Staphylococcus aureus virulence genes are identified, thereby allowing the identification of novel anti-bacterial agents that target these virulence genes and their products, and the provision of novel S. aureus mutants useful in vaccines.

The invention relates generally to the identification of genes responsible for the virulence of Staphylococcus bacteria, thereby allowing the identification of new anti-bacterial agents that target these virulence genes and their products and the provision of novel S. aureus mutants useful in vaccines.
The staphylococci, of which Staphylococcus aureus is the most important human pathogen, are hardy, gram-positive bacteria that colonize the skin of most humans. Staphylococcal strains that produce coagulase are designated S. aureus; other clinically important coagulase-negative staphylococci are S. epidermidis and S. saprophyticus. When the skin or mucous membrane barriers are disrupted, staphylococci can cause localized and superficial infections that are commonly harmless and self-limiting. However, when staphylococci invade the lymphatics and the blood, potentially serious complications may result, such as bacteremia, septic shock, and serious metastatic infections, including endocarditis, arthritis, osteomyelitis, pneumonia and abscesses in virtually any organ. Certain strains of S. aureus produce toxins that cause skin rashes, food poisoning, or multisystem dysfunction (as in toxic shock syndrome). S. aureus and S. epidermidis together have become the most common cause of nosocomial non-urinary tract infection in U.S. hospitals. They are the most frequently isolated pathogens in both primary and secondary bacteremias and in cutaneous and surgical wound infections. See generally Harrison""s Principles of Internal Medicine, 13th ed., Isselbacher et al., eds., McGraw-Hill, New York (1994), particularly pages 611-617.
Transient colonization of the nose by S. aureus is seen in 70 to 90 percent of people, of which 20 to 30 percent carry the bacteria for relatively prolonged periods of time. Independent colonization of the perineal area occurs in 5 to 20 percent of people. Higher carriage rates of S. aureus have been documented in persons with atopic dermatitis, hospital employees, hospitalized patients, patients whose care requires frequent puncture of the skin, and intravenous drug abusers.
Infection by staphylococci usually results from a combination of bacterial virulence factors and a diminution in host defenses. Important microbial factors include the ability of the staphylococcus to survive under harsh conditions, its cell wall constituents, the production of enzymes and toxins that promote tissue invasion, its capacity to persist intracellularly in certain phagocytes, and its potential to acquire resistance to antimicrobials. Important host factors include an intact mucocutaneous barrier, an adequate number of functional neutrophils, and removal of foreign bodies or dead tissue.
Cell wall components of S. aureus include a large peptidoglycan complex that confers rigidity on the organism and enables it to survive under unfavorable osmotic conditions, a unique teichoic acid linked to peptidoglycan, and protein A, which is found both attached to peptidoglycan over the outermost parts of the cell and released in soluble form. Proteins designated femA and femB are involved in the formation of cell wall peptidoglycan pentaglycine cross-bridges and are factors in methicillin resistance. [Berger-Bachi et al., Mol. Gen. Genet., 219:263-269 (1989).] S. aureus also has specific receptors for laminin and fibronectin that may mediate the organism""s spread through the bloodstream to other tissues. Both peptidoglycan and teichoic acid are capable of activating the complement cascade via the alternative pathway. S. aureus also appears to activate tissue factor in the coagulation pathway.
Certain enzymes produced by S. aureus may play a role in virulence. Catalase degrades hydrogen peroxide and may protect the organism during phagocytosis. Coagulase is present in both soluble and cell-bound forms and causes plasma to clot by formation of thrombin-like material. The high correlation between coagulase production and virulence suggests that this substance is important in the pathogenesis of staphylococcal infections, but its precise role as a determinant of pathogenicity has not been determined. Many strains also produce hyaluronidase, an enzyme that degrades hyaluronic acid in the connective tissue matrix and that may promote spreading of infection. A trypsin-like protease from some strains enhances influenza virus infection by proteolytic cleavage of the viral precursor hemagglutinin into its active fragments and may contribute to the morbidity of such coinfections.
S. aureus produces numerous extracellular exotoxins that have been implicated in disease processes. The exfoliatin toxins A and B, the staphylococcal enterotoxins, and the toxic shock syndrome toxin, TSST-1, belong to the growing family of microbial superantigens that activate T cells and monocytes/macrophages, resulting in the production of cytokines that mediate local or systemic effects depending on the amount of toxin formed, the immune status of the host, and the access of the toxin to the circulation. The exfoliatin toxins mediate the dermatologic manifestations of the staphylococcal scalded-skin syndrome and bullous impetigo. These toxins cause intraepidermal cleavage of the skin at the stratum granulosum, leading to bullae formation and denudation. Seven distinct enterotoxins (A, B, C1, C2, C3, D, and E) have been implicated in food poisoning due to S. aureus. These toxins enhance intestinal peristalsis and appear to induce vomiting by a direct effect on the central nervous system. Toxic shock syndrome (TSS) is most commonly mediated by TSST-1, which is present in 5 to 25 percent of clinical isolates of S. aureus. TSS is also mediated less frequently by enterotoxin B and, rarely, enterotoxin C1.
S. aureus produces other toxins whose role in virulence is incompletely understood. Four different red blood cell hemolysins, which are designated alpha, beta, gamma, and delta toxins, have been identified. Alpha toxin also causes necrosis of the skin when injected subcutaneously into animals, while delta toxin also inhibits water absorption in the intestines and may play a role in the acute watery diarrhea seen in some cases of staphylococcal infection. Leukocidin lyses granulocyte and macrophage membranes by producing membrane pores permeable to cations.
The agr, xpr, sae and sar genes have been identified as being involved in the regulation of staphylococcal exotoxins. See U.S. Pat. No. 5,587,228 and International Patent Publication Nos. WO 96/10579 and WO 97/11690. Of interest is the report in WO 97/11690 of screening for inhibitors of these regulatory systems.
Staphylococci can invade the skin or mucosa through plugged hair follicles and sebaceous glands or areas traumatized by burns, wounds, abrasions, insect bites, or dermatitis. Staphylococci often colonize prosthetic devices and intravenous catheters; S. aureus infection of the vascular access site is a major cause of morbidity and death among patients on hemodialysis. Colonization and invasion of the lungs may occur with endotracheal intubation, or when the lungs"" clearance mechanisms are depressed, e.g., after viral infections, after aspiration, or in patients with cystic fibrosis. Mucosal damage to the gastrointestinal tract following cytotoxic chemotherapy or radiotherapy predisposes to invasion from that site.
Once the skin or mucosa have been breached, local bacterial multiplication is accompanied by inflammation, neutrophil accumulation, tissue necrosis, thrombosis and fibrin deposition at the site of infection. Later, fibroblasts create a relatively avascular wall about the area. When host mechanisms fail to contain the cutaneous or submucosal infection, staphylococci may enter the lymphatics and the bloodstream. Common sites of metastatic spread include the lungs, kidneys, cardiac valves, myocardium, liver, spleen, bones and brain.
Bacteremia due to S. aureus may arise from any local infection, at either extravascular (cutaneous infections, burns, cellulitis, osteomyelitis, arthritis) or intravascular foci (intravenous catheters, dialysis access sites, intravenous drug abuse). Rarely, patients with bacteremia die within 12 to 24 hours with high fever, tachycardia, cyanosis, and vascular collapse. Disseminated intravascular coagulation may produce a disease mimicking meningococcemia. Commonly, the disease progresses more slowly, with hectic fever and metastatic abscess formation.
A major complication of S. aureus bacteremia is endocarditis. S. aureus is the second most common cause of endocarditis and the most common cause among drug addicts. The disease is typically acute, with high fever, progressive anemia, and frequent embolic and extracardiac septic complications. Valve ring and myocardial abscesses are common. The mortality rate is 20 to 30 percent.
Staphylococcal scalded-skin syndrome (SSSS) is a generalized exfoliative dermatitis that is a complication of infection by exfoliatin toxin-producing strains of S. aureus. The disease typically occurs in newborns (Ritter""s disease) and in children under the age of five. A scarlatiniform rash begins in the perioral area, becomes generalized over the trunk and extremities, and finally desquamates. The disease may consist of rash alone (staphylococcal scarlet fever), or large, flaccid bullae develop that may be localized (more common in adults) or generalized. The bullae burst, resulting in red, denuded skin resembling a burn. Most adults with SSSS are immunosuppressed or have renal insufficiency. Blood cultures are frequently positive, and mortality is significant.
Toxic shock syndrome (TSS) is a multisystem disease mediated by toxins (generally TSST-1, and less frequently enterotoxins B and C1) produced by certain strains of S. aureus. It was first described in children, but in 1980 became epidemic among young women, with onset during menstruation. The diagnosis of TSS is based on clinical criteria that include high fever, a diffuse rash that desquamates on the palms and soles over the subsequent one or two weeks, hypotension that may be orthostatic, and evidence of involvement in three or more organ systems. Such involvement commonly includes gastrointestinal dysfunction (vomiting or diarrhea), renal or hepatic insufficiency, mucous membrane hyperemia, thrombocytopenia, myalgias with elevated creatine phosphokinase (CK) levels, and disorientation with a normal cerebrospinal fluid examination. The mortality rate of TSS is three percent.
S. aureus causes approximately three percent of community-acquired bacterial pneumonias. This disease occurs sporadically except during influenza outbreaks, when staphylococcal pneumonia is relatively more common, although still less frequent than pneumococcal pneumonia. Primary staphylococcal pneumonia in infants and children frequently presents with high fever and cough. Multiple thin-walled abscesses are seen on the chest X-ray, and empyema formation is common. In older children and healthy adults, staphylococcal pneumonia is generally preceded by an influenza-like respiratory infection. Onset of staphylococcal involvement is abrupt, with chills, high fever, progressive dyspnea, cyanosis, cough, pleural pain, and sometimes bloody sputum. Staphylococcal pneumonia is seen more frequently in patients with cystic fibrosis, in intubated patients in intensive care units and in debilitated patients who are prone to aspiration.
S. aureus is responsible for the majority of cases of acute osteomyelitis. Although the disease is most common in people under the age of 20, it is becoming increasingly prevalent in adults over 50, particularly with involvement of the spine. A primary portal of entry is frequently not identified, although many patients give a history of preceding trauma to the involved area. Once established, infection spreads through the bone to the periosteum or along the marrow cavity. Rarely, the joint capsule is penetrated, producing pyogenic arthritis. Osteomyelitis in children may present as an acute process beginning abruptly with chills, high fever, nausea, vomiting, and progressive pain at the site of bony involvement.
S. aureus causes 1 to 9 percent of cases of bacterial meningitis and 10 to 15 percent of brain abscesses. Most commonly, the bacteria are spread from a focus outside the central nervous system, typically from infective endocarditis, by extension from a paraspinal or parameningeal abscess, or by nosocomial infection following neurosurgical procedures. Over 50 percent of epidural abscesses are due to S. aureus; up to half of these cases may be associated with vertebral osteomyelitis. Patients present with either acute or chronic back pain, usually with low-grade fever and malaise. The onset of radicular pain is an ominous sign that the disease may progress to neurologic dysfunction and ultimate paralysis.
