examination of cyanogen bromide generated cleavage fragments of murine SAA2.1 purified by reverse phase HPLC showed murine SAA2.11-16 to have a profound effect inhibiting ACAT activity in a dose-dependent manner. In contrast, murine SAA2.124-103 exhibited no inhibitory effect on ACAT activity (Ely et al. (2001) Amyloid 8:169-181).
Murine SAA2.1 has also been shown to stimulate hepatic, macrophage, and pancreatic cholesterol esterase activities in vitro (Lindhorst et al. (1997) Biochim. Biophys. Acta 1339:143-154; Ely et al. (2001) Amyloid 8:169-181; Tam et al. (2002) J. Lipid Res. 43:1410-1420). This effect was shown to reside in the 80 residue COOH-terminal region of murine SAA2.1 liberated by cyanogen bromide cleavage (Ely et al. (2001) Amyloid 8:169-181). This 80 residue region comprises residues 24-103 of murine SAA2.1.
The ability of HDL-SAA and liposomes containing murine SAA2.1 to cause a marked reduction of acyl CoA:cholesterol acyl transferase activity and enhancement of cholesterol efflux activity was confirmed in macrophages in culture (Tam et al. (2002) J. Lipid Res. 43:1410-1420). Intravenous injection of [3H]-cholesterol-loaded macrophages into inflamed mice has also been reported to result in a 3- to 3.5-fold increase in the amount of radiolabeled cholesterol released into the plasma when compared to similarly treated un-inflamed control animals (Tam et al. J. Lipid Res. 2002 43:1410-1420). In this study, macrophage cholesterol efflux was shown to be coupled to the ATP-binding cassette transporter, ABCA1, which is an important protein for the initial step of the reverse cholesterol transport pathway. Furthermore, [3H]-cholesterol-laden macrophages, when pre-treated with HDL-SAA2.1 (murine) in tissue culture and then injected into un-inflamed mice, rapidly released their cholesterol into the plasma (Tam et al. (2002) J. Lipid Res. 43:1410-1420). This result was not observed when macrophages were treated with HDL alone.
Thus, isoforms SAA1.1 and SAA2.1 are up-regulated during inflammation; they are evolutionarily conserved; and they are predominantly associated with HDL and HDL's established role in the reverse cholesterol transport pathway (Lindhorst et al. (1997) Biochim. Biophys. Acta 1339:143-154; Kisilevsky, R. (1991) Med. Hypotheses 35: 337-341; Kisilevsky, R. et al. (1996) Amyloid 3: 252-260; and Kisilevsky, R. and Subrahmanyan, L. (1992) Lab. Invest. 66: 778-785).
U.S. Pat. No. 5,318,958 discloses methods of potentiating the release and collection of macrophage cholesterol in vivo by administering an effective amount of HDL bound to a ligand having serum amyloid A affinity for HDL. A preferred ligand of this method taught in this patent is serum amyloid A itself.
U.S. Pat. No. 6,004,936 describes similar methods to U.S. Pat. No. 5,318,958. However, in the method claimed in U.S. Pat. No. 6,004,936, the ligand having serum amyloid affinity is not bound to HDL prior to administration. This patent teaches that preferred ligands having serum amyloid affinity are non-amyloidogenic isoforms of serum amyloid A such as SAA2.1.

Compositions and methods for treating atherosclerosis


An electronic module has a rear surface-exposed conductor 10, 20, 30 having a rear surface-exposed part 12, 22, 32 whose rear surface is exposed; an electronic element 15, 25 provided on a front surface of the rear surface-exposed conductor 10, 20, 30; and a connector 60 configure to connect the rear surface-exposed conductor 10, 20, 30 and the electronic element 15, 25 or two rear surface-exposed conductors 10, 20, 30 each other. A groove 150 is provided on the front surface of the rear surface-exposed conductor 10, 20, 30. The sealing part 90 is provided with a press hole or a press impression 110, 120, 130 used to press the rear surface-exposed conductor 10, 20, 30. In an in-plane direction, a center portion of the press hole or the press impression 110, 120, 130 is provided on the side opposite to the connector 60 or the electronic element 15, 25 with respect to the groove 150.

