The present invention relates to versatile linkers for tethering a molecule to a solid support, e.g., for tethering a monomer, oligomer or polymer to a solid support, which are stable to a wide range of reaction conditions, but can be cleaved under well-defined conditions, thereby liberating the molecule from the solid support. Preferably, the linkers are used to tether to the solid support unprotected, partially-protected or fully-protected monosaccharides or oligosaccharides, or unprotected, partially-protected or fully-protected glycoconjugates. The linkers of the present invention may be used to tether to solid supports building blocks useful in the assembly of libraries of other types of small molecules. The present invention also relates to a molecule or plurality of molecules tethered to the solid support via a linker or linkers of the present invention. The present invention also relates to processes for synthesizing molecules, e.g., monomers, oligomers or polymers, on a solid support, wherein a starting material in the synthesis of the molecule, intermediates in the synthesis of the molecule, and the molecule itself are tethered to the solid support during the process via one of the linkers of the present invention. In certain processes of the present invention, the molecule is liberated from the solid support by cleavage of the linker of the present invention.

Nucleic acids, proteins and polysaccharides are three major classes of biopolymers. While the first two systems are principally linear assemblies, polysaccharides are structurally more complex. This structural and stereochemical diversity results in a rich content of xe2x80x9cinformationxe2x80x9d in relatively small molecules. Nature further xe2x80x9cleveragesxe2x80x9d the structural versatility of polysaccharides by their covalent attachment (i.e., xe2x80x9cconjugationxe2x80x9d) to other biomolecules such as isoprenoids, fatty acids, neutral lipids, peptides or proteins. Oligosaccharides in the form of glycoconjugates mediate a variety of events including inflammation, immunological response, metastasis and fertilization. Cell surface carbohydrates act as biological markers for various tumors and as binding sites for other substances including pathogens.
Moreover, many physiologically important recognition phenomena involving carbohydrates have been discovered in recent years. Lectins, proteins which contain carbohydrate recognition domains, have been identified. Prominent members of the calcium dependent (C-type) lectin family (Drickamer, K. Curr. Opin. Struct. Biol. 1993, 3, 393) are the selectins which play a crucial role in leukocyte recruitment in inflammation. Bevilacqua, M. P.; Nelson, R. M. J. Clin. Invest. 1993, 91, 379. Members of the C-type lectin superfamily have been described on NK cells and Ly-49, NKR-P1 and NKG2 constitute group V of C-type lectins. While many lectins have been purified and cloned, their ligands have not been identified due to the heterogeneous nature of carbohydrates.
The recognition that interactions between proteins and carbohydrates are involved in a wide array of biological recognition events, including fertilization, molecular targeting, intercellular recognition, and viral, bacterial and fungal pathogenesis, underscores the importance of carbohyrates in biological systems. It is now widely appreciated that the oligosaccharide portions of glycoproteins and glycolipids mediate certain recognition events between cells, between cells and ligands, between cells and the extracellular matrix, and between cells and pathogens. See, e.g., U.S. Pat. No. 4,916,219 (describing oligosaccharides with heparin-like anticomplement activity).
These recognition phenomena may be inhibited by oligosaccharides having the same sugar sequence and stereochemistry found on the active portion of a glycoprotein or glycolipid involved in the recognition phenomena. The oligosaccharides are believed to compete with the glycoproteins and glycolipids for binding sites on the relevant receptor(s). For example, the disaccharide galactosyl-xcex2-1-4-N-acetylglucosamine is believed to be one component of the glycoproteins which interact with receptors in the plasma membrane of liver cells. Thus, to the extent that they compete with moieties for cellular binding sites, oligosaccharides and other saccharide compositions have the potential to open new horizons in pharmacology, diagnosis, and therapeutics.
The growing appreciation of the key roles of oligosaccharides and glycoconjugates in fundamental life sustaining processes has stimulated a need for access to usable quantities of these materials. Glycoconjugates are difficult to isolate in homogeneous form from living cells since they exist as microheterogeneous mixtures. The purification of these compounds, when possible, is at best tedious and generally provides only very small amounts of the compounds. The travails associated with isolation of oligo- and poly-saccharides and glycoconjugates from natural sources present a major opportunity for the development and exploitation of chemical synthesis. See, e.g., U.S. Pat. Nos. 4,656,133; 5,308,460; 5,514,784; and 5,854,391 (describing representative means of glycosylating saccharides and peptides).
Intense work is ongoing on the further development of the use of biologically-active oligosaccharides within a number of different fields, including novel diagnostics and blood typing reagents; highly specific materials for affinity chromatography; cell specific agglutination reagents; targeting of drugs; monoclonal antibodies, e.g., against cancer-associated reagents; as an alternative to antibiotics, based on the inhibition with specific oligosaccharides of the attachment of bacteria and viruses to cell surfaces; and stimulation of the growth of plants and protection of them against pathogens. Additionally, a considerable future market is envisaged for fine chemicals based on biologically-active carbohydrates.
As stated above, due to the difficulties associated with purification of glycoconjugates and oligosaccharides from natural sources, chemical synthesis may be the only way to procure sufficient amounts of these structures for detailed biochemical and biophysical studies. Additionally, combinatorial carbohydrate libraries hold great potential for the identification of carbohydrate-based ligands to cellular receptors. Identification of these molecules will open many new avenues for the development of diagnostic tools and therapeutic agents.
The invention of solid phase peptide synthesis by Merrifield 35 years ago dramatically influenced the strategy for the synthesis of these biopolymers. The preparation of structurally defined oligopeptides (Atherton, E.; Sheppard, R. C. Solid phase peptide synthesis: A practical approach; IRL Press at Oxford University Press: Oxford, England, 1989, pp 203) and oligonucleotides (Caruthers, M. H. Science 1985, 230, 281) has benefited greatly from the feasibility of conducting their assembly on various polymer supports. The advantages of solid matrix-based synthesis, in terms of allowing for an excess of reagents to be used and in the facilitation of purification are now well appreciated. However, the level of complexity associated with the synthesis of an oligosaccharide on a polymer support dwarfs that associated with the other two classes of repeating biooligomers. First, the need to differentiate similar functional groups (hydroxyl or amino) in oligosaccharide construction is much greater than the corresponding needs in the synthesis of oligopeptides or oligonucleotides. Furthermore, in these latter two cases, there is no stereoselection associated with construction of the repeating amide or phosphate bonds. In contrast, each glycosidic bond to be fashioned in a growing oligosaccharide ensemble constitutes a new locus of stereogenicity.
Combinatorial chemistry has been used in the synthesis of large numbers of structurally distinct molecules in a time and resource-efficient manner. Peptide, oligonucleotide, and small molecule libraries have been prepared and screened against receptors or enzymes to identify high-affinity ligands or potent inhibitors. These combinatorial libraries have provided large numbers of compounds to be screened against many targets for biological activity. Every pharmaceutical company now devotes a major effort to the area of combinatorial chemistry in order to develop new lead compounds in a rapid fashion.
The development of protocols for the solid support synthesis of oligosaccharides and glycopeptides requires solutions to several problems. Of course, considerable thought must be addressed to the nature of the support material. The availability of methods for attachment of the carbohydrate from either its xe2x80x9creducingxe2x80x9d or xe2x80x9cnon-reducingxe2x80x9d end would be advantageous. Also, selection of a linker which is stable during the synthesis, but can be cleaved easily when appropriate, is critical. A protecting group strategy that allows for high flexibility is desirable. Most important is the matter of stereospecific and high yielding coupling reactions.
Combinatorial carbohydrate libraries hold a tremendous potential with regard to therapeutic applications. The key role complex oligosaccharides play in biological processes such as inflammation, immune response, cancer and fertilization makes them highly attractive therapeutic targets. The ability to create true oligosaccharide libraries has the potential to trigger a revolution in the area of biopharmaceuticals.
The generation of combinatorial carbohydrate libraries will facilitate the rapid identification of ligands to many carbohydrate binding proteins which are involved in a variety of important biological events including inflammation (Giannis, A. Angew. Chem. Int. Ed. Engl. 1994, 33, 178), immune response (Ryan, C. A. Proc. Natl. Acad. Sci. U.S.A. 1994, 91, 1) and metastasis (Feizi, T. Curr. Opin. Struct. Biol. 1993, 3, 701). Analogs of ligands can help to define important lectin-ligand interactions. Non-natural ligands can be powerful inhibitors of carbohydrate-protein binding and will facilitate the study of cascade-like events involving such interactions. Furthermore, inhibitors of carbohydrate-lectin binding are potential candidates for a variety of therapeutic applications.
Moreover, the development of an automated oligosaccharide synthesizer holds great potential to influence glycobiology just as the peptide synthesizer impacted protein research. A reliable strategy for the solid support synthesis of oligosaccharides depends to a great extent on the choice of the linker which is used to anchor the first building block to the polymeric matrix.
A set of versatile linkers is described which are stable to a wide range of reaction conditions but can be cleaved in several ways to produce free oligosaccharides, fully-protected oligosaccharide building blocks and novel glycoconjugates. Furthermore, these linkers may be used to attach to solid supports building blocks useful in the assembly of libraries of other types of small molecules.
In certain embodiments, the present invention relates to versatile linkers for tethering a molecule to a solid support, e.g., for tethering a monomer, oligomer or polymer to a solid support, which are stable to a wide range of reaction conditions, but can be cleaved under well-defined conditions, thereby liberating said molecule from the solid support. In preferred embodiments, the linkers of the present invention are used to tether to the solid support unprotected, partially-protected or fully-protected monosaccharides or oligosaccharides, or unprotected, partially-protected or fully-protected glycoconjugates. In other embodiments, the linkers of the present invention may be used to tether to solid supports building blocks useful in the assembly of libraries of other types of small molecules. In certain embodiments, the present invention relates to a molecule or plurality of molecules tethered to the solid support via a linker or linkers of the present invention.
In certain embodiments, the present invention relates to processes for synthesizing molecules, e.g., monomers, oligomers or polymers, on a solid support, wherein a starting material in the synthesis of said molecule, intermediates in the synthesis of said molecule, and said molecule itself are tethered to the solid support during the process via one of the linkers of the present invention. In certain processes of the present invention, the molecule is liberated from the solid support by cleavage of the linker of the present invention.
The invention described herein is expected to enable the automated synthesis of oligosaccharides and glycoconjugates in much the same fashion that peptides and oligonucleotides are currently assembled. The ability to synthesize defined biologically important glycoconjugates will be far reaching with many direct applications to biomedical questions. Opportunities for the application of the present invention include the development of automated oligosaccharide synthesis machines.

