A coherence shifting interferometry system determines the thickness or refractive index of an optically transmissive medium. The system utilizes an optical energy source for generating a source beam. The source beam is divided into two beams by reflecting one portion of the beam off of a reflective surface, e.g., a front surface, of an optically transmissive test sample having the unknown thickness or refractive index. A second portion of the beam is transmitted through the sample and reflected off a refractive interface in the sample, e.g., a back surface. The two reflected beams are combined into a composite beam. The composite beam is then directed to a Mach-Zehnder type interferometer, where a portion of the combined beam traverses a fixed optical path length, and another portion traverses a variable optical path length. The variable optical path length is adjusted to realign portions of the two beams and cause interference, and the amount of adjustment provides a measure of the unknown thickness or refractive index.

Optical interferometry is a measurement 10 technique that exploits the wave nature of light to produce extremely accurate measurements and provides excellent resolution without requiring any physical contact with the object being examined. Optical interferometry has been used to determine surface textures, shapes, distances, the speed of light through different media, and indices of refraction.
Optical interferometry is based on the phenomenon that two coherent light waves which are brought together (superimposed) behave similarly to water waves rippling through a pond. If the crest of one wave coincides with the crest of another wave, the waves reinforce one another in what is referred to as constructive interference. If the crest of one wave coincides with the trough of another wave, the waves cancel each other out. This canceling process is referred to as destructive interference. Several wave disturbances arriving at a point simultaneously result in a disturbance that is the vector sum of each of the separate disturbances.
The Michelson interferometer is a well known device that uses interferometry to make extremely precise measurements. One common embodiment of this device, shown in FIG. 1A includes a partially mirrored surface 200, which serves as a beam splitter, to divide a beam of monochromatic light into two beams 204 and 208 that are directed to travel in different directions. Monochromatic light is light having one color, and hence, one wavelength. One divided beam 204 reflects off a flat reference mirror 206 back to the beam splitter. The other divided beam 208 reflects off of a surface 210 being studied and returns back to the beam splitter where the two divided beams are recombined in an output beam 212. Recombining optical waves that are out of phase partially or totally cancel one another out. Optical waves that are in phase reinforce each other. The combined beams produce a pattern of alternating light and dark regions known as an interference pattern.
The difference in the lengths of the overall paths taken by each of the split beams is encoded in the interference pattern. For example, a difference between the distance traversed by the two beams equal to one wavelength (one-half wavelength up and back) results in the recombined wave going through one bright-dark-bright cycle. A dark region is created whenever the round-trip path along one arm, or beam path, of the interferometer increases or decreases by one-quarter wavelength with respect to the other arm since the total increased distance traveled by one of the split beams both up and back is one-half the wavelength (two times one-quarter wavelength) of the monochromatic beam.
Because the resolution of the Michelson interferometer is approximately one-half of the wavelength of the monochromatic light beam, optical interferometry provides very precise distance measurements.
Another type of interferometer is a Mach-Zehnder interferometer, shown in FIG. 1B. A Mach-Zehnder interferometer typically uses light 214 from a coherent light source that is split into two optical beams 216 and 218. The first split, or divided, beam can be used as a reference and traverses an optical path of fixed length. The other divided beam can be guided along an optical path having a variable length. The beams are subsequently recombined to produce an output beam 220 having an interference pattern. The length of the variable optical path length may be lengthened or shortened to achieve a desired relation between the two beams.
Optical interferometry has many practical applications. In the field of quality control, optical interferometry is used to measure distances and surface textures. Techniques for measuring surface texture using interferometry are described in Robinson, G.M., et al., "Optical Interferometry of Surfaces," Scientific American, pages 66-71, July 1991. The techniques described in that article are particularly suitable for measuring the surface textures of products such as photographic film, magnetic tape, and computer diskettes. Interferometric techniques that measure surface texture are also used to measure the degree of wear on products such as bearings.
Optical measurement techniques are well suited to physical dimension measurements in which accuracies on the order of microns are required. One common technique for optical measurements of physical dimensions is optical triangulation. Optical triangulation involves directing two laser beams from an electro-optic sensor unit towards a surface of interest, with the laser beams originating at points separated by several inches, and intersecting at the surface. As the distance between the sensor unit and surface of interest changes, the angle of one laser beam must be changed to keep the beam intersection point at the surface. The angle of this laser beam therefore provides an indication of the distance between the sensor head and surface of interest.
If two sensor heads are positioned on opposite sides of an object, the thickness of the object is determined by subtracting the distance readings of each sensor from the known separation between the two sensor heads. Thickness measurements by optical triangulation thus requires that a sensor be located on each side of an object under test. However, many structures having a thickness to be measured are formed on supporting structures, thus precluding access to one side of such structures. Therefore, optical triangulation is not suitable for applications lacking access to both sides of such structures.
Present optical triangulation systems can attain distance measurement accuracies of .+-.0.0001 inches (.+-.2.5 microns). When two such devices are used to determine thickness, the accuracy limits of each sensor combine to yield a total thickness accuracy limit of .+-.0.00014 inches (.+-.3.5 microns). However, there are plastic product manufacturing applications where greater thickness accuracies, up to .+-..00003 (.+-.1 .mu.m), are required.
There are many other applications where it is necessary to measure the thickness of optically transmissive (transparent and translucent) media, associated with, for example, lenses, video tape, audio tape, photographic films, coatings, compact disks, laminated structures, and supported structures. However, optical triangulation methods cannot measure the thicknesses of transparent or translucent objects because the sensor requires a visible illuminated spot, from diffuse reflection of the irradiating beams, on the surface of interest. This spot is measured by an optical sensor and as the surface distance changes, the beam angle of one of the beams is changed to maintain a single spot. However, transparent or translucent surfaces do not reflect enough light to provide a detectable spot.
Therefore, there is a need for a system capable of measuring the thickness of optically transparent or translucent media. A further need exists for a thickness measurement system that can be used when only one side of the object having the thickness to be measured is accessible. Still a further need exists for a thickness measurement system and method that can determine thicknesses of optically transmissive media with accuracies greater than .+-.0.00014 inches (.+-.3.5 microns).

Measurement system using optical coherence shifting interferometry

Science Applications International Corporation


Supporting disks are provided for a supporting disk bearing for open-end spinning rotors, which supporting disks have an essentially cylindrical running surface defined by encircling border edges. The running surface is provided with a helically shaped groove comprising at least two turns. The starting point of the groove as well as the end thereof have an eased runout which is at a distance to the respective border edge.

This application claims the priority of foreign application No. 197 12 916.1 filed in Germany on Mar. 27, 1997, the disclosure of which is expressly incorporated by reference herein.
The present invention relates to a supporting disk for a supporting disk bearing of an open-end spinning rotor comprising a damping, essentially cylindrical running surface defined by encircling border edges, which running surface comprises a helically shaped groove having a beginning and an end.
The U.S. Pat. No. 5,551,226 teaches that in the case of the known supporting disk of this type, the beginning and end of the helically shaped groove are located on the border edges which define the running surface. As during operation, the open-end spinning rotor exerts its greatest load on the border edges, the danger exists that the edges of the damping running surface, which is made of plastic, will crumble as a result of the interruptions on the surface caused by the groove.
German published patent application 33 42 768 teaches a similar supporting disk, in which the helically shaped groove has only one thread. The groove extends to both border edges, so that here also the same danger as in the above mentioned prior art exists.
One purpose of both known helically shaped grooves is to prevent the running surfaces heating up excessively during operation thus preventing that in the center of the running surface heat damage which could destroy the running surface.
It is an object of the present invention to eliminate the risk of the supporting disks of the above mentioned type crumbling at the border edges.
This object has been achieved in accordance with the present invention in that the beginning and the end of the helical groove are provided with an eased runout, which is located at a distance from the respective border edge.
The beginning and end of the groove are thus still located according to the present invention on the cylindrical running surface, whereby at both points the groove gradates over from the groove base into the running surface. The helically shaped groove thus does not extend to the extremely loaded border edges, so that damage of the running surface, in the form of crumbling, is prevented at these points.
For this purpose, the two eased runouts are located diametrically opposed to one another. Thus imbalances, which could be caused by the gradation of the helically shaped groove into the running surface, are avoided.
The groove is advantageously provided with two and a half helical turns. This is a good compromise between two loads which are not compatible, namely operational temperature and surface pressure. From experience, it is known that, with a higher number of thread turns, the heat build-up of the running surface is reduced. On the other hand, the stress bearing area of the running surface is reduced by the number of thread turns, which results in a higher surface pressure by the open-end rotor.
In order that the whole width of the running surface can be used to a large extent, it is further provided that the distance of the eased runouts from the respective border edges measures approximately 1 mm.

Supporting disk for a supporting disk bearing of an open-end spinning rotor and method of making same


A hydraulically damped multi-chamber hydraulic bushing is disclosed, which is capable of being loaded perpendicular to its axis. The bushing comprises two supporting sleeves which surround one another and are braced against each other by way of at least one spring element of an elastic material. The spring element at least partially surrounds the inner supporting sleeve, and at least two chambers--which are filled with an incompressible fluid--are arranged diametrically opposite one another. The chambers are fluidly interconnected through at least one connecting port. The chambers are formed by the outer supporting sleeve and a movable chamber wall that is turned toward the inner supporting sleeve. Each chamber wall has a central area that is resistant to expansion and an edge area with a rolling-diaphragm-type design, and is formed in one piece with the spring element. During use, the central areas and the inner supporting sleeve define a gap, which extends in the axial direction and has a maximum radial width of 1 mm.