Antimicrobial resistance by staphylococci favors their persistence in the hospital environment. Over 90 percent of both hospital and community strains of S. aureus causing infection are resistant to penicillin. This resistance is due to the production of xcex2-lactamases enzymes; the genes for these enzymes are usually carried by plasmids. Infections due to organisms with such acquired resistance can sometimes be treated with penicillinase-resistant xcex2-lactam antimicrobial agents. However, the true penicillinase-resistant S. aureus organisms, called methicillin-resistant S. aureus (MRSA), are resistant to all the xcex2-lactam antimicrobials as well as the cephalosporins. MRSA resistance is chromosomally mediated and involves production of an altered penicillin-binding protein (PBP 2a or PBP 2xe2x80x2) with a low binding affinity for xcex2-lactams. MRSA frequently also have acquired plasmids mediating resistance to erythromycin, tetracycline, chloramphenicol, clindamycin, and aminoglycosides. MRSA have become increasingly common worldwide, particularly in tertiary-care referral hospitals. In the United States, approximately 5 percent of hospital isolates of S. aureus are methicillin-resistant.
Thus, there continues to exist a need for new agents useful for treating bacterial infections, particularly those caused by antibiotic-resistant bacteria, and for methods of identifying such new agents. Such methods ideally would identify agents that are unrelated to existing antimicrobials and that target different aspects of staphylococcal invasion of and replication in the host, compared to existing antimicrobials.
The present invention relates generally to the identification of genes responsible for the virulence of Staphylococcus bacteria, thereby allowing the identification of new anti-bacterial agents that target these virulence genes and their products and the provision of novel S. aureus mutants useful in vaccines.
According to one aspect of the present invention, methods are provided for identifying anti-bacterial agents that target the function of staphylococcal virulence genes or gene products. Such methods include assaying potential agents for the ability to interfere with expression of virulence gene products represented by the DNA sequences set forth in any one of SEQ ID NOS: 1 through 94, or assaying potential agents for the ability to interfere with the function of a bacterial protein encoded in whole or in part by a DNA sequence set forth in any one of SEQ ID NOS: 1 through 94 or the complementary strand thereof, followed by identifying agents that are positive in such assays.
The use of a number of different assays is contemplated according to this aspect of the invention. When the function of the virulence gene product is known or predicted by sequence similarity to a known gene product, potential inhibitors can be screened in enzymatic or other types of assays keyed to the function of the gene product. When the virulence gene product is known or predicted by sequence similarity to a known gene product to interact with another protein or nucleic acid, inhibitors of this interaction can be screened directly in binding assays or using the two-hybrid assay. Other assays may be used when a ligand for the virulence gene product is not known, including two-hybrid screening assays that identify gene products that interact with target protein, assays that identify ligands of target protein through measuring of direct binding of test ligand to target protein, and assays that identify ligands of target proteins through affinity ultrafiltration with ion spray mass spectroscopy/HPLC methods or other physical and analytical methods.
In another aspect of this invention, methods are provided for assaying potential agents for the ability to interfere with expression of or function of virulence gene products, wherein the virulence genes encoding these products are obtainable by identification through signature-tagged mutagenesis as defined herein and exemplified in Example 1.
According to a further aspect of this invention, novel anti-bacterial agents identified by the methods described herein are provided, as well as methods for treating a subject suffering from infection with staphylococci involving administration of such novel anti-bacterial agents. In particular, agents that interfere with the expression of virulence gene products include anti-sense polynucleotides that are complementary to the virulence gene sequences. Agents that interfere with the function of virulence gene products include variants of virulence gene products, ligands of these virulence gene products and variants thereof, and enzyme inhibitors (where the product is an enzyme).
Yet a further aspect of this invention provides Staphylococcus aureus organisms containing a functional mutation in a gene represented by any one of SEQ ID NOS: 1 through 94, said functional mutation resulting in a reduction in virulence of the organism. Also contemplated are vaccine compositions comprising such mutated S. aureus organisms, optionally comprising a suitable adjuvant and a pharmaceutically acceptable diluent or carrier.
Numerous additional aspects and advantages of the invention will become apparent to those skilled in the art upon consideration of the following detailed description of the invention which describes presently prepared embodiments thereof.
xe2x80x9cVirulence genes,xe2x80x9d as used herein, are genes whose function or products are required for successful establishment and/or maintenance of bacterial infection in a host animal. Thus, virulence genes and/or the proteins encoded thereby are involved in pathogenesis in the host organism, but may not be necessary for growth in vitro. Since antibiotics are typically screened in vitro, identification of these in vivo virulence genes provides a means for discovering new antimicrobials with different targets and mechanisms of action compared to existing antibiotics. There may be 50 to 100 virulence genes in S. aureus [see Groisman and Ochman, Trends Microbiol. Sci., 2:289-294 (1984) (discussing Salmonella virulence genes); Muhldorfer and Hacker, Microb. Pathogenesis, 16:171-181 (1994) (discussing E. coli virulence genes].
xe2x80x9cSignature-tagged mutagenesis,xe2x80x9d as used herein, is a method generally described in International Patent Publication No. WO 96/17951, incorporated herein by reference, and includes, for example, a method for identifying S. aureus genes required for virulence in a murine model of bacteremia. In this method, each insertional mutation carries a different DNA signature tag which allows mutants to be differentiated from each other. The tags comprise 40-bp variable central regions flanked by invariant xe2x80x9carmsxe2x80x9d of 20-bp which allow the central portions to be co-amplified by polymerase chain reaction (PCR). Tagged mutant strains are assembled in microtitre dishes, then combined to form the xe2x80x9cinoculum poolxe2x80x9d for infection studies. At an appropriate time after inoculation, bacteria are isolated from the animal and pooled to form the xe2x80x9crecovered pool.xe2x80x9d The tags in the recovered pool and the tags in the inoculum pool are separately amplified, labelled, and then used to probe filters arrayed with the different tags representing the mutants in the inoculum. Mutants with attenuated virulence are those with tags that give hybridization signals when probed with tags from the inoculum pool but not when probed with tags from the recovered pool.
Signature-tagged mutagenesis allows a large number of insertional mutant strains to be screened simultaneously in a single animal for loss of virulence. Screening thirteen pools of 96 mutant S. aureus strains resulted in the identification of 50 strains with reduced virulence, many of which were confirmed to be attenuated in virulence by subsequent analysis of individual mutants. The nucleotide sequences of the regions flanking the transposon insertion points of these mutants were analyzed by searching DNA and protein sequence databases to identify the genes inactivated by the insertion of the transposon.
On the basis of these searches many of the virulence genes may be grouped into different classes. The first class encodes proteins involved in cell surface metabolism (e.g., P2C73, P11C29, P13C83, P9B65, P10B89). Both femA and femB, which are involved in the formation of cell wall peptidoglycan pentaglycine cross bridges, were identified as virulence genes. Mutant P2C73 contains a transposon insertion in a previously unknown gene that shares significant similarity to femB. Mutant P14C15 contains a transposon insertion in a gene whose product is similar to aspartate semialdehyde dehydrogenase (Asd) from different bacteria, with the highest level of similarity to Asd from Bacillus subtilis. Asd is a key enzyme in the biosynthesis of methionine, threonine, isoleucine, lysine and diaminopimelic acid (DAP), which is an important component of cell wall peptidoglycan.
The second class encodes enzymes involved in cellular biosynthetic pathways (e.g., P9B74, P5C4, P9B66, P14C15, P13B26, P7C18, P15C31, P10B18, P6B18, P10B66, P10C34, P12C3). Deduced protein products of two genes (mutants P7C18 and P13B26) show strong similarity to B. subtilis LysA and ThrB. These enzymes, like Asd, are involved in aspartate metabolism. LysA is diaminopimelate decarboxylase, which converts diaminopimelate to lysine, and ThrB phosphorylates homoserine before conversion into threonine. Transposon insertions were also obtained in genes homologous to Methanococcus jannaschii trpA, Lactococcus lactis trpB and L. lactis trpD. These genes encode enzymes of the tryptophan biosynthetic pathway: the alpha chain of tryptophan synthetase, the beta chain of tryptophan synthetase, and anthranilate phosphoribosyltransferase, respectively. The gene mutated in P15C31 is a homolog of L. lactis purL encoding phosphoribosylformylglycinamidine decarboxylase, an enzyme of the purine biosynthetic pathway. Mutant P9B66 contains an insertion in a gene whose product is similar to peptide methionine sulphoxide reductases.
A third class of genes are those encoding components of the TCA cycle (e.g., mutants P4C27, P4C52, P10B2, P10C20, P12C32). Strains P10B2 and P12C32 carry mutations in genes for a subunit of the oxoglutarate dehydrogenase complex and aconitase, respectively.
The fourth class is composed of genes whose products are similar to a oligopeptide transport proteins of the ATP-binding cassette (ABC) transporter superfamily (e.g., mutants P7C26, P10C15, P5C3, P11C66, P5C34). Oligopeptide transport is important for peptide utilization and the proteolytic system in lactococci. In Group A streptococci, Opp proteins are involved not only in obtaining nutrients but also in adherence, protease production and processing of secreted proteins.
The fifth class of genes are involved in cellular regulatory and repair processes (e.g., mutants P4C15, P13B74, P13C72, P10B30, P6C63, P14B25). Mutant P4C15 and P6C63 contain insertions in S. aureus MarR/LuxR-like regulatory proteins. MarR and LuxR belong to a family of transcription regulators and these MarR/LuxR-like proteins likely have a similar function in S. aureus. In Streptococcus pneumoniae, Neisseria gonorrhoeae and Escherichia coli this enzyme helps to maintain surface adhesins in their functional oxidative state. Mutant P10B30 is associated with a transposon insertion in a gene with a product similar to the ATP-dependent Clp protease of E. coli. The Clp stress response system for intracellular protein degradation is widely conserved in bacteria and components of the system are important for virulence of Listeria monocytogenes and S. typhimurium. Mutants P13B74 and P13C72 have stem-loop termination sequences which possibly function in transcription termination.
The identification of these genes as virulence genes renders them useful in methods of identifying new anti-bacterial agents according to the present invention. Such methods include assaying potential agents for the ability to interfere with expression of virulence gene products represented by the DNA sequences set forth in any one of SEQ ID NOS: 1 through 94 (i.e., the genes represented by DNA sequences of SEQ ID NOS: 1 through 94 encode the virulence gene product, or the DNA sequences of SEQ ID NOS: 1 through 94 are adjacent to the gene encoding the virulence gene product, or are involved in regulation of expression of the virulence gene product), or assaying potential agents for the ability to interfere with the function of a bacterial protein encoded in whole or in part by a DNA sequence set forth in any one of SEQ ID NOS: 1 through 94 or the complementary strand thereof, followed by identifying agents that are positive in such assays. Polynucleotides and polypeptides useful in these assays include not only the genes and encoded polypeptides as disclosed herein, but also variants thereof that have substantially the same activity as the wild-type genes and polypeptides. xe2x80x9cVariants,xe2x80x9d as used herein, includes polynucleotides or polypeptides which contain one or more deletions, insertions or substitutions, as long as the variant retains substantially the same activity of the wild-type polynucleotide or polypeptide. With regard to polypeptides, deletion variants are contemplated to include fragments lacking portions of the polypeptide not essential for biological activity, and insertion variants are contemplated to include fusion polypeptides in which the wild-type polypeptide or fragment thereof have been fused to another polypeptide.