Linkers for synthesis of oligosaccharides on solid supports


An electric shaver (1) is provided comprising a housing (2), a shaving head (5) comprising a shaving foil (7) and a cutter assembly (8) with cutter lamellae (19), and a motor (23) of the rotary type as well as a transmission mechanism (31) acting between the motor (23) and the cutter assembly (8) in order to convert the rotation of the motor shaft (30) into a reciprocating movement to the drive a cutter assembly (8), the motor (23) having a nominal operating speed between 8000 and 10,000 revolutions per minute, wherein the motor (23) is covered with a sheath (32) of an elastic material, the transmission mechanism (31) comprises at least one balancing mass (52), and each of the cutter lamellae (19) of the cutter assembly (8) is provided with an indentation (57, 58) in each of its two side faces (55, 56) which are each bounded by a cutting edge (53, 54), which indentation extends parallel to the cutting edge (53, 54), and exhibits a cutting angle (.beta.) smaller than 90.degree. at both its cutting edges (53, 54).

Such a shaver as defined in the opening paragraph is known, for example from DE-PS 27 49 936. The shaver employs a motor whose nominal operating speed is not specified in DE-PS 27 49 936. The nominal operating speed is to be understood to mean the speed of the motor when it drives the cutter assembly which cooperates with the shaving foil while no shaving takes place, i.e. while no hairs are severed. In most prior-art electric shavers the nominal operating speed of the motors used in these shavers lies at the most in a range of from 6500 to 7500 revolutions per minute. Such a maximum nominal operating speed, which is in a directly proportional relation to the oscillating frequency of the cutter assembly, relatively limits the shaving performance attainable with these shavers, which in its turn depends directly on the oscillating frequency of the cutter assembly. Recently some shavers have become known whose motors have a nominal operating speed in excess of the range of 6500 to 7500 r.p.m. in order to improve the shaving performance but in which no or inadequate steps have been taken to reduce noise and vibrations, so that these shavers are comparatively noisy in operation and produce comparatively strong vibrations. In a way comparable with these shavers the motor of the shaver known from DE-PS 27 49 936 is also secured directly in the housing of the shaver without any noise-reduction measures, so that in particular at a higher motor speed an undamped transmission of structure-borne noise from the motor to the housing is possible and occurs. Moreover, in the shaver known from DE-PS 27 49 936 the motor shaft of the motor can drive a transmission mechanism in which no measures have been taken to reduce vibrations of the shaver housing which originate in this mechanism, so that in particular at a higher motor speed comparatively strong housing vibrations may occur. Finally, the shaver in accordance with DE-PS 27 49 936 has a cutter assembly with conventional smooth-walled cutter lamellae, in which no steps have been taken to improve the cutting action and to reduce the housing vibrations and shaving-foil vibrations caused by the shaving actions and the shaving noise produced by such vibrations, so that in particular at a higher motor speed these vibrations van be comparatively strong and the shaving noise produced thereby can be comparatively loud.


Novel heterocyclic alkaloids were isolated from a marine sponge. These compounds, and derivatives thereof, are useful antifungal and antitumor compounds. The novel compounds have the following structures: ##STR1## wherein R.sub.1 =H, alkyl, alkenyl, aryl, benzyl, acyl, benzoyl, or alkali metal;. R.sub.2 and R'.sub.2 =O, S, NOX, or NNHX, wherein X is alkyl or aryl.