1. Field of the Invention
The present invention relates to a hydraulically damped multi-chamber hydraulic bushing, which is capable of being loaded perpendicular to its axial direction. The bushing comprises two supporting sleeves, one surrounding the other. The sleeves are braced against each other by means of at least one spring element of an elastic material, whereby the spring element partially surrounds the inner supporting sleeve, and whereby the outer supporting sleeve and the spring element define at least two diametrically opposed chambers. The chambers are arranged perpendicular to the axial direction, are filled with an incompressible fluid, and are fluidly interconnected through at least one connecting port.
2. Description of the Related Art
European Published Patent Application 0 326 504 discloses a hydraulic bushing. The hydraulic bushing in this patent is formed with a plurality of fluid-filled chambers, which are fluidly interconnected through a passage. In this bushing, however, the insulation of high-frequency, small-amplitude vibrations is not satisfactory because of the small isolation surface necessitated by the type of construction used. Moreover, because the supporting body contacts the fluid over a large surface, high-frequency vibrations are transmitted to the fluid and lead to vorticity and to an increase in the dynamic spring rate. Furthermore, this hydraulic bushing has a comparatively complicated construction and consists of many individual parts, so that many separate operations are required during assembly. In this bushing the damping of low-frequency vibrations is also not very satisfactory. Finally, dissimilar pump conditions occur in the working chambers and as a result, the volume increase must be taken up in the chamber walls, causing them to expand. Thus, the output volume and attainable damping are adversely affected.

Vibration damper with axial caps and diaphragm-edge areas


Mammalian adenylyl cyclases are functionally expressed in yeast cells. The yeast cells may be used to screen for inhibitors or activators of the adenylyl cyclase, or of a regulator of adenylyl cyclase which is functionally co-expressed in the yeast cell. Methods of identifying such inhibitors, activators and regulators are also disclosed.

