A melanoma associated antigen known as gp100. Furthermore, peptides derived from the antigen are described. Gp100 and its peptides can be used in vaccines for the treatment of melanoma. Another aspect of the invention is host cells capable of expressing gp100 for the gp100-derived peptides. Furthermore, tumor infiltrating lymphocytes (TIL""s) specifically recognizing gp100 are described, as are vaccines with these TIL""s. Also disclosed are diagnostics for the detection of melanoma and for the monitoring of vaccination.

The present invention is concerned with cancer treatment and diagnosis, especially with a melanoma associated antigen, epitopes thereof, vaccines against melanoma, tumor infiltrating T lymphocytes recognizing the antigen and diagnostics for the detection of melanoma and for the monitoring of vaccination.
Tumor cells may emancipate themselves from restrictive growth control by oncogene activation, and/or by the inactivation of tumor suppression genes. The course of tumor progression proceeds by a series of gradual, stepwise changes in different xe2x80x98unit characteristicsxe2x80x99, i.e. phenotypic traits, many of which are known to be determined or at least influenced by the altered expression of defined oncogenes and/or tumor suppressive genes. Emancipation of the cell from immunological host restriction may follow multistep pathways similar to the emancipation from growth control.
A problem often encountered in cancer immunotherapy is the lack of immunogenicity of the tumor. This escape of the immune control system can be understood on basis of phenotype differences encountered in neoplastic cells (differences found in Burkitt""s lymphoma cells according to Klein, G. and Boon, T., Curr. Opinion in Immunol. 5, 687-692, 1993):
decreased ability to process and present antigens;
decreased ability to stimulate autologous T cells;
complete downregulation of immunogenic proteins associated with transformed cells;
no or low expression of leukocyte adhesion molecules or other accessory molecules; and
selective downregulation of certain MHC class I and class II alleles.
MHC Class I/II antigens are often downregulated in solid tumors. This may affect all class I/II antigens, or only part of them. Viral and cellular peptides that can sensitize appropriate target cells for cytotoxic T lymphocyte mediated lysis may fail to do so when produced in cells with a low level of expression of MHC class I antigen. Cytotoxic sensitivity may be induced, at least in some cases by raising the level of MHC class I/II antigen expression by interferon xcex3 and tumor necrosis factor xcex1.
However, during the stepwise changes from normal to tumor tissue tumor-associated antigens appear. These antigens can be exposed through various mechanisms:
they can be molecules that are masked in some way during normal cell development, but where the neoplastic change induces removal of the masking protection for the immunosystem;
deletion of some molecules from the plasma membrane may alter the profile of adjacent molecules in a given membrane patch, and thus, in effect generate a new profile that might become immunogenic to the host;
a membrane alteration accompanying neoplastic transformation may expose new, previously hidden regions of a molecule, or may result in addition of new structural features to an existing molecule.
shedding and disintegration of tumor cells may expose the immune system to nuclear, nucleolar, or cytoplasmic components that are normally hidden in the cell.
The characteristics of tumor-associated antigens are very much dependent on the origin of the tumor carrying them. The existence of antigens associated with animal tumors was documented in the last century, and the antigenic character of human cancers has been well established, primarily through recent studies with monoclonal antibodies.
Attempts to isolate and chemically characterize these antigens have encountered serious difficulties, many having to do with a lack of reagents suitable for precipitation of the antigen-bearing molecules from a solution.
Like many other stimuli, the tumor-associated antigens activate not one but a whole set of defense mechanismsxe2x80x94both specific and unspecific, humoral and cellular. The dominant role in in vivo resistance to tumor growth is played by T lymphocytes. These cells recognize tumor-associated antigens presented to them by antigen presenting cells (APC""s), and will be activated by this recognition, and upon activation and differentiation, attack and kill the tumor cells. A special class of these sort of lymphocytes is formed by the tumor infiltrating lymphocytes (TIL""s) which can be found in solid tumors.
It has already been suggested (EP 147,689) to activate T lymphocytes with an antigenic substance linked to an insoluble carrier in vitro and then to administer these activated lymphocytes to a tumor patient.
Conventional chemotherapy is relatively ineffective in the treatment of patients with metastasic melanoma, and approximately 6000 patients die of this disease in the United States each year.
Rosenberg et al. (New Eng. J. Med. 319(25), 1676-1681, 1988) have shown the beneficial effect of immunotherapy with autologous TIL""s and interleukin-2 (IL-2) in melanoma patients.
This therapy constitutes of resection of the tumor deposit, isolation of the TIL""s, in vitro expansion of the TIL""s and infusion into the patient under concurrent treatment of high and toxicity inducing doses of IL-2.
The TIL""s used by Rosenberg are directed to and able to recognize melanoma-associated antigens.
It has been our goal to isolate such a melanoma-associated antigen in order to be able to use the antigen and/or its epitopes for the development of an immunotherapy for melanoma patients.
Melanoma antigens have already been described by Old, L. (1981) who identified 6 antigenic glycoproteins and 3 glycolipids occurring in 120 melanoma cell lines.
Also vaccines with melanoma antigens have been described: in U.S. Pat. Nos. 5,030,621 and 5,194,384 a polyvalent vaccine has been made by culturing melanoma cells and subsequent isolation of excreted melanoma-specific antigens from the culture medium.
Some specific antigens have already been proposed for therapy and diagnosis of melanoma type of cancer: the peptide p97 has been disclosed in U.S. Pat. Nos. 5,262,177 and 5,141,742, while a 35 kD protein has been mentioned in EP 529,007.
We now have found a melanoma-associated polypeptide, characterized in that it comprises the aminoacid sequence of SEQ ID NO: 2.
This melanocyte lineage-specific antigenic polypeptide (also mentioned gp100) is recognized by the monoclonal antibody NKI-beteb, which antibody has proven suitable for diagnostic purposes. The antigens recognized by this antibody are intracellular proteins of approximately 10 kd (gp 10) and 100 kd (gp100). The latter is also detectable in a culture medium of melanoma cells (Vennegoor, C. et al, Am. J. Pathol. 130, 179-192, 1988). It has also been found that the gp100 antigen reacts with other melanoma-specific antibodies such as HMB-50 (described by Vogel, A. M. and Esclamado, R. M., Cancer Res. 48, 1286-1294, 1988) or HMB-45 (described by Gown, A. M. et al., Am. J. Pathol. 123, 195-203, 1986). Since the proteins reacting with these monoclonal antibodies have been shown te be glycosylated in melanoma cells, differences have been found in mobility when analyzed by SDS-PAGE.
Although this gp100 antigen is predominantly expressed intracellularly, it has now been established that it is a suitable immunogenic antigen, because it has been demonstrated that these intracellular proteins can be processed and presented as peptides in the context of MHC molecules to cells of the immune system. In fact, tumor infiltrating lymphocytes derived from tumors of melanoma patients have been found which react with the antigen.
Therefore, the gp100 polypeptide is a potential target for cellular responses against carcinoma and thus a suitable subject for therapy and diagnosis in melanoma patients.
Gp100 is a type I transmembrane protein, which has a threonine-rich domain containing repetitive amino acid sequences present in the middle of the protein (amino acids 309-427). This threonine-rich domain, which may be subjected to extensive O-linked glycosylation, is preceded by a histidine-rich region (amino acids 182-313) and followed by a cysteine-rich domain (amino acids 475-566). Based on hydrophobicity plot analysis (Kyte, J. and Doolittle, R. F., 1982), a single transmembrane domain bordered by charged residues is present in the carboxy-terminal part (amino acids 591-611) of gp100. The predicted cytoplasmic domain is 45 amino acids long. Five putative N-linked glycosylation sites are present, consistent with gp100 being a glycoprotein.
The term xe2x80x9cpolypeptidexe2x80x9d refers to a molecular chain of amino acids, does not refer to a specific length of the product and if required can be modified in vivo or in vitro, for example by glycosylation, amidation, carboxylation or phosphorylation; thus inter alia peptides, oligopeptides and proteins are included within the definition of polypeptide.
Of course, functional derivatives as well as fragments of the polypeptide according to the invention are also included in the present invention. Functional derivatives are meant to include polypeptides which differ in one or more amino acids in the overall sequence, which have deletions, substitutions, inversions or additions. Amino acid substitutions which can be expected not to essentially alter biological and immunological activities, have been described. Amino acid replacements between related amino acids or replacements which have occurred frequently in evolution are, inter alia Ser/Ala, Ser/Gly, Asp/Gly, Asp/Asn, Ile/Val (see Dayhof, M. D., Atlas of protein sequence and structure, Nat. Biomed. Res. Found., Washington D.C., 1978, vol. 5, suppl. 3). Based on this information Lipman and Pearson developed a method for rapid and sensitive protein comparison (Science 227, 1435-1441, 1985) and determining the functional similarity between homologous polypeptides.
Functional derivatives which still show immunological activity towards the monoclonal antibody NKI-beteb or HMB-50 or HMB-45 are included within the scope of this invention.
Furthermore as functional derivatives of these peptides are also meant peptides derived from gp100 which are able to induce target cell lysis by tumor infiltrating lymphocytes.
In addition, with functional derivatives of these peptides are also meant addition salts of the peptides, amides of the peptides and specifically the C-terminal amides, esters and specifically the C-terminal esters and N-acyl derivatives specifically N-terminal acyl derivatives and N-acetyl derivatives.
The polypeptides according to the invention can be produced either synthetically or by recombinant DNA technology. Methods for producing synthetic polypeptides are well known in the art.
The organic chemical methods for peptide synthesis are considered to include the coupling of the required amino acids by means of a condensation reaction, either in homogenous phase or with the aid of a so-called solid phase. The condensation reaction can be carried out as follows:
a) condensation of a compound (amino acid, peptide) with a free carboxyl group and protected other reactive groups with a compound (amino acid, peptide) with a free amino group and protected other reactive groups, in the presence of a condensation agent;
b) condensation of a compound (amino acid, peptide) with an activated carboxyl group and free or protected other reaction groups with a compound (amino acid, peptide) with a free amino group and free or protected other reactive groups.
Activation of the carboxyl group can take place, inter alia, by converting the carboxyl group to an acid halide, azide, anhydride, imidazolide or an activated ester, such as the N-hydroxy-succinimide, N-hydroxy-benzotriazole or p-nitrophenyl ester.
The most common methods for the above condensation reactions are: the carbodiimide method, the azide method, the mixed anhydride method and the method using activated esters, such as described in The Peptides, Analysis, Synthesis, Biology Vol. 1-3 (Ed. Gross, E. and Meienhofer, J.) 1979, 1980, 1981 (Academic Press, Inc.).
Production of polypeptides by recombinant DNA techniques is a general method which is known, but which has a lot of possibilities all leading to somewhat different results. The polypeptide to be expressed is coded for by a DNA sequence or more accurately by a nucleic acid sequence.
It has been found that the amino acid sequence of gp100 closely resembles the amino acid sequence of the already known melanoma-associated peptide pMel17, disclosed in Kwon, B. S. (1991).