The virulence genes may be cloned by PCR, using S. aureus genomic DNA as the template. For ease of inserting the gene into expression vectors, PCR primers are chosen so that the PCR-amplified gene has a restriction enzyme site at the 5xe2x80x2 end preceding the initiation codon ATG, and a restriction enzyme site at the 3xe2x80x2 end after the termination codon TAG, TGA or TAA. If desirable, the codons in the gene are changed, without changing the amino acids, according to E. coli codon preference described by Grosjean and Fiers, Gene, 18:199-209 (1982), and Konigsberg and Godson, Proc. Natl. Acad. Sci. (USA), 80:687-691 (1983). Optimization of codon usage may lead to an increase in the expression of the gene product when produced in E. coli. If the gene product is to be produced extracellularly, either in the periplasm of E. coli or other bacteria, or into the cell culture medium, the gene is cloned without its initiation codon and placed into an expression vector behind a signal sequence. For example, cloning and expression of the femA gene is described in Example 3 below.
To simplify the protein purification process, a purification tag may be added either at the 5xe2x80x2 or 3xe2x80x2 end of the gene coding sequence. Commonly used purification tags include a stretch of six histidine residues (U.S. Pat. Nos. 5,284,933 and 5,310,663), a streptavidin-affinity tag described by Schmidt and Skerra, Protein Engineering, 6:109-122 (1993), a FLAG peptide [Hopp et al., Biotechnology, 6:1205-1210 (1988)], glutathione S-transferase [Smith and Johnson, Gene, 67:31-40 (1988)], and thioredoxin [LaVallie et al., Bio/Technology, 11:187-193 (1993)]. To remove these peptide or polypeptides, a proteolytic cleavage recognition site may be inserted at the fusion junction. Commonly used proteases are factor Xa, thrombin, and enterokinase.
Proteins are produced in any number of well-known prokaryotic or eukaryotic expression systems using known promoters, vectors, and hosts. Any suitable host cell may be used for expression of the gene product, such as E. coli, other bacteria, including Bacillus and S. aureus, yeast, including Pichia pastoris and Saccharomyces cerevisiae, insect cells, or mammalian cells, including CHO cells, utilizing suitable vectors known in the art. Proteins may be produced directly or fused to a peptide or polypeptide, and either intracellularly or extracellularly by secretion into the periplasmic space of a bacterial cell or into the cell culture medium. Secretion of a protein requires a signal peptide (also known as pre-sequence); a number of signal sequences from prokaryotes and eukaryotes are known to function for the secretion of recombinant proteins. During the protein secretion process, the signal peptide is removed by signal peptidase to yield the mature protein.
The virulence gene products produced by the methods described above are used in high throughput assays to screen for inhibitory agents. The sources for potential agents to be screened are chemical compound libraries, fermentation media of Streptomycetes, other bacteria and fungi, and cell extracts of plants and other vegetations. For proteins with known enzymatic activity, assays are established based on the activity, and a large number of potential agents are screened for ability to inhibit the activity. For proteins that interact with another protein or nucleic acid, binding assays are established to measure such interaction directly, and the potential agents are screened for ability to inhibit the binding interaction.
Alternatively, such binding interactions are evaluated indirectly using the yeast two-hybrid system described in Fields and Song, Nature, 340:245-246 (1989), and Fields and Sternglanz, Trends in Genetics, 10:286-292 (1994), both of which are incorporated herein by reference. The two-hybrid system is a genetic assay for detecting interactions between two proteins or polypeptides. It can be used to identify proteins that bind to a known protein of interest, or to delineate domains or residues critical for an interaction. Variations on this methodology have been developed to clone genes that encode DNA-binding proteins, to identify peptides that bind to a protein, and to screen for drugs. The two-hybrid system exploits the ability of a pair of interacting proteins to bring a transcription activation domain into close proximity with a DNA-binding domain that binds to an upstream activation sequence (UAS) of a reporter gene, and is generally performed in yeast. The assay requires the construction of two hybrid genes encoding (1) a DNA-binding domain that is fused to a protein X, and (2) an activation domain fused to a protein Y. The DNA-binding domain targets the first hybrid protein to the UAS of the reporter gene; however, because most proteins lack an activation domain, this DNA-binding hybrid protein does not activate transcription of the reporter gene. The second hybrid protein, which contains the activation domain, cannot by itself activate expression of the reporter gene because it does not bind the UAS. However, when both hybrid proteins are present, the noncovalent interaction of protein X and protein Y tethers the activation domain to the UAS, activating transcription of the reporter gene. When the virulence gene product (protein X, for example) is already known to interact with another protein or nucleic acid (protein Y, for example), this assay can be used to detect agents that interfere with the interaction of X and Y. Expression of the reporter gene is monitored as different test agents are added to the system; the presence of an inhibitory agent results in lack of a reporter signal.
When the function of the virulence gene product is unknown and no ligands are known to bind the gene product, the yeast two-hybrid assay can also be used to identify proteins that bind to the gene product. In an assay to identify proteins that bind to protein X (the target protein), a large number of hybrid genes each containing a different protein Y are produced and screened in the assay. Typically, Y is encoded by a pool of plasmids in which total cDNA or genomic DNA is ligated to the activation domain. This system is applicable to a wide variety of proteins, and it is not even necessary to know the identity or function of protein Y. The system is highly sensitive and can detect interactions not revealed by other methods; even transient interactions may trigger transcription to produce a stable mRNA that can be repeatedly translated to yield the reporter protein.
Other assays may be used to search for agents that bind to the target protein. One such screening method to identify direct binding of test ligands to a target protein is described in U.S. Pat. No. 5,585,277, incorporated herein by reference. This method relies on the principle that proteins generally exist as a mixture of folded and unfolded states, and continually alternate between the two states. When a test ligand binds to the folded form of a target protein (i.e., when the test ligand is a ligand of the target protein), the target protein molecule bound by the ligand remains in its folded state. Thus, the folded target protein is present to a greater extent in the presence of a test ligand which binds the target protein, than in the absence of a ligand. Binding of the ligand to the target protein can be determined by any method which distinguishes between the folded and unfolded states of the target protein. The function of the target protein need not be known in order for this assay to be performed. Virtually any agent can be assessed by this method as a test ligand, including, but not limited to, metals, polypeptides, proteins, lipids, polysaccharides, polynucleotides and small organic molecules. For example, use of femA in this method of screening for potential ligands is described in Example 3 below.
Another method for identifying ligands for a target protein is described in Wieboldt et al., Anal. Chem., 69:1683-1691 (1997), incorporated herein by reference. This technique screens combinatorial libraries of 20-30 agents at a time in solution phase for binding to the target protein. Agents that bind to the target protein are separated from other library components by centrifugal ultrafiltration. The specifically selected molecules that are retained on the filter are subsequently liberated from the target protein and analyzed by HPLC and pneumatically assisted electrospray (ion spray) ionization mass spectroscopy. This procedure selects library components with the greatest affinity for the target protein, and is particularly useful for small molecule libraries.
The inhibitors/binders identified by the initial screens are evaluated for their effect on virulence in in vivo mouse models of S. aureus infections. Models of bacteremia, endocarditis, septic arthritis, soft tissue abscess or pneumonia may be utilized. Inhibitors/binders that interfere with bacterial virulence are capable of preventing the establishment of an infection or reversing the outcome of an infection once it is established.
The identification of S. aureus virulence genes also provides for microorganisms exhibiting reduced virulence, which are useful in vaccines. Such microorganisms include the S. aureus mutants generated in Example 1 below and other S. aureus mutants containing at least one functional mutation in a gene represented by any one of SEQ ID NOS: 1 through 94. The reduced virulence of these organisms and their immunogenicity may be confirmed by administration to a subject. While it is possible for an avirulent microorganism of the invention to be administered alone, one or more of such mutant microorganisms are preferably administered in a vaccine composition containing suitable adjuvant(s) and pharmaceutically acceptable diluent(s) or carrier(s). The carrier(s) must be xe2x80x9cacceptablexe2x80x9d in the sense of being compatible with the avirulent microorganism of the invention and not deleterious to the subject to be immunized. Typically, the carriers will be water or saline which will be sterile and pyrogen free. The subject to be immunized is a subject needing protection from a disease caused by a virulent form of S. aureus.
Any adjuvant known in the art may be used in the vaccine composition, including oil-based adjuvants such as Freund""s Complete Adjuvant and Freund""s Incomplete Adjuvant, mycolate-based adjuvants (e.g., trehalose dimycolate), bacterial lipopolysaccharide (LPS), peptidoglycans (i.e., mureins, mucopeptides, or glycoproteins such as N-Opaca, muramyl dipeptide [MDP], or MDP analogs), proteoglycans (e.g., extracted from Klebsiella pneumoniae), streptococcal preparations (e.g., OK432), Biostim(trademark) (e.g., 01K2), the xe2x80x9cIscomsxe2x80x9d of EP 109 942, EP 180 564 and EP 231 039, aluminum hydroxide, saponin, DEAE-dextran, neutral oils (such as miglyol), vegetable oils (such as arachis oil), liposomes, Pluronic(copyright) polyols or the Ribi adjuvant system (see, for example GB-A-2 189 141). Recently, an alternative adjuvant consisting of extracts of Amycolata, a bacterial genus in the order Actinomycetales, has been described in U.S. Pat. No. 4,877,612. Additionally, proprietary adjuvant mixtures are commercially available. The adjuvant used will depend, in part, on the recipient organism. The amount of adjuvant to administer will depend on the type and size of animal. Optimal dosages may be readily determined by routine methods.
The vaccine compositions optionally may include pharmaceutically acceptable (i.e., sterile and non-toxic) liquid, semisolid, or solid diluents that serve as pharmaceutical vehicles, excipients, or media. Any diluent known in the art may be used. Exemplary diluents include, but are not limited to, polyoxyethylene sorbitan monolaurate, magnesium stearate, methyl- and propylhydroxybenzoate, talc, alginates, starches, lactose, sucrose, dextrose, sorbitol, mannitol, gum acacia, calcium phosphate, mineral oil, cocoa butter, and oil of theobroma.
The vaccine compositions can be packaged in forms convenient for delivery. The compositions can be enclosed within a capsule, sachet, cachet, gelatin, paper or other container. These delivery forms are preferred when compatible with entry of the immunogenic composition into the recipient organism and, particularly, when the immunogenic composition is being delivered in unit dose form. The dosage units can be packaged, e.g., in tablets, capsules, suppositories or cachets.
The vaccine compositions may be introduced into the subject to be immunized by any conventional method including, e.g., by intravenous, intradermal, intramuscular, intramammary, intraperitoneal, or subcutaneous injection; by oral, sublingual, nasal, anal, vaginal, or transdermal delivery; or by surgical implantation, e.g., embedded under the splenic capsule or in the cornea. The treatment may consist of a single dose or a plurality of doses over a period of time.
It will be appreciated that the vaccine of the invention may be useful in the fields of human medicine and veterinary medicine. Thus, the subject to be immunized may be a human or an animal, for example, cows, sheep, pigs, horses, dogs and cats, and poultry such as chickens, turkeys, ducks and geese.