CROSS REFERENCE TO RELATED APPLICATIONS
A commonly owned application, U.S. Ser. No. 08/322,137, filed Oct. 13, 1994, incorporated by reference herein, relates to use of engineered yeast cells in screening for substances which modulate the activity of a mammalian surrogate of a yeast pheromone system protein, e.g., the yeast pheromone receptor, a G protein-coupled receptor.
1. Field of the Invention
The invention relates inter alia, to expression of a mammalian adenylyl cyclase in yeast, the transformed yeast cells, and their use, e.g., in identifying potential inhibitors or activators of the mammalian adenylyl cyclase, or of other proteins which are natively or artificially coupled to the mammalian adenylyl cyclase in the engineered yeast cell.
2. Description of the Background Art
Signal Transduction
In some instances, for a drug to cure a disease or alleviate its symptoms, the drug must be delivered to the appropriate cells, and trigger the proper "switches." The cellular switches are known as "receptors." Hormones, growth factors, neurotransmitters and many other biomolecules normally act through interaction with specific cellular receptors. Drugs may activate or block particular receptors to achieve a desired pharmaceutical effect. Cell surface receptors mediate the transduction of an "external" signal (the binding of a ligand to the receptor) into an "internal" signal (the modulation of a pathway in the cytoplasm or nucleus involved in the growth, metabolism or apotosis of the cell).
In many cases, transduction is accomplished by the following signaling cascade:
An agonist (the ligand) binds to a specific protein (the receptor) on the cell surface. PA1 As a result of the ligand binding, the receptor undergoes an allosteric change which activates a transducing protein in the cell membrane. PA1 The transducing protein activates, within the cell, production of so-called "second messenger molecules." PA1 The second messenger molecules activate certain regulatory proteins within the cell that have the potential to "switch on" or "off" specific genes or alter some metabolic process.
This series of events is coupled in a specific fashion for each possible cellular response. The response to a specific ligand may depend upon which receptor a cell expresses. For instance, the response to adrenalin in cells expressing .alpha.-adrenergic receptors may be the opposite of the response in cells expressing .beta.-adrenergic receptors.
The above "cascade" is idealized, and variations on this theme occur. For example, a receptor may act as its own transducing protein, or a transducing protein may act directly on an intracellular target without mediation by a "second messenger".
Signal Transduction Through G Proteins
Signals initiated by a variety of mammalian hormones and neurotransmitters are received by seven transmembrane domain receptors in the plasma membrane of cells and are transduced to intracellular effectors via heterotrimeric G proteins. Many different G proteins are known to interact with receptors. G protein signaling systems include three components: the receptor itself, a GTP-binding protein (G protein), and an intracellular target usually a protein.
The cell membrane acts as a switchboard. Messages arriving through different receptors can produce a single effect if the receptors acts on the same type of G protein. On the other hand, signals activating a single receptor can produce more than one effect if the receptor acts on different kinds of G proteins, or if the G proteins can act on different effectors.
The heterotrimeric G protein is composed of a guanine nucleotide-binding a subunit together with a tight complex of .beta. and .gamma. subunits. In their resting state, the G proteins, which consist of alpha (.alpha.), beta (.beta.) and gamma (.gamma.) subunits, are complexed with the nucleotide guanosine diphosphate (GDP) and are in contact with receptors. When a hormone or other first messenger binds to receptor, the receptor changes conformation and this alters its interaction with the G protein. This spurs the a subunit to release GDP, and the more abundant nucleotide guanosine tri-phosphate (GTP), replaces it, activating the G protein. The G protein then dissociates to separate the .alpha. subunit from the still complexed beta and gamma subunits. The free G.alpha. and the G.beta..gamma. subunits both may be capable of influencing the activity of specific effector molecules (e.g., the enzymes adenylyl cyclase, cyclic GMP phosphodiesterase (PDE), phospholipase C, phospholipase A.sub.2, and selected ion channels). The effector (which is often an enzyme) in turn converts an inactive precursor molecule into an active "second messenger," which may diffuse through the cytoplasm, triggering a metabolic cascade. After a few seconds, G protein signalling is terminated with the hydrolysis of GTP to GDP through the intrinsic GTPase activity of the G.alpha. subunit and the subsequent reassociation of G.alpha.-GDP with G.beta..gamma. to form the inactive heterotrimer. This reassociation is driven by the high affinity of GDP-bound G.alpha. for G.beta..gamma..
Hundreds, if not thousands, of receptors convey messages through heterotrimeric G proteins, of which at least 17 distinct forms have been isolated. Although the greatest variability has been seen in the a subunit, several different .beta. and .gamma. structures have been reported. There are, additionally, several different G protein-dependent effectors.
The study of microorganisms indicates that the development of G protein signal transduction pathways arose early in the evolution of eukaryotic cells. G protein regulatory function is intrinsic to the response to mating pheromones in yeast (Whiteway et al. 1989) and the development of the cellular slime mold Dictyostelium discoideum is controlled by G protein-mediated responses to cAMP (Devreotes 1989).
The Role of Adenylyl Cyclase in G-Protein-Mediated Signal Transduction
Adenylyl cyclase is among the best studied of the effector molecules which function in mammalian cells in response to activated G proteins. Activation of adenylyl cyclase occurs when signals transduced from specific cellular receptors result in the release of GTP-bound G.alpha.s. G.alpha.s ("s" denotes stimulatory) was originally identified as a regulator of adenylyl cyclase activity in mutant S49 cells which lacked adenylyl cyclase activity. G.alpha.s-GTP stimulated adenylyl cyclase activity in those cyc.sup.- cells (Northup et al. (1980) Proc. Natl. Acad. Sci. USA 77, 6516-6520). The production of cAMP can be stimulated by pure GTP-.gamma.S-bound G.alpha.s (GTP-.gamma.S is a non-hydrolyzable form of the nucleotide). Activation of cyclase by GTP-bound G.alpha.s is reversed by excess G.beta..gamma.; inhibition is assumed to occur as an inactive G protein heterotrimer re-forms.
Molecules which signal through receptors that interact with another class of G proteins, G.alpha.i (including G.alpha.i1, G.alpha.i2, and G.alpha.i3), mediate inhibition of adenylyl cyclase. Upon agonist binding to Gi-coupled receptors, both activated G.alpha.i protein and the released G.beta..gamma. complex appear to be capable of inhibiting the activity of adenylyl cyclase [Taussig et al. (1993) Science 261, 218-221]. The G.beta..gamma. complex may inhibit the enzyme's activity by reforming a heterotrimer with free G.alpha.s, thereby sequestering that stimulatory molecule (Gilman (1984) Cell 36, 577-579). In addition, the G.alpha.i subunit may directly inhibit adenylyl cyclase activity (Taussig et al. (1993) Science 261, 218-221.) A third mechanism for the negative regulation of adenylyl cyclase involves direct inhibition by the G.beta..gamma. complex. Purified type 1 adenylyl cyclase has been shown to be directly inhibited by .beta..gamma. subunits (Taussig et al. (1993) J. Biol. Chem. 268, 9-12).
Cyclic nucleotides play an important role in the regulation of a multitude of cellular activities. The synthesis of adenosine 3', 5'-cyclic phosphate (cyclic adenosine monophosphate or cAMP) is catalyzed by adenylyl cyclase, an enzyme which, in mammalian cells, is an integral membrane protein. Cyclic AMP is a second messenger which acts in response to cellular signals through a specific protein kinase (cAMP-dependent protein kinase or protein kinase A) to phosphorylate target molecules, e.g., other protein kinases or proteins involved in transport or cellular morphology. Through stimulation of the kinase, intracellular cAMP mediates many of the effects of hormones in the regulation of cellular metabolism and cell growth. Cyclic AMP is hydrolyzed by several phosphodiesterases (PDE) and can be actively secreted from some cell types, presumably via a specific transporter, or sequestered from the cytoplasm via transporters present in the membranes of intracellular organelles.
In vertebrate cells, adenylyl cyclase is regulated by heterotrimeric G proteins [Gilman (1984) Cell 36, 577-579] while in yeast, RAS proteins regulate adenylyl cyclase [Toda et al. (1985) Cell 40, 27-36; Broek et al. (1985) Cell 41, 763-769]. In turn, the activity of both the heterotrimeric G proteins and RAS proteins are controlled by the forms of guanine nucleotides to which they are bound.
While most adenylyl cyclases are found associated with the plasma membrane, certain forms of the enzyme expressed in bacteria are cytosolic, as is a mammalian enzyme found in testis. Peripheral membrane adenylyl cyclases are expressed in E. coli (Aiba et al. 1984) and in S. cerevisiae (Kataoka et al. 1985). The adenylyl cyclase encoded by the ACG gene of Dictyostelium appears to have a single transmembrane domain (Pitt et al. 1992). A second adenylyl cyclase gene from Dictyostelium (ACA) (Pitt et al. 1992), the Drosophila rutabaga gene (Levin et al. 1992), and the six full-length cDNAs encoding mammalian adenylyl cyclases that have been cloned to date code for integral membrane proteins.
Yeast Pheromone System Proteins and Their Metabolic Function
Haploid yeast cells are able not only to grow vegetatively, but also to mate to form a diploid cell. The two mating types ("sexes") of haploid cells are designated a and .alpha.. The a cells produce the dodecapeptide a-factor, and the .alpha. cells, the tridecapeptide .alpha.-factor. Because a-factor and .alpha.-factor elicit a mating response in the yeast cell of the opposite "sex", they are called "pheromones". These pheromones, as well as other proteins specifically involved in the production or transport of, or response to, pheromones, are considered "pheromone system proteins".
The gene encoding a-factor pheromone, like the .alpha.-factor receptor gene, is an a cell-specific gene; a cell-specific genes are only expressed in a cells. The gene encoding .alpha.-factor pheromone, like the a-factor receptor gene, is an .alpha. cell-specific gene; .alpha. cell-specific genes are only expressed in a cells. Other yeast genes belong to a haploid-specific gene set and are expressed in haploid cells (a cells or .alpha. cells) but not in diploid (a/.alpha.) cells. In addition, there exists a diploid cell-specific gene set, including those genes involved in sporulation.
In eukaryotic cells, RNA polymerase II promoters contain a specific sequence (the TATA box) to which the transcription factor TFIID (TATA binding protein or TBP) binds. An active transcription initiation complex includes TFIID, accessory initiation proteins, and RNA Pol II. As in higher eukaryotic cells, the TATA box is an essential control sequence in yeast promoters. Yeast TATA-box-binding protein (TBP) was identified by its ability to substitute in function for mammalian TFIID [Buratowski et al., Nature 334, 37 (1988); Cavallini et al., Nature 334, 77 (1988)]. With only a few apparent exceptions [transcription of some glycolytic enzyme genes, see Struhl, Mol. Cell. Biol. 6, 3847 (1986) and Ogden et al., Mol. Cell Biol. 6, 4335 (1986)] transcription of yeast genes requires the proximal TATA box element and TFIID binding for initiation of transcription. Also required for efficient transcription are gene-specific activator proteins; the precise mechanism whereby these gene-specific regulatory proteins influence transcription has not been completely elucidated.
MCM1p (encoded in the MCM1 gene) is a non-cell-type-specific transcription factor in yeast. MCM1p acts alone or in concert with other regulatory proteins to control expression of a- and .alpha.-cell specific genes. Yeast mating type loci encode the regulatory proteins that contribute to the control of cell type-specific expression. These proteins are Mata1p (encoded by the MATa gene) and Mat.alpha.1p and Mat.alpha.2p (encoded by the MAT.alpha. locus). MCM1p activates transcription of a-specific genes by binding to an upstream activation sequence (UAS) located in the control region of a-specific genes. Mat.alpha.1p and MCM1p interact to enhance each other's binding to specific UAS binding sites to activate .alpha.-cell-specific gene transcription in .alpha.-cells. Mat.alpha.2p associates with MCM1p to repress a-specific gene transcription in .alpha.-cells. In diploid (a/.alpha.) cells, Mat.alpha.1p and Mat.alpha.2p associate to repress the transcription of haploid-specific genes. The Mat.alpha.1p/Mat.alpha.2p regulatory entity is found only in diploid cells.
Yeast contain two genes encoding the .alpha.-factor pheromone, MF.alpha.1 and MF.alpha.2. Analysis of yeast bearing mutations in these sequences indicates that MF.alpha.1 gives rise to the majority of .alpha.-factor produced by cells. Expression occurs at a higher level from MF.alpha.1 than from MF.alpha.2 (Kurjan, Mol. Cell. Biol. 5, 787 (1985). The MF.alpha.1 gene of yeast encodes a 165 aa precursor protein containing an 85 aa leader sequence at the N-terminus. The leader includes a 19 aa signal sequence and a 66 aa sequence which contains sites for the addition of three oligosaccharide side chains (Kurjan and Herskowitz, Cell 39, 933 (1982); Singh et al. Nuc. Acids Res. 11, 4049 (1983); Julius et al. Cell 36, 309 (1984). Four tandem copies of the 13 aa .alpha.-factor are present in the C-terminal portion of the precursor; 6-8 aa spacer peptides precede the .alpha.-factor sequences (see FIG. 2).
After translocation of the nascent .alpha.-factor polypeptide to the ER, the signal sequence is cleaved from the precursor protein to yield pro-.alpha.-factor (Waters et al. J. Biol. Chem. 263, 6209 (1988). The core N-linked carbohydrate is added to three sites in the N-terminus of pro-.alpha.-factor (Emter et al. Biochem. Biophys. Res. Commun. 116, 822 (1983); Julius et al. Cell 36, 309 (1984); Julius et al. Cell 37, 1075 (1984). Additional glycosylation occurs in the Golgi prior to cleavage of pro-.alpha.-factor by the KEX2 endopeptidase. This enzyme cleaves within each of the spacer repeats leaving a Lys-Arg sequence attached to the C-terminus of .alpha.-factor peptide (Julius et al. Cell 37, 1075 (1984). The Lys-Arg sequence is removed by the action of the KEX-1 protease (Dmochowska et al. Cell 50, 573 (1987). The additional spacer residues present at the N-terminus of .alpha.-factor peptide are removed by the dipeptidyl aminopeptidase encoded by STE13 (Julius et al. Cell 32, 839 (1983). Four .alpha.-factor peptides are released from each precursor protein via the proteolytic processing outlined above and the mature .alpha.-factor is secreted from the cell.
Precursors of the 12 aa mature a-factor peptide are encoded in the MFa1 and MFa2 genes and are 36 aa and 38 aa residues, respectively (for schematic of MFa1 gene see FIG. 5). The precursors contain one copy of a-factor and the products of the two genes differ in sequence at one amino acid. The two forms of a-factor are produced in equal amounts by a cells (Manney et al. in Sexual interactions in eukaryotic microbes, p21, Academic Press, New York (1981).
Processing of a-factor entails a process that differs in every detail from that of .alpha.-factor. The processing of a-factor begins in the cytosol and involves the farnesylation of the C-terminal cysteine residue near the carboxyl terminus (-CVIA) by a farnesyl transferase (Schafer et al. Science 245, 379 (1989); Schafer et al. Science 249, 1133 (1990). The .alpha. and .beta. subunits of the farnesyl transferase are encoded by the RAM2 and RAM1 genes, respectively (He et al. Proc. Natl. Acad. Sci. 88, 11373 (1991). Subsequent to farnesylation is the proteolytic removal of the three amino acids that are C-terminal to the modified cysteine by a membrane-bound endoprotease. Next, the carboxy-terminal farnesylated cysteine residue is modified further: the carboxyl group is methylated by the product of the STE14 gene. STE14p is a membrane-bound S-farnesyl-cysteine carboxyl methyl transferase (Hrycyna et al. EMBO. J. 10, 1699 (1991). The mechanisms of the N-terminal processing of a-factor have not been elucidated. After processing of the precursors is complete, mature a-factor is transported to the extracellular space by the product of the STE6 gene (Kuchler et. al. EMBO J. 8, 3973 (1989), an ATP-binding cassette (ABC) transporter.