Considerable research and resources have been devoted to oncology and antitumor measures including chemotherapy. While certain methods and chemical compositions have been developed which aid in inhibiting, remitting, or controlling the growth of tumors, new methods and antitumor chemical compositions are needed. The prevention and control of fungi is also of considerable importance to man, and much research has been devoted to development of antifungal measures.
It has been found that some natural products and organisms are potential sources for chemical molecules having useful biological activity of great diversity. Marine life has been the source for the discovery of compounds having varied biological activities. Some of the United States patents which have issued for such inventions are as follows: U.S. Pat. No. 4,548,814 for didemnins, having antiviral activity, were isolated from a marine tunicate; U.S. Pat. No. 4,729,996 discloses compounds, having antitumor properties, that were isolated from marine sponges Teichaxinella morchella and Ptilocaulis walpersi; U.S. Pat. No. 4,808,590 discloses compounds, having antiviral, antitumor, and antifungal properties, isolated from the marine sponge Theonella sp.; and U.S. Pat. No. 4,737,510 discloses compounds, having antiviral and antibacterial properties, isolated from the Caribbean sponge Agelas conifera. Clearly, marine sponges have proved to be a source of biological compounds, and a number of publications have issued disclosing organic compounds derived from marine sponges, including Scheuer, P. J. (ed.) Marine Natural Products, Chemical and Biological Perspectives, Academic Press, New York, 1978-1983, Vol. I-V; Faulkner, D. J., (1984) Natural Products Reports 1:551-598; Natural Products Reports (1986) 3:1-33; Natural Products Reports (1987) 4:539-576; J. Am. Chem. Soc. (1985) 107:4796-4798.
The subject invention concerns novel heterocycle alkaloids. Heterocycle compounds of marine tunicate origin have previously been described in Kobayashi, J., J. Chang, Y. Ohizumi, Y. Hirata, T. Sasaki, T. Ohta and S. Nozoe (1988) Tetrahedron Letters 29:1177-1180. Other heterocycle compounds of marine ascidian origin have been described in Bloor, S., F. J. Schmitz (1987) J. Amer. Chem. Soc. 109:6134; and de Guzman, F. S., F. J. Schmitz (1989) Tetrahedron Letters 30:1069. The present invention, utilizing sponges as a source material has provided the art with a new class of biologically active compounds and new pharmaceutical compositions useful as antitumor and antifungal agents. Subsequent to the filing of our parent application describing the novel compounds and their biological activities claimed here, Schmitz et al. have also reported the isolation of these compounds from marine organisms (Schmitz, F. J., F. S. DeGuzman, M. Bilayet Hossain, D. van der Helm [1991] J. Org. Chem. 56:804-808).
Other advantages and further scope of applicability of the present invention will become apparent from the detailed descriptions given herein; it should be understood, however, that the detailed descriptions, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent from such descriptions.

Bioactive heterocycle alkaloids and methods of use


The invention provides a process to form a “first composition comprising a first ethylene/α-olefin interpolymer and a second ethylene/α-olefin interpolymer,” said process comprising polymerizing a first mixture comprising ethylene, an α-olefin, and optionally a polyene, in a stirred tank reactor to form a first ethylene/α-olefin interpolymer, and transferring at least some of the first ethylene/α-olefin interpolymer to a loop reactor, and polymerizing, therein, a second mixture comprising ethylene, an alpha-olefin, and optionally a polyene, in the presence of the first ethylene/α-olefin interpolymer, to form the “first composition comprising the first ethylene/α-olefin interpolymer and the second ethylene/α-olefin interpolymer.” The invention also provides a polymerization reactor configuration comprising at least the following: a stirred tank reactor followed by a loop reactor.

There is a need to polymerize high molecular weight ethylene/alpha-olefin interpolymers at reduced costs, while minimizing the load (solvent) on the devolatilization system of the polymerization and isolation process. Polymerization processes of the art are described in the following references: U.S. Pat. Nos. 7,910,637; 6,329,471; 8,143,352; 6,686,419; 7,223,822; 8,178,625; 6,329,477; 7,223,822; 7,122,606; 7,449,530; 7,220,806; 8,431,657; 8,735,517; 8,088,867; 8,562,915; 9,102,824; U.S. Provisional 62/097,257 (filed Dec. 29, 2014) and U.S. Publication 2007/0004875. However, as discussed above, there remains a need to prepare high molecular weight ethylene/alpha-olefin interpolymers at reduced costs, while minimizing the devolatilzation load (solvent) on the reactor system. These needs have been met by the following invention.

Process to form ethylene/alpha-olefin interpolymers


A lighting device, particularly a motor vehicle light, having a primary reflector (2) composed of a hollow body that defines a pair of pointed elements (4, 5), which face opposite directions, between which a luminous source (1) is placed; and a secondary reflector composed of an elongated element (3) with steps (6), at whose centre the primary reflector (2) is placed with its relative pointed elements (4, 5) facing the steps (6).

The present invention relates to lighting devices of the type that comprise a luminous source, a primary reflector and a secondary reflector.
The purpose of this invention is to implement a lighting device of the type described above, which is especially designed for use inside a motor vehicle light, more specifically, inside the xe2x80x9cthird stop-lightxe2x80x9d, and has a simple, compact structure, which is nevertheless capable of providing high performance.
According to the invention, the lighting device is essentially characterized in that:
the primary reflector is composed of a hollow body that defines a pair of ellipsoid-shaped pointed elements, which face in opposite directions to each other, between which a luminous source is placed,
said luminous source consists of a filament lamp,
the secondary reflector comprises an elongated element with steps, at whose centre said primary reflector is placed with the above pointed elements facing the steps.
As explained previously, according to the invention, the shape of the lighting device makes it especially suitable for application to the rear light, or xe2x80x9cthird stop-lightxe2x80x9d of motor vehicles. In this application, the lighting device is normally made complete by a transparent plate for diffusing the luminous beam produced by the primary and the secondary reflectors.

Lighting device, particularly a motor vehicle light


The present invention is a method and system which can be applied to supply a constant bit rate link with digital information from a packet or cell based link, such as an Asynchronous Transfer Mode (ATM) link. The present invention automatically provides compensation information to a buffer supplying the constant bit rate link with information. The compensation is not used if packets or cells are made available in the buffer.