Conventionally, electronic modules used for inverter circuits and relay circuits used in vehicles such as automobiles are known. Japanese Patent No. 5067679 discloses an electronic module having a power supply terminal, an output terminal, and a ground terminal. In such an electronic module, it is desirable to increase a heat dissipation efficiency from an electronic element. Therefore, it is conceivable to expose the rear surface of a conductor having a terminal part from a sealing part such as mold resin.
However, as described above, there is a case where the rear surface of the conductor is exposed, and a resin may wrap around the rear surface side of the conductor when resin sealing is performed.
In this respect, it is conceivable to press the conductor, but it is also conceivable that the conductor floats up on the side opposite to the side pressed when the conductor is pressed, and the resin wraps around the back side of the floated conductor.

Electronic module with a groove and press hole on the surface of a conductor

SHINDENGEN ELECTRIC MANUFACTURING CO., LTD.


Methods are provided for readily and efficiently determining resonant frequencies that can be used therapeutically or beneficially, for debilitation of specific types of genomic materials, including DNA and/or RNA, genes, and gene sections. The methods can be used in a variety of circumstances related to various human and animal diseases and conditions. Methods allow determination of therapeutic resonant frequencies for use in various media having different refractivities. Therapeutic or beneficial resonance frequencies thus determined are adapted for use with currently available frequency-emitting devices by shifting resonant frequencies to electromagnetic ranges capable of generation by such devices.

Resonant frequency therapy (RFT) is a non-invasive treatment that has been reported to offer significant relief to sufferers of a variety of ailments and medical conditions. The use of RFT for human and animal therapeutic purposes began in the early 1900's, and experienced accelerated development through the research of Royal Rife and his associates in the 1930's and afterward.
Using new microscope technology he developed, Rife discovered that plasma waves could be used to transmit radio and audio frequencies, which were tuned to the frequencies of specific microorganisms, and that each microorganism responded to its unique frequencies. For example, Rife found that staphylococcus, streptococcus, microorganisms associated with tuberculosis, typhoid, and leprosy, as well as cancer particles, and other disease-causing agents succumbed when exposed to certain frequencies peculiar to each organism or particle. See, Siedel, R. E., and M. E. Winter, The New Microscopes, Smithsonian Annual Report 1944, pp. 193-200.
Using the principles of Rife's discoveries, various researchers developed devices for emitting frequencies designed to treat a range of diseases and conditions. For example, Dr. Abraham Ginsberg used an apparatus which produced intermittent bursts of high energy in the short wave spectrum. Ginsberg's modality was found to stimulate the reticuloendothelial system without undesirably heating tissue. Using his device, Ginsberg reported successfully treating patients with various clinical conditions, including chronic Staphylococcus infections, acute inflammatory middle ear, chronic ulcerative colitis, bronchitis, rheumatoid arthritis, gout, flu, and thrombophlebitis, among others. See, Cominole, B., Clinical Impressions and Speculations on the Use of High-Frequency Pulsed Energy, The Dr. Abraham J. Ginsberg Foundation for Medical Research Symposium, Jun. 29, 1959.
Research utilizing resonant frequencies and therapeutic modalities implementing such frequencies have proliferated over the past ten years. A recent example of the use of resonant frequency therapy is the Christchurch Resonant Frequency Therapy Centre in Dunedin, New Zealand. While the Centre emphasizes that resonant frequency therapy is not intended to replace treatment regimens and medication prescribed by physicians, it does report successful treatment of a range of clinical conditions, including arthritis, tinnitis, blood pressure, cataracts, headaches, shingles, and psoriasis. Arthritis patients report particular success with pain reduction and greater mobility. See The Christchurch Press, Frequency Therapy Offers Relief, Independent Newspapers Limited, Oct. 28, 1999.
Thus, the use of audio, radio, and light waves to inhibit microbial growth and to treat diseases and affected tissue is well known in the art. Effective therapeutic resonant frequencies have been identified through various means. Trial and error approaches with resonant frequencies have been used to obtain therapeutic responses. Devices for applying electromagnetic energy to living tissue are disclosed, for example, in U.S. Pat. Nos. 3,876,373, 4,524,079, and 5,091,152. Effective resonant frequencies have also been identified through the use of frequency scanning with electronic devices capable of detecting a frequency response from a bacterial, viral, and/or tissue sample. Such devices for detecting frequency response are disclosed, for example, in U.S. Pat. Nos. 5,552,274, 5,981,182, and 6,004,257. Thus, there exists a need for more efficient and accurate methodology than trial and error, to determine therapeutic resonant frequencies for specific target materials, such as microorganisms.
Therapeutic resonant frequencies may be used to inhibit, or debilitate, and/or stimulate a biophysical event. The efficacy of such frequencies, whether for stimulation or for debilitation, depends to some extent on the type of frequency delivery system used, including variables such as power levels, waveform, harmonic content of the wave, and other factors. Once therapeutic resonant frequencies are determined, the user must choose which devices and delivery systems are most effectively used in conjunction with those frequencies. To increase efficacy, an easier, quicker, and more accurate way of determining therapeutic resonant frequencies is needed.
Despite both historical and increasing recent interest in use of resonant frequency therapy, mechanism(s) of action underlying the use of known therapeutic resonant frequencies is not fully understood. While it is recognized that some type of resonance phenomenon debilitates or destroys microorganisms, the biophysical and/or biochemical mechanism(s) associated with use of specific resonant frequencies and that lead to microbial inhibition are not completely known.
Before now, there has never existed a methodology that links effective therapeutic resonant frequencies to a biophysical or biochemical event, process, or structure. The electronic scanning devices and methods currently commercially available provide no explanation or insight regarding which physical structure or process is influenced by the frequencies used.
There is a need for methodology to more readily and efficiently influence genomic materials, by more precisely and efficiently determining therapeutic resonant frequencies that can be easily and accurately adjusted to ranges used by currently available devices. It is to these perceived needs that the present invention is directed.