In normal S. cerevisiae (budding yeast) a cells, the .alpha.-factor binds the G protein-coupled membrane receptor STE2. The G protein dissociates into the G.sub..alpha. and G.sub..beta..gamma. subunits, and the G.sub..beta..gamma. binds an unidentified effector, which in turn activates a number of genes. STE20, a kinase, activates STE5, a protein of unknown function. STE5 activates STE11 kinase, which stimulates STE7 kinase, which induces the KSS1 and/or FUS3 kinases. These switch on expression of the transcription factor STE12. STE12 stimulates expression of a wide variety of genes involved in mating, including FUS1 (cell fusion), FAR1 (cell-cycle arrest), STE2 (the receptor), MFA1 (the pheromone), SST2 (recovery), KAR3 (nuclear fusion) and STE6 (pheromone secretion). Other genes activated by the pathway are CHS1, AG.alpha.1, and KAR3. The multiply tandem sequence TGAAACA has been recognized as a "pheromone response element" found in the 5'-flanking regions of many of the genes of this pathway.
One of the responses to mating pheromone is the transient arrest of the yeast cell in the G1 phase of the cell cycle. This requires that all three G1 cyclins (CLN1, CLN2, CLN3) be inactivated. It is believed that FUS3 inactivates CLN3, and FAR1 inhibits CLN2. (The product responsible for inactivating CLN1 is unknown).
The growth arrest is terminated by a number of different mechanisms. First, the .alpha.-factor receptor is internalized following binding of the pheromone, resulting in a transient decrease in the number of pheromone binding sites. Second, the C-terminal tail of the receptor is phosphorylated consequent to ligand binding, resulting in uncoupling of the receptor from the transducing G proteins. Third, pheromone-induced increases in expression of GPA1p (the G.alpha.-subunit of the heterotrimeric G protein) increase the level of the .alpha. subunit relative to the G.sub..beta. and G.sub..gamma. subunits, resulting in reduction in the level of free G.sub..beta..gamma. and consequent inactivation of the pheromone response pathway. Additional mechanisms include induction of the expression of SST2 and BAR1 and phosphorylation of the .alpha. subunit (perhaps by SVG1).
Signaling is inhibited by expression of a number of genes, including CDC36, CDC39, CDC72, CDC73, and SRM1. Inactivation of these genes leads to activation of the signaling pathway.
A similar pheromone signaling pathway may be discerned in .alpha. cells, but the nomenclature is different in some cases (e.g., STE3 instead of STE2).
Other yeast also have G protein-mediated mating factor response pathways. For example, in the fission yeast S. pombe, the M factor binds the MAP3 receptor, or the P-factor the MAM2 receptor. The dissociation of the G protein activates a kinase cascade (BYR2, BYR1, SPK1), which in turn stimulates a transcription factor (STE11). However, in S. pombe, the G.alpha. subunit transmits the signal, and there are of course other differences in detail.
Expression of Foreign Proteins in Yeast Cells
A wide variety of foreign proteins have been produced in S. cerevisiae, that remain in the yeast cytoplasm or are directed through the yeast secretory pathway (Kingsman et al. TIBTECH 5, 53 (1987). These proteins include, as examples, insulin-like growth factor receptor (Steube et al. Eur. J. Biochem. 198, 651 (1991), influenza virus hemagglutinin (Jabbar et al. Proc. Natl. Acad. Sci. 82, 2019 (1985), rat liver cytochrome P-450 (Oeda et al. DNA 4, 203 (1985) and functional mammalian antibodies (Wood et al. Nature 314, 446 (1985). Use of the yeast secretory pathway is preferred since it increases the likelihood of achieving faithful folding, glycosylation and stability of the foreign protein. Thus, expression of heterologous proteins in yeast often involves fusion of the signal sequences encoded in the genes of yeast secretory proteins (e.g., .alpha.-factor pheromone or the SUC2 [invertase] gene) to the coding region of foreign protein genes.
A number of yeast expression vectors have been designed to permit the constitutive or regulated expression of foreign proteins. Constitutive promoters are derived from highly expressed genes such as those encoding glycolytic enzymes like phosphoglycerate kinase (PGK1) or alcohol dehydrogenase I (ADH1) and regulatable promoters have been derived from a number of genes including the galactokinase (GAL1) gene. In addition, supersecreting yeast mutants can be derived; these strains secrete mammalian proteins more efficiently and are used as "production" strains to generate large quantities of biologically active mammalian proteins in yeast (Moir and Davidow, Meth. in Enzymol. 194, 491 (1991).
Heterologous G protein-coupled receptors have been functionally expressed in S. cerevisiae. Marsh and Hershkowitz, (Cold Spring Harbor Symp., Quant. Biol., 53: 557-65 (1988)) replaced the S. cerevisiae STE2 with its homologue from S. Kluyven. More dramatically, a mammalian beta-adrenergic receptor and G.alpha. subunit have been expressed in yeast and found to control the yeast mating signal pathway. King, et al., Science, 250: 121-123 (1990).
Duke University, WO92/05244 (Apr. 2, 1992) describes a transformed yeast cell which is incapable of producing a yeast G protein .alpha. subunit, but which has been engineered to produce both a mammalian G protein .alpha. subunit and a mammalian receptor which is "coupled to" (i.e., interacts with) the aforementioned mammalian G protein .alpha. subunit. Specifically, Duke reports expression of the human beta-2 adrenergic receptor (h.beta.AR), a seven transmembrane receptor (STR), in yeast, under control of the GAL1 promoter, with the h.beta.AR gene modified by replacing the first 63 base pairs of coding sequence with 11 base pairs of noncoding and 42 base pairs of coding sequence from the STE2 gene. (STE2 encodes the yeast .alpha.-factor receptor). Duke co-expressed a rat G protein .alpha. subunit in the same cells, yeast strain 8C, which lack the cognate yeast protein. Duke found that the modified h.beta.AR was functionally integrated into the membrane, as shown by studies of the ability of isolated membranes to interact properly with various known agonists and antagonists of h.beta.AR. The ligand binding affinity for yeast-expressed h.beta.AR was said to be nearly identical to that observed for naturally produced h.beta.AR. Ligand binding resulted in G protein-mediated signal transduction. Duke did not co-express a mammalian adenylyl cyclase in these cells.
Expression of Heterologous Adenylyl Cyclases in Yeast
African trypanosomes are protozoan parasites which are able to evade host immune defenses by altering their surface glycoproteins. The variable antigenicity is accomplished by sequential expression of genes encoding coat proteins. The variable surface glycoprotein genes (VSG) are transposed from silent regions to active, telomere-linked expression sites. Additional open reading frames (ORFs) termed the Expression Site Associated Genes (ESAGs), are found at these expression sites. ESAG4, cloned from Trypanosoma brucei, contains a sequence which is homologous to S. cerevisiae adenylyl cyclase [Pays et al. (1989) Cell 57, 835-845]. In addition, an ESAG from Trypanosoma equiperdum (eESAG4c), which is homologous to the ESAG4 of T. brucei, has been shown to encode an adenylyl cyclase which will complement an adenylyl cyclase (cyr-1) deletion mutant of S. cerevisiae [Ross et al. (1991) EMBO J. 10, 2047-2053].
The eESAG4c ORF contains sequence with homology to both S. cerevisiae and S. pombe adenylyl cyclases [Kataoka et al. (1985) Cell, 43, 493-505; Yamawaki-Kataoka et al. (1989) PNAS 86, 5693-5697; Young et al. (1989) PNAS 86, 7989-7993]. The region that is conserved between trypanosomes and yeast is within the yeast adenylyl cyclase catalytic domain and exhibits a sequence identity on the order of 50%. The eESAG4c sequence is approximately 40% identical to that of bovine brain adenylyl cyclase type 1 (Krupinski et al. (1990) Science 244, 1558-1562). The protein predicted by the eESAG4c sequence bears an N-terminal sequence that encodes a putative transmembrane domain flanking the sequence that is homologous to the adenylyl cyclase catalytic domain.
Also identified within the ESAG of Trypanosoma equiperdumare sequences which bear homology to a "leucine-rich repeat" gene family (Takahashi et al. (1985) PNAS 82, 1906-1910; Lopez et al. (1988) PNAS 85, 2135-2139; Hashimoto et al. (1988) Cell 52, 269-279). Proteins encoded by members of this family are involved in diverse functions, however, the repeat sequences are believed to be involved in membrane association and in protein-protein interactions. In S. cerevisiae the repeat domain of adenylyl cyclase is required for regulation of the enzyme by RAS proteins and for the association of the enzyme with the plasma membrane [Colicelli et al. (1990) Mol. Cell. Biol. 10, 2539-2543; Field et al. (1990) Science 247, 464-467; Mitts et al. (1990) Mol. Cell. Biol. 10, 3873-3883.] Also within the ESAG are sequences with limited homology to nucleotide binding domains [Florent et al. (1991) Mol. Cell. Biol. 11, 2180-2188] that have been hypothesized to have a regulatory function in trypanosomes analogous to that of Ras in yeast. Neither the leucine-rich repeat region nor the nucleotide binding domain were included in the sequences that complemented the yeast cyr deletion mutants [Ross et al. (1991) EMBO J. 10, 2047-2053]. Ross et al. (1991) speculated that the lack of these potential regulatory sequences would account for the much greater adenylyl cyclase activity exhibited by cyr-1 deletion mutants expressing eESAGc than was seen in yeast expressing the endogenous CYR gene from plasmids.
In addition to ESAG4, cloned from the VSG region of Trypanosoma brucei, at least three other genes cloned from T. brucei, GRESAG 4.1, 4.2 and 4.3, bear sequence homology to eukaryotic adenylate and guanylate cyclases (Alexandre et al. (1990) Mol. Biochem. Parasitol. 43, 279-288). ["GRESAG" indicates Genes Related to Expression Site Associated Genes.] It has been demonstrated that both ESAG 4 and GRESAG 4.1 can complement a S. cerevisiae adenylyl cyclase deletion mutant, cyr1.DELTA.. The trypanosome cyclases associate with the yeast membrane fraction, differ in their response to Ca.sup.2+, and do not appear to be properly regulated in yeast [Paindavoine et al. (1992) Mol. Cell. Biol. 12, 1218-1225].
Thus, the heterogenous adenylyl cyclases that have been shown to exhibit activity, although unregulated, in yeast are derived from trypanosome species. The trypanosome cyclase genes lie in regions near sequences encoding leucine-rich motifs with homology to a regulatory domain of yeast adenylyl cyclase. This suggests that proteins which derive from the two different trypanosome sequences may interact to form a regulatory complex. This could be analogous to the situation in Saccharomyces cerevisiae where activity of adenylyl cyclase is controlled through the interaction of the enzyme with regulatory RAS proteins. The homologies of sequence and regulation between the yeast and trypanosome enzymes appear to have favored the complementation of yeast deleted for adenylyl cyclase with sequences encoding the trypanosome enzyme.
Attempts to Express Mammalian Adenylyl Cyclases in Yeast
Previous attempts by other laboratories to express mammalian adenylyl cyclase in yeast were unsuccessful. Ronald Taussig, working in the laboratory of Alfred Gilman at the University of Texas Southwestern Medical Center, attempted to express mammalian type 1 adenylyl cyclase in Saccharomyces cerevisiae (personal communication). The protocol used by Taussig involved rescue of cyc cells by transformation with mammalian type 1 adenylyl cyclase; the metric of cyclase activity was growth of the test cells on forskolin-containing medium. Forskolin is known to bind directly to and to stimulate adenylyl cyclase types 1-6 in mammalian cells. Taussig was unable to detect enzyme activity in cyc cells transformed with the mammalian enzyme, i.e., he was unable to detect growth of transformed cells on forskolin-containing medium.
Expression of Mammalian Adenylyl Cyclases in Dictyostelium Discoideum.
The mammalian type 2 cyclase has been functionally expressed, by means not publicly disclosed, in the primitive eukaryote Dictyostelium discoideum [personal communication from P. Devreotes cited in Iyengar (1993)]. The structure of one of the two adenylyl cyclase genes that have been isolated from Dictyostelium, ACA, is predicted to be structurally analogous to the mammalian cyclases in that it is also an integral membrane protein [Pitt et al. 1992]. In addition, Dictyostelium can express eight G.alpha. subunits, each bearing approximately 45% sequence homology to mammalian G.alpha. proteins [Hadwiger et al. 1991; Wu and Devreotes 1991]. The lack of success in Gilman's laboratory at expressing a functional mammalian type 1 adenylyl cyclase in yeast, and the successful expression of the mammalian enzyme in Dictyostelium, indicate that differences in the transduction of signal to this enzyme exist between yeast and the higher eukaryotes. Furthermore, those differences must be taken into consideration in any attempt to recapitulate a signal transduction pathway with mammalian adenylyl cyclase in yeast.
Peptide Libraries
Peptide libraries are systems which simultaneously display, in a form which permits interaction with a target, a highly diverse and numerous collection of peptides. These peptides may be presented in solution (Houghten 1991), or on beads (Lam 1991), chips (Fodor 1991), bacteria (Ladner U.S. Pat. No. 5,223,409), spores (Ladner U.S. Pat. No. '409), plasmids (Cull 1992) or on phage (Scott, Devlin, Cwirla, Felici, Ladner '409). Many of these systems are limited in terms of the maximum length of the peptide or the composition of the peptide (e.g., Cys excluded). Steric factors, such as the proximity of a support, may interfere with binding. Usually, the screening is for binding in vitro to an artificially presented target, not for activation or inhibition of a cellular signal transduction pathway in a living cell. While a cell surface receptor may be used as a target, the screening will not reveal whether the binding of the peptide caused an allosteric change in the conformation of the receptor.
Ladner, U.S. Pat. No. 5,096,815 describes a method of identifying novel proteins or polypeptides with a desired DNA binding activity. Semi-random ("variegated") DNA encoding a large number of different potential binding proteins is introduced, in expressible form, into suitable host cells. The target DNA sequence is incorporated into a genetically engineered operon such that the binding of the protein or polypeptide will prevent expression of a gene product that is deleterious to the cell under selective conditions. Cells which survive the selective conditions are thus cells which express a protein which binds the target DNA. While it is taught that yeast cells may be used for testing, bacterial cells are preferred. The interactions between the protein and the target DNA occur only in the cell, not in the periplasm, and the target is a nucleic acid, not a protein.
Substitution of random peptide sequences for functional domains in cellular proteins permits some determination of the specific sequence requirements for the accomplishment of function. Though the details of the recognition phenomena which operate in the localization of proteins within cells remain largely unknown, the constraints on sequence variation of mitochondrial targeting sequences and protein secretion signal sequences have been elucidated using random peptides (Lemire et al., J. Biol. Chem. 264, 20206 (1989) and Kaiser et al. Science 235, 312 (1987), respectively).
Yeast have been engineered to express foreign polypeptide variants to be tested as potential antagonists of mammalian receptors. Libraries encoding mutant glucagon molecules were generated through random misincorporation of nucleotides during synthesis of oligonucleotides containing the coding sequence of mammalian glucagon. These libraries were expressed in yeast and culture broths from transformed cells were used in testing for. antagonist activity on glucagon receptors present in rat hepatocyte membranes (Smith et al. 1993).
All references cited in this specification are hereby incorporated by reference. No admission is made that any reference constitutes prior art.