1. Field of the Invention
The invention relates to telecommunications, and in particular to the processing of cells containing voice information to account for lost cells.
2. Description of the Prior Art
Transmission based on cell or packet technology is well known. In these systems, a sequence of information to be transmitted is broken into discreet segments called cells or packets (these will be referred to as cells hereinafter, as is known, cells are typically fixed length and packets are typically variable length). Typically, the cell information is referred to as the payload which contains user information bits and is given a header which includes network information bits to aid in identifying, checking, and routing the cell. The cells can then be transmitted across a network independently of other cells in the information sequence. Those skilled in the art are familiar with the advantages of such systems. One of the prominent systems is known as Asynchronous Transfer Mode (ATM). ATM employs a 53 byte cell which includes a 5 byte header and a 48 byte payload.
A major challenge currently facing the industry is the transmission of voice information using ATM cells. Various protocols have been developed for converting voice information into the ATM cell format and then back into a digital voice signal. These protocols are known as ATM Adaption Layers (AALs).
Digital voice information is commonly transmitted on the well known 64 kbit/second DS0 circuit. The DS0 bit stream has interface problems with ATM systems. As the voice cells egress from the ATM network, they are converted and placed on a DS0 circuit. The DS0 circuit requires 64,000 bits of information each second, but each cell only transmits 384 bits (48 bytes) of payload. Multiple cells must arrive and be processed each second to feed the DS0 circuit.
A DS0 circuit requires information from a new cell every 6 milliseconds. The 6 millisecond timing constraint derives from the 4 KHz voice bandwidth which is sampled at the Nyquist rate of 8000 samples/second for digital voice transmission. Each sample requires 8 bits (one byte) in current Pulse Code Modulation (PCM) techniques. As a result, a PCM voice signal produces 8000 bytes per second. Since ATM uses a 48 byte payload, 167 cells per second are required (8000/48=167) to transmit the PCM voice signal. Inverting this number shows that a cell must be processed every 6 milliseconds to transmit voice information from an ATM link to a DS0 link. In other words, Each cell can carry 6 milliseconds of voice information, and a cell must be processed every 6 milliseconds when the full payload is used to avoid gaps in the continuous DS0 voice signal.
Cells are sometimes lost during transmission or arrive at the egress point out of sequence. A problem is how to detect these cells and compensate for them. A delay in cell arrival greater than 6 milliseconds means there is no cell to transfer to the DS0--this is a lost cell condition. Also, mis-sequenced cells may arrive and the problem exists of how to detect and correct the sequence.
Although compensation for lost cells may not be critical for true voice communications due to the small amount of information lost, voice band modems experience significant problems given a 6 millisecond loss of information. The use of voice band modems has increased dramatically in recent years due to the proliferation of Personal Computer (PC) based communications over standard telephone lines.
Various attempts have been made to solve these problems. One solution is based on a sequence number which is placed in the payload of the cell. (ATM cells have no sequence number in the header) The sequence number of the cell can be used at the remote end to detect missing cells, and to provide the proper cell sequence. This system is lacking in that it wastes payload capacity carrying the sequence number. The system also requires additional logic to create and identify the sequence number, and additional buffering to realign the cells. The use of a sequence number also introduces delay because multiple cells must arrive before the sequence numbers can re-aligned.
Another solution is based on a detection window within the cell arrival interval. The cell arrival interval is the amount of time a cell has to arrive before a missing cell condition is created. For a DS0 circuit, cells carrying a 48 bytes of voice information must arrive every 6 milliseconds. The 6 milliseconds is the cell arrival window. If the payload size were to change, i.e from 48 bytes to 32 bytes, the interval would shrink because the DS0 would require cells more frequently.
Within the cell arrival interval, a cell detection window is calculated. Current calculations for the 6 millisecond interval put the cell detection window at 1 millisecond. If a cell arrives during the 1 millisecond detection window, it is assumed to be the correct cell. If no cell arrives during the detection window, a cell is assumed to be lost and compensation is then triggered. If a cell arrives during the interval, but outside of the detection window, the cell is assumed to be mis-sequenced and is discarded.
This system is also lacking in that it requires the calculation of a cell detection window. The window would need to be re-calculated for different cell arrival intervals. The system also requires a separate clock to time the detection window. Additional logic is required to detect if cells arrive in or out of the detection window and to trigger compensation based on this determination.
Lost cell compensation requires providing a replacement cell for the lost cell. The replacement cell is typically a white noise cell. A problem exists with using a white noise cell since an ATM cell only contains 48 bytes, there is not enough information to create a true white noise signal which includes energy spread statistically among multiple frequencies. A 48 byte white noise signal becomes a tone when repeated over and over.
Mis-sequenced cell compensation usually entails re-sequencing the cells through a buffer arrangement. Another technique is to discard mis-sequenced cells.
One system has used a sequence number to place cells in sequential addresses of a buffer. The cells are read out of the buffer in sequence. Silence data is written to each address after the address is read. In that way, cells which do not arrive are automatically replaced with silence data at that address. This system still has deficiencies in that a sequence number is used and additional buffering and logic are required. Additionally, the problem of providing white noise in the restricted cell payload is not addressed.
At present, a system is needed that converts cells containing voice information into a digital voice signal and compensates for lost and mis-sequenced cells with a minimum of logic and processing.


There is provided an improved image reader. In the image reader, light from an object is converted into an analog signal by a photoelectric converter. The signal is quantized by an A/D converter on the basis of top and bottom reference voltages to produce a digital picture signal. A digital picture signal corresponding to a dark current flowing to the photoelectric converter is stored in a memory as a dark current data. Also a digital picture signal corresponding to the optical strength of predetermined level is stored in another memory as a white level data. Then, the top and bottom reference voltages are produced which correspond to the top and bottom quantization-threshold values on the basis of the dark current and white level data, respectively. The top and bottom reference voltages are then respectively applied to the A/D converter to produce the digital picture signal without signal distortion.

The present invention relates to an improved image reader.
An image reader produces a picture signal by converting light from an object into an electrical signal for each picture element by means of a photoelectric conversion device. However, the picture signal thus produced includes a distortion component (shading) caused by non-uniformity of the optical strength from a light source, variation of the optical strength-distribution from an optical system and differences of sensitivity between elements constituting the photoelectric conversion device.
An image reader in which a photoelectrically converted picture signal is converted into a digital siganl by an A/D (analog-to-digital) converter has adopted a method for eliminating the shading as follows.
First, light reflected at a standard white board is photoelectrically converted into an analog signal. The analog signal is converted into a digital signal to be stored in a compensation memory prior to image-reading.
Then, a picture signal is produced by actual image-reading, digitized and multiplied by the signal stored previously in the compensation memory corresponding outputs from each element or picture element of the image reader to compensate for the shading.
This system requires a digital multiplier for shading-compensation. Furthermore, when the number of quantized bits is large, the number of digits required for calculation increases greatly. This results in the circuit configuration of the system becoming complex.
Moreover, the photoelectric conversion device has a problem for the accuracy of analog-to-digital conversion due to the dark current if a picture signal with a large gradation is to be produced or if the number of quantized bits is to be made large.


A disk drive system including one or more platters, each platter supporting at least one recording surface, where the platters are aligned about a common central axis. The platters spin about the common central axis. A recording head is associated with each recording surface. An actuator mechanism couples to each recording head to move the recording head into proximity with selected portions of the recording surface in response to received commands. At least two replicates of data are stored in at least two data storage areas such that any one of the at least two replicates can be accessed to service a data access request.