The amino acid differences between gp100 and Pmel17 consist of substitutions at amino acid position 274 (T-C/PRO-LEU) and 597 (C-G/ARG-PRO) and a stretch of 7 amino acid absent in gp100 at position 587. A single nucleotide difference at position 762 (C-T) does not result in an amino acid substitution. Gp100 is also 80% homologous to a putative protein deduced from a partial cDNA clone (RPE-1) isolated from a bovine retinal cDNA library (Kim, R. Y. and Wistow, G. J., 1992) and 42% homologous to a chicken melanosomal matrix protein, MMP115 (Mochii, M., 1991). See also FIG. 2.
Also part of the invention is the nucleic acid sequence comprising the sequence encoding the gp100 polypeptide.
Preferably the sequence encoding gp100 is the sequence of SEQ ID NO:1.
As is well known in the art, the degeneracy of the genetic code permits substitution of bases in a codon resulting in another codon still coding for the same amino acid, e.g. the codon for the amino acid glutamic acid is both GAT and GAA. Consequently, it is clear that for the expression of a polypeptide with an amino acid sequence shown in SEQ ID NO:2 use can be made of a derivate nucleic acid sequence with such an alternative codon composition thereby differring from the nucleic acid sequence shown in SEQ ID NO:1.
xe2x80x9cNucleotide sequencexe2x80x9d as used herein refers to a polymeric form of nucleotides of any length, both to ribonucleic acid (RNA) seqeuences and to desoxyribonucleic acid (DNA) seqeuences. In principle this term refers to the primary structure of the molecule. Thus, this term includes double and single stranded DNA, as well as double and single stranded RNA, and modifications thereof.
The nucleotide sequence of gp100 contains 2115 base pairs (bp) and terminates with a poly (A) tract of 15 nucleotides which is preceded by the consensus polyadenylation sequence AATAAA SEQ. ID. NO. 31. An open reading frame (ORF) extending from nucleotide 22 through 2007 is present in gp100 DNA. This ORF starts with an ATG codon within the appropriate sequence context for translation initiation and codes for a protein of 661 amino acids. The amino-terminal 20 amino acids fit all criteria for signal sequences, including a potential cleavage site after ALA at position 20 (xe2x88x921), which would indicate that mature gp100 contains 641 amino acids (approximately 70 kD).
The most striking difference between gp100 and Pmel17 cDNAs is the inframe deletion of 21 bp in gp100 cDNA (FIG. 2). Comparison of the nucleotide sequence of genomic DNA with the sequence of gp100 cDNA revealed the presence of an intron (102 bp) just at the position of the 21 bp insertion in Pmel17 cDNA. The exon/intron boundaries nicely fit the consensus 5xe2x80x2 donor and 3xe2x80x2 acceptor splice site sequences (Padgett, 1986). In the genomic DNA, the sequence comprising the additional 21 bp in Pmel17 cDNA is located directly upstream of the 3xe2x80x2 cleavage site used to generate gp100 RNA and is preceded by an alternative 3xe2x80x2 acceptor splice site. Whereas the gp100-specific 3xe2x80x2 acceptor splice site fits the consensus sequence, the Pmel17-specific 3xe2x80x2 acceptor splice site appears to be sub-optimal in that it lacks a pyrimidine-rich region. Sub-optimal RNA processing sites are present in many alternatively processed messenger RNA precursors and have been implicated to function in regulation of alternative RNA processing (reviewed by Green, M. R., 1991). Collectively, these data prove that the transcripts corresponding to gp100 and Pmel17 cDNAs are generated by alternative splicing of a single primary transcript.
A further part of the invention are peptides, which are immunogenic fragments of the gp100 polypeptide.
Immunogenic fragments are fragments of the gp100 molecule, which still have the ability to induce an immunogenic response, i.e. that it is either possible to evoke antibodies recognizing the fragments specifically, or that it is possible to find T lymphocytes which have been activated by the fragments.
As has been said above it has been known that the immunogenic action of tumor associated antigens is often elicited through a T cell activating mechanism (Townsend, A. R. M. and Bodmer, H., Ann. Rev. Immunol. 7, 601-624, 1989). Cytotoxic T lymphocytes (CTLs) recognizing melanoma cells in a T cell receptor (TCR)-dependent and MHC-restricted manner have been isolated from tumor-bearing patients (reviewed by Knuth, A., 1992). Brichard et al. (1993) have shown that a peptide derived from tyrosinase, an other melanocyte-specific antigen, is recognized by a CTL clone.
It is known that the activation of T cells through the MHC molecule necessitates processing of the antigen of which short pieces (for example 8-12 mers) are presented to the T lymphocyte.
The immunogenic oligopeptides located in the gp100 sequence form also part of the invention.
We have found immunogenic peptide sequences of the gp100 sequence which are not only able to bind with the MHC I molecule, but which also have been demonstrated to recognize tumor infiltrating lymphocytes which have been isolated from a melanoma patient.
Several peptides have been found: the peptides having the amino acid sequences V-L-P-D-G-Q-V-I-W-V (SEQ ID NO:5), M-L-G-T-H-T-M-E-V (SEQ ID NO:24), R-L-M-K-Q-D-F-S-V (SEQ ID NO:25), (V)-(W)-(K)-T-W-G-Q-Y-W-Q-V-(L) (SEQ ID NO:10) and L-L-D-G-T-A-T-L-R-L (SEQ ID NO:4) have been found to bind to the MHC HLA-A2.1 molecule. In addition, the latter two peptides are recognized by anti-melanoma cytotoxic T lymphocytes in the context of HLA-A2.1.
Preferably these peptides are flanked by non-related sequences, i.e. sequences with which they are not connected in nature, because it has been found that such flanking enhances the immunogenic properties of these peptides, probably through a better processing and presentation by APC""s.
Another part of the invention is formed by nucleotide sequences comprising the nucleotide sequences coding for the above mentioned peptides.
Next to the use of these sequences for the production of the peptides with recombinant DNA techniques, which will be exemplified further, the sequence information disclosed in the sequence listings for gp100 or its epitopes can be used for diagnostic purposes.
From these sequences primers can be derived as basis for a diagnostic test to detect gp100 or gp100-like proteins by a nucleic acid amplification technique for instance the polymerase chain reaction (PCR) or the nucleic acid sequence based amplification (NASBA) as described in U.S. Pat. No. 4,683,202 and EP 329,822, respectively.
With PCR large amounts of DNA are generated by treating a target DNA sequence with oligonucleotide primers such that a primer extension product is synthesized which is separated from the template using heat denaturation and in turn serves as a template, resulting in amplification of the target sequence. When RNA is to be amplified with PCR the RNA strand is first transcribed into a DNA strand with the aid of reverse transcriptase.
With the aid of NASBA large amounts of single stranded RNA are generated from either single stranded RNA or DNA or double stranded DNA. When RNA is to be amplified the ssRNA serves as a template for the synthesis of a first DNA strand by elongation of a first primer containing a ssRNA polymerase recognition site. The formed DNA strand in turn serves as the template for the synthesis of a second, complementary, DNA strand by elongation of a second primer, resulting in a double stranded active RNA-polymerase promoter site, and the second DNA serves as a template for synthesis of large amounts of the first template, the ssRNA, with the aid of RNA polymerase.
Detection of the amplified nucleotide sequence is established by hybridizing a complementary detection probe to the amplified nucleic acid. This probe can be labelled and/or immobilized on a solid phase.
Detection of the label can be performed through methods known in the art. Detection of nucleic acids bound through the probe to the solid phase can be done by compounds capable of selective detection of nucleic acids.
As said before the nucleotide sequences can be used for the production of gp100 or one of its epitopes with recombinant DNA techniques. For this the nucleotide sequence must be comprised in a cloning vehicle which can be used to transform or transfect a suitable host cell.
A wide variety of host cell and cloning vehicle combinations may be usefully employed in cloning the nucleic acid sequence. For example, useful cloning vehicles may include chromosomal, non-chromosomal and synthetic DNA sequences such as various known bacterial plasmids, and wider host range plasmids such as pBR 322, the various pUC, POEM and pBluescript plasmids, bacteriophages, e.g. lambda-gt-Wes, Charon 28 and the M13 derived phages and vectors derived from combinations of plasmids and phage or virus DNA, such as SV40, adenovirus or polyoma virus DNA (see also Rodriquez, R. L. and Denhardt (1988); Lenstra, 1990).
Useful hosts may include bacterial hosts, yeasts and other fungi, plant or animal hosts, such as Chinese Hamster Overy (CHO) cells or monkey cells and other hosts.
Vehicles for use in expression of the peptides will further comprise control sequences operably linked to the nucleic acid sequence coding for the peptide. Such control sequences generally comprise a promoter sequence and sequences which regulate and/or enhance expression levels. Furthermore an origin of replication and/or a dominant selection marker are often present in such vehicles. Of course control and other sequences can vary depending on the host cell selected.
Techniques for transforming or transfecting host cells are quite known in the art (see, for instance, Maniatis et al., 1982 and 1989).
It is extremely practical if, next to the information for the peptide, also the host cell is co-transformed or co-transfected with a vector which carries the information for an MHC molecule to which said peptide is known to bind. Preferably the MHC molecule is HLA-A2.1, HLA-A1 or HLA-A3.1, or any other HLA allele which is known to be present in melanoma patients. HLA-A2.1 is especially preferred because it has been established (Anichini A., 1993) that melanoma cells carry antigens recognized by HLA-A2.1 restricted cytotoxic T cell clones from melanoma patients.
Host cells especially suited for the expression of gp100 are the murine EL4 and P8.15 cells. For expression of gp100 human BLM cells (described by Katano, M., 1984) are especially suited because they already are able to express the MHC molecule HLA-A2.1.
Gp100 or any of its peptides or their nucleotide sequences mentioned above can be used in a vaccine for the treatment of melanoma.
In addition to an immunogenically effective amount of the active peptide the vaccine may contain a pharmaceutically acceptable carrier or diluent.
The immunogenicity of the peptides of the invention, especially the oligopeptides, can be enhanced by cross-linking or by coupling to an immunogenic carrier molecule (i.e. a macromolecule having the property of independently eliciting an immunological response in a patient, to which the peptides of the invention can be covalently linked).
Covalent coupling to the carrier molecule can be carried out using methods well known in the art, the exact choice of which will be dictated by the nature of the carrier molecule used. When the immunogenic carrier molecule is a protein, the peptides of the invention can be coupled, e.g. using water soluble carbodiimides such as dicyclohexylcarbodiimide, or glutaraldehyde.
Coupling agents such as these can also be used to cross-link the peptides to themselves without the use of a separate carrier molecule. Such cross-linking into polypeptides or peptide aggregates can also increase immunogenicity.
Examples of pharmaceutically acceptable carriers or diluents useful in the present invention include stabilizers such as SPGA, carbohydrates (e.g. sorbitol, mannitol, starch, sucrose, glucose, dextran), proteins such as albumin or casein, protein containing agents such as bovine serum or skimmed milk and buffers (e.g. phosphate buffer).
Optionally, one or more compounds having adjuvant activity may be added to the vaccine. Suitable adjuvants are for example aluminium hydroxide, phosphate or oxide, oil-emulsions (e.g. of Bayol F(R) or Marcol 52(R)), saponins or vitamin-E solubilisate.
The vaccine according to the present invention can be given inter alia intravenously, intraperitoneally, intranasally, intradermally, subcutaneously or intramuscularly.
The useful effective amount to be administered will vary depending on the age and weight of the patient and mode of administration of the vaccine.
The vaccine can be employed to specifically obtain a T cell response, but it is also possible that a B cell response is elicited after vaccination. If so, the B cell response leads to the formation of antibodies against the peptide of the vaccine, which antibodies will be directed to the source of the antigen production, i.e. the tumor cells. This is an advantageous feature, because in this way the tumor cells are combatted by responses of both immunological systems.