Methods for determining therapeutic resonant frequencies


Described embodiments automatically and dynamically generate and update resource templates for deployable resources, utilizing composable parts that may be dynamically replaced. Common portions of the templates may be easily defined once and reused multiple times, reducing risk of error and ensuring stability and cross-resource compatibility. Each dynamic template may have replaceable parts, as well as replaceable parameters and values within the replaceable parts, as well as in common portions of the template. The replacement properties and values may be determined at runtime or during deployment, ensuring that configurations are always current.

Virtual computing environments, sometimes referred to as hosted environments, hosted desktops or servers, cloud computing environments, virtual processing environments, virtual storage environments, deployable resources, or by similar terms, allow remote execution and processing of computationally intensive tasks and control over deployable services. These environments may include virtual machines, virtual storage devices, virtual networks, databases, web applications, servers of different types, or any other type and form of deployable resource. These environments may be dynamically deployed (e.g. instantiating virtual machines or servers, beginning execution of the hosted environment, etc.) or undeployed (e.g. stopping or terminating execution of the hosted environment, etc.).
Resources are typically deployed according to configuration files or templates with entries specifying various features and parameters of the resource, such as names, addresses, group memberships, types, or other such features. Particularly for large organizations, these templates may be complex and numerous. Maintaining and updating templates may require significant time and effort, and inadvertent typographical errors in a template may result in drastic consequences for the organization, including data loss or lack of stability.

Systems and methods for synthesizing resource templates using composable parts


Disclosed herein are methods for automatically adjusting a speed of a vehicle capable of allowing the vehicle to arrive at a destination within a time desired by a driver in consideration of a desired arrival time to the destination, a distance to the destination, a current traffic volume, or the like. A method for adjusting a speed of a vehicle in a system for adjusting a speed of a vehicle may include a driver of the vehicle setting a destination, and the system for adjusting a speed of a vehicle searching a path to the destination and calculating a distance to the destination and a current traffic volume to calculate an arrival time to the destination. The method may further include the driver setting a desired arrival time to the destination, and the system for adjusting a speed of a vehicle deciding whether or not the vehicle arrives at the destination within the desired arrival time.