1. Field of the Invention
The present invention relates, in general, to mass storage devices, and, more particularly, to software, systems and methods for implementing high performance disk drives using self-mirroring within the drive.
2. Relevant Background
Computing systems are used to manipulate and store large quantities of data used in commerce, government and research. While the instruction rate at which data processors can perform has increased dramatically over recent years, the input/output (I/O) rate at which data can be supplied to processors has not increased proportionately. Accessing mass data storage systems remains a critical bottleneck limiting the overall performance of many computing systems.
Mass data storage systems strive to provide high storage density with low latency access to the stored data. Disk drive manufacturers continue to provide increasingly higher densities of data storage in storage devices using magnetic, optical, magneto-optical and other storage technologies. Advancements in hard disk magnetic recording technology are enabling the doubling of disk capacity every year.
Disk drives typically comprise one or more disks or platters having a recording media formed on one or more of the disk surfaces. A read/write head attached to an actuator mechanism moves across the disk between inner and outer diameters while the platters spin. Read channel electronics read and write data onto the disk surfaces as well as reading formatting, control and servo data recorded on the disk surface. Servo control electronics coupled to the read channel and the actuator mechanisms use servo control data and high-resolution servo data (e.g., servo bursts) recorded on the media surface to locate particular tracks and align the head over the centerline of selected tracks in response to read/write commands.
By carefully controlling spin speed and actuator movement, data can be accessed with great precision. Although capacity improvements in early disk drives came from adding more platters within a single enclosure, more recently improvements arise from decreasing the track size and recording rate to improve the areal density of the disk drive system. Continuing improvements in media properties, read channel electronics, and servo control systems are expected to lead to continuing higher areal densities.
However, hard disk performance in terms of reduced latency is only improving 15% to 30% a year. Latency in a disk drive system is primarily determined by the seek time and the rotational latency. Other contributing factors to latency, including various processing time required to process commands, tend to be minor. Seek time refers to the time required to position a read/write head over a selected track on a disk drive surface. Rotational latency refers to the time required for the desired location on a track to rotate under the read/write head and is largely a factor of spin speed of the rotating media.
There is a continuing need for higher performance hard disk drives. To achieve high performance customers use multiple drives in RAID (redundant array of independent disks) configurations such as RAID level 0 and RAID level 1 mirroring. Mirroring allows a data set to be spread out or "striped" across many disk drives or spindles so that latency associated with accessing each drive can be overlapped. When choosing to spread data across many spindles customers often choose smaller capacity drives over larger capacity drives driven by overall solution cost and the total capacity needed. There is a trend today that individual drive capacity is larger than what customers need.
While mirroring across multiple drives has improved performance in multiple-drive systems, until now, only a few strategies have existed to reduce latency within the drive itself. The mechanics of the head and actuator mechanisms limit the acceleration and speed at which a head can be moved between inner and outer diameters of the disk. Hence, seek latency is difficult to reduce. However, smaller disks and "short-stroking" (i.e., causing the actuator to move over less than all of the disk surface) have been used to lessen seek latency. In the case of solutions like short-stroking, disk capacity is simply discarded to improve performance. It would be preferable to implement a system that could use this capacity productively to improve performance rather than simply throwing it away.
Rotational latency is conventionally addressed by increasing the rotational speed of the platters. In recent years spin speed has increased to 7200 rpm in consumer disk drives, while 10,000 rpm and 15,000 rpm drives are available in enterprise class drives. However, increasing spin speed has several drawbacks. Higher spin speeds create more vibration, oscillation, and media flutter making it correspondingly difficult to precisely position the head during read/write operations. Also, the read channel and servo electronics must be able to handle the increased data rate. Moreover, faster spinning drives require more power making them impractical for many portable electronic devices. As a result, increasing the spin speed of drives by itself is not expected to provide adequate performance improvements in the near future. Hence, a need exists to provide the benefits of higher spin speeds disk drives without the associated drawbacks that increase cost and reduce performance.

Self-mirroring high performance disk drive

Hewlett-Packard Development Company, L.P.


The let-off warp beam coupling device has a let-off warp beam rotatably and detachably mounted on a specified position of a weaving machine and a beam gear provided on the weaving machine so as to be opposite to a shaft end of a beam shaft of the let-off warp beam for rotating the let-off warp beam. A driven coupling member is provided so as to be secured at the shaft end of the beam shaft of the let-off warp beam, and has a plurality of projections provided on a circle positioned on a plane perpendicular to the axial direction of the beam shaft. Each projection projects in the axial direction of the beam shaft and has a side face tapered in the axial direction. A driving coupling member is also provided on the beam gear side so as to be opposite to the driven coupling member with a specified spacing therebetween. It has a plurality of projections provided on a circle positioned on a plane perpendicular to the axial direction of the beam shaft.

The present invention relates to a let-off warp beam coupling device used when a let-off warp beam is coupled to or uncoupled from a weaving machine for coupling and uncoupling an end of a rotating shaft of the let-off warp beam and a beam gear. The beam gear is positioned on the weaving machine side and provided movably in the axial direction of the beam gear.
An example of a conventional let-off warp beam coupling device for coupling and uncoupling a shaft-end of a let-off warp beam and a beam gear on the side of a weaving machine is as shown in FIG. 1. In the device, on a surface around one end of a beam shaft 2 of a let-off warp beam 1, there was formed a spline portion 3, to which a beam gear 4 was fitted in coaxial. The beam gear 4 is provided on the side of the weaving machine for driving the let-off warp beam 1 through the beam shaft 2 and has a fitting bore 5 at the center with grooves 4a formed inside so as to be fitted to the spline portion 3. The beam gear 4 is provided so as to be movable backward and forward with respect to the end of the beam shaft 2 of the let-off warp beam 1 by a back and forth driving mechanism 6. This allows the grooves 4a of the fitting bore 5 of the beam gear 4 to be engaged and disengaged with the spline portion 3.
There was also another type of the let-off warp beam coupling device as shown in FIG. 2. The device is provided with a clutch 8 comprising a channel 6 provided at an end surface of the beam shaft 2 of the let-off warp beam 1 and a protruded strip 7 provided so as to be freely fitted to the channel 6. The protruded strip 7 of the clutch 8 is provided integrally with the beam gear 4 on the side of a weaving machine with its center on the axis of the beam gear 4. It is movable together with the beam gear 4 backward and forward with respect to the beam shaft 2 by the back and forth driving mechanism 6.
In each of the conventional devices, there were problems such as misalignment of the axis of the beam shaft 2 of the let-off warp beam 1 with the axis of the beam gear 4 or the back and forth driving mechanism 6. Backlash produced between the spline portion 3 and the grooves 4a of the fitting bore 5, and rattling produced between the channel 6 and the protruded strip 7 of the clutch 8. In addition, this failed to transmit accurately the rotating motion of a pinion, which is in a driving mechanism for driving the beam gear 4 provided on the weaving machine side, to the let-off warp beam 1. As a result, variation was caused in the tension of the warp fed from the let-off warp beam.
In view of the foregoing, it is an object of the present invention to provide a let-off warp beam coupling device which can accurately transmit a driving motion without any variation factors such as backlash and misalignment.


The invention relates to analyzing and controlling collection of liquid eluate output from a separation process, in particular by use of a measure of suspended material in the eluate based on a light scattering detection method. Exemplary embodiments include a method of controlling collection of a sample of a liquid eluate output from a separation process. The method includes exposing the liquid eluate to light from a light source; detecting light from the light source scattered by suspended material in the eluate at a detector; and beginning and ending collection of the sample when a measure of the suspended material derived from the detected scattered light enters and leaves a predetermined range.