Both immunological systems will even be more effectively triggered when the vaccine comprises the peptides as presented in an MHC molecule by an antigen presenting cell (APC). Antigen presentation can be achieved by using monocytes, macrophages, interdigitating cells, Langerhans cells and especially dendritic cells, loaded with one of the peptides of the invention. Loading of the APC""s can be accomplished by bringing the peptides of the invention into or in the neighbourhood of the APC, but it is more preferable to let the APC process the complete gp100 antigen. In this way a presentation is achieved which mimicks the in vivo situation the most realistic. Furthermore the MHC used by the cell is of the type which is suited to present the epitope.
An overall advantage of using APC""s for the presentation of the epitopes is the choice of APC cell that is used in this respect. It is known from different types of APC""s that there are stimulating APC""s and inhibiting APC""s.
Preferred are the listed cell types, which are so-called xe2x80x98professionalxe2x80x99 antigen presenting cells, characterized in that they have co-stimulating molecules, which have an important function in the process of antigen presentation. Such co-stimulating molecules are, for example, B7, CTLA-4, CD70 or heat stable antigen (Schwartz, 1992).
Fibroblasts, which have also been shown to be able to act as an antigen presenting cell, lacks these co-stimulating molecules.
It is also possible to use cells already transfected with a cloning vehicle harbouring the information for gp100 and which are cotransfected with a cloning vehicle which comprises the nucleotide sequence for an MHC class I molecule, for instance the sequence coding for HLA A2.1, HLA A1 or HLA A3.1. These cells will act as an antigen presenting cell and will present gp100-fragments in the MHC class I molecules which are expressed on their surface. It is envisaged that this presentation will be enhanced, when the cell is also capable of expressing one of the above-mentioned co-stimulating molecules, or a molecule with a similar function. This expression can be the result of transformation or transfection of the cell with a third cloning vehicle having the sequence information coding for such a co-stimulating molecule, but it can also be that the cell already was capable of production of co-stimulating molecules.
In stead of a vaccine with these cells, which next to the desired expression products, also harbour many elements which are also expressed and which can negatively affect the desired immunogenic reaction of the cell, it is also possible that a vaccine is composed with liposomes which expose MHC molecules loaded with peptides, and which, for instance, are filled with lymphokines. Such liposomes will trigger a immunologic T cell reaction.
By presenting the peptide in the same way as it is also presented in vivo an enhanced T cell response will be evoked. Furthermore, by the natural adjuvant working of the, relatively large, antigen presenting cells also a B cell response is triggered. This B cell response will a.o. lead to the formation of antibodies directed to the peptide-MHC complex. This complex is especially found in tumor cells, where it has been shown that in the patient epitopes of gp100 are presented naturally, which are thus able to elicit a T cell response. It is this naturally occurring phenomenon which is enlarged by the vaccination of APC""s already presenting the peptides of the invention. By enlarging not only an enlarged T cell response will be evoked, but also a B cell response which leads to antibodies directed to the MHC-peptide complex will be initiated.
The vaccines according to the invention can be enriched by numerous compounds which have an enhancing effect on the initiation and the maintenance of both the T cell and the B cell response after vaccination.
In this way addition of cytokines to the vaccine will enhance the T cell response. Suitable cytokines are for instance interleukines, such as IL-2, IL-4, IL-7, or IL-12, GM-CSF, RANTES, tumor necrosis factor and interferons, such as IFN-.
In a similar way antibodies against T cell surface antigens, such as CD2, CD3, CD27 and CD28 will enhance the immunogenic reaction.
Also the addition of helper epitopes to stimulate CD4+ helper cells or CD8+ killer cells augments the immunogenic reaction. Alternatively also helper epitopes from other antigens can be used, for instance from heat shock derived proteins or cholera toxin.
Another part of the invention is formed by usage of gp100 reactive tumor infiltrating lymphocytes (TIL""s). In this method the first step is taking a sample from a patient. This is usually done by resection of a tumor deposit under local anaesthesia. The TIL""s present in this specimen are then expanded in culture for four to eight weeks, according to known methods (Topalian, S. L. et al., 1987). During this culture the TIL""s are then checked for reactivity with gp100 or one of the epitopes derived from gp100. The TIL""s which recognize the antigen are isolated and cultured further.
The tumor infiltrating lymphocytes, reactive with gp100, which are obtained through this method, form also of the invention. One such TIL cell line, designated TIL 1200, has been found which specifically reacts with gp100 and its epitopes. This TIL 1200 cell line also expresses the MHC molecule HLA-A2.1. Furthermore expression of TCR xcex1/xcex2, CD3 and CD8 by this cell line has been demonstrated. Furthermore TIL 1200 recognizes transfectants expressing both HLA-A2.1 and gp100.
This TIL 1200 and other TIL""s recognizing gp100 are suited for treatment of melanoma patients. For such treatment TIL""s are cultured as stated above, and they are given back to the patients by an intravenous infusion. The success of treatment can be enhanced by pre-treatment of the tumor bearing host with either total body radiation or treatment with cyclophosphamide and by the simultaneous administration of interleukin-2 (Rosenberg, S. A. et al., 1986).
The TIL""s infused back to the patient are preferably autologous TIL""s (i.e. derived from the patient""s own tumor) but also infusion with allogenic TIL""s can be imagined.
A further use of the TIL""s obtained by the method as described above is for in vivo diagnosis. Labelling of the TIL""s, for instance with 111In (Fisher, 1989) or any other suitable diagnostic marker, renders them suited for identification of tumor deposits in melanoma patients.
Another part of the invention is formed by the T cell receptor (TCR) expressed by gp100 reactive CTLs. As is well known in the art, the TCR determines the specificity of a CTL. Therefore, the cDNA encoding the TCR, especially its variable region, can be isolated and introduced into T cells, hereby transferring anti-tumor activity to any T cell. Especially introduction of such a TCR into autologous T cells and subsequent expansion of these T cells, will result in large numbers of CTL suitable for adoptive transfer into the autologous patient.
Also cells harbouring this T cell receptor can be used for vaccination purposes.
A vaccine can also be composed from melanoma cells capable of expression of gp100. It is possible to isolate these cells from a patient, using anti-gp100 antibodies, such as NKI-beteb, but is also possible to produce such melanoma cells from cultured melanoma cell lines, which either are natural gp100-producers or have been manipulated genetically to produce gp100. These cells can be irradiated to be non-tumorogenic and infused (back) into the patient. To enhance the immunologic effect of these melanoma cells it is preferred to alter them genetically to produce a lymphokine, preferably interleukine-2 (IL-2) or granulocyte-macrophage colony stimulation factor (GM-CSF). Gp100+ melanoma cells can be transfected with a cloning vehicle having the sequence coding for the producti on of IL-2 or GM-CSF.
Infusion of such a vaccine into a patient will stimulate the formation of CTL""s.
Another type of vaccination having a similar effect is the vaccination with pure DNA, for instance the DNA of a vector or a vector virus having the DNA sequence encoding the gp100 antigen or peptides derived therefrom. Once injected the virus will infect or the DNA will be transformed to cells which express the antigen or the peptide(s).
Antibodies to any gp100 peptide, including antibodies to (V)-(W)-(K)-T-W-G-Q-Y-W-Q-V-(L) SEQ ID NO:10, and L-L-D-G-T-A-T-L-R-L SEQ ID NO:4, are also part of the invention.
Monospecific antibodies to these peptides can be obtained by affinity purification from polyspecific antisera by a modification of the method of Hall, R. et al. (1984). Polyspecific antisera can be obtained by immunizing rabbits according to standard immunisation schemes.
Monospecific antibody as used herein is defined as a single antibody species or multiple antibody species with homogeneous binding characteristics for the relevant antigen. Homogeneous binding as used herein refers to the ability of the antibody species to bind to ligand binding domain of the invention.
The antibody is preferably a monoclonal antibody, more preferably a humanised monoclonal antibody.
Monoclonal antibodies can be prepared by immunizing inbred mice, preferably Balb/c with the appropriate protein by techniques known in the art (Kxc3x6hler, G. and Milstein C., 1975). Hybridoma cells are subsequently selected by growth in hypoxanthine, thymidine and aminopterin in an appropriate cell culture medium such as Dulbecco""s modified Eagle""s medium (DMEM). Antibody producing hybridomas are cloned, preferably using the soft agar technique of MacPherson (1973). Discrete colonies are transferred into individual wells of culture plates for cultivation in an appropriate culture medium. Antibody producing cells are identified by screening with the appropriate immunogen. Immunogen positive hybridoma cells are maintained by techniques known in the art. Specific anti-monoclonal antibodies are produced by cultivating the hybridomas in vitro or preparing ascites fluid in mice following hybridoma injection by procedures known in the art.
It is preferred to use humanized antibodies. Methods for humanizing antibodies, such as CDR-grafting, are known (Jones, P. T. et al., 1986). Another possibility to avoid antigenic response to antibodies reactive with polypeptides according to the invention is the use of human antibodies or fragments or derivatives thereof.
Human antibodies can be produced by in vitro stimulation of isolated B-lymphocytes, or they can be isolated from (immortalized) B-lymphocytes which have been harvested from a human being immunized with at least one ligand binding domain according to the invention.
Antibodies as described above can be used for the passive vaccination of melanoma patients. A preferred type of antibodies for this kind of vaccine are antibodies directed against the above-mentioned peptides presented in connection with the MHC molecule. To produce these kind of antibodies immunization of peptides presented by APC""s is required. Such an immunization can be performed as described above. Alternatively, human antibodies to peptide-MHC complexes can be isolated from patients treated with a vaccine consisting of APC""s loaded with one of said peptides.
The antibodies, which are formed after treatment with one of the vaccines of the invention can also be used for the monitoring of said vaccination. For such a method serum of the patients is obtained and the antibodies directed to the peptide with which has been vaccinated are detected. Knowing the antibody titre from this detection it can be judged if there is need for a boost vaccination.
Specific detection of said antibodies in the serum can be achieved by labelled peptides. The label can be any diagnostic marker known in the field of in vitro diagnosis, but most preferred (and widely used) are enzymes, dyes, metals and radionuclides, such as 67Ga, 99mTc, 111In, 113mIn, 123I, 125I or 131I.
The radiodiagnostic markers can be coupled directly to the peptides of the invention or through chelating moieties which have been coupled to the peptide directly or through linker or spacer molecules. The technique of coupling of radionuclides to peptides or peptide-like structures is already known in the field of (tumor) diagnostics from the numerous applications of labelled antibodies used both in in vivo and in in vitro tests.
Direct labelling of peptides can for instance be performed as described in the one-vial method (Haisma, 1986). A general method for labelling of peptides through chelators, with or without linker or spacer molecules, has for instance been described in U.S. Pat. Nos. 4,472,509 and 4,485,086. Chelators using a bicyclic anhydride of DTPA have been disclosed in Hnatowich, D. J. et al. (1983). Coupling through diamide dimercaptide compounds has been disclosed in EP 188,256.