Functional expression of mammalian adenylyl cyclase in yeast


Detergent or personal care compositions comprising capsules, which in turn comprise a hydrophobic continuous phase, the latter comprising an isotropic mixture of a thermoplastic-type co-polymer and hydrocarbon oil. Preferably, the capsules further comprise an oil-soluble benefit agent. Preferably, the compositions are liquid, especially aqueous-based compositions. In the preferred embodiment, the capsules are transparent or translucent.

In many articles of commerce, particularly consumer products, it is desirable to separate certain ingredients, yet have them disposed in a common container. Separation is particularly beneficial where one or more ingredients have negative interactions with each other. For example, in laundry detergents, enzymes are useful in removing stains but it is also best to separate them from other constituents, such as sources of alkalinity and surfactants, especially anionic surfactants like linear alkylbenzene sulfonates or alkyl sulfates. Bleaches, vitamins, perfumes, vegetable oils, plant extracts and ceramides are further examples of ingredients that sometimes need to be separated from the rest of the composition.
A known technique for separating ingredients in a common container includes encapsulation. Encapsulation technology is well known for different applications. Generally, encapsulation includes a medium that surrounds at least one component and thereby provides a barrier between the "encapsulated" component and other components. The barrier is typically temporary and is designed to break down and release the encapsulated material at a desired time, such as at a particular temperature, upon reaction or dissolution with chemicals, or due to mechanical stress. Methods of encapsulation include coacervation, liposome formation, granulation, coating, emulsification, atomization and spray-cooling.
WO 92/20771 discloses particles having a substantially anhydrous core comprising a matrix polymer containing active ingredient, a layer of hydrophobic oil around the core and a shell of polymer around the oil layer. The process for making the particles includes the steps of dispersion in oil of particles of a matrix polymer containing active ingredient, dispersing this dispersion into the aqueous solution and causing a solid polymer shell to form around the droplets of the matrix particles.
Norbury et al. (U.S. Pat. Nos. 4,976,961 and 5,013,473) disclose the addition of a polymer (which may be a polystyrene-polybutadiene-polystyrene block copolymer, e.g. Kraton® 1107) or a wax to an oil phase to increase the viscosity of the oil. The oil-polymer phase forms a core of the Norbury capsule; shell material is formed from another polymer.
Falholt et al. (U.S. Pat. No. 4,906,396, UK 2 186 884, and EP 0 273 775) disclose a protective enzyme system wherein enzymes are dispersed in a hydrophobic material which is an organopolysiloxane oil or a high molecular weight hydrocarbon including a solid or waxy material. Especially preferred are said to be hydrophobic liquids which had been stabilized by suspending therein hydrophobic solid particles such as hydrophobic silica.
Mitchnick et al. (U.S. Pat. No. 5,733,531) disclose a cosmetic sun-blocking composition containing particles which include a matrix and a UV-attenuating compound incorporated into the matrix. A preferred encapsulating matrix comprises wax. Certain polymeric materials may also be employed.
Ratuiste et al. (U.S. Pat. No. 5,589,370) discloses a process for encapsulation of sensitive materials, the capsule containing an oil dispersion holding an active and an outer polymer shell surrounding the oil dispersion. Tsaur et al. (U.S. Pat. No. 5,441,660 and U.S. Pat. No. 5,434,069) also disclose a capsule containing an oil dispersion containing an active and the polymer shell surrounding the dispersion. Another patent by Tsaur et al. (U.S. Pat. No. 5,498,378) discloses wax capsules containing a structuring agent which may be a hydrophobic silica, a hydrocarbon material and organophilic clay. An example of a high molecular weight hydrocarbon is given as a hydrocarbon rubber or elastomers.
Morrison et al. (U.S. Pat. No. 5,879,694) discloses transparent stiff gel candles comprising a hydrocarbon oil and a block co-polymer of a thermoplastic rubber (e.g., Kraton® series of polymers).
Despite numerous capsules in the prior art, a problem remains to produce a commercially attractive capsule which is stable—the encapsulated ingredient should not leach out of the capsule upon storage (especially important is the stability of capsules in liquid compositions)—but should release the protected ingredient with ease during normal use.
An additional challenge is that the capsules need to be manufactured with relative ease. For instance, in some prior art capsules the melt temperature of the encapsulating material may damage the encapsulated material during the encapsulation process. In the case of perfume encapsulation, for example, many perfumes are essential oils which are volatile, and thus can particularly benefit from low temperature processing. Yet, typical encapsulating ingredients, e.g. wax, have a high melting temperature.
A further problem with capsules for consumer products is the manufacture of either transparent or colored capsules. The encapsulating material, e.g. wax, is typically opaque. In such capsules the opaque shell obscures the color; neither can the transparency be attained. Yet, it is frequently desirable to produce a transparent or colored capsule to increase the appeal of the consumer product.

This patent is subject to a terminal disclaimer.

Detergent or personal care composition with oil capsules

Unilever Home & Personal Care USA, Division of Conopco, Inc.


A method of manufacturing an organic light-emitting element is provided. A first layer is formed above a substrate, and exhibits hole injection properties. A bank material layer is formed above the first layer using a bank material. Banks are formed by patterning the bank material layer, and forming a resin film on a surface of the first layer by attaching a portion of the bank material layer to the first layer. The banks define apertures corresponding to light-emitters. The resin material is the same as the bank material. A functional layer is formed by applying ink to the apertures that contacts the resin film. The ink contains an organic material. The functional layer includes an organic light-emitting layer. A second layer is formed above the functional layer and exhibits electron injection properties. The hole injection properties of the first layer are degraded by applying electrical power to an element structure.

Organic light-emitting elements, which have been researched and developed in recent years, are light-emitting elements that utilize light-emitting phenomenon of a solid fluorescent substance (For example, see Patent Literature 1). Organic light-emitting elements are manufactured by the following steps.
First, a reflective anode, which is a metal film, is formed on a substrate with an interlayer insulation film therebetween. Next, banks made of insulating material are formed to surround the edges of the reflective anode. Then, a hole injection layer and a light-emitting layer, which are organic films, are formed in the apertures defined by the banks. Here, as a constituent material of the hole injection layer, conductive polymer such as PEDOT/PSS (a mixture of polythiophene and polystyrene sulfonic acid) has conventionally been used.
Next, a cathode, which is a transparent conductive film, is formed on the light-emitting layer, and thus an organic light-emitting element is formed.

Method of manufacturing an organic light-emitting element, organic light-emitting element, display panel, and display device


The present disclosure provides a display panel comprising pixel units. Each of the pixel units has one or more first domains and one or more second domains. Each of the pixel units comprises a pixel electrode. The pixel electrode comprises first branches disposed in the one or more first domains at a first angle, and second branches disposed in the one or more second domains at a second angle. Every one or more first branches and every one or more second branches are staggered to form a zigzag structure between every first and second domains.

A pixel unit of a current display panel comprises a plurality of domains. A linear backbone is disposed between two adjacent domains. First branches and second branches are respectively disposed on two opposite sides of the linear backbone and are symmetrically disposed with respect to the linear backbone. Such an arrangement may cause a large width of dark lines at the linear backbone so that a current display panel has a large width of dark lines, which results in poor display quality.