Biotherapeutic drugs, which may be defined as being derived from living organisms, and includes both proteins and DNA, comprise an important class of medicines that have attracted significant interest from both academic and commercial environments Examples of such drugs include monoclonal antibodies (mABs), Human growth hormone (hGH), insulin and glucagon. In the last decade in particular, a remarkable increase in the research, development and clinical use of biotherapeutic drugs for the treatment of a wide range of diseases, including various cancers, rheumatoid arthritis and multiple sclerosis, has been observed. Many of the currently approved mAB-based drugs (as approved by the US Food and Drug Administration) have been approved in the past five years.
Strict regulatory guidelines require that biotherapeutic drug products are clinically safe. This can significantly lengthen the time before a drug becomes available on the market. Product-specific requirements are, in part, satisfied through full characterisation of the molecular properties, solution behaviour and stability of the biotherapeutic drug formulation and, ultimately, by the demonstrable manufacture of a pure, homogeneous product.
One of the main challenges associated with the production of pure and homogenous biotherapeutic drugs is the formation of aggregates. An aggregate may be defined as an assembly of n monomers (where n>1), a monomer being defined as the most basic, non-divisible, unit of an intact protein molecule. Although natural aggregates, in the form of dimers, trimers and other higher order associations, can occur in some proteins and are essential for proper physiological function, for example in Thyroglobulins, Superoxide distumases and Aldolases; for mABs and other protein-based drugs the formation of aggregates is not required for drug function and instead can adversely affect their role as biotherapeutic agents. The immunogenic potency, i.e. the degree to which a molecule (biological or otherwise) is able to elicit an immunological response, of large protein aggregates that may be made up of several million protein molecules has been widely documented, for example via draft FDA guidance on immunogenicity assessment for therapeutic products issued in 2013 (available from www.fda.org). Whilst potency is a desired attribute for any therapeutic drug, the inherent heterogeneity of aggregate structures is problematic from manufacturing and quality control (QC) perspectives. Presently, the currently limited understanding of aggregate formation does not allow for the controlled and reproducible assembly of homogenous high-order aggregates and, thus, the intensity of immunogenic response cannot be accurately correlated with such molecules. Consequently, FDA guidelines currently recommend that that the presence of aggregates in biotherapeutic drug products are minimised, not only at manufacture but for the duration of the product's shelf-life.
The primary experimental strategy for the production of a highly pure biotherapeutic product centres on the physical separation, or otherwise known as fractionation, of the protein monomer from its aggregated counterparts within a mixture. Fractionation is principally achieved through liquid chromatography, a technique that uses a liquid flow through specifically functionalised columns to influence the elution of protein molecules by exploiting a range of molecular properties, such as size, binding affinity and electrostatic charge. Since the monomer and aggregates do not share the same molecular properties, their elution from functionalised columns is temporally differentiated, and isolation of the purified monomer is possible. Clearly, the analytical power in this technique is only truly realised when it is coupled with a detector that can monitor and visualise the fractionated species eluting over time. Examples of various techniques that are either currently used, or could be used, as liquid chromatography detectors are summarised in Table 1 on the following page.
TABLE 1A summary of properties of various detectors coupled with existing liquidchromatography systems.Used tocontrolfractionDetectorPropertyInformationAdvantageDisadvantagecollectorUltraviolet (UV)Absorbance ofConcentrationInexpensive; widelyNo identity information;Ylight at a specificif detector isapplicable for proteindoesn't respond towavelengthcalibrated &analysisstructural changesRefractive IndexRefraction of lightextinctionInexpensiveSignal instability with bufferY(RI)compared tocoefficientchanges or environmentalreferenceknowncondition e.g. salt gradientsand temperatureFluorescenceEmission of lightInexpensive; sensitiveRequires fluorescent probe;Y1at a specificsmall linear range due towavelengthsensitivityMassMass to chargeMolecularIdentification of elutingExpensive; destructive; notY2Spectrometryratiomassspecies positivelycompatible with excipients;(MS)confirmedrequires compatible buffer;cannot work at formulationconcentrations; ionisationcan disrupt weaklyassociated; automateddetection of intact proteinsnot availableNuclear MagneticProbing3D structureDetailed structuralExpensive; Large footprint;NResonanceelectromagneticinformationLong analysis times; Labour(NMR)field of nucleiintensive; Requireswith NMR-activeexperienced person for dataspininterpretation; non-continuous detection;mainly used for smallmolecules; requireddeuterated solvents andsample modification; notcompatible with formulationexcipientsElementalAbsorbance ofConcentrationSpecific way to followDestructive; Usually used inNanalysislight at discreteof elementelution of targetconjunction with(AAS/ICP-OES)wavelengths byunder studyprotein if metalconcentration detector;free atoms in abindingunless metal stoichiometrygaseous stateknown then protein speciescannot be identifiedEvaporative lightLight scattered byMassRelatively inexpensiveNot applicable for proteinYscatteringmolecule in gasapplications as techniquephaserequires gas phaseNotes:1See for example U.S. Pat. No. 5,541,4202See R. Moritz and N. O'Reilly, J Biomol Tech. 2003 June; 14(2): 136-142
Ultraviolet (UV) detectors are common amongst detectors coupled with liquid chromatography systems for the purpose of profiling protein mixtures for isolation or purification of their components. This is largely due to their cost-effectiveness and wide applicability for protein samples. UV detectors capitalise on a natural and quantifiable phenomenon common to all proteins arising from chromophores: functional groups that are able to absorb light at specific wavelengths. In proteins, for example, the peptide bond absorbs light at 215 nm and the aromatic group on certain amino acids absorbs at 280 nm. No modification is therefore required to be made to the proteins to determine their presence under UV light, and analyses can occur with the protein unmodified.
Signal responses from refractive index (RI) and fluorescence detectors are also quantifiable, but their use in protein purification applications is limited due to signal instability with changes in buffer components and the need for a fluorescent probe, respectively. Concentration information is inherent in the data from all three types of detectors, but no further information confirming the identity of the fractionated species can be extracted from such measurements. Such confirmation would require the implementation of additional offline methods, which contribute significantly to the total analysis and handling times and, correspondingly, decrease efficiency and profitability.
To provide identity information, mass spectrometry (MS) and nuclear magnetic resonance (NMR) technologies in tandem with liquid chromatography systems may be used. However, the additional information obtained through these techniques comes at a price, including increased equipment and running costs and prolonged analysis times. MS and NMR also suffer from sample-specific issues, where the excipients used in biopharmaceutical formulations are frequently incompatible with either technique, as are the concentrations of the protein within. Whether MS accurately reflects the species contained within the original solution is another critical issue for consideration in biopharmaceutical analyses. Since this technique relies on the ionisation of charged groups, the likelihood of observing a given species is directly correlated with the accessibility of these charged sites—which may or may not be the same for two aggregate particles of the same mass—and, furthermore, the ionisation itself may also cause dissociation of weakly associated molecules.
Following successful fractionation of a biopharmaceutical or protein/aggregate mixture, the next consideration is isolation of the fractionated protein monomer. UV, RI, fluorescence and mass spectrometer technologies that facilitate the isolation of fractionated species by controlling a fraction collector are commercially available. This signal-based fraction collection is largely accomplished through user-set parameters that define specific conditions during which a particular fraction should be collected. Such conditions may for example relate to changes in peak slope or absolute signal above a threshold value. The disadvantage of using UV- or RI-directed fraction collection for biopharmaceutical mixtures is that these detectors lack the ability to discriminate between a protein monomer and its aggregates. For example, there is no discernable difference in the signal response for 10 individual monomers or an aggregate composed of 10 monomers. This means that the efficiency of the peak fractionation, i.e. the ability to identify where peaks begin and end in both baseline resolved and unresolved situations, is solely based on the ability to detect changes in the intensity of the resultant peaks during real-time measurements of the phenomenon under observation.
More selective fraction collection based on species identity is possible with mass spectrometry, where a given fraction is collected based on the detection of a particular mass. However, it is important to remember that when looking at intact proteins that the observed species are presented as a mass to charge ratio; therefore, as far as the MS detector is concerned a species of mass 100 and charge of +10 would be the same as a species of mass 10 and charge of +1. No real-time deconvolution of such data is currently possible. Even when looking at peptide fingerprints, such selective fractionation excludes all information from other molecules and as a result does not allow the user to effectively minimise contamination of the fraction from other closely eluting species. Similar drawbacks are applicable for selective fractionation based on fluorescence or elemental analyses.
Ensuring a high purity product at maximum yield is a specific manufacturing challenge faced by the biopharmaceutical industry, on both small and large scales, which is required to meet stringent regulatory demands whilst maintaining profitability. Concomitantly, the demand for technologies that detect, characterise and quantify aggregates, both during and post-manufacture, has increased and manufacturers of scientific instrumentation have duly responded. The benefits of using detectors for online monitoring in polymer manufacture have been recognised and implemented with the optimisation tool ACOMP (automatic continuous online monitoring of polymerisation reaction, as described for example in U.S. Pat. Nos. 6,052,184 and 6,653,150). This tool aims to increase efficient production of a polymerisation reaction by adjusting variables, such as pressure, temperature and reagent concentrations, based on the feedback from real-time analyses of several parameters. However, no technology currently available offers real-time, signal-based fraction collection or identity-based selective fractionation. All existing technologies for analysis of protein/aggregate mixtures require some degree of user intervention for data processing, and none currently allow for automated, software-driven identification of monomer and aggregate peaks or corresponding fractions.
It is an object of the invention to address one or more of the above mentioned problems.