Melanoma associated antigenic polypeptide, epitopes thereof and vaccines against melanoma


Provided are a method of growing carbon nanotubes and a carbon nanotube device. The method includes: depositing an aluminum layer on a substrate; forming an insulating layer over the substrate to cover the aluminum layer; patterning the insulating layer and the aluminum layer on the substrate to expose a side of the aluminum layer; forming a plurality of holes in the exposed side of the aluminum layer to a predetermined depth; depositing a catalyst metal layer on the bottoms of the holes; and growing the carbon nanotubes from the catalyst metal layer.

Priority is claimed to Korean Patent Application No. 2004-12537, filed on Feb. 25, 2004, in the Korean Intellectual Property Office, the disclosure of which is incorporated herein in its entirety by reference.
1. Field of the Invention
The present invention relates to a method of horizontally growing carbon nanotubes (CNTs) and a device having the same, and more particularly, to a method of horizontally growing CNTs of uniform diameter from holes in an alumina template and a device having the same.
2. Description of the Related Art
The electric and mechanical properties of CNTs have been intensively studied to apply to various devices and applications. CNTs can be formed by vertically and horizontally growing methods. CNTs formed using the vertically growing method are used as an electron emission source in a field emission display (FED). Also, such CNTs may be used as a field emission source in a backlight device for a liquid crystal display (LCD), for example.
Horizontally grown CNTs may be applied to inter-connectors, Micro Electro Mechanical System (MEMS) sensors, horizontal field emitter tips, and the like.
In the prior art, an electric field is applied between two electrodes between which a patterned catalyst is interposed so as to horizontally grow CNTs. In such a horizontally growing method, it is not easy to accurately control the magnitude of a catalyst. Thus, it is difficult to uniformly control the diameter of CNTs. Also, yield of the CNTs is low and an electric field must be applied.
U.S. Pat. No. 6,515,339 discloses a method of horizontally growing CNTs from catalysts which are formed in a shape of nanodots, nanowires, or stripes, between which a predetermined space is formed, and on which a vertical growing preventing layer is disposed. However, in this method, it is difficult to uniformly control the diameter of the CNTs.

Method of horizontally growing carbon nanotubes and device having the same


The bolt has, in sequence along a bolt axis: a threaded portion, a thread run-out portion, a shank, and a head, the threaded portion having a thread with a given root radius and a given depth, the thread run-out portion connected to the shank via a thread run-out fillet having a thread run-out fillet radius, the thread run-out fillet radius being between two and six times the thread root radius.

Bolts are sometimes the only suitable option for clamping small size rotors having low bore radii. The head of the bolt is on one side of the bore, the shank of the bolt extends inside the bore, and the threaded portion extends out the other side of the bore, to which a nut is secured in a "thru bolt" configuration. Thru bolts can provide satisfactory access for torquing/untorquing. Bolt weight, nut weight, tightening torques, distance between the bolt's axis and the rotation axis, and rotation speed can be high, which can result in significantly high stresses in the bolt which can limit the bolt life. Lightweight alloys for the bolt have been used to achieve acceptable low-cycle fatigue (LCF) life for the discs, but there are limits in the advantages gainable solely by alloy selection since some properties or features of lightweight materials are typically traded off for their lighter weight.
Accordingly, there remains room for improvement in addressing LCF life of bolts, especially in the context of high stress rotary environments.


Provided is an endoscopic surgical device includes: a sheath having a pipe for delivering a liquid and inserted into a channel of an endoscope; a pair of forceps components disposed so as to protrude from a distal end cover of the sheath; an operable member that is connected to the forceps components and opens and closes the forceps components; and a flow-path formation member positioned at proximal end side rather than the forceps components and is disposed in a rotatable manner about a central axis of the sheath together with the forceps components. The flow-path formation member includes a recess disposed in a direction orthogonal to an opening-closing direction of the forceps components and recessed radially inward. The recess forms a gap between the flow-path formation member and an inner peripheral surface of the distal end cover so as to constitute a liquid delivery hole.

A known endoscopic surgical device includes a forceps section that is provided at a distal end of a sheath to be inserted into a channel of an endoscope and that is to be opened and closed by a wire. This endoscopic surgical device delivers a liquid guided via a flow path within the sheath toward a target site from a liquid delivery lumen offset toward the lateral side of the forceps (for example, see Japanese Unexamined Patent Application, Publication No. 2005-224426).


An invention is provided for dynamic cache allocation in a solid state drive environment. The invention includes partitioning a cache memory into a reserved partition and a caching partition, wherein the reserved partition begins at a beginning of the cache memory and the caching partition begins after an end of the reserved partition. Data is cached starting at a beginning of the caching partition. Then, when the caching partition is fully utilized, data is cached the reserved partition. After receiving an indication of a power state change, such as when entering a sleep power state, marking data is written to the reserve partition. The marking data is examined after resuming the normal power state to determine whether a deep sleep power state was entered. When returning from a deep sleep power state, the beginning address of valid cache data within the reserve partition is determined after resuming a normal power state.