The statements in this section merely provide background information related to the present disclosure and may not constitute prior art.
Recently, in accordance with the rapid development of a vehicle related technology, a vehicle system significantly improving convenience of a driver has been commercialized in various patterns. As a typical function of convenience functions of a vehicle, there is a smart cruise control (SCC) system, which is a convenience apparatus providing freedom in a longitudinal direction to the driver by automatically driving and braking the vehicle so that a speed of the vehicle is maintained at a speed set by the driver.
In addition, as a convenience function sensing a driving path, there is a lane departure warning (LDW) system, which is a kind of vision system using a camera sensor and is configured to recognize a lane of a road using a camera to give the driver a warning when the vehicle is about to depart from the lane due to sleepiness or carelessness of the driver.
The above-mentioned two systems, which are systems that have been currently commercialized, have been mounted as driver convenience or safety systems in high class vehicles.
In addition, recently, a technology of providing a function of allowing the driver to set a predetermined speed or constantly maintaining a road speed limit to the driver has also appeared.
Further, a technology of allowing the driver to set a desired time in which he/she will arrive at a destination in order to arrive at the destination within a specified time or allowing the vehicle to automatically adjust a speed of the vehicle by reflecting a traffic volume, or the like has been required.


A communication signal is communicated between an implantable medical device including an implant transceiver and an external unit including an external unit transceiver. At least one of the transceivers includes a receiver capable of sampling a communication channel for the communication signal at times based on a macro sampling interval and a micro sampling interval. The sampling includes a series of micro samples. The duration of the series of micro samples is less than the macro sampling interval.

Implantable medical devices (IMDs) provide therapies and monitor a wide variety of physiological events. With the increased uses of IMDs has also come the need for improved methods of communicating with and between IMDs.
Conventionally, communication with IMDs has been with magnetic field communication systems. Such systems, however, are generally only capable of communicating over very short distances, on the order of a few inches. As a result, a magnetic head of a programmer (or other external device) needs to be placed near to the IMD for communication to occur. More recently, radio frequency (RF) based communication systems have been developed for use with IMDs. RF communication provides a number of benefits over magnetic field communication systems, including much greater communication distances. However, conventional RF communication systems consume more battery power than magnetic field communication systems, thus impacting the service life of the IMD battery.
Accordingly, there is a need to improve RF receiver efficiency and inter-IMD communication modalities to conserve battery life.
RF communication may generally be divided into two categories: synchronous and asynchronous. Synchronous communication is conducted at scheduled times. However, in synchronous communication systems, the internal clocks of two communicating devices are prone to drift over time. As more time elapses, the internal clocks become increasingly out of sync, such that neither device can precisely detect when the other device will commence communication. To compensate for this drift, one or both of the devices must stay in an "on" mode. During that time, energy is consumed while no communication is effected.
In an asynchronous communication system, transmission occurs at random times. Because it is impractical to maintain the receiver on at all times, asynchronous communication systems utilize sampling methods in which the receiver is repeatedly turned on for brief periods to check for a transmission signal and turned on fully when the signal is detected. The more often the receiver is turned on, the faster the response time of the receiver. However, more energy is required. To guarantee that data will be received, the transmitter transmits a preamble for at least as long as the time interval between samples prior to transmitting a message. Once the preamble is detected, the receiver remains on until the message is received. As a result, energy is consumed by the receiver while receiving the preamble, a time in which no valuable communication is taking place.

Implantable medical device communication system with macro and micro sampling intervals


A method and structure for developing a photographic image from a camera, filtering the image, and developing the image. Filtering of an image entails separating the image into a luminance channel and chrominance channels, alternating the luminance channel by saturation and hue to produce the altered luminance channel and recombining the chrominance channels and the altered luminance channel. The hue controls the color of filtering and the saturation controls the strength of the filtering. Prior to separating and converting the linear exposure is converted to a logarithmic space. This conversion of linear exposure to logarithmic space allows the luminance""s channel to be altered independently from the chrominance channels. The alteration creates a filtered RGB to YCrCb matrix and applying the matrix to the logarithmic space produces a filtered logarithmic space. The invention recombines the YCrCb to RGB matrix to the filtered logarithmic space.