Shenzhen China Star Optoelectronics Semiconductor Display Technology Co., Ltd.


Synthetic biomaterials are provided with irreversibly bound amino diphosphonate, polyphosphonate, or other anticalcification agent to prevent in vivo calcification. Such biomaterials include biocompatible elastomers such as polyurethane and/or polydimethylsiloxane, and the like which are intended for invasive, or in-dwelling use in a human or animal body. Illustratively, reaction conditions utilizing bi- or polyfunctional epoxides result in epoxide bridge incorporation of the anticalcification agent to the biomaterial elastomer.

This invention relates generally to materials which are resistant to in vivo calcification, and more particularly, to calcification-resistant biomaterials, suitable for implantation in a living being, comprising a synthetic biocompatible polymer to which an anticalcification agent(s) is bound by stable, irreversible covalent bonds.
More than 100,000 cardiac valve prostheses are placed in patients each year. Frequently, valve replacement surgery is the only means of treating cardiac valve disease. Currently used replacement valves include mechanical valves which may be composed entirely of a synthetic polymeric material such as polyurethane; bioprosthetic valves derived from bovine pericardium or porcine aortic valves; and aortic homografts.
Use of mechanical valves is frequently complicated by thrombosis and tissue overgrowth leading to valvular failure. Calcification is the most frequent cause of the clinical failure of bioprosthetic heart valves fabricated from porcine aortic valves or bovine pericardium. Human aortic homograft implants have also been observed to undergo pathologic calcification involving both the valvular tissue as well as the adjacent aortic wall albeit at a slower rate than the bioprosthetic heart valves. Pathologic calcification leading to valvular failure, in such forms as stenosis and/or regurgitation, necessitates re-implantation. Therefore, the use of bioprosthetic heart valves and homografts has been limited because such tissue is subject to calcification.
Pathologic calcification also further complicates the use of synthetic vascular grafts and other artificial heart devices, such as ventricular assist systems, because its affects the flexibility of the synthetic polymers used to produce the devices.
The mechanism for pathological calcification of cardiovascular tissue is not understood. Generally, the term "pathologic calcification" refers to the deposition of calcium phosphate mineral salts in association with a disease process. Calcification may be due to host factors, implant factors, and extraneous factors, such as mechanical stress. There is some evidence to suggest that deposits of calcium are related to devitalized cells, and in particular, cell membranes, where the calcium pump (Ca.sup.+2 -Mg.sup.+2 -ATPase) responsible for maintaining low intracellular calcium levels is no longer functioning or is malfunctioning. Calcification has been observed to begin with an accumulation of calcium and phosphorous, present as hydroxyapatite, which develops into nodules which can eventually lead to valvular failure.
Research on the inhibition of calcification of bioprosthetic tissue has focussed on tissue pretreatment with either detergents or diphosphonates. Both of the aforementioned compounds tend to wash out of the bioprosthetic tissue with time due to blood-material interactions. Thus, these treatments merely delay the onset of the inevitable calcification process. To date, long-term prevention of calcification has been an unattainable result. Accordingly, there is a need for a means of providing long-term calcification resistance for bioprosthetic or synthetic heart valves and other implantable, or in-dwelling, devices which are subject to in vivo pathologic calcification.
Systemic use of anticalcification agents, such as diphosphonates, results in significant side effects on bone, and overall, growth. Site specific therapy offers treatment with low regional drug levels and minimal side effects.
It is, therefore, an object of this invention to provide biomaterials for implantation in a mammal which have increased resistance to in vivo pathologic calcification.
It is another object of this invention to provide biomaterials for implantation in a mammal which have a long-term, or prolonged, resistance to in vivo pathologic calcification.
It is also an object of this invention to provide biomaterials for implantation in a mammal which have localized calcification inhibition and, hence, avoid the toxic side effects associated with systemic administration of anticalcification agents.
It is a further object of this invention to provide new synthetic biomaterial elastomers with increased utility due to retention of flexibility over an extended period of time.
It is additionally an object of this invention to provide a method of fabricating and/or treating biomaterials for implantation in a mammal to render the biomaterials resistant to in vivo pathologic calcification.
It is yet a further object of this invention to provide a novel method of covalently bonding anticalcification agents, specifically polyphosphonates or other anticalcification agents bearing functionalities capable of epoxide derivatization, to biomaterials.
It is also another object of this invention to provide a novel method of irreversibly binding polyphosphonates to synthetic biomaterials for permanent calcification inhibition.


A biosensor has a micrometer sized antibody substrate surface which may include patterned single or multiple antibody substrates. The substrate is capable of detecting multiple chemical species simultaneously and/or continuously monitoring a single chemical species.

1. Field of the Invention
The present invention relates micrometer scaled mono-analyte and multi-analyte substrates used in biosensors, including immunosensors, for environmental monitoring and medical diagnostic purposes.
2. Brief Description of the Related Art
Antibody substrates for use in biosensors or immunosensors have been described in U.S. Pat. Nos. 5,858,801 and 6,235,541, both to Robert A. Brizzolara and entitled "Patterning Antibodies on a Surface", the disclosure of each patent is herein incorporated by reference.

Patterned, micrometer-sized antibody features

The United States of America as represented by the Secretary of the Navy


This syringe for dispensing multiple doses of a liquid medicament one at a time has a cylindrical barrel for containing the medicament. At one end of the barrel is a single outlet opening. A plunger in the barrel discharges the medicament. A syringe cap rotatable mounted on the barrel has guide means restraining the cap against axial movement. Ratchet means on the cap and barrel permit only unidirectional rotation of the cap and sequentially retain the cap in equally spaced detent positions on the barrel. Nozzles on top of the cap have passages for ejecting the medicament. Each passage terminates in a hole in the top of the cap. Only one nozzle at a time communicates with the barrel vis the outlet opening in each detent position of the cap on the barrel.

1. Field of the Invention
This invention concerns the art of syringes; and more specifically this invention concerns syringes used for multiple injections of a medicinal fluid from a single syringe; and more particularly, this invention concerns a syringe having multiple nozzles arranged to permit only one nozzle at a time to inject medicinal fluid.
2. Description of the Prior Art
Heretofore it has been conventional to employ a syringe with a single nozzle to inject medicinal fluid into an animal's teats. This has proven very objectionable, since the use of the same syringe nozzle for all four teats of an animal, such as a cow, where one or more teats may be infected, exposes the animal to infection of all teats as the syringe is used upon each of the teats in succession. When for reasons of economy or lack of time, a single syringe with a single nozzle is used to treat more than one animal, many animals in a herd can be infected by the single nonsterile infected syringe. This is extremely undesirable practice. The lack or loss of sterility of the syringe when it has been used only once constitutes a positive health hazard which must be avoided.

Syringe for administering sequentially multiple doses of a medicament


A method and a device are provided for generating a multi-pulse laser beam including at least three laser beams, from a single pulse laser beam oscillated from a single laser beam source and a method and an apparatus using the pulse generator for laser beam machining with a two-wavelength multi-pulse laser beam suitable for fine-patterning a multi-layered circuit board formed of metal thin films and resin layers or ceramic bodies. One pulse laser beam from the laser beam source is separated into linear polarized laser beams in first and second directions by a polarization beam splitter, with the beam in the first direction being outputted as a first pulse beam of the multi-pulse beam. The beam in the second direction is incident to a combination of an optical fiber or a glass plate and a 1/4 wavelength plate, and is delayed and non-linearly polarized there to become a delayed and non-linearly polarized laser beam. The laser beam is again incident to the polarization beam splitter, where the delayed non-linearized pulse beam separated in the first direction is outputted as a second pulse beam of the multi-pulse laser beam. The delayed non-linearized pulse beam separated in the second direction is passed again through the above-described delay and non-linearization loop to generate a third pulse beam.

The present invention relates to a multi-pulse laser beam generation method and a device therefor and a laser beam machining method and an apparatus using a multi-pulse laser beam, and more particularly to a method and a device for converting one laser-pulse beam into two-wavelength multi-pulse laser beams and a laser beam machining method and an apparatus using a two-wavelength multi-pulse laser beam.
As a conventional multi-pulse laser beam generator, a delayed pulse laser beam generator has been disclosed in JP-A-62-123788 (NEC) laid open on Jun. 16, 1987. The delayed laser beam pulse generator includes first and second polarizers of similar characteristics for transmitting or reflecting an input laser beam of circular polarization to split the input laser beam into P-polarized and S-polarized beams, a polarization rotation device disposed between these polarizers for rotating by 90 degrees the polarized laser beam of the P-polarization or S-polarization reflected by the first polarizer, and a total reflection mirror for forming an optical path for a laser beam, and converts a laser beam of circular polarization into a laser beam of linear polarization having two mutually orthogonal axes with a delay time, to thereby provide a delayed pulse laser beam. In such a case, pulse beams to be outputted to the outside of the device are limited to two kinds of beam. The prior art device does not use a half-mirror to prevent loss of laser beam energy.
A laser beam machining is suitable for application to a multi-layered board including metal thin layers like conductor wiring layers. It is general that drilling is applied to provide holes on a printed circuit board laminated with metal thin layers and resin layers.
A drilling method to provide holes has been widely distributed because the method has an advantage of batch processing a laminated board. However, as the circuit structure on the printed circuit board has become highly packed a drilling diameter has also become smaller, raising a problem of poor durability of holed printed circuit boards. Particularly, in the case of a large circuit board used for a large computer, a lot of drilling work is required and it becomes essential to carry out periodical changing of drills for avoiding occurrence of faults on a single circuit board due to damage of the circuit board by a poor drilling work.
A laser beam machining method has recently been studied to replace the above-mentioned drilling method. The laser beam machining method not only eliminates poor circuit boards, but also can cope with providing holes of small diameters such as about 0.3 mm which is difficult to achieve by the mechanical drilling work. Further, the laser beam machining method has an advantage of not applying physical force to a printed circuit board because of its non-contact processing. However, it is difficult to carry out an efficient processing work of a laminated board by using only one laser beam of one wavelength obtained by one laser unit, because a laminated board including laminated layers of different materials such as metal thin films and resin layers has different absorption coefficients of laser beams. Accordingly, two laser beams of two different wavelengths obtained from two different laser units must be used to solve this problem. Examples of this type of device are disclosed in JP-A-01-266983 laid open on Oct. 24, 1989, JP-A-63-273587 laid open on Nov. 10, 1988 and JP-A-01-273684 laid open on Nov. 1, 1989. Techniques relating to laser beam machining are laid open by the JP-A-54-120498, JP-A-63-136546 and JP-A-63-154280.
While the above-described prior art techniques are processing methods taking advantage of characteristics of different laser wavelengths, they have a problem that an overall structure of a device becomes complex and is large-scaled because two completely different types of laser units are used in the system.