1. Field of the Invention
This invention relates generally to disk caching, and more particularly to dynamic cache allocation in a solid stage drive environment.
2. Description of the Related Art
Today, mobile computing allows individuals the freedom to utilize the processing power of computers anywhere it is needed. Using mobile computing devices, such as laptops and notebooks, users are able to take their projects with them whenever they leave their normal computing environment such as their home or office. To enable such freedom, power consumption by these mobile computing devices must be reduced to enable longer operating times away from permanent power sources.
One solution for power conservation is the use of various power states based on computing use. For example, when a system is idle for a predetermined period of time, the system can enter a standby, or sleep power state. In the sleep power state, the system turns off power to peripherals, such as the keyboard, mouse and the hard drive. However, to save the current state of the computing device, power is still supplied to the RAM, enabling the system to hold the current state in memory. Upon an event indicating the end of the idle period, such as a keyboard use or mouse use, the system resumes from the sleep power state, supplying power back to peripherals and resuming normal operation.
However, although much reduced, the sleep power state still consumes power in order to save the system state in RAM. To further save power, a system can enter a hibernation power state, wherein the system state is saved to non-volatile memory. When entering the hibernation state, the contents of the RAM is saved to non-volatile memory and power is turned off in the entire system. Upon resuming, the RAM contents is loaded back into RAM from the non-volatile memory and normal operation continues.
Although the hibernation power state saves power, resuming from the hibernation state requires an increased period of time over resuming from a sleep power state, particularly when the non-volatile memory used to save the system state is a hard disk drive. To address this issue, Intel Fast Flash Standby (iFFS) has been developed. iFFS saves the system state on a solid state drive (SSD) and further introduces a sleep state prior to entering an iFFS-hibernation state. Together, these improvements decrease the time required to resume after the system has been idle and power savings modes have been entered.
FIG. 1 is a block diagram showing a prior art iFFS based computer system 100. The iFFS based computer system includes a central processing unit (CPU) in communication with system RAM 104 and an iFFS non-volatile memory 106. The iFFS non-volatile memory 106 is an SSD that has the same storage capacity as the RAM 104 and is dedicated to storage of the RAM contents upon hibernation. When the system 100 is idle for a predetermined period of time, the system enters a sleep power mode, wherein power is still supplied to the RAM 104, however all other peripherals are turned off.
After a predetermined period of time, if the system 100 remains idle it enters an iFFS-hibernation power state. When the system 100 enters the iFFS-hibernation power state, the contents of the RAM 104 is written to the iFFS non-volatile memory 106 and power is turned off in the entire system 100. Later, upon resume, the RAM data stored in the iFFS non-volatile memory 106 is loaded back into the RAM 104 and the system 100 resumes normal operation. Since the iFFS non-volatile memory 106 is an SSD, the period of time need to complete resume is greatly decreased.
Caching has long been used in storage environments to enhance the performance of slower storage devices, such as disk drives. In caching, a smaller and faster storage medium is utilized to temporarily store and retrieve frequently used data, while the larger and typically slower mass storage medium is used for long term storage of data. However, using the SSD of the iFFS non-volatile memory can lead to data loss for the cache and/or wasted SSD storage.
In view of the foregoing, there is a need for systems and methods that allow SSD based caching in an iFFS environment. The methods should allow full iFFS functionality without wasting expensive SSD memory. Moreover, the methods should provide a means to fully utilized memory for caching, yet account for any data loss as a result of iFFS based hibernation use.

Dynamic cache allocation in a solid state drive environment


A method for determining a critical reduction in shaft speed in a work machine having at least one monitored shaft driven by a crankshaft of an engine includes determining an actual rotational speed of the crankshaft; determining an actual rotational speed of a first monitored shaft; determining a filtered rotational speed of the first monitored shaft based on an algorithm that utilizes the actual rotational speed of the first monitored shaft, the actual rotational speed of the crankshaft, and a reference rotational speed of the crankshaft; comparing the filtered rotational speed to the actual rotational speed of the first monitored shaft; determining whether the actual rotational speed of the first monitored shaft deviates from the filtered rotational speed by a predetermined amount; and activating an alarm if the actual rotational speed of the first monitored shaft deviates from the filtered rotational speed by the predetermined amount.

One type of work machine is an agricultural combine. Agricultural combines are engine powered machines that harvest, thresh, separate and clean an agricultural crop. The resulting clean grain is stored in a grain tank located on the combine. The clean grain can then be transported from the grain tank to a truck, grain cart or other receiving bin by an unloading auger.
Agricultural combines have several rotating shafts (e.g., rotor, clean grain elevator, etc.) that are monitored for changes in speed. Typically, a large table is maintained that indicates the allowed speed range for each shaft based on machine type and other customer settings. When the shaft speed falls below the value as defined in this table, a low shaft speed alarm is activated to notify the operator.
However, with this approach, with every change in machine settings and options, the table becomes more difficult to maintain. Also, an operator may experience the occasional nuisance alarm when nothing is wrong.

Method for determining a critical reduction in shaft speed in a work machine


A transaction center including a transaction processing platform having an automatic call distribution system for queuing customer transaction requests, for choosing an appropriate service agent according to information gathered about the transaction requests, and for determining when the appropriate service agent is available. Also provided is a transaction request server for receiving and storing the transaction requests. In operation, the message is first received and stored by the transaction request server. Next, the presence of the message is detected and in response thereto a signal is sent to the transaction processing platform. Finally, the message is sent to the appropriate service agent in response to a signal from the transaction processing platform indicating that the appropriate service agent is available to service the transaction request.

Traditionally, business has been conducted as a store front operation where customers had to be physically present at a store to conduct business. Customers may also have been able to request items for future delivery by calling the store or by writing a letter or filling out an order form which was later mailed to the store.
In recent years, however, there has been a tremendous shift in the way business is conducted. The telephone has become the new "front door" to businesses. As the use of 800 telephone numbers proliferated, customers had an alternative and most often more convenient way to conduct business. A physical presence at the store was no longer needed, only the ability to communicate was necessary.
In response to the growth in the use of the telephone by customers, call center technology was developed. Traditional call centers consist solely of customers trying to reach service agents by telephone. The calls are placed into a queue where they wait until a service agent is available to service them. In a call center, the private branch exchange, or PBX, is provided with what is commonly known as an automatic call distribution, or ACD, system. ACD systems are well known in the art and generally comprise customer administrable software that governs what happens to a call between arrival at the call center and the time the call is answered and processed by a service agent. The ACD system may, for instance, allow the caller to hear ringing, music or announcements about the company while waiting in the queue. In addition, the ACD system may perform functions of which the caller is not aware. These functions include using information gathered about the caller to access caller records or invoices which will later be used by the service agent, and routing the call to an appropriate available agent based on a variety of factors relating to the customer request or the service agents. Such factors include, for example, the geographical origin of the call indicated by the telephone number from which the call came, whether the call is a sales or service call, whether there should be a priority assigned to the call based on the caller, the type of skills necessary to properly service the request and the relative workloads of the service agents which may include when a particular agent is available to handle a call. The information may also include information such as the number that the customer called in order to obtain service, as in the case where a business uses separate numbers for sales and service. This information may be gathered in a variety of ways. For example, the caller may be prompted to enter information such as an account number, social security number or zip code, or may be prompted to choose from a menu of options. Also, some information is inherent in each call, such as the area code of the origin of the call, and can be gathered without the knowledge of the caller. Typically, the type of information used to route the calls is selectable and programmable by the organization utilizing the call center and the calls are ultimately routed based upon a set of programmable rules which utilize the information.
In addition, the PBX is typically provided with monitoring and reporting software that is able to monitor and report quantitative metrics relating to the customer requests, the individual service agents and the call center as a whole. An example of such software is Lucent Technologies, Inc.'s Call Management System, or CMS, and Basic Call Management System, or BCMS, software. For instance, this software can provide quantitative metrics relating to real time information such as the number of calls currently waiting in the call center queue and how long each call has been waiting. Also, the software can provide quantitative metrics relating to statistical information, such as the number of calls handled by the call center in a given day and the average process time of each call, and historical information, such as the volume of calls handled in a given day in a given year. Furthermore, quantitative metrics relating to the performance of individual service agents, such as the number of calls handled in a day and the average process time of each, can be gathered and reported. These exemplary quantitative metrics are provided for illustrative purposes only, and it should be understood that a large variety of other quantitative metrics could be measured as desired by a particular user.
Keeping up with the rising volume of calls can be costly and inefficient. Thus, in order to provide quality service in a cost effective manner, businesses developed alternative routes for customers to communicate their requests. Voice mail technology was provided to callers as an alternative to waiting in the queue for an available service agent. With the voice mail option, a caller can leave a message stating his or her request which is processed and serviced by a service agent at a later time. Other alternatives include media such as the facsimile, electronic mail via subscriber networks and the Internet, regular mail, such as a letter or a purchase order, video, and live multi-media requests from sources such as the Internet. Also, in some situations, the customer does not make the request at all, but instead their equipment does, as is the case with burglar or fire alarms. The requests can also originate from within the business operating the call center, as in the case where automatic subscription renewals are to be generated and sent to customers.
These alternatives provide the benefit of reducing the number of direct telephone calls that a business, and its service agents, must handle. Another benefit is that a customer is not forced to wait on the telephone in a queue in real time. Instead, the customer has left a proxy, in the form of a facsimile, a voice mail, a letter, an electronic mail message or video message, which is awaiting service. These alternatives, however, also have a price. Fragmenting customer service among these many media may result in an inability to monitor and report the performance of the call center for some or all of the media. Therefore, the business is unable to obtain a total picture of service quality. Also, when service requests come in by facsimile, regular mail, electronic mail, and the like, they are typically kept in a central location where they are either picked up by the service agents or manually distributed to the service agents. Thus, because humans are involved in the process of getting these types of service requests to the service agents, the more difficult request may be passed over or put aside. Also, often times these types of service requests are handled during service agent off-time, rather than on a first come, first serve basis. Accordingly, the true time it takes to handle these requests cannot be accurately tracked. Finally, it takes time for the request to get to the service agents in this manner and, in the meantime, customers may get tired of waiting and call in anyway.

Transaction center for processing customer transaction requests from alternative media sources


A filter candle apparatus is provided comprising an elongated mesh pack defining an axial opening therethrough and having an end cap fixedly secured to one end and an end fitting fixedly secured to the opposite end, the end fitting defining an opening therethrough which is coaxial with the axial opening of the mesh pack, a hollow core tube defining a plurality of openings therein positioned within the axial opening of the mesh pack and being removably slidable therethrough, the core tube having an end member surrounding one end thereof that engage the end fitting of the mesh pack, a ring-shaped bearing recessedly held within and by the end fitting of the mesh pack and engaging the core tube proximate the end member, and an outer guard defining apertures therein adjacently surrounding the mesh pack.