A four-wire electrostatic actuator including: a stator having a substrate with two surfaces and at least four main-movement linear electrodes separately provided on one surface of the substrate and arranged in parallel at regular intervals; and a movable element disposed on the stator. The first and third main-movement linear electrodes of the four main-movement linear electrodes are supplied with rectangular wave signals or sine wave signals with reversed phases. The second and fourth main-movement linear electrodes are supplied with rectangular wave signals or sine wave signals with reversed phases. The two signals inputted to the adjacent two electrodes are shifted from each other by a quarter of a period with identical strength. The stator further includes a plurality of one-side auxiliary-movement linear electrodes on one side of the four main-movement linear electrodes, the auxiliary-movement linear electrodes being extended perpendicularly to the four main-movement linear electrodes.

A conventional electrostatic actuator includes a stator provided with electrodes and a movable element having a resistance element such as a film. For such an electrostatic actuator, a driving method is used with multiphase signals.
Unfortunately, in such an electrostatic actuator, a movable element linearly moving on a stator may meander or laterally deviate due to in-plane variations in friction resistance or foreign matters.
To address this problem, a conventional electrostatic actuator is provided with a guide that prevents meandering and a lateral deviation of a movable element. However, a friction resistance between the guide and the movable element may interfere with a movement of the movable element, and friction between the guide and the movable element may cause an unnecessary electric charge. Moreover, the movable element is regulated by the position of the guide but a restoring force to the center is not applied to the movable element.
Thus, a conventional electrostatic actuator has voltage supply lines on both sides of electrodes along the direction of movement of a movable element (for example, JP03-65081). With this configuration, a moment component perpendicular to the direction of movement of the movable element is fixed by the electrostatic force of the two voltage supply lines on both sides of the electrodes. Thus, the occurrence of a rotation moment on the movable element is suppressed, preventing meandering of the movable element.
In the conventional electrostatic actuator; unfortunately, the movable element is attracted to the voltage supply lines by the electrostatic force of the voltage supply lines, increasing the friction resistance of the movable element. Since the electrostatic force of the voltage supply lines does not contain a moment component that restores the movable element to the center of a stator, a restoring force to the center of the stator is not applied to the movable element and the movable element keeps moving while being deviated from the center of the stator.

Four-wire electrostatic actuator and stator


An optical/electrical hybrid substrate is provided. The optical/electrical substrate includes: a wiring substrate; an optical waveguide disposed on the wiring substrate and configured to transmit an optical signal; a mirror support bonded onto the wiring substrate with an adhesive and being made of glass or silicon; and a mirror which reflects the optical signal and which is formed on an inclined surface of the mirror support.

1. Technical Field
The present disclosure relates to an optical/electrical hybrid substrate and its manufacturing method. More particularly, the disclosure relates to an optical/electrical hybrid substrate which includes a wiring substrate, an optical waveguide disposed on the wiring substrate, and a mirror for reflecting an optical signal, as well as to its manufacturing method.
2. Related Art
In recent years, with the increase in the speed of the information communication, light has come to be used as an information communication medium in place of an electrical signal. In such an optical communication field, it is necessary to convert an optical signal into an electrical signal and vice versa and to perform various kinds of processing such as modulation on light. To meet this requirement, optical/electrical hybrid substrates for such conversion processing are being developed.
FIG. 1 is a sectional view of the related-art optical/electrical hybrid substrate.
Referring to FIG. 1, the related-art optical/electrical hybrid substrate 200 includes a wiring substrate 201, an optical waveguide 202, mirrors 203 and 204, cladding members 206, an adhesive 207, a light-emitting element 208, a light-receiving element 209, and underfill resins 211 and 212.
The wiring substrate 201 includes a core substrate 215, through vias 216, upper traces 218, solder resist layers 219 and 223, solders 221, and lower traces 222.
The through vias 216 are provided to penetrate through the core substrate 215. The upper traces 218 are formed on a top surface 215A of the core substrate 215 and a top surface of the through vias 216. The top traces 218 have connection surfaces 218A to which a terminal 236 of the light-emitting element 208 and a terminal 238 of the light-receiving element 209 are connected. The solder resist layer 219 is formed on the top surface 215A of the core substrate 215 so as to cover parts of the top traces 218. The solder resist layer 219 has openings 219A through which the connection surfaces 218A are exposed when the openings 219A are not filled up. The solders 221 are formed in the openings 219A. The solders 221 serve to fix the terminal 236 of the light emitting element 208 and the terminal 238 of the light-receiving element 209 to the respective top traces 218.
The lower traces 222 are formed on a bottom surface 215B of the core substrate 215 and the bottom surface of the through vias 216. The lower traces 222 are electrically connected to the top traces 218 via the through vias 216, respectively. The lower traces 222 have connection surfaces 222A on which external connection terminals (not shown) are formed. The solder resist layer 223 is formed on the bottom surface 215B of the core substrate 215 so as to cover parts of the bottom traces 222. The solder resist layer 223 has openings 223A through which the connection surfaces 222A are exposed.
The optical waveguide 202 has a first cladding layer 226, a core 227, and a second cladding layer 228. The core 227, which serves to transmit an optical signal, is formed on the first cladding layer 226. The core 227 is made of a material having a larger refractive index than the first cladding layer 226 and the second cladding layer 228. The second cladding layer 228 is formed over the first cladding layer 226 so as to cover the core 227. The optical waveguide 202 has grooves 231 and 232 through which the core 227 is exposed when the grooves 231 and 232 are not filled up. The groove 231 is a V-shaped groove and has a inclined surface 231A on which the mirror 203 is formed. The inclined surface 231A is inclined so as to form a certain angle (45°, for example) with the top surface of the solder resist layer 219. The groove 232 is a V-shaped groove and has a inclined surface 232A on which the mirror 204 is formed. The inclined surface 232A is inclined so as to form a certain angle (45°, for example) with the top surface of the solder resist layer 219. The cladding members 206 fill the grooves 231 and 232 in which the mirrors 203 and 204 are formed, respectively.
The optical waveguide 202 in which the mirrors 203 and 204 and the cladding members 206 are formed is bonded to the top surface of the solder resist layer 219 with the adhesive 207.
The light-emitting element 208 has a light-emitting portion 235 and the terminal 236. The light-emitting element 208 is disposed on the wiring substrate 201 in such a manner that the light-emitting portion 235 is opposed to that part of the mirror 203 which is located in the core 227. The terminal 236 is formed on the connection surface 218A of the left-hand top trace 218. The terminal 236 is fixed to the left-hand top trace 218 with the solder 221.
The light-receiving element 209 has a light-receiving portion 237 and the terminal 238. The light-receiving element 209 is disposed on the wiring substrate 201 in such a manner that the light-receiving portion 237 is opposed to that part of the mirror 204 which is located in the core 227. The terminal 238 is formed on the connection surface 218A of the right-hand top trace 218. The terminal 238 is fixed to the right-hand top trace 218 with the solder 221.
The underfill resin 211 is formed so as to fill the space between the light-emitting element 208 and each of the wiring substrate 201 and the optical waveguide 202. The underfill resin 212 is formed so as to fill the space between the light-receiving element 209 and each of the wiring substrate 201 and the optical waveguide 202. The underfill resins 211 and 212 are made of an optically transparent resin.
FIGS. 2-8 show a manufacturing process of the above-described related-art optical/electrical hybrid substrate 200.
The manufacturing method of the related-art optical/electrical hybrid substrate 200 will be described below with reference to FIGS. 2-8. First, in a step of FIG. 2, a wiring substrate 201 is formed by a known method. Then, in a step of FIG. 3, a core 227 and a second cladding layer 228 are placed on a first cladding layer 226 in this order.
Then, in a step of FIG. 4, a groove 231 having a inclined surface 231A and a groove 232 having a inclined surface 232A are formed by processing the structure of FIG. 3 with a dicer. Then, in a step of FIG. 5, mirrors 203 and 204 are formed by forming metal films on the inclined surfaces 231A and 232A, respectively.
Then, in a step of FIG. 6, the groove 231 formed with the mirror 203 and the groove 232 formed with the mirror 204 are filled with respective cladding members 206. Then, in a step of FIG. 7, the optical waveguide 202 (the structure of FIG. 6) which is formed with the mirrors 203 and 204 and the cladding members 206 is bonded to the top surface of the solder resist layer 219 of the wiring substrate 201 shown in FIG. 2.
Subsequently, in a step of FIG. 8, the solders 221 are melted, a terminal 236 of a light-emitting element 208 and a terminal 238 of a light-receiving element 209 are fixed to the connection surfaces 218A of the upper traces 218, respectively, and underfill resins 211 and 212 are formed. The optical/electrical hybrid substrate 200 is thus completed (see e.g., JP-A-2000-304953).
As described above, in the related-art optical/electrical hybrid substrate 200, the grooves 231 and 232 in which the mirrors 203 and 204 are to be formed are formed in the optical waveguide 202 and then the mirrors 203 and 204 are formed on the inclined surfaces 231A and 232A of the grooves 231 and 232.
However, there is a problem in that it is difficult to form, with high accuracy, the inclined surfaces 231A and 232A at the certain angle (45°, for example) in the optical waveguide 202 in which the core 227 and the second cladding layer 228 are formed on the first cladding layer 226 in this order. This results in a problem that when an optical signal is transmitted via the mirrors 203 and 204 which are formed on the inclined surfaces 231A and 232A of the grooves 231 and 232, the optical signal suffers a large transmission loss.