Multi-pulse laser beam generation method and device and laser beam machining method and apparatus using multi-pulse laser beam


An example method of providing a solid texture map to a graphics processing unit (GPU) includes dividing a tile of renderable content into a plurality of partitions. The method also includes determining that a set of partitions of the plurality of partitions is a solid color. The method further includes generating a solid texture map indicating that the set of partitions of the plurality of partitions is a solid color. The method also includes providing access to the solid texture map to a GPU.

Computing devices may be equipped with one or more high-performance graphics processing units (GPUs) that provide high performance with regard to computations and graphics rendering. Computing devices may use a GPU to accelerate the rendering of graphics data for display. Examples of such computing devices may include a computer workstation, mobile phones (e.g., smartphones), embedded systems, personal computers, tablet computers, and video game consoles.
Rendering generally refers to the process of converting a three-dimensional (3D) graphics scene, which may include one or more 3D graphics objects, into two-dimensional (2D) rasterized image data. In particular, GPUs may include a 3D rendering pipeline to provide at least partial hardware acceleration for the rendering of a 3D graphics scene. The 3D graphics objects in the scene may be subdivided by a graphics application into one or more 3D graphics primitives (e.g., points, lines, triangles, patches, etc.), and the GPU may convert the 3D graphics primitives of the scene into 2D rasterized image data.

Bandwidth reduction using texture lookup by adaptive shading


A telecommunication system for transmission along power supply lines of modulated information signals having a selected carrier frequency. The system includes a plurality of communication devices connected along the supply lines, each including a microcontroller and a modem. The modem includes an interface module which establishes a link to the power supply lines for modulated information signals, a transmission module which contains a digital modulator followed a digital/analog converter, and a receiving module. The receiving module includes a digital filter connected between an analog/digital converter and a digital demodulator. The digital filter is adapted to the carrier frequency of modulated information signals present on the power supply lines, whereby the modem can operate at any selected frequency over a broad frequency range.

1. Field of the Invention
The present invention relates to a telecommunication system for transmission of information signals along power supply lines, the system comprising communication devices which each of include a microcontroller and a modem, said modem comprising an interface module which establishes a link to the power supply lines for information signals. The modem also comprises a transmission module which contains a digital modulator followed by a digital/analog converter whose output is connected to the interface module, the modem finally comprising a receiving module which contains an analog/digital converter whose input is connected to the interface module, followed by a digital demodulator.
2. Description of the Prior Art
A modem comprised in such a communication device is known from U.S. Pat. No. 4,510,611. Such known modem used for transmitting and receiving data on power supply lines comprises an interface module, a transmission module and a receiving module. The transmission module of such a device performs digital modulation of the transmit signal in the ASK mode (Amplitude Shift Keying), which consists of transmitting a fixed carrier signal at a nominal frequency when the signal to be modulated is a "1", and stopping the transmission when the signal to be modulated is a "0". The range of the authorized frequencies for this type of transmission runs from 60 to 140 Khz. The receiving module of such a device comprises an autocorrelator which checks the integrity of the received signal. This signal may have been jammed either by the presence of other information signals transmitted on the line, or by the turning on of powerful apparatus such as, for example, a washing machine, which causes a sudden variation of current in the power supply lines. If the integrity of the signal is questioned by the autocorrelator, an information signal is sent to a microcontroller which controls the modem, which microcontroller has the task of sending a signal through the modem's transmission module to the device that has initially transmitted the signal of doubtful integrity in order to request a retransmission, and this is done until the signal reaches the modem unaffected. However, this procedure lacks flexibility and may cause considerable delays in communications, particularly in the case where parasitic or, in terms of electromagnetic noise, polluting phenomena are lasting and/or frequent.

Telecommunication system and modem for transmission of modulated information signals over power supply lines


A painting robot outer arm includes a non-conductive housing mounting a color changer on the outside and a paint canister on the inside connected by an isolating paint transfer line. The paint canister is releasably attached to a piston ram and drive motor by a quick disconnect coupling. The paint supply is isolated from the applied high voltage by cleaning and drying the transfer line. The rate of filling of the canister can be controlled in response to sensed torque applied by a drive motor moving a piston in the paint canister.

The present invention relates generally to robotic painting systems and in particular to a robotic apparatus for applying electrically conductive paint to the external surfaces of automotive vehicle bodies.
Prior art paint booths are well known. A typical prior art paint booth, used to paint the exterior surfaces of vehicle bodies in both continuous conveyance and stop station systems, includes an enclosure housing a plurality of paint applicators. In one configuration, the applicators are mounted on an inverted U-shaped support structure that includes two vertical supports, one on either side of the path of travel of the bodies, connected at their tops by a horizontal support. This support structure is used to paint the top surfaces of the body and the horizontal beam can be fixed or can have an additional degree of freedom to move along the top of the vehicle body being painted. Another painting device is used in the same painting zone to paint the sides of the body and generally does not have the capability to move laterally along the length of the body. Disadvantages of this type of painting apparatus include lack of flexibility to provide optimized standoff distance between the body surface and the applicator along with inefficient use of the allotted painting cycle time. In the case of the top surface painting machine, the paint applicators are mounted on a common beam: therefore, the distance between each paint applicator and the surface to be painted varies with the contours of the vehicle body. In the case of the side painting machine, the paint applicators do not move transverse to the path of the vehicle body. They can only paint the portion of the body that is in front of the applicator leaving a good portion of the available cycle time unused.
A more recent alternative to the support structure has been floor-mounted robots disposed along the sides of the painting booth. The robots mount either spray guns or rotary applicators (bell machines) for directing atomized paint toward the vehicle body.
While rotary applicators have advantages over spray guns, there are some associated disadvantages. The prior art floor mounted robots, especially bell machines, are inherently very costly and limit visual access to the spray booth. The bell machines require more bells for the same throughput due to limited orientation capability. The additional bells use more paint per vehicle due to per bell paint waste during color changing. Prior art floor mounted robots also require significant booth modification when installed in existing paint booths, increasing installation time and cost, and require more booth length and width. The rail axis of floor mounted robots requires doors at both ends of the booth. The waist axis of the floor mounted robot requires an additional safety zone at the ends of the spray booth and the rail cabinets of the floor mounted robots encroach into the aisle space. Floor mounted robots also require frequent cleaning due to the down draft of paint overspray causing paint accumulation on the robot arm and base, which results in higher maintenance and cleaning costs.
The prior art bell zone machines also lack flexibility. Additional and more flexible robot zones are required because the prior art machines unable to reach substantially all paintable surfaces on one side of the body and, therefore, have limited backup capability for an inoperative painting machine. Additional robot zones are also used to provide backup capability for the less flexible prior art painting machine.
It is desirable, therefore, to provide a painting apparatus and a painting system that utilizes robots in an efficient and cost-effective manner that minimizes paint waste, occupies little space (length and width) in the paint booth and can be installed in existing paint booths without requiring significant booth modification. It is also desirable to provide a painting apparatus wherein one painting robot is able to reach substantially all paintable surfaces on one side of the article to provide backup capability in the case of an inoperative robot.
Due to the conductivity of the waterborne paint, it is necessary to electrically isolate the grounded bulk paint supply system from a charged local dispensing canister and spray application system. In the prior art, the bell applicator, canister, canister drive, electrostatic cascade, and docking interface were all integrated into a single unit mounted on the robot wrist as shown in the U.S. Pat. No. 5,293,911 and the U.S. Pat. No. 5,367,944. Such an applicator had the following shortcomings:
1) The applicator is heavy, expensive, and subject to damage via collision with objects in the painting booth.
2) The applicator docking with a docking station must occur in a fixed booth position and therefore limits process flexibility.
3) The docking process takes cycle time as the robot must travel to and from the docked position. The canister filling can not start until the applicator reaches the docked position.
4) The docking hardware is expensive and unique to waterborne systems.
Another method to isolate the bulk paint supply system from the charged paint dispensing canister is to clean and dry the paint transfer line between the supply system and the canister. The cleaning and drying of the transfer line between the bulk supply system and the paint dispensing canister has not been successfully implemented in an automotive type painting system (rapid color changing on a continuous conveyance type system). There are several reasons why this type of system has not been used in the past. These shortcomings include:
1) The time to clean and dry the line and provide high voltage isolation exceeds the allotted dwell time between the vehicle bodies being painted.
2) The high voltage cannot be easily contained and thus the isolation system is compromised (arcing and pin holing). The electrostatic erosion can burn holes in the transfer line, the dispensing system, the supply line to the applicator, or the waste collection lines. The resultant down time would not be readily accepted for this type of application.
3) The amount of waste that is left in the paint transfer line is excessive when compared to other means of isolation.
4) The acceptable voltage limit does not provide high painting transfer efficiency as compared to other means of voltage isolation.


The invention relates to a measuring arrangement for measuring the density of a product by means of microwaves. The measuring arrangement comprises a first microwave resonator, from which, in operation, microwaves enter a product area, and a device for compensating for environmental influences and disturbance variables affecting the measuring signal of the first microwave resonator. According to the invention, the compensation device comprises a second microwave resonator, which is shielded towards the product area with respect to microwave radiation.