A converter for raising a weak input DC voltage of 0.3 volts or less to an output voltage has a blocking oscillator and a field-effect transistor included in the oscillator. The transistor having a base earthed is of the enhancement type activated by a gate-source voltage equal to or lower than the input, and the oscillator is activated responding to the gate-source voltage, so that pulses are generated as the output. Typically, the oscillator is composed of a resistor and a condenser connected in series through a lead and between an earth and an input line, a transformer whose secondary winding is connected to transistor's gate and the lead, with a primary winding being connected to the input line and the transistor's drain, and a diode earthed and having a cathode connected to the gate. An electromotive force induced through the secondary winding when the transistor is inactivated has positive and negative poles directed to the condenser and the gate, respectively.

The present invention relates to a DC-to-DC converter for converting a DC input into a DC output of a voltage different from that of the input, and more particularly relates to a DC-to-DC converter adapted to convert a weak input of 0.3 V or less into an intensive output of a raised voltage.
A variety of solar batteries and thermal batteries are now available as the power source for electric instruments or apparatuses. Each of these batteries includes a generating device in which an electromotive force will be produced when irradiated with light or when heated. In order to amplify a very weak voltage of such an electromotive force, two or more generating devices are connected one to another in series, or alternatively they are made larger in size. An amplified output from the generating devices will further be intensified in voltage, using a converter if necessary.
The series connection of generating devices to form an array thereof is however somewhat disadvantageous in that even if only one or some of them is or are not irradiated with light or comes or come out of order, they would fail as a whole to generate an output at a desired level of voltage. On the other hand, an electric circuit or the like in which the large-sized generating device is incorporated as a power source would be rendered larger in size and more expensive.


A complementary acoustic charge transport circuit element comprises first and second buried channels. Each of the channels is comprised of a piezoelectric semiconductor and each channel has a source through which charge is injected and a drain through which charge is extracted. A transducer propagates an acoustic wave through each channel and the propagated waves transport the charge between the sources and the drains. A source and/or a drain of one channel is connected in parallel with the corresponding souce and/or drain of the other channel. The waves are complementary at the interconnected ones of the sources and/or the drains.

An acoustic charge transport device ("ACT device") is a novel analog semiconductor device which is particularly useful as a delay line and for performing various signal processing functions. An ACT device overcomes many of the problems of the prior charge-coupled devices ("CCD") because the ACT device processes signals at extremely high speeds while avoiding the necessity of clock driver and certain other circuitry. An ACT device achieves its high processing speed because a surface acoustic wave ("SAW") propagates through a channel buried within the piezoelectric semiconductor substrate and the wave transports the charge through the channel. The charge is transported in discrete identifiable packets which move at the speed of the wave.
Transportation of charge through the buried channel produces an output at the drain (output contact) which is in the form of sharp current pulses. These current pulses originate from the drain only when the SAW voltage under the drain is positive, because the electron packets are carried in the positive half cycles of the SAW voltage. Therefore, it is possible to monitor the pulses after transport through the channel and to determine the quantity of charge that was transported.
The voltage-to-charge conversion process occurring at the ACT device source (input contact) is one wherein the amount of charge injected into each packet is proportional to the input voltage at specific sample points in time. In the ACT device, these sample points tend to occur at the instants in time when the positive peaks of the SAW potential are under the source. Hence, the input section of the ACT device operates as an automatic time domain sampler having a sampling rate equal to the SAW frequency.
In theory, an ideal sampler would produce a sample (charge packet) which is proportional to the analog input voltage at specific exact instants in time. The achievement of the ideal sampler, however, requires a physical process or circuit element which can turn on and off, or switch, infinitely fast. Actual physical devices have finite switching times which cause the sample amplitude to be proportional to a weighted average of the input signal over the switching time. This switching time is referred to as the sampling aperture. The primary impact of a finite sampling aperture is to limit the precision with which the sample can be formed. This, in turn, tends to limit the highest frequency of input signal which can be unambiguously sampled. The sampling aperture of the input sampling mechanism of the ACT device is very short, and is substantially equal to approximately one tenth (1/10) of the SAW period. Hence, in principle, the ACT device input is capable of high speed voltage-to-charge conversion over a wide bandwidth of input frequencies, up to approximately ten times the SAW frequency.
Prior art structures have not been able to take full advantage of the high speed input properties of the ACT device. The prior structures limited the sampling rate to the SAW frequency, even though the input charge injection process is capable of injecting charge at a rate substantially equal to approximately ten times the SAW frequency. In analog signal processing applications, the limitation of the sampling rate at the SAW frequency also limits the signal bandwidth obtained in the devices to one half of the SAW frequency, because of the Nyquist criteria.
In view of the above, it can be seen that there is a need for a structure permitting maximum utilization of the high speed processing capabilities of the ACT device. The disclosed invention is a novel arrangement of at least two ACT devices, so that the waves are complementary at specific points, in a manner which overcomes the limitation of the SAW frequency sample rate. In addition, the Nyquist limited signal bandwidth is correspondingly extended in certain embodiments.
The complementary ACT device ("C-ACT device") is a circuit element which combines two or more similar ACT devices by the parallel connection of the sources and/or the drains thereof. The SAWs of the devices are complementary in phase with respect to the connected sources and/or drains, so that the input sampling time intervals and/or the output detection intervals are staggered, thereby permitting input sampling and output detection at a rate equal to N times the SAW frequency, where N is the number of devices which are connected. The invention may be realized by externally interconnecting discrete devices, or by integrating the required multiple ACT transport channels on one chip in a parallel configuration. In the latter case, which is particularly useful for achieving the required accurate signal balance between channels, the composite circuit element forms a new device with terminal properties considerably different from those obtained from a prior art ACT device.