This invention relates generally to filtration devices, and more particularly to filter media material for polymer filtration.
Various apparatuses exist within the prior art relating to filter media material for polymer filtration. Such apparatuses utilize a metal filtration media in order to obtain the desired characteristics for polymer filtration. Typical properties desired for such filtration relate to strength, temperature, chemical resistance and media migration. Various structures of metal filtration media can be utilized in apparatuses designed for polymer filtration, and numerous factors can be considered when evaluating the various media.
There are various types of filter elements utilizing different types of filter media including flat filters, candle and basket filters, and disc filters. In the art of filter candles, it has been common in the past to provide some of the filtration components as an integral unit in order to prevent leakage therebetween. As a result, cleaning of the filtration components is quite difficult, and a problem with a specific component of the filtration device means replacement of all components which are an integral part thereof.
In light of the prior art filtration devices, there exists much room for improvement in the art of filter elements for polymer filtration.


Apparatus and systems may operate to provide a first reactant as a gas that flows under reduced atmospheric pressure to interact with a surface, such as a tool body surface, the interaction confined to a passage within the tool body, wherein the passage includes the surface and extends without interruption from an entrance end of the passage to an exit end of the passage. Additional activity may include providing a second reactant as a gas under the reduced atmospheric pressure, subsequent to the first reactant, to interact with the surface of the tool body; and repeated provision of the first and second reactants until a selected coating thickness on the surface is formed. Additional apparatus, systems, and methods are disclosed.

Understanding the structure and properties of geological formations can reduce the cost of drilling wells for oil and gas exploration. Measurements made in a borehole (i.e., down hole measurements) are typically performed to attain this understanding, to identify the composition and distribution of material that surrounds the measurement device down hole. To obtain such measurements, a variety of sensors housed in down hole tools are used. Since the down hole environment is relatively harsh (e.g., as compared to a laboratory), these tools are often subject to corrosion and other destructive influences.

Controlled coating apparatus, systems, and methods


A material information detection apparatus, a phantom information recording apparatus, and operation methods for the apparatuses are provided. The material information detection apparatus includes a database configured to store phantom information obtained by projecting a multi-energy X-rayon a phantom comprising a plurality of materials, an input device configured to receive a plurality of pieces of object information per energy level, the object information obtained by projecting a multi-energy X-ray on an analysis object, and a detector configured to detect information related to materials constituting the analysis object based on the phantom information and the plurality of pieces of object information per energy level.

1. Field
One or more exemplary embodiments of the following description relate to a technology of detecting information on materials of an analysis object.
2. Description of the Related Art
For quantitative medical imaging or quantitative imaging using a multi-energy X-ray such as a dual energy X-ray, components of an object of imaging, in other words, an analysis object, should be calculated.
When imaging an analysis object such as, for example, a human body using the multi-energy X-ray, a plurality of materials of the analysis object may be separated based on a combination according to a mixture ratio and a thickness, that is, a quantity, of the plurality of materials. Calibration refers to a process of finding relationships between an X-ray image and the combination based on the mixture ratio and the thickness of the plurality of materials.
To increase the accuracy of calibration with respect to a phantom including a plurality of materials, calibration should be performed with respect to all possible mixture ratios and thicknesses.

Information detection apparatus, phantom information recording apparatus and operation method thereof


The present invention provides a strain detector improved in output characteristic, in which no internal stress is generated in strain resistance elements even with it fixed on a fixing member. The strain detector is configured in that a detection hole is provided nearly at the center of a insulating substrate, a fixing hole is formed at the position of peripheral end, a portion around the detection hole is pressed by a pressing member, one holding member and the other holding member are provided with fixing holes opposing to the fixing holes of the insulating substrate, and the peripheral end of the insulating substrate is fixed by the fixing member inserted. Accordingly, the detection hole in the insulating substrate is peripherally displaceable in a vertical direction with respect to one holding member and the other holding member.

A conventional strain detector of this type has a configuration as shown in FIG. 19 and FIG. 20.
FIG. 19 is a bottom view of a conventional strain detector, and FIG. 20 is a perspective view of the strain detector. In FIG. 19 and FIG. 20, insulating substrate 1 is formed from elastic material, which is disposed at the bottom of pressing member 2. Also, four strain resistance elements 3 are provided at the bottom of the insulating substrate 1. The strain resistance elements 3 are electrically connected to a pair of power electrodes 4, a pair of output electrodes 5, and a pair of GND electrodes 6, thereby forming a bridge circuit.
The operation of a conventional strain detector having such a configuration will be described in the following discussion.
As shown in FIG. 21, when a pressing force is applied by pressing member 2 to the top nearly at the center of insulating substrate 1 after fixing a conventional strain detector on fixing member 7 by using male screws 8, a bending moment is generated in insulating substrate 1 due to the pressing force. Due to the bending moment, a bending moment is also generated in each of four strain resistance elements 3 disposed at the bottom of insulating substrate 1. And, when a bending moment is generated in strain resistance element 3, it causes the resistance value of strain resistance element 3 to change. Accordingly, the change of the resistance value is outputted from a pair of output electrodes 5 to an external computer (not shown), and thereby, the pressing force applied to insulating substrate 1 is measured.
Such a conventional strain detector is, for example, disclosed in Japanese Laid-Open Patent H8-87375.

Strain detector having a displaceable insulating substrate

Matsushita Electric Industrial Co., Ltd.


First a standard image to which a link is defined is displayed. If a user operates a input device and if the viewpoint enters into the first link boundary, and if the information indicating that a resume operation is required does not exist the first frame of moving image data is read out and displayed on the display device as a still image. If the viewpoint enters into the second link boundary, playing back and displaying of the moving image and is started. If the viewpoint moves out from the second link boundary, a frame displayed when the playback of the moving image is suspended is displayed as a still image and the resume point is stored. If the viewpoint moves out of the first link boundary, a standard image is displayed.

Home entertainment systems capable of playing back moving images as well as running game programs have been proposed. In home entertainment systems, a GPU generates three-dimensional images using polygons (see, for example, patent document No. 1).
Meanwhile, technology capable of the enlargement or reduction of a displayed image or the movement of the image in the upward, downward, leftward, or rightward directions, is proposed, by using tile images of a plurality of resolutions generated from a digital image such as a high-definition photo. In this image processing technology, the size of an original image is reduced into a plurality of stages to generate images of different resolutions so as to represent the original image in a hierarchical structure, where the image in each layer is divided into one or more tile images. Normally, the image with the lowest resolution is composed of one tile image. The original image with the highest resolution is composed of the largest number of tile images. An image processing device is configured to enlarge or reduce a displayed image such that an enlarged view or reduced view is presented efficiently by switching a currently-used tile image to a tile image of a different layer.

Information processing device and information processing method


The present invention provides a system and method for automatically combining image and audio data to create a multimedia presentation. In one embodiment, audio and image data are received by the system. The audio data includes a list of events that correspond to points of interest in an audio file. The audio data may also include an audio file or audio stream. The received images are then matched to the audio file or stream using the time. In one embodiment, the events represent times within the audio file or stream at which there is a certain feature or characteristic in the audio file. The audio events list may be processed to remove, sort or predict or otherwise generate audio events. Images processing may also occur, and may include image analysis to determine image matching to the event list, deleting images, and processing images to incorporate effects. Image effects may include cropping, panning, zooming and other visual effects.

With the developments of computers, digital cameras and the Internet, a new medium for displaying digital images has emerged. Images from digital cameras and other sources are often sent through email, displayed on the Internet, and presented as slide shows. Typically, slide show applications allow a user to choose between viewing images individually, proceeding to the next image upon user input, or viewing images as a slide show where they are consecutively displayed automatically at a constant rate. In a few of these applications, an audio file may be played while simultaneously viewing the slide show.
Though relatively easy to create, these unsynchronized multimedia presentations generated by slide show applications that display images at a constant rate do not retain viewer interest very well. Currently, any synchronization between the image display and audio must be generated manually in these applications. This manual synchronization requires an unreasonably large amount of time to parse information from the audio file and inspect the images manually. What is needed is a system and method for generating a multi-media presentation that automatically synchronizes an audio file and set of images.

Automatic generation of multimedia presentation


The dispenser dispenses tickets such as instant-winner lottery tickets from a strip in which the individual tickets are delineated by perforation lines. The dispenser includes a separator to tear the tickets apart before issuing them from the machine. The separator preferably is rotary and has a dull helical blade which rotates to contact the ticket strip adjacent a perforation line, and presses against the strip to tear the tickets apart along the perforation line at a point which progresses across the strip along the perforation line. Preferably, a bar code reader is provided for reading codes on tickets in the dispensing machine. The bar code reader is used in reading information from the tickets to initialize the dispenser control system and in accounting for the sale of tickets, and in the verification of winning tickets being sold. A game is provided in which the concept is that a jackpot is built up using the detection of each ticket sold. The mechanism is simplified by using a separator member and transport drive structures which span a plurality of different channels or bins of a multi-channel dispenser. This provides a very compact, simple, secure and low-cost mechanism which is used both in stand-alone ticket vending machines, and in counter-top attended dispensers.