1. Field of the Invention
The invention relates generally to the field of digital image processing and more particularly to luminance processing that is based upon color dependence.
2. Description of the Related Art
Color filters are used in black and white photography to modify contrast in the captured image. For example, yellow filters are used in outdoor black and white photography to darken the sky and reduce haze. However, when the same filters are applied to color photography an undesirable yellow color cast can result.
Further, the filters used in photography are conventionally applied to the image globally. In other words, a certain color (or combination color) filter is applied to the entire image. While the filter may change one portion of the image in a desired fashion, there is a possibility that another portion of the image will be adversely affected. Therefore, conventional filters always present a trade-off between desirable and undesirable effects. The invention overcomes these problems as discussed below.
The invention includes a method for developing a photographic image from a digitized color image by filtering the image and developing the image. Filtering of the image entails separating the image into a luminance channel and chrominance channels, altering the luminance channel by modifying the red, green and blue coefficients used to generate the luminance channel based upon a filter color to produce the altered luminance channel and recombining the chrominance channels and the altered luminance channel. The hue controls the color of filtering and the saturation controls the strength of the filtering. Prior to separating and converting the linear exposure is converted to a logarithmic space. This conversion of linear exposure to logarithmic space allows the luminance""s channel to be altered independently from the chrominance channels. The alteration creates a filtered RGB to YCrCb matrix and applying the matrix to the logarithmic space produces a filtered logarithmic space. The invention returns to the RGB space by applying the YCrCb to RGB matrix to the filtered logarithmic space.
Alternatively, the system for developing a photographic image includes a filter applied to the image, a divider adapted to separate the image into luminance and chrominance channels, a saturation control and a hue control adapted to alter the luminance channel and to produce an altered luminance channel, and an adder adapted to recombine the chrominance channels and the altered luminance channel. The hue control changes the color of the filter and the saturation control changes the strength of the filter. Further, a converter is adapted to convert the image into a linear exposure and the linear exposure to a logarithmic space. The converter allows the luminance channel to be altered independently from the chrominance channels. The saturation input and the hue input produce a filtered RGB to YCrCb matrix and the invention applies the matrix to a logarithmic space to produce a filtered logarithmic space. The adder applies YCrCb to RGB matrix to the filtered logarithmic space.
With the invention, color filters can be synthesized for a digitized color image and applied to the image to modify the contrast without imparting a color cast. Further, the invention allows unrenderable portions of the image that are too bright or too dark to be modified based upon chrominance so as to bring the unrenderable portions into a mid-tone range and make them renderable.
Using the invention, the filter applied (e.g., the calculation of the luminance channel) could optionally be a function of the color coordinates described by the chrominance, the pixel""s location in the image, the proximity to other colors/hues, or results from other image analysis/image understanding algorithms. Thus, the invention allows different filters to be selectively applied to different portions of the image. For example, the calculation of blue regions may suppress blue to darken a blue sky, while a flesh tone region may use a different filter to alter contrast differently in those regions.


A method for printing a label on a keypad, including the steps of providing a substrate plate, spraying ink on the substrate plate to form a label layer, solidifying the label layer, spraying ink around the label layer to form a primer layer, and solidifying the primer layer.

1. Technical Field
The present disclosure relates to a key labeling method, and more particularly, to a method used for printing a label on a keypad.
2. Description of Related Art
A commonly used key labeling method for printing a label on a keypad includes the following two steps. Providing a keypad substrate plate and forming a plurality of primer layers on the keypad substrate plate with screen printing technology; etching the primer layers to form a label with laser etching technology on the keypad substrate plate to finish the process of printing a label on the keypad.
However, the commonly used key labeling method has a relatively high manufacturing cost and is time-consuming because of the laser etching step. A second commonly used key labeling method is provided to decrease the manufacturing cost. The second commonly used key labeling method includes one step, printing a label layer and a plurality of the primer layers on the keypad substrate plate simultaneously. The laser etching step is thereby omitted, efficiently decreasing the manufacturing cost. However, when using the second commonly used key labeling method, the labeled layer and the primer layers easily mix with each other thereby obtaining a deformed and distorted label on the keypad substrate plate.
Therefore, there is room for improvement in the art.