Optical/electrical hybrid substrate and method of manufacturing the same


A mobile station UE of the present invention is a mobile station that communicates with a radio base station using equal to or greater than two carriers, wherein the equal to or greater than two carriers include a first carrier and a second carrier, the mobile station including: a first communicating unit configured to perform communication with the first carrier; and a second carrier measuring unit configured to perform measurement of the second carrier, wherein, in a case where a measurement gap for measuring the second carrier is set, the first communicating unit is configured to perform communication with the first carrier without considering the measurement gap when the second carrier is activated, and not to perform communication with the first carrier in the measurement gap when the second carrier is not activated.

As a successor of a WCDMA (Wideband Code Division Multiplexing Access) system, an HSDPA (High-Speed Downlink Packet Access) system, and an HSUPA (High-Speed Uplink Packet Access) system, an LTE (Long Term Evolution) system has been considered and standardized by 3GPP (The 3rd Generation Partnership Project), which is a standardization organization of WCDMA.
Furthermore, as a successor of the LTE system, an LTE-advanced system is under consideration by 3GPP. The requirements for the LTE-advanced system are summarized in the non-patent document 1.
As one of the requirements in the LTE-advanced system, an agreement is reached that carrier aggregation is applied. When carrier aggregation is applied, a mobile station UE can receive downlink signals simultaneously using plural carriers or transmit uplink signals simultaneously using plural carriers. Each carrier used in carrier aggregation is referred to as a "component carrier".
The plural component carriers are categorized into a primary component carrier as a main carrier and one or more secondary component carriers other than the primary component carrier.
When a mobile station UE performs communications always using the primary component carrier and the secondary component carriers, a problem arises that power consumption becomes higher in proportion to the number of component carriers. As used herein, communicating using the primary component carrier and the secondary component carriers includes usual data transmission and reception, cell search or measurement on the respective carriers, and radio link monitoring.
For example, the cell search includes establishing synchronization in downlink using downlink synchronization signals in a serving cell and an adjacent cell. Since cell search is the processing for detecting a destination cell while a mobile station UE is moving, the mobile station UE periodically needs to perform cell search. For example, the measurement includes measuring received power (more specifically, RSRP (Reference Signal Received Power) or the like) of reference signals in a serving cell and an adjacent cell. It should be noted that the combined processing of cell search and measurement may be referred to as "measurement". The radio link monitoring includes measuring radio quality (more specifically, SIR (Signal-to-Interference Ratio)) of reference signals in a serving cell, determining whether the SIR is above a predetermined threshold, and determining that the serving cell is in out-of-synchronization when the SIR is below the predetermined threshold. The processing associated with cell search, measurement, and radio link monitoring and their performance definitions are described in non-patent documents 2 and 3, for example.
In order to address the problem of power consumption, it is considered that control of activation/de-activation is applied in the secondary component carrier, for example. For example, on a secondary component carrier in a de-activation state, the mobile station UE does not perform usual data transmission and reception and reduces the frequencies of cell search, measurement, and radio link monitoring, thereby saving the battery. The processing of de-activation on a secondary component carrier is performed when the amount of data to be communicated is small, for example.
In addition, in the LTE system, a measurement gap is defined in order to perform measurement on a carrier with a different frequency or a carrier for a different radio communication system (non-patent document 4). The length of the measurement gap is defined as 6 ms and its periodicity is defined as 40 ms or 80 ms, for example. During the measurement gap, the mobile station UE suspends communications in a serving cell and performs measurement of a carrier of a different frequency or a carrier of a different radio communication system. In this case, communication with the serving cell is stopped, throughput of communication with the serving cell deteriorates.

Mobile station, radio base station and communication control method


A locking device for quarter turn valves avoids the inadvertent, unauthorized or careless manipulation of quarter turn valves from a desired position thereby minimizing, if not eliminating, the likelihood for damage or injury in an industrial or other setting as a result of the unauthorized opening/closing of the valve. The locking device includes a shroud which is adapted to be placed over the upper end of the stem and adjacent portion of the handle on the valve. The shroud has a capping portion which securely engages a mounting pad of the valve and an upwardly extending channel designed to at least partially surround a portion of the handle. A hole is provided in the channel and aligns with a hole in an adjacent portion of the handle when the shroud is placed on the valve. As such, the post of a lock is easily inserted through the aligned holes. The locking device according to this invention advantageously prevents the unauthorized manipulation of the handle and the resulting rotation of the valve member to/from the open/closed positions.

This invention relates to locking systems in general, and specifically to a locking arrangement to secure valves against intentional and unintentional adjustment.
A wide variety of types of valves are currently used in a vast array of applications. One common type of valve is known as a quarter turn valve. Quarter turn valves are used in many industrial applications, particularly in the food and beverage, pulp and paper, pharmaceutical, specialty chemical, and fine chemical industries. Quarter turn valves offer numerous advantages, including the ability to be turned on/off more quickly than linear (or multi-turn) valves such as gate valves. Quarter turn valves are often used in industrial applications because they can be turned on and left open until the required amount of fluid has passed through the valve. Conversely, quarter turn valves are maintained in a closed position until actuated to allow the fluid to pass there through.
Quarter turn valves are one type of rotary motion valve in that they have a component that rotates to allow or shut off flow through the valve. The valve member within a quarter turn valve may be a butterfly, ball, plug or other operating member to alternately permit and inhibit flow of the fluid through the valve. Quarter turn valves may be a two-way valve in which the rotary valve member blocks or permits fluid flow through a single inlet port and single outlet port. Alternatively, a multi-port quarter turn valve allows for flow into or out of multiple inlet and/or outlet ports. The ports on quarter turn valves may be flanged, threaded or welded ports for connection to appropriate pipes conveying the fluid to and from the valve.
The rotary valve member within a quarter turn valve is typically connected by a stem projecting from the valve body. Manually actuated quarter turn valves typically have an elongate handle mounted to the stem. The handle is rotated through an arc of generally 90° for adjusting the valve member to and between the open and closed positions. However, other rotary motion valves within the broad category of quarter turn valves often include rotary valves that rotate more than or less than 90°. Therefore, as used herein, the term "quarter turn valve" is defined as any valve that includes a rotary valve member movable between open and closed positions.
As previously discussed, quarter turn valves are commonly used for general purpose and industrial applications. Their popularity is due in large part to lower maintenance requirements, economical operation, ease of installation and continued savings throughout their operational life as compared to other types of valves. Nevertheless, it is generally desirable to secure any valve and, in particular, quarter turn valves in a desired orientation (open/closed). Unfortunately, tampering with industrial operations is often a great temptation to certain people, or organizations, by unauthorized manipulation of the valves to interrupt or interfere with industrial productions. Alternatively, unauthorized manipulation of quarter turn valves may not be a premeditated act intended to cause damage but may simply be the result of inadvertent misuse or innocent mistakes. Nevertheless, occupational health and safety organizations, such as OSHA and other work place and industrial oversight organizations, advocate more control over the operation of quarter turn valves. The ability to securely lock quarter turn valves in a desired position avoids many, if not all, of the above-described problems. By so doing, potentially catastrophic and/or injurious conditions can be avoided if the adjustment of quarter turn valves in industrial and other settings is limited to authorized operators only.
However, known techniques for locking or disabling quarter turn valves have proven to be deficient. Standard pad locks having a U-shaped clevis or shackle cannot be conveniently and securely installed on quarter turn valves to inhibit unauthorized operation. Moreover, such locking mechanisms offer convenient access for someone to saw or otherwise dismantle the exposed shackle portion of the lock.
Alternatively, removal of the handle or manual actuator on such valves may temporarily inhibit someone from unauthorized operation of the valve but does not prevent manipulation of the valve with a similar user-provided handle or other tool. Moreover, handle removal by the authorized user for later valve manipulation is inconvenient, inefficient and troublesome.
Therefore, a need exists for a secure, effective and easily implemented device to prevent the unauthorized, inadvertent or otherwise undesired manipulation of quarter turn valves.


An integrated prefabricated furniture system is provided for fitting-out a building room of the type having a generally open plan interior and having at least one utility source. The furniture system includes a plurality of horizontal beam segments interconnected in an end-to-end fashion to form a rigid overhead framework configured to be positioned above the floor surface. A plurality of overhead support columns each have an upper portion rigidly connected with the overhead framework, and a lower portion shaped to support the overhead framework on a floor in a freestanding fashion at a predetermined elevation above average user height. Each of the overhead support columns includes an interior support structure in a generally X-shaped plan-form defining vertical raceways between adjacent legs of the cross-section. Each support column also includes four vertical edges forming substantially planar faces therebetween, of which at least one of the vertical edges has a plurality of slots for supporting office furniture items and office accessories.

The present invention relates to furnishings, and in particular to a furniture system that is particularly adapted to support group activities in open plans, and the like.
Open office plans are well known in the art, and generally comprise large, open floor spaces in a building that are furnished in a manner that is readily reconfigurable to accommodate the ever changing needs of a specific user, as well as the divergent requirements of different tenants. One arrangement typically used for furnishing open plans includes movable partition panels that are detachably interconnected to partition off the open space into individual workstations and/or offices. Some such partition panels are configured to receive hang-on furniture units, such as worksurfaces, overhead cabinets, shelves, etc., and are generally known in the office furniture industry as "systems furniture." Another arrangement for dividing and/or partitioning open plans includes modular furniture arrangements in which a plurality of differently shaped, freestanding furniture units are interconnected in a side-by-side relationship, with upstanding privacy screens attached to at least some of the furniture units to create individual, distinct workstations and/or offices.
Such prior art partitioning arrangements create relatively permanent, multifunction workstations for the users, which workstations are required to support both individual work activities, as well as some types of group activities, such as inter-office conferences, and the like. However, these types of conventional workstation arrangements are not particularly adapted to support workers engaged in group work, such as self-managing teams, or others involved in team problem solving techniques, wherein a relatively large number of workers from different disciplines, such as engineering, design, manufacturing, sales, marketing, purchasing, finance, etc., meet together as a group to define and review issues, set general policy, and then break out into a number of smaller sub-groups or individuals to resolve those specific problems relating to their particular discipline. Team projects typically have a rather specific objective and are of a limited duration, such that the individual workers are temporarily assigned to the group for the life of the project, and are then reassigned to a new group when the project is completed. Group work is steadily gaining importance as a way of improving productivity and time-to-market, thereby emphasizing the need to support such activities more efficiently and effectively.
Conventional conference rooms, meeting halls, and the like have heretofore been required to handle such group meetings, but are typically expensive to construct and maintain, and are not usually considered an efficient use of space in open plan environments. When such conventional rooms are constructed in rented office space, they become permanent leasehold improvements, which must be depreciated over a lengthy time period, and can not be readily removed upon the expiration of the lease. The reconfiguration of such spaces is quite messy, and very disruptive to conducting day-to-day business. Furthermore, with conventional conference room arrangements, breakout meetings among the various sub-groups of workers often prove inconvenient, since the workstations of the participant workers are seldom located in close proximity to the conference room.
Another objective of furnishings from modern office environments is to promote the establishment of an optimum balance between worker privacy and worker interaction. Throughout a given workday, an office worker normally oscillates between interaction with others and time spent alone. Each such worker actively seeks out or avoids others based upon their ever changing task, objectives, and goals. Furnishings can serve to help these workers better regulate involvement with or isolation from coworkers. For example, full height offices are known for privacy. Their surrounding walls and door provide privacy by consistently controlling unwanted distractions, but often limit opportunities for spontaneous interaction. On the other hand, open offices precipitate an awareness of coworkers. Furniture and partition based workstations encourage participation and convenient access, but often lack sufficient controls for individual quiet work. Both private workspace and convenient access to coworkers for the completion of work involving group or team efforts is quite important to the overall success of such projects.
The use of displays to communicate information to large groups in office environments and the like, is also generally well-known, and includes such devices as marker boards, tackable surfaces, electronic displays, reflective projector screens, etc. Such displays are normally incorporated into conventional style conference rooms, meeting halls, and other similar facilities. However, as previously noted, such conventional meeting spaces are typically expensive, and are not usually considered a cost effective use of floor space in most modern offices. Rather, modern office layouts are typically of the open plan type, and do not include large, conventional types of conference rooms.
Information displays in modern, open plan workstations, such as those created by movable partition panels, modular furniture, or the like, are usually quite limited, and not particularly adapted to support workers engaged in group or team problem solving techniques. Due to the inherent nature of group problem solving techniques, the effective display of information is quite important to the effective management of the team's human resources.
Office space users are experiencing increasing needs for improved utilities support at each workstation or work setting. These "utilities," as the term is used herein, encompass all types of resources that may be used to support or service a worker, such as communications and data used with computers and other types of data processors, telecommunications, electronic displays, electrical power, and physical accommodations, such as lighting, signage, security, sound masking, and the like. For example, modern offices for highly skilled "knowledge workers" such as engineers, accountants, stock brokers, computer programs, etc., are typically provided with multiple pieces of very specialized computer and communications equipment that are capable of processing information from numerous local and remote data sources to assist in solving complex problems. Such equipment has very stringent power and signal requirements, and must quickly and efficiently interface with related equipment at both adjacent and remote locations. Many other types of high technology equipment and facilities are also presently being developed which will need to be accommodated in the workplaces of the future.
In order to gain increased efficiency in the use of expensive office real estate, attempts are now being made to try to support highly paid knowledge workers with modular furniture systems in open office settings, instead of conventional private offices. However, in order to ensure peak efficiency of such knowledge workers, the work settings must be equipped with the various state-of-the-art utilities and facilities. Since such work settings must be readily reconfigurable to effectively meet the ever-changing needs of the user, the distribution and control of utilities throughout a comprehensive open office plan has emerged as a major challenge to the office furniture industry. The inherent nature of modular furniture system, which permits them to be readily reconfigurable into different arrangements, makes it very difficult to achieve adequate utility distribution and control.
Hence, utility distribution and control are fast becoming one of the major issues in office fit-out and furniture. Changing technology is creating greater demands on power and signal distribution networks. Routing power and signal distribution below the floor or in furniture systems often ends up in complex idiosyncratic systems which are difficult to manage or change. There is presently an oversupply of office space and furniture systems which do not properly respond to or support change. Many older buildings do not have adequate utility capabilities and the cost of conventional renovations or improvements often renders the same in practice. Even relatively new buildings can quickly be rendered obsolete by the fast paced changes in modern technology. The refurnishing of existing building space is therefore a concern which must be addressed by furniture systems.


A device that includes a pixel array and a sample and hold circuit configured to provide sampled current to the pixel array wherein the sample and hold circuit includes a first transistor, a capacitor and a pair of current mirrors. The pair of current minors are connected to the first transistor and wherein the capacitor is connected to a drain of the first transistor. The sample and hold circuit is configured to sample an input current during a sampling phase to provide a sampled current, wherein the sampling includes: providing the input current to the pair of current mirrors and allowing the capacitor to be charged to the capacitor voltage that is a function of the input current; store, during a hold phase, the capacitor voltage; wherein the capacitor voltage forces an output stage of the sample and hold circuit to output a current that is substantially equal to the sampled current; and utilize the pair of current mirrors to force the gate, source and drain voltages of the first transistor to be equal to each other during the hold phase.

Digital cameras include a two-dimensional pixel arrays. A single pixel can include one or more photo-detectors as well as multiple transistors. Typical photo-detectors include photodiodes, photo transistors, photo-gates, hole accumulation diodes, pinned diodes, avalanche diodes, buried accumulation and transfer layer devices.
Various prior art pixels are known. The most commonly used pixels are either CCD pixels or CMOS pixels. Prior art CMOS pixels and two dimensional CMOS arrays are illustrated in the following U.S. patents which are incorporated herein by reference: U.S. Pat. No. 6,777,660 of Lee, U.S. Pat. No. 6,762,401 of Lee, U.S. Pat. No. 6,567,495 of Harada, U.S. Pat. No. 6,750,912 of Tennant et al., U.S. Pat. No. 6,697,111 of Kozlowski et al., U.S. Pat. No. 6,665,013 of Fossum et al., U.S. Pat. No. 6,587,142 of Kozlowski et al., U.S. Pat. No. 6,538,245 of Kozlowski, U.S. Pat. No. 6,532,040 of Kozlowski et al., U.S. Pat. No. 5,892,540 of Kozlowski et al., U.S. Pat. No. 5,238,276 of Dhuse et al., U.S. Pat. No. 6,326,230 of Pain et al., and European patent application EP1881692A2 titled "Methods for reading a pixel and for writing to a pixel and a device having pixel reading capabilities and pixel writing capabilities" which is incorporated herein by reference.
The pixels that form the pixel array are usually arranged in columns. Pixels that belong to the same pixel column may share various control and feedback signals.
The pixel columns are usually activated in a pipeline manner—while some pixel columns are reset other pixel columns are read ad yet other pixel columns undergo an integration phase. Typically, there is a relatively long time gap between the reset phase of a certain pixel column to a read phase of that pixel column.
Reference voltages are provided to pixels and to pixel readout circuits during the reset, read and integration phases. These reference voltages can affect the outcome of the reset and read phases, especially when applying correlated double sampling techniques.
Highly stable reference voltage sources are relatively expensive and their complexity can increase as a function of the number of pixel columns. Unstable reference voltage sources can output an unstable reference voltage that can vary between a reset phase of a pixel column to a read phase of that pixel column.
There is a need to provide efficient methods to provide reference signals to groups of pixels and to pixel readout circuits.

Device and method for providing a reference signal


The present invention relates to newly identified nucleic acid molecules and polypeptides present in normal and neoplastic ovarian cells, including fragments, variants and derivatives of the nucleic acids and polypeptides. The present invention also relates to antibodies to the polypeptides of the invention, as well as agonists and antagonists of the polypeptides of the invention. The invention also relates to compositions containing the nucleic acid molecules, polypeptides, antibodies, agonists and antagonists of the invention and methods for the use of these compositions. These uses include identifying, diagnosing, monitoring, staging, imaging and treating ovarian cancer and noncancerous disease states in ovarian, identifying ovarian tissue, monitoring and identifying and/or designing agonists and antagonists of polypeptides of the invention. The uses also include gene therapy, production of transgenic animals and cells, and production of engineered ovarian tissue for treatment and research.

Cancer of the ovaries is the fourth-most common cause of cancer death in women in the United States, with more than 23,000 new cases and roughly 14,000 deaths predicted for the year 2001. Shridhir, V. et al., Cancer Res. 61(15): 5895-904 (2001); Memarzadeh, S. & Berek, J. S., J. Reprod. Med. 46(7): 621-29 (2001). The incidence of ovarian cancer is of serious concern worldwide, with an estimated 191,000 new cases predicted annually. Runnebaum, I. B. & Stickeler, E., J. Cancer Res. Clin. Oncol. 127(2): 73-79 (2001). Unfortunately, women with ovarian cancer are typically asymptomatic until the disease has metastasized. Because effective screening for ovarian cancer is not available, roughly 70% of women diagnosed have an advanced stage of the cancer with a five-year survival rate of 25-30%. Memarzadeh, S. & Berek, J. S., supra; Nunns, D. et al., Obstet. Gynecol. Surv. 55(12): 746-51. Conversely, women diagnosed with early stage ovarian cancer enjoy considerably higher survival rates. Werness, B. A. & Eltabbakh, G. H., Int'l. J. Gynecol. Pathol. 20(1): 48-63 (2001). Although our understanding of the etiology of ovarian cancer is incomplete, the results of extensive research in this area point to a combination of age, genetics, reproductive, and dietary/environmental factors. Age is a key risk factor in the development of ovarian cancer: while the risk for developing ovarian cancer before the age of 30 is slim, the incidence of ovarian cancer rises linearly between ages 30 to 50, increasing at a slower rate thereafter, with the highest incidence being among septagenarian women. Jeanne M. Schilder et al., Hereditary Ovarian Cancer: Clinical Syndromes and Management, in Ovarian Cancer 182 (Stephen C. Rubin & Gregory P. Sutton eds., 2d ed. 2001).
With respect to genetic factors, a family history of ovarian cancer is the most significant risk factor in the development of the disease, with that risk depending on the number of affected family members, the degree of their relationship to the woman, and which particular first degree relatives are affected by the disease. Id. Mutations in several genes have been associated with ovarian cancer, including BRCA1 and BRCA2, both of which play a key role in the development of breast cancer, as well as hMSH2 and hMLH1, both of which are associated with hereditary non-polyposis colon cancer. Katherine Y. Look, Epidemiology, Etiology, and Screening of Ovarian Cancer, in Ovarian Cancer 169, 171-73 (Stephen C. Rubin & Gregory P. Sutton eds., 2d ed. 2001). BRCA1, located on chromosome 17, and BRCA2, located on chromosome 13, are tumor suppressor genes implicated in DNA repair; mutations in these genes are linked to roughly 10% of ovarian cancers. Id. at 171-72; Schilder et al., supra at 185-86. hMSH2 and hMLH1 are associated with DNA mismatch repair, and are located on chromosomes 2 and 3, respectively; it has been reported that roughly 3% of hereditary ovarian carcinomas are due to mutations in these genes. Look, supra at 173; Schilder et al., supra at 184, 188-89.
Reproductive factors have also been associated with an increased or reduced risk of ovarian cancer. Late menopause, nulliparity, and early age at menarche have all been linked with an elevated risk of ovarian cancer. Schilder et al., supra at 182. One theory hypothesizes that these factors increase the number of ovulatory cycles over the course of a woman's life, leading to "incessant ovulation," which is thought to be the primary cause of mutations to the ovarian epithelium. Id.; Laura J. Havrilesky & Andrew Berchuck, Molecular Alterations in Sporadic Ovarian Cancer, in Ovarian Cancer 25 (Stephen C. Rubin & Gregory P. Sutton eds., 2d ed. 2001). The mutations may be explained by the fact that ovulation results in the destruction and repair of that epithelium, necessitating increased cell division, thereby increasing the possibility that an undetected mutation will occur. Id. Support for this theory may be found in the fact pregnancy, lactation, and the use of oral contraceptives, all of which suppress ovulation, confer a protective effect with respect to developing ovarian cancer. Id.
Among dietary/environmental factors, there would appear to be an association between high intake of animal fat or red meat and ovarian cancer, while the antioxidant Vitamin A, which prevents free radical formation and also assists in maintaining normal cellular differentiation, may offer a protective effect. Look, supra at 169; Reports have also associated asbestos and hydrous magnesium trisilicate (talc), the latter of which may be present in diaphragms and sanitary napkins. Id. at 169-70.
Current screening procedures for ovarian cancer, while of some utility, are quite limited in their diagnostic ability, a problem that is particularly acute at early stages of cancer progression when the disease is typically asymptomatic yet is most readily treated. Walter J. Burdette, Cancer: Etiology, Diagnosis, and Treatment 166 (1998); Memarzadeh & Berek, supra; Runnebaum & Stickeler, supra; Werness & Eltabbakh, supra. Commonly used screening tests include biannual rectovaginal pelvic examination, radioimmunoassay to detect the CA-125 serum tumor marker, and transvaginal ultrasonography. Burdette, supra at 166.
Pelvic examination has failed to yield adequate numbers of early diagnoses, and the other methods are not sufficiently accurate. Id. One study reported that only 15% of patients who suffered from ovarian cancer were diagnosed with the disease at the time of their pelvic examination. Look, supra at 174. Moreover, the CA-125 test is prone to giving false positives in pre-menopausal women and has been reported to be of low predictive value in post-menopausal women. Id. at 174-75. Although transvaginal ultrasonography is now the preferred procedure for screening for ovarian cancer, it is unable to distinguish reliably between benign and malignant tumors, and also cannot locate primary peritoneal malignancies or ovarian cancer if the ovary size is normal. Schilder et al., supra at 194-95. While genetic testing for mutations of the BRCA1, BRCA2, hMSH2, and hMLH1 genes is now available, these tests may be too costly for some patients and may also yield false negative or indeterminate results. Schilder et al., supra at 191-94.
The staging of ovarian cancer, which is accomplished through surgical exploration, is crucial in determining the course of treatment and management of the disease. AJCC Cancer Staging Handbook 187 (Irvin D. Fleming et al. eds., 5th ed. 1998); Burdette, Supra at 170; Memarzadeh & Berek, supra; Shridhar et al., supra. Staging is performed by reference to the classification system developed by the International Federation of Gynecology and Obstetrics. David H. Moore, Primary Surgical Management of Early Epithelial Ovarian Carcinoma, in Ovarian Cancer 203 (Stephen C. Rubin & Gregory P. Sutton eds., 2d ed. 2001); Fleming et al. eds., supra at 188. Stage I ovarian cancer is characterized by tumor growth that is limited to the ovaries and is comprised of three substages. Id. In substage IA, tumor growth is limited to one ovary, there is no tumor on the external surface of the ovary, the ovarian capsule is intact, and no malignant cells are present in ascites or peritoneal washings. Id. Substage IB is identical to A1, except that tumor growth is limited to both ovaries. Id. Substage IC refers to the presence of tumor growth limited to one or both ovaries, and also includes one or more of the following characteristics: capsule rupture, tumor growth on the surface of one or both ovaries, and malignant cells present in ascites or peritoneal washings. Id.
Stage II ovarian cancer refers to tumor growth involving one or both ovaries, along with pelvic extension. Id. Substage IIA involves extension and/or implants on the uterus and/or fallopian tubes, with no malignant cells in the ascites or peritoneal washings, while substage IIB involves extension into other pelvic organs and tissues, again with no malignant cells in the ascites or peritoneal washings. Id. Substage IIC involves pelvic extension as in IIA or IIB, but with malignant cells in the ascites or peritoneal washings. Id.
Stage III ovarian cancer involves tumor growth in one or both ovaries, with peritoneal metastasis beyond the pelvis confirmed by microscope and/or metastasis in the regional lymph nodes. Id. Substage IIIA is characterized by microscopic peritoneal metastasis outside the pelvis, with substage IIIB involving macroscopic peritoneal metastasis outside the pelvis 2 cm or less in greatest dimension. Id. Substage IIIC is identical to IIIB, except that the metastasis is greater than 2 cm in greatest dimension and may include regional lymph node metastasis. Id. Lastly, Stage IV refers to the presence distant metastasis, excluding peritoneal metastasis. Id.
While surgical-staging is currently the benchmark for assessing the management and treatment of ovarian cancer, it suffers from considerable drawbacks, including the invasiveness of the procedure, the potential for complications, as well as the potential for inaccuracy. Moore, supra at 206-208, 213. In view of these limitations, attention has turned to developing alternative staging methodologies through understanding differential gene expression in various stages of ovarian cancer and by obtaining various biomarkers to help better assess the progression of the disease. Vartiainen, J. et al., Int'l J. Cancer, 95(5): 313-16 (2001); Shridhar et al. supra; Baekelandt, M. et al., J. Clin. Oncol. 18(22): 3775-81.
The treatment of ovarian cancer typically involves a multiprong attack, with surgical intervention serving as the foundation of treatment. Dennis S. Chi & William J. Hoskins, Primary Surgical Management of Advanced Epithelial Ovarian Cancer, in Ovarian Cancer 241 (Stephen C. Rubin & Gregory P. Sutton eds., 2d ed. 2001). For example, in the case of epithelial ovarian cancer, which accounts for ˜90% of cases of ovarian cancer, treatment typically consists of: (1) cytoreductive surgery, including total abdominal hysterectomy, bilateral salpingo-oophorectomy, omentectomy, and lymphadenectomy, followed by (2) adjuvant chemotherapy with paclitaxel and either cisplatin or carboplatin. Eltabbakh, G. H. & Awtrey, C. S., Expert Op. Pharmacother. 2(10): 109-24. Despite a clinical response rate of 80% to the adjuvant therapy, most patients experience tumor recurrence within three years of treatment. Id. Certain patients may undergo a second cytoreductive surgery and/or second-line chemotherapy. Memarzadeh & Berek, supra.
From the foregoing, it is clear that procedures used for detecting, diagnosing, monitoring, staging, prognosticating, and preventing the recurrence of ovarian cancer are of critical importance to the outcome of the patient. Moreover, current procedures, while helpful in each of these analyses, are limited by their specificity, sensitivity, invasiveness, and/or their cost. As such, highly specific and sensitive procedures that would operate by way of detecting novel markers in cells, tissues, or bodily fluids, with minimal invasiveness and at a reasonable cost, would be highly desirable.

Compositions and methods relating to ovarian specific genes and proteins


A cell contains a device for inserting a stack of electrodes and holding it in place. This device enables a stack of electrodes to be inserted and held in place in a container having a base and receiving a lid, the stack having two opposite plane faces and the container having two opposite plane walls. The device comprises two tapes connected by a central blade and is disposed between the plane faces of the stack and the plane walls of the container. The two tapes form elastic cylindrical sectors, advantageously parabolic in shape, with the concave side facing outwards when fitted.

1. Field of the Invention
The invention concerns a cell containing a device for inserting a stack of electrodes and holding it in place in a container having a base and receiving a lid, said stack having two opposite plane faces and said container having two opposite plane walls.
It can be a cell in which the stack of electrodes comprises a flattened coil inserted into a pot to which the last negative electrode turn must be connected.
2. Description of the Prior Art
Disposing spacers, possibly conductive spacers, between the electrodes and the container is known in itself (cf. for example US-2 842 607). Spacers with ribs or other raised patterns intended among other things to compress the stack of electrodes in order to optimize its performance in terms of power are also known in themselves (cf. for example EP-0 666 608), but this pressure is not constant, varying in particular with deformation of the stack (bulging consequent upon discharge cycles). The same problem arises in a locating and insertion device known in itself comprising two tapes connected by a central blade and adapted to be disposed between the plane faces of the stack (coil) and the plane walls of the container (pot).
The aim of the invention is to solve this problem by proposing a cell containing a device for applying a pressure to the stack of electrodes that is more uniform in time than in prior art devices.

Cell containing a device for inserting a stack of electrodes and holding in place


A hydrogen-absorbing alloy electrode for an alkaline storage cell, the electrode including a hydrogen-absorbing alloy expressed by a composition formula ReBxMy, wherein Re is at least one element selected from a group consisting of rare earth elements and alkali earth elements, B is boron, and M is at least one element selected from a group consisting of Ni, Co, Mn, Al, Cr, Fe, Cu. Sn, Sb, Mo, V, Nb, Ta, Zn, Zr and Ti. The alloy consists essentially of an RM.sub.y main metal phase, and a subordinate boron-containing phase, such as a compound phase of boron and a IV-a, V-a, or VI-a group metal.

(1) Field of the Invention
This invention relates to a hydrogen-absorbing alloy electrode used as a negative electrode of an alkaline storage cell.
(2) Description of the Prior Art
Conventional storage cells include nickel-cadmium or other alkaline cells and lead cells. Drawing current attention is a metal oxide-hydrogen alkaline storage cell comprising a hydrogen-absorbing alloy electrode, which can realize lightness, large capacity and high energy density. Used for such an electrode is a hydrogen-absorbing alloy such as LaNi.sub.5 (disclosed in U.S. Pat. No. 4,004,943), LaNi.sub.4 Co, or LaNi.sub.4.8 Fe.sub.0.2. LaNi.sub.4 Co and LaNi.sub.4.8 Fe.sub.0.2 have both been developed from LaNi.sub.5. Also having been developed for use in the above electrode are hydrogen-absorbing alloys including Misch metal (Mm), which is a mixture of rare earth elements such as La, Ce, Pr, Nd and Sm (Japanese Patent Publication Kokai No. 62-20245).
A negative electrode using a hydrogen-absorbing alloy reacts on a surface of the hydrogen-absorbing alloy and absorbs hydrogen generated while the cell is charged into the alloy. If the hydrogen-absorbing alloy used for the negative electrode absorbs an especially large amount of hydrogen and so is excellent in functioning as a catalyst, the negative electrode obtains a high energy density. Moreover, a cell having such an electrode is completely compatible with a nickel-cadmium cell having a cadmium negative electrode since both of the cells have the similar charge/discharge potential.
The cell employing the hydrogen-absorbing alloy as the negative electrode has an energy density which is approx, 1.5 times as high as that of a nickel-cadmium cell.
The hydrogen-absorbing alloys which have been developed so far have an ability of absorbing and desorbing a large amount of hydrogen, namely, has a high energy density, in the room temperature, where a cell is usually used. However, these alloys result in low initial charge/discharge efficiency and thus in insufficient electrochemical capacity on the initial stage of charge/discharge cycles. More specifically, an electrode using such a hydrogen-absorbing alloy is easy to charge from the initial stage, but the process of diffusing hydrogen from the inside to the outside of the alloy is a rate-determining step and so the polarity is changed with hydrogen remaining in the alloy. This phenomenon deteriorates the discharge efficiency.

Multiphase hydrogen-absorbing alloy electrode for an alkaline storage cell


A system for a transmission of several additional pieces of information by a single modification of a speed signal. In addition to the modification of the speed signal in the area near the wheel (modified speed sensor), the system provides the special evaluation of the speed signal, modified, at a distance from the wheel (controller). In addition, the system naturally also includes the combination of the special speed sensor and the controller.

Measuring the speeds of rotation of the vehicle wheels for control of the braking force, drive force and/or driving dynamics of a motor vehicle in open or closed loop is known. To do this in conventional manner, various methods (e.g. Hall or magneto-resistive sensors) are used. In addition, measuring the wear of the brake pad of a vehicle is known in that, for example, contact pins are embedded at a specific depth of the brake pads, which trigger a contact upon actuation of the brake when the brake pad is worn to this depth.
For example, the article "Integrierte Hall-Effekt-Sensoren zur Positions-und Drehzahlerkennung" (Integrated Hall Effect Sensors for Position and Speed Recognition) of the journal "elektronik industries," 7-1995, pp. 29-31, describes active sensors for use in the motor vehicle for anti-lock braking, traction control, engine and transmission open-loop and closed-loop control systems. Such sensors supply two current levels in a two-wire circuit which are converted into two voltage levels by a measuring resistor in an appropriate controller.
In addition to the Hall effect sensors, the use of magneto-resistive sensors is also known for speed recording, e.g., from the article "Neue, alternative Lösungen für Drehzahlsensoren im Kraftfahrzeug auf magnetoresisitiver Basis," (New Alternative Solutions for Speed Sensors based on Magnetoresistance of the Motor Vehicle), VDI Reports No. 509, 1984.
German Patent No. 26 06 012 (U.S. Pat. No. 4,076,330) describes a special common arrangement for detecting the wear on a brake pad and for detecting the wheel speed. To do this, the brake pad wear detected and the wheel speed detected by an inductively operating sensor are supplied via a common signal line to an analyzer. This is achieved in that the wheel speed sensor is completely or partially short-circuited in response to a detected brake pad wear.
Other systems as described, for example, in German Patent No. 43 22 440, require at least two signal lines between one wheel unit and the analyzer for detecting the speed and the brake pad wear on a wheel and a wheel brake, respectively.
In the speed detection method mentioned above, it is known that the air gap between the rotating ring gear and the actual sensor element has a considerable influence on the quality of the speed signal. Reference is made in this respect to e.g., German Patent Application No. 32 01 811.
The above-mentioned information (for example, brake pad wear and air gap/signal quality) is generally detected near the wheel and evaluated in a control unit mounted at a distance from the wheel. To do this, the information must be transmitted to the controller.
German Patent Application No. 196 18867.9 describes how to modify a rotational-speed signal in various specifiable ways for transmitting additional information (excessive brake pad wear, air gap that is too large/defective signal quality). The modification is carried out in different ways depending on the additional information to be transmitted. Making the different modifications of the speed signal requires a certain amount of effort.
The object of the present invention is to implement a very simple and reliable transmission of the speed signal and other information.

System for changing and/or evaluating a speed signal


A new Impatiens plant particularly distinguished by double, rose-shaped, red flowers, semi-compact, upright, outwardly spreading and mounded plant habit, free flowering, good basal branching, and moderately vigorous growth habit, is described.

The present invention comprises a new and distinct cultivar of Impatiens, botanically known as Impatiens walleriana, and hereinafter referred to by the cultivar name 'Silte Reeda'. The new cultivar originated from a controlled self pollination made in January 2003 in Andijk, The Netherlands. The parent was the proprietary red-flowered Impatiens plant 'ID03-14-4' (unpatented). The seeds produced by the controlled self pollination were sown in April 2003. A single plant selection was chosen for further evaluation and for asexual propagation in July 2003.
The new cultivar was created in 2003 and has been asexually reproduced repeatedly by vegetative cuttings and tissue culture in Andijk, The Netherlands and Gilroy, Calif. over a three and a half year period. The plant has also been trialed at Gilroy, Calif., Andijk, The Netherlands and Hillscheid, Germany. The present invention has been found to retain its distinctive characteristics through successive asexual propagations.
Plant Breeder's Rights for this cultivar were applied for with the European Union on Nov. 13, 2006.