This invention relates to dispensers, vending machines and methods, and particularly to dispensers, vending machines and methods for dispensing tickets from strips in which the tickets are delineated from one another by lines of weakness, such as perforation lines. Preferably, the dispensers are used to dispense "scratch-off" or "instant-winner" lottery tickets.
Various types of machines have been provided for dispensing and vending "instant-winner" or "scratch-off" type lottery tickets. Such tickets are characterized by being printed in long strips in which each ticket is delineated from the others by perforation lines.
The tickets often are stored in fan-fold form in a vending machine, and are issued in response to the insertion of money by the customer. The tickets are characterized by being printed on relatively heavy stock which is relatively stiff, but still flexible.
In some of the vending machines, the customer tears the tickets off of the strip by pulling on the ticket as it extends from the machine. In other such machines, the machine bursts each ticket apart from the rest and dispenses it separately.
In the type of dispenser in which tickets are burst free from the strip, there are several significant problems.
One of the main problems is that the separating mechanism currently used is relatively large, complex, and expensive to build and maintain.
Another problem with such prior dispensers is that the most common of such dispensers burst each and every ticket free from the strip. This tends to thwart the desires of some customers who would prefer to receive a string of connected tickets.
A further problem is with the loading of tickets into a multi-bin lottery ticket dispensing machine. The service representative usually is required to read certain information off of the tickets in a batch to be loaded into one bin, and load the information into the microprocessor controller of the vending machine by use of a keypad or the like. This is time-consuming, laborious, and error-prone.
Another problem with such systems is that winning tickets usually require verification. Ticket verification usually is performed when the customer carries a winning ticket to a clerk in a store, who then inserts it into a machine which reads the code on the back of the ticket and checks with a central computer to ascertain that the ticket so identified is, indeed, a winner, and to verify the winning amount. When this verification is complete, the holder can be paid the winnings.
Although this procedure minimizes certain kinds of errors and fraud, it does not detect a ticket which has come into the possession of the holder by means other than its actual dispensation from a vending machine.
Another problem with instant-winner gaming tickets is that a relatively large variety of different games are developed in order to keep the ticket buyers' interest. This creates additional costs for the lottery ticket issuing organization, requires more dispensing bins per vending machine, and/or more vending machines to dispense the multitude of games.
It is another problem with ticket vending systems that they do not provide accounting for all of the tickets dispensed by the vending machines in the system.


Methods and systems for the conversion of hydrocarbon feedstocks, in particular, naphtha feedstocks, into a hydrocarbon product stream containing a high yield of high-octane gasoline and chemicals products. In particular, the conversion takes place over a series of functionally distinctive catalyst beds, at least one of which includes a modified zeolitic catalyst comprising a zeolite, a transition metal, and optionally a binder. Systems provided include a hydrocarbon feed stream, which may be full-range naphtha, a hydrocarbon product stream, and a plurality of functionally distinctive catalyst beds arranged in series, wherein at least one of the catalyst beds comprises a modified zeolitic catalyst. A hydrocarbon feed stream may be conveyed through the plurality of functionally distinctive catalyst beds, producing an intermediate hydrocarbon stream between each, under conditions effective to convert the hydrocarbon feed stream to a hydrocarbon product stream comprising high-octane gasoline, xylenes, benzene, and/or toluene.

This application relates to methods and systems for the conversion of hydrocarbon feedstocks, in particular, naphtha feedstocks, into a hydrocarbon product stream containing a high yield of high-octane gasoline and/or chemicals products (e.g., benzene, toluene, and xylenes). In particular, the conversion takes place over a series of functionally distinctive catalyst beds, at least one of which includes a modified zeolitic catalyst as described herein, configured to subject a hydrocarbon feed stream to a preferred order of reforming reactions.
Naphtha reforming has been an important refining process for decades, generating hydrogen, chemicals feedstock (benzene, toluene, xylenes, which are also known as BTX), and high-octane gasoline. A typical naphtha feedstock will contain paraffins, olefins, naphthenes, aromatics, and isomers thereof. To reform a typical naphtha feedstock into gasoline and/or BTX, a reforming catalyst converts these molecules into aromatics. Isoparaffins may be present in gasoline as well, though isoparaffins contribute much less than aromatics to the octane rating and do not contribute to BTX yield at all. Paraffins and naphthenes contribute even less to octane rating. To carry out the necessary reforming reactions, reforming catalysts typically include a metal (e.g., platinum) to dehydrogenate paraffins and naphthenes (to form olefins and aromatics, respectively) and an acid function to catalyze the ring closure of the olefins and isomerization of remaining paraffins. However, paraffin-to-aromatic conversion is not particularly favored by conventional reforming catalysts and conversion is often slow and/or incomplete.
Metal-doped chlorided alumina is the most common reforming catalyst; however, chlorided alumina has its limitations. For example, chlorided alumina catalysts tend to favor paraffin isomerization at the expense of paraffin dehydrocyclization, which is undesirable for the reasons noted above. Metal-doped chlorided alumina, however, is good at dehydrogenation. Therefore, feedstocks best suited for chlorided alumina catalysts are typically limited to those having a low paraffin content and a higher naphthene content.
Chlorided alumina is also intolerant to chemical impurities in a feedstock. In particular, basic nitrogen in a feedstock will react with chloride loaded onto a chlorided alumina catalyst, effectively stripping the catalyst of its acidity (and activity), but also forming a compound (chloramine) that is corrosive to the reforming system. Finally, but not least importantly, chlorided alumina catalysts are susceptible to deactivation by coking, particularly under operating conditions most suited to forming a high-octane product stream. As coke accumulates, it blocks active sites for performing the reforming reactions, leading to reduced product yields.
Zeolitic catalysts may be modified to favor paraffin-to-aromatic conversion; however, such zeolites are also particularly good at converting naphthenes into paraffins. Thus, feedstocks best suited for zeolitic catalysts include as little naphthenes as possible. Additionally, much of the reforming catalytic activity in a zeolitic catalyst takes place in its pores. Mass diffusion of hydrocarbons from a hydrocarbon feed stream into and out of the pores dictate the catalytic activity and selectivity. As such, larger molecules are difficult to convert as their size excludes them from entering a pore. Thus, in addition to limiting the naphthene content, suitable feedstocks for zeolitic catalysts preferably have smaller hydrocarbons that easily diffuse in and out of the pores of a zeolite. However, the smallest hydrocarbons (e.g., C1-C5 hydrocarbons) are not readily converted to aromatics and their presence in a feedstock does not lend itself to formation of a high-octane product stream. Thus, C6-C7 feedstock (as opposed to a full-range C4-C12 feed) is most preferred. A C6-C7 feedstock, in turn, produces a hydrocarbon product stream that is primarily benzene and toluene products.
What is needed is a reforming system that that can effectively convert hydrocarbon feedstocks having varying paraffin and naphthenic content as well as varying sizes of hydrocarbons, into a high-octane hydrocarbon product stream and/or a product stream having high BTX yield.

Naphtha reformer yield using modified zeolitic catalysts

EXXONMOBIL RESEARCH AND ENGINEERING COMPANY


A grinding apparatus optimizes grinding efficiency within the grinding chamber by providing grinding action over a 360.degree. range encompassed by a set of breaker bars and one or more arcuate screening elements. Arcuate screening elements have an inside radius that identically matches the inside radius of the breaker bars. The screening elements act as grinding mechanisms as well as more conventional filtering mechanisms due to increased grinding capacity when the inside radial surface of the screening elements and the inside radial surface defined by the breaker bars are equidistant from the axis of a rotational shaft that supports a plurality of circumferentially spaced hammers.

1. Field of the Invention
This invention relates to grinding apparatus. More particularly, this invention relates to a grinding apparatus that facilitates highly efficient grinding of granular and large particle products as well as tough fiber dry grain products such as barely and waste gypsum.
2. Description of the Prior Art
Present product grinding and comminuting apparatus are known in the art for reducing the size of materials such as food products, chemicals, rubbers, resins, garbage (food waste), waste-paper, wood chips, waste fiber (cloth, gypsum), plastics, glass, metal chips or the like. Conventional grinding/comminuting apparatus such as that disclosed in U.S. Pat. No. 4,129,260, issued Dec. 12, 1978 to Baker, entitled Garbage Disposal, and U.S. Pat. No. 3,973,735, issued Aug. 10, 1976 to Ito et al., entitled Apparatus For Pulverizing And Sorting Municipal Waste, typically include a grinding chamber with high speed rotating beaters/hammers that tear, shred, slash, cut and grind one or more desired products to a desired particle size as the desired product(s) are forced between the rotating beaters/hammers and a set of breaker bars and to a very limited extent, also between the rotating beaters/hammers and one or more screening elements. Conventional screening elements for grinding and comminuting apparatus such as disclosed in U.S. Pat. No. 5,526,988, issued Jun. 18, 1996 to Rine, entitled Comminuting Apparatus With Tangentially Directed Discharge, are generally well known items and are currently in wide-spread use. These screening elements may include herringbone slots, round perforations, cross slot perforation screens, and jump gap screens in combination with a drum-type structure such as disclosed in U.S. Pat. No. 4,076,177, issued Feb. 28, 1978 to Hirayama et al., entitled Pulverizing Method And Apparatus, to form a grinding/comminuting apparatus for feeding the constituents axially there through so that the resulting pulverized constituents can be discharged at different axial positions depending upon the progress of pulverization and the selected screening elements. A significant disadvantage accompanying conventional grinding/comminuting apparatus known and used in the art is reduced grinding/comminuting efficiency due to the different radial distance between the axial centerline of the rotating shaft used to rotate the beaters/hammers and the working surfaces of the screening elements as compared with the working surfaces of the beaters/hammers. For example, known grinding/comminuting apparatus of the type using breaker bars and screening elements use the breaker bars to accomplish nearly all grinding/comminuting action of constituents that ultimately pass through the aforesaid screening elements. Constituents that do not immediately pass through the screening elements continue to be pulverized between the beaters/hammers and the breaker bars until sufficiently reduced to a size that is sufficiently small to allow the filtering process to continue to completion. It is readily apparent from the above, that the grinding/comminuting action that occurs in a drum type apparatus is one of the important factors in providing a highly efficient pulverizing process.