A snap-fit joint comprises plastic male and female members. The male member has a flexible wall shaped along its length to define a terminal anchor portion and a relatively narrower neck portion defined by opposing curved indentations of the wall. The opposing curved indentations taper inwardly from a top to a bottom, such that the distance between opposing curved indentations of the wall is least at the bottom of the wall and greatest at the top. The female member comprises a socket corresponding in shape to that of the male member. As the male member is inserted into the socket, the curved indentations of the male member are flexed inwardly until the male member is seated in the socket.

Various types of joints are known for interconnecting two or more separate parts. In the realm of plastic products more particularly, numerous snap-fit joints have heretofore been devised, although all are attended by various drawbacks. There is, accordingly, a continued need for a snap-fit joint for plastic frame elements, as well as frames formed thereby, that are at once simple in their manufacture and construction yet characterized by acceptable strength.

Snap-fit joint for plastic frame elements and frames formed thereby


Devices and methods for selecting network nodes in a cellular communication network. By dropping fixed connections between network nodes of different hierarchy levels, for instance, a network node of an access network has to make a selection among different core network nodes. The selected core network node requires service information on the access network node and the area serviced by the access network node. Devices and methods are disclosed for selecting network nodes and for providing service information. The devices can be implemented in the network centrally or in a distributed manner.