Complementary acoustic charge transport device and method


A presser bar 11 is provided at its lower end with a fabric presser foot 13 and a nut 16. The fabric presser foot 13 is individually moved on the presser bar 11 independent of a reciprocating motion of the presser bar 11. Due to an action of a spring 14 the presser foot 13 is resiliently pressed against a fabric 40 to be sewn to reduce a noise while a needle is associated with a hook or shuttle 26.

The present invention relates to a fabric presser foot device for a sewing machine, wherein a presser foot movable in a vertical plane is mounted on a lower end of a presser bar.
Generally, a sewing machine is used to sew two superposed fabrics or leathers to be stitched together with a needle taking up a thread. This type of the machine is well known, for example, by Japanese Patent Publication (KOKOKU) No. 41080/1983 wherein a needle bar is arranged in parallel with a fabric presser bar in such a manner that the needle bar is moved upward and downward by synchronizing with a rotation of a main shaft of the machine and the fabric presser bar is movable on the needle bar. Disposed is a coil spring between a frame of the machine and the top of the fabric presser bar so as to press a presser foot formed on a lower end of the fabric presser bar against the fabrics or leathers to be sewn. In this case, when the needle is moved downward toward its dropping position, the coil spring is compressed and the biasing force of this compressed coil spring is exerted upon the fabrics or leathers to be sewn via the fabric presser foot which is moved downward for pressure contact with the fabric on a needle plate.
It is noted in a prior art that the fabric presser foot is forced to be dropped by the biasing force of the coil spring until it comes into collision with the fabric on the needle plate while the moving needle is stuck into the fabric. This collision is repeated at a high frequency so that undesired noise emits.
To eliminate such the noise Japanese Patent Publication (KOKOKU) No. 38239/1981 and Utility Model Publication (KOKOKU) No. 8855/1985 propose a fabric presser foot device as shown in FIGS. 7 and 8, wherein a main shaft 100 of a sewing machine is connected through a link 101 with a presser bar 102 having a fabric presser foot 103 at its lower end. A rotation of the main shaft 100 allows the presser bar 102 to be moved upward and downward so that a stroke of the fabric presser foot 103 with respect to a needle plate 105 is predetermined to reduce the noise.
However, a problem normally arises when the fabric of a hard material is sewn, because a weight of the presser bar 102 having the fabric presser foot 103 and a biasing force of a spring 104 are directly transmitted to the needle plate 105 through the fabric and the noise is not reduced to a comfortable level or magnitude.

Fabric presser foot device of a sewing machine


This invention relates to a pharmaceutical composition for treatment of a bone disease comprising, as an active ingredient, a protein comprising an extracellular cysteine-rich domain, which is from the Frizzled receptor selected from the group consisting of mammalian animal-derived Frizzled 1, Frizzled 2, and Frizzled 7 and has activity of increasing bone mass, bone density, and/or bone strength, or a mutant of such domain having sequence identity of 85% or higher to the amino acid sequence of the domain and having activity of increasing bone mass, bone density, and/or bone strength, or a vector comprising a nucleic acid encoding the protein.

A super-aging society has arrived, the number of people with osteoporosis has increased, and bone fractures resulting therefrom have come to constitute a serious issue of concern at a societal level. In particular, patients with femoral neck fractures and vertebral body fractures become bedridden, which causes significant deterioration of the quality of life thereof, and the social, medical, and econimc burdens caused by care and hospital treatment have increased (Tosteson, A. N., et al., Osteoporos Int., 12, 1042-1049, 2001; and Yoh, K., et al., J. Bone Miner. Metab., 23, 167-173, 2005). It has also been discovered in recent years that osteoporosis is significantly associated with mortality in old age (Nguyen, N. D., et al., J. Bone Miner. Res., 22, 1147-1154, 2007; and Muraki, S., et al., J. Bone Miner. Metab., 24, 100-104, 2006). Under such circumstances, prevention and treatment of osteoporosis have become critical objectives to be achieved. Osteoporosis (i.e., a pathological condition where bone mass is reduced while the rate of the amount of the bone matrix to the amount of the mineralized bone matrix is held) is classified as primary osteoporosis or secondary osteoporosis. The former type is a pathological condition heretofore referred to as postmenopausal osteoporosis or senile osteoporosis. The latter type is a pathological condition caused by changes in bone metabolism resulting from other diseases, and such osteoporosis is classified based on the cause thereof, such as osteoporosis caused by endocrine, nutritional/metabolic, inflammatory, immobile, drug-induced, hematologic, congenital, or other diseases. According to the above classification, examples of causes for secondary osteoporosis include: endocrine causes, such as hyperparathyreosis, hyperthyreosis, hypogonadism, Cushing's syndrome, somatotropin deficiency, diabetes, Addison's disease, and calcitonin deficiency; nutritional/metabolic causes, such as chronic degenerative diseases, emaciation, serious liver diseases (primary biliary cirrhosis, in particular), gastric resection, scorbutus, malabsorption syndrome (including celiac disease), hypophosphatemia, chronic renal disease, hypercalciuria, hemochromatosis, amyloidosis, mast cell tumor, ingestion of excess sodium, insufficient calcium intake, and hypervitaminosis D or A; inflammatory causes, such as articular rheumatism, periarticular bone disease (elevated bone resorption induced by proinflammatory cytokines), and sarcoidosis; immobility-related causes, such as systemic, bed rest, paralysis, local, and post-fracture causes; drug-induced causes, such as with the use of steroids (steroids are extensively used for inflammatory diseases as immunosuppressive agents; examples of diseases treated with the use of steroids include collagen diseases, asthma, inflammatory bowel diseases, and in the case of organ transplantation, and bone loss is a serious side effect of such therapy), methotrexate, heparine, warfarin, anticonvulsant agents, lithium, and tamoxifen; blood-disease-induced causes, such as multiple myeloma, lymphoma, leukaemia, hemophilia, and chronic hemolytic diseases; congenital causes, such as dysosteogenesis, Marfan's syndrome, Kleinfelter's syndrome, congenital erythropoetic porphyria, and cystic fibrosis; and other disease-induced causes, such as with chronic obstructive lung diseases, hepatic failure, renal diseases, articular