A PBIST architecture is described. A data path circuit is configured for bit-to-associated bit comparisons of expected result data read from a tile with the expected result data read from result memory. The data path circuit is configured to write a first type of failure indication to first failure memory responsive to a data 0 being read from the result memory and a data 1 being read from the tile for a bit-to-associated bit comparison failure. The data path circuit is further configured to write a second type of failure indication to second failure memory responsive to a data 1 being read from the result memory and a data 0 being read from the tile for the bit-to-associated bit comparison.

As is known, integrated circuits use built-in self-test ("BIST") circuitry for testing. However, using BIST circuitry for testing memory cells, such as for example thinly-capacitively-coupled thyristor ("TCCT") memory cells, has heretofore not provided sufficient programmability and granularity.
Accordingly, it would be desirable and useful to provide programmable BIST ("PBIST") circuitry for testing TCCT memory cells with enhanced granularity.

Programmable built-in self-test architecture


The invention relates generally to the use of pharmaceutical compositions to increase milk production alone or in combination with certain biological active ingredients. Specifically, the method relates to the use of pharmaceutical compositions that will act on the feedback of the intrinsic regulatory pathway in the mammalian mammary gland. The present invention provides for as a method of increasing bovine milk production as well as a method of correcting certain human lactation abnormalities. Preferably, the compounds used in the methods of the present invention are one or more active agents capable of inhibiting peripheral aromatic amino acid decarboxylase (AADC) enzymes, peripheral tryptophan hydroxylase (TPH) enzymes, peripheral serotonin (5-HT) enzymes, or a combination of enzymes thereof.

Basically, lactation is regulated by a dynamic balance between endocrine stimulation and feedback inhibition from local "milk stasis" signals (Wilde, C. J., Knight, C. H. & Flint, D. J. Control of milk secretion and apoptosis during mammary involution. J. Mammary Gland Biol Neoplasia 4, 129-136. (1999)). Milk stasis feedback may contribute to lactational insufficiency in women, and is an important limiting factor in the production of milk in dairy animals. Various serotonin receptor genes are expressed in mammary cells, and therefore the components of an autocrine/paracrine serotonin network are present in the mammary gland. The present invention provides for a method that disrupts serotonin synthesis and signaling causing elevated milk protein gene expression and milk yield. Therefore, the present invention provides for methods for manipulating milk production in women, and in non-human animals.
Serotonin, or 3-(β-aminoethyl)-5-hydroxyindole (5-hydroxytryptophan, or "5-HT") is a neurotransmitter in the central nervous system which is known to play an important role in the pathogenesis of affective illness. Serotonin acts through specific cellular receptors that comprise at least 15 individual subtypes. These receptors can be grouped into at least four families based on structural and functional characteristics. The 5-HT1, 5-HT2 and 5-HT4 families are G-protein coupled receptors linked to enzymatic and electrical effector systems, while the 5-HT3 receptor is a gated ion channel.
The biosynthesis of 5-HT in the brain proceeds from the uptake of dietary tryptophan by the neuron, followed by its conversion to 5-hydroxytryptophan (abbreviated 5-HTP) by an enzyme called tryptophan hydroxlyase. 5-HTP is decarboxylated to 5-HT by an enzyme called L-aromatic amino acid decarboxylase (AADC). When administered to animals or humans, 5-HTP is rapidly decarboxylated by an enzyme called L-amino acid decarboxylase also commonly called peripheral decarboxylase. Carbidopa is an inhibitor of L-aromatic amino acid decarboxylase. Carbidopa does not enter the brain, and thus administration of carbidopa selectively blocks the conversion of 5-HTP to 5-HT in the periphery, but not the brain.
It has now been discovered that administration of various serotonin agonists and antagonists is effective in increasing milk production as well as a method of correcting certain mammalian lactation abnormalities.
Accordingly, in a preferred embodiment, the present invention is directed to the administration of active agents capable of inhibiting peripheral aromatic amino acid decarboxylase (AADC) enzymes, peripheral tryptophan hydroxylase (TPH) enzymes, peripheral serotonin (5-HT) enzymes, or a combination thereof for increasing milk production as well as for correcting certain mammalian lactation abnormalities.


An article for framing a visual work includes a framing member, the rear surface of which has first and second adhesive portions. A transparent sheet adheres to the first portion and a bifurcated cling film adheres to the second adhesive portion. A pocket is formed between the sheet and the film for receiving the visual work. A method for making an article for framing a visual work includes providing a framing member having a rear surface and adhering a transparent sheet on the first adhesive portion of such surface. A cling film is mounted on a second adhesive portion of the rear surface and in a manner such that the cling film overlays the transparent sheet. The cling film is bifurcated by slitting it, thereby providing an access slot for inserting the visual work between the sheet and the film.

It is common to employ articles of any of many available types to display and protect visual works such as art, pictures, photographs, certificates and the like. Perhaps traditional picture frames are the most common articles used for these purposes. The reasons people display visual works are equally varied. They are reminders of personal successes, loved ones or past events which are pleasurable to recall.
With the already-substantial and growing popularity of photography, it is increasingly common for people to display postcards and informal candid photos of friends, pets, or scenery and to change the displayed work with some frequency. Not all articles used for such purposes are configured with frequency of change and ease-of-use in mind.
For example, U.S. Pat. No. 3,341,961 (Shanks) discloses a picture frame in which a picture is retained by adhesive applied to the rear side of the frame. Whenever adhesive is used and a picture retained thereby is removed, there is a possibility that adhesive will stick to the displayed surface of the picture. If the adhesive is very "aggressive," there is even the possibility that attempted removal will tear the picture. In addition, positioning the picture may be more difficult when adhesive retention is used. And the frame disclosed in the Shanks patent includes no overlay to protect the front of the picture.
The picture mount disclosed in U.S. Pat. No. 3,987,569 (Chase) also uses adhesive to secure a picture. The mount incorporates a feature whereby the picture may be properly aligned before securing it to the adhesive.
A type of picture mount is disclosed in U.S. Pat. No. 3,707,053 (Itano) and includes a rigid front panel made of, e.g., glass, and an apertured panel behind the front panel to receive a picture. A back panel cooperates with the other panels and "captures" the picture therebetween. Stud-like connectors are used to hold the assembly together. The mount totally protects the picture at the front, rear and sides. While this mount is probably satisfactory for its intended purpose, it seemingly would have a cost (and therefore selling price) which is unacceptably high for casual purchase and casual display of visual works.
A framing article and method which address shortcomings and disadvantages of the prior art would be an important advance in the field.


A moisture sensing apparatus includes an oscillator for supplying an AC signal to a driven ring of a sensor having a pad inside the ring, and a ground plate around the ring. An amplifier forms part of a temperature-compensated, precision rectifier, and is connected to the pad. A first diode connected to the output of the amplifier in a feedback loop of the amplifier, where the amplifier nullifies temperature variation effects upon the diode, and supplies a rectified signal which is a function of moisture content of material in contact with the sensor. A second diodes is connected between the amplifier output and other input of the amplifier. The sensor has a rounded configuration to avoid electrostatic field fringing effects, and a passive impedance with both resistive and capacitive members is connected in parallel with the sensor to provide a functional baseline for the rectified signal.

The present invention relates generally to the field of moisture detectors, and, in particular, to a novel mechanism and method of signal processing for sensing of moisture in building construction materials, including wood, concrete, gypsum, roofing felt, various flooring and other materials.
Although the approach illustrated is intended for use in moisture sensing by surface contact with a measurement sample, this is the preferred embodiment for the invention, with variations possible in sensor shape and configuration, and electronic circuitry described herein, known to those skilled in the art to be considered as integral to the spirit and scope of this disclosure.
It is an object of this disclosure to illustrate novelty of the design approach and reading stability offered by the same.
Many approaches to capacitive moisture measurement have been utilized over a substantial number of years.
Virtually all rely upon creation of an electrostatic field within the sample undergoing moisture measurement, where the alternating current potential component between two electrodes within the field is measured. Such an arrangement is described in U.S. Pat. No. 3,967,197.
Alternatively, some designs utilize a three electrode configuration, where a form of voltage division occurs between a driven and common or ground electrode, and a receiving electrode via the induced electrostatic field. Often this alternative geometry is implemented in the form of a center receiving electrode, surrounded by a co-planar driven or transmitting ring, in turn surrounded by a coplanar ground plane surface. Such an arrangement is described in U.S. Pat. No. 5,486,815, which specifically utilizes a rectangular arrangement of the stated three electrode implementation. Other approaches use a multiplicity of electrodes for specific measurement purposes.
Electronic circuitry for sensor excitation and reading of sensor output generally consists of an oscillator for excitation, and a diode detector for development of a DC voltage related to peak, average, or other parameter which relates the AC component to a DC value required for display purposes, be the display analog or digital. Commonly the diode detector is temperature compensated to an extent in order to minimize accuracy degradation due to the well known phenomenon of diode voltage drop dependence upon temperature. In silicon diodes, this effect is approximately −2.2 millivolts per degree Celsius. In virtually all instances, another diode or PN junction of a bipolar transistor is used for compensation, but due to circuit design constraints, it cannot be employed in such a way that the current versus time profile is exactly the same as the measurement rectification diode.
While the subject profile in each of the diodes or other PN junction devices can be quite similar, the profiles cannot be exactly the same, or no way will exist for extraction of a DC signal from the receiving electrode of the sensor. Dissimilarities between diode current profiles, while small, become meaningful due to the need for implementing temperature compensation involving very small signals. In some instances of instrument operation, there are differences of only several millivolts in sensor signal output when sensing moisture content of certain relatively dry materials. Under such conditions, temperature of the measurement instrument may meaningfully influence moisture reading accuracy. Indeed, some design approaches totally ignore the degrading effects of diode temperature. One example is found in U.S. Pat. No. 4,733,166, where a diode 82 in FIG. 4 thereof, serves as an RF amplitude detector, but is not voltage drop or temperature compensated.
A need remains for an improved moisture detector, particularly one with improved temperature independence and improved moisture signal sensitivity.