The invention relates to devices and methods for selecting network nodes for a cellular communication network. It further relates to methods and devices for modifying information scored in said devices for selecting.
In cellular communication networks of the so-called "third generation" new architectures become possible. Said architectures are to allow simplified network adaptations co changing requirements. An important aspect of the new architectures is that so far fixed hierarchic connections between the network nodes are replaced by dynamic hierarchic connections. Fixed connections exist in GSM (Global System for Mobile communication) networks, for instance, between base station controllers and radio base stations or base station subsystems and mobile services switching centers, where each mobile services switching center cooperates with predefined base station subsystems.
Communication networks can be split into core networks and access networks and service networks.
The change to dynamic connections affects the cooperation within core networks and access networks as well as between them.
A core network consists of at least one network node, which offers a telecommunication service or administration service, a so-called core network node. The tasks of a core network are, for example, the switching of connections, the mobility management, the charging data acquisition and the storage and administration of subscriber data. Nodes of a core network are, for example, mobile services switching centers, GPRS (General Packet Radio Service) support nodes, home location registers and service control nodes.
An access network is a network enabling at least one subscriber to have access to a core network. Common tasks of an access network are the administration of access resources such as radio frequency channels, the measurement of connection qualities and the cooperation with a core network. Nodes of an access network are, for instance, base station controllers, radio base stations and radio network controllers.
Service networks comprising network nodes for supporting services surpassing the switching of connections are regarded in the present application as a part of the core network. Nodes of said networks are regarded as core network nodes.
One example is the cooperation between core network nodes and access network nodes in networks being constructed according to a new architecture, for instance, so-called "third generation networks". The cooperation between said nodes is handled more flexibly. Thus, no longer only one core network node can cooperate with a plurality of access network nodes, a so-called 1:N relation, but now rather a plurality of access network nodes can cooperate with a plurality of core network nodes in a so-called N:M relation. The technical report of the Third Generation Partnership Project 3G TR 23.922 version 1.0.0, published in October 1999, describes the introduction of cellular communication networks, wherein the connections between network nodes are based on the internet protocol. The internet protocol realizes the dynamic connections between network nodes. A press release by the company Ericsson of Feb. 1, 2000 describes the improved flexibility of this architecture. The architecture makes it possible to react on requests such as, for instance, temporary inhomogeneous traffic load, more flexibly.
It is prior art technology, that by means of the fixed connections between the network nodes, specific network nodes are allocated to each network node within a communication network for cooperating purposes.
U.S. Pat. No. 5,557,029 describes a system and a method for selecting a network node for handling a call in a network with said fixed connections. However, the system and the method are adapted to select a network node in a network with said fixed relation and are not capable of performing a selection in a network with an n:m relation between access network nodes and core network nodes.
Thus, the problem is avoided that a selection has to be made from a plurality of network nodes, for instance, with connection setups, location updates or handovers. This problem arises only with the dynamic allocation of network nodes to each other. Another difficulty resides in the provision of service information. According to the prior art technologies said information are stored in the respective network nodes, in which they are needed. To maintain this solution means—in the case of dynamic allocation—that each network node has to store the service information of all network nodes, with which it can cooperate.
Service information of a network node comprise, data on the service area and the routing area being serviced by the network node, as well as information on the node as such. This is, for instance, the emergency call center, which is switched in the routing area in the case of an emergency, the billing origin and the call origin, both being geographical data on a caller, wherein the first data is required for drafting the bill and the second data is required for processing the call, and local call restrictions, if a subscriber is allowed to phone only in a certain area. The Standard GSM 03.08 version 7.2.0 for handling subscriber data, published in 1998, describes location information and local call restrictions.
The multiple storage of service information of all network nodes brings about the disadvantage that unnecessarily large quantities of data are stored and that said information have to be kept consistent with a large amount of work.
In accordance therewith it is the object of the invention to provide methods and devices for an improved cooperation between network nodes in a cellular communication system.
Moreover, it is an object of the invention to develop methods for providing network nodes with service information on additional network nodes, if required also on service areas or routing areas, in a simplified manner.
It is an advantage that available processing capacities can be optimally exploited. A more even utilization of network nodes is achieved.
It is particularly advantageous that network nodes independently verify internal states and, if required, independently send messages for updating the corresponding entries in selection devices and configuration databases. This facilitates the registration of new nodes and allows a high updating of the data by involving only a small extent of administrative work.
It is moreover advantageous that the well-known data base technology is used. This reduces development costs and avoids downtimes caused by development errors.
Especially advantageous is the enabling or disabling of the selection of nodes by operator commands or settings.
Equally advantageous is the use of a configuration database which enables the provision of service information with a high data consistency and with a small extent of administrative work. Especially advantageous is the automatic updating of the data in a selection device by network nodes, which enables the automatic integration of new network nodes, or of network nodes which had been defect in the meantime, in a communication network.
The storage of free capacities proves to be an advantage. This enables a deliberate selection of network nodes.
Moreover, it proves to be an advantage if only data from network nodes with free capacities are stored. This reduces the number of data records, which have to be administered and searched through in a selection device.
Particularly advantageous is the consideration of distances between network nodes which avoids unnecessarily far connections within a network, thereby saving network capacities.
Furthermore advantageous is the storage of preferred allocations of network nodes in a selection device, which avoids unnecessary accesses to the node capacity memory and possible distance calculations.
Also advantageous is a service information memory. It enables the provision of service information for network nodes in a simple manner.
Further advantageous is the storage of service information in a network node, which avoids unnecessary transmissions of service information and service information requests thereby reducing the data traffic through the network. Equally an advantage is the sending of service information, e.g. by means of a selection device, which reduces the signaling work as no data have to be requested, and it ensures the availability of information in due time.
Another advantage is the automatic sending of service information by a configuration data base, which combines the advantage of the punctual availability of the service information with the advantage of the small extent of administrative work for a central facility.
Further advantageous is the modification of data stored in a node capacity memory for a group of network nodes. By this the selection of several nodes can be enabled or disabled by a single action. This reduces the number of necessary actions for a modification.
Another advantage is the sending of a list of access network nodes after a modification. By this user equipment gets the latest information about valid nodes that can be contacted. This avoids unnecessary attempts to contact invalid nodes or increases the choice of nodes to contact.
Especially advantageous is the contacting of a preferred network node prior to sending a network node request. This reduces the number of network node requests to be processed.
Especially advantageous is the storage of a list of preferred network nodes. This reduces the number of network node requests and thus reduces the data traffic through the network.

Methods and devices for optimal selection of core network nodes in a cellular communication network