The invention relates to a measuring arrangement for measuring the density of a product by means of microwaves.
Certain known devices comprise a first microwave resonator, from which, in operation, microwaves enter a product area. When measuring density using such a measuring arrangement, the measuring variables used, in particular the resonant frequency and the width of the resonance curve, are dependent on different environmental influences and disturbance variables in the first resonator. In order to compensate for the influences, such as the temperature of the resonator, on the systematic measuring error, it is known to provide a temperature sensor at a specific point of the first resonator and to carry out a computational correction by means of the measured temperature. Since the temperature in the first resonator is determined only at one point, the accuracy of the compensation in the first resonator is limited, especially in the case of a spatially non-homogeneous and/or time-variant temperature distribution.
Generally speaking, to compensate, for example, for the effects of temperature, it is known to use different materials, in the present case for the first resonator, having temperature coefficients of opposite signs. Apart from increased manufacturing costs, however, the transition points between the different materials in particular can have an adverse effect in microwave technology. The use of special alloys and composite materials having thermal expansion coefficients close to or equal to zero is also known. These too involve increased manufacturing costs.
It is an aim of the invention to provide an apparatus and method of the kind mentioned above in which the measuring error caused by different environmental influences and by internal and external disturbance variables in the first resonator can be reduced.

Microwave measuring arrangement for product density measurement


Melanoma is a solid tumor that is notoriously resistant to chemotherapy, and its incidence is rapidly increasing. Recently, several signaling pathways have been demonstrated to contribute to melanoma tumorigenesis, including constitutive activation of MAP kinase, Akt and stat-3. The activation of multiple pathways may account in part for the difficulty in treatment of melanoma. In a recent screen of compounds, we found that an organopalladium complex showed significant antiproliferative activity against melanoma cells. This complex, tris(dibenzylideneacetone)dipalladium (Tris DBA), has activity against B16 murine and A375 human melanoma in vivo. Tris DBA inhibits several signaling pathways including activation of MAP kinase, Akt, stat-3 and S6 kinase activation. Tris(dibenzylideneacetone)dipalladium is thus a novel compound that is a member of a class of noble metal complexes with potential antitumor activity. Further preclinical evaluation of TrisDBA and related complexes is warranted.

Melanoma is a common solid tumor notorious for its high rate of metastasis and resistance to chemotherapy and radiation. Several factors may account for the resistance of melanoma to current therapies. First, melanomas are derived from melanocytes, specialized neural crest cells that are specialized to produce melanin. The production of melanin results in the generation of toxic reactive oxygen species and cytotoxic phenol derivatives, thus melanocytes are equipped with mechanisms to resist these insults. Recently, the microopthalmia gene (MITF), which is a master transcriptional switch of melanocytes, has been shown to possess antiapoptotic activity and is found in metastatic lesions at a high frequency (25) (26). Second, multiple signaling pathways are activated in melanoma. B-raf is mutated in many melanomas, resulting in constitutive activation of MAP kinase signaling (1, 27). N-ras is also mutated frequently in melanoma, resulting in activation of MAP kinase, phosphoinositol-3 kinase/akt signaling, and S6 kinase activation (28, 29). While B-raf and constitutive MAP kinase activation is sufficient to cause transformation of melanocytes into melanoma (3, 4), other signal transduction events are frequently observed in B-raf mutant melanomas, such as loss of the tumor suppressor PTEN (29, 30). The consequences of PTEN loss is activation of phosphoinositol-3 kinase/akt activation.
Multiple regimens have been tried for the treatment of locally advanced and metastatic melanoma. Initial trials several decades ago used agents such as hydroxyurea, and more recent agents used against melanoma include dacarbazine and platinum based therapies including cisplatin and carboplatin. Other therapies, including biochemotherapy, have included interleukin-2 infusion and infusion of lymphocytes which are present in melanoma lesions and have been expanded ex vivo (31). All of these therapies have had modest success in a minority of patients, but with significant toxicity, including pulmonary leak syndrome (32-36). Currently, interferon alpha is employed in high risk patients, and prolonged therapy results in a 10% long term survival benefit.
Targeted therapies have been attempted in melanoma. Sorafenib was developed as a B-raf inhibitor based upon the observation that B-raf mutation is common in melanoma. However, results from initial trials of sorafenib in melanoma have been disappointing (15). Everolimus has also been tried against human melanoma, and has not been successful as a single agent (37). Current knowledge of signaling may provide an explanation of why previous therapies have failed. Phosphoinositol-3 kinase activation has been shown to mediate against extrinsic pathways of apoptosis, which include apoptosis due to TRAIL, TNF alpha, and interferons (38). Monotherapies of these cytokines may be frustrated in the face of phosphoinositol-3 kinase activation. Similarly, apoptosis induced by tumor infiltrating lymphocytes may be frustrated by phosphoinositol-3 kinase activation. Phosphoinositol-3 kinase also activates VEGF expression, and in addition to stimulating angiogenesis, VEGF inhibits dendritic cell function, impairing immune responses to melanoma (39-43).
Targeting MAP kinase as monotherapy in melanoma is clearly insufficient to eliminate melanoma in most patients. MAP kinase is activated in a majority of human melanomas, including those that lack B-raf mutation (3). In a previous study of human melanomas, we demonstrated that a subset of advanced melanomas had decreased MAP kinase activation, implying that additional signaling pathways are operative in vivo. Further support of this hypothesis is our previous finding that treatment of EBV-induced Burkitt's lymphomas with antioxidants resulted in compensatory MAP kinase activation (17). It is likely that treatment of melanoma patients with sorafenib results in compensatory activation of non-MAP kinase pathways. Similarly, mTOR inhibition due to rapamycin and derivatives has been shown to result in compensatory Akt activation (44).
Tris-DBA has the advantage that it inhibits several pathways required for melanoma tumorigenesis, including MAP kinase activation, phosphoinositol-3 kinase/akt activation, stat-3 activation, and S6 kinase activation. These pathways are involved in virtually all forms of human cancer, as well as autoimmune disorders, such as lupus, eczema (atopic dermatitis), asthma, psoriasis and arthritis. While no drug is likely to be completely effective as monotherapy in melanoma, Tris-DBA is well tolerated systemically in mice, and has a novel profile of action compared with other clinically used chemotherapeutic agents. Its ability to inhibit phosphoinositol-3 kinase activation may enhance the activity of cytokines which require akt inactivation for optimal activity, and may enhance the activity of other chemotherapeutic agents. Tris-DBA may also enhance the activity of sorafenib by blocking compensatory signaling that may occur in vivo due to MAP kinase blockade. Our studies provide a rationale for the further investigation of Tris-DBA in the treatment of malignant melanoma.

Palladium complexes inhibit N-myristoyltransferase activity in vitro and cancer growth in vivo


A pressure sensor is composed of a cylindrical connector case firmly connected to a housing and a pressure-detecting chamber formed between the connector case and the housing. The pressure-detecting chamber is confined by a metal diaphragm, an outer peripheral portion of which is firmly held at a portion connecting the connector case and the housing. A relatively thick ring plate is connected to a mounting surface of the housing, and the outer peripheral portion of the metal diaphragm is sandwiched between the ring plate and a pushing plate placed on and connected to the ring plate. Thus, the pressure-detecting chamber containing a sensing element therein is surely sealed even if the housing is made of a material, such as aluminum, which is different from that of the metal diaphragm.

1. Field of the Invention
The present invention relates to a pressure sensor having a metal diaphragm responsive to pressure to be measured.
2. Description of Related Art
An example of a pressure sensor having a metal diaphragm responsive to pressure is disclosed in JP-A-7-243926. The essence of the pressure sensor is illustrated in FIG. 5 attached hereto. This pressure sensor is a diaphragm-seal type pressure sensor having a connector case J1 and a housing J2 connected to each other for forming a pressure-detecting chamber J5 therebetween. A bottom end of the connector case J1 is inserted into a mounting space J4 of the housing J2, and an upper end of a wall forming the mounting space J4 is staked against a shoulder of the connecting case J1. A ring-weld J9, to which an outer peripheral portion of a metal diaphragm J3 is connected, is sandwiched between the connector case J1 and the housing J2.
A sensing element J6 is disposed in the pressure-detecting chamber J5 enclosed by the metal diaphragm J3. The pressure-detecting chamber J5 is filled with encapsulated liquid J8 for transferring pressure imposed on the metal diaphragm J3 to the sensing element J6. A portion connecting the connector case J1 and the housing J2 is sealed with an O-ring J7. The pressure sensor is connected to a pipe or a block containing fluid therein by a screw formed at a bottom portion of the housing J2. A pressure introduced into the pressure sensor is imposed on the metal diaphragm J3 and detected by the sensing element J6.
To prevent fluid introduced into the pressure sensor from leaking through a portion connecting the ring-weld J9 and the housing J2, the ring-weld J9 and the housing J2 are made of a same material and welded together at a contacting portion J10. In order to obtain a firm welding strength between the ring-weld J9 and the housing J2, both have to be made of the same material, such as carbon steel S15C or SUS.
On the other hand, the pipe or the block to which the pressure sensor is connected is often made of a material such as aluminum. If the housing J2 made of carbon steel is connected to a pipe made of aluminum, a potential difference is generated between two different materials contacting each other. Such a potential difference may be as high as 0.5 volts. It is highly probable that corrosion by battery-effect will occur in the pipe made of aluminum. To avoid such corrosion, it is conceivable to make the housing J2 with the same material, such as aluminum, as the pipe to which the pressure sensor is connected. However, it becomes considerably difficult to weld the ring-weld J9 made of SUS or the like to the housing J2 made of aluminum. This is because the melting point of SUS is considerably higher than that of aluminum. Further, it is very difficult to change the material of the metal diaphragm J3 and the ring-weld J9 to other materials because the material such as SUS or the like is most suitable for these parts to obtain high reliability and strength of the welded portion.

Pressure sensor having a metal diaphragm responsive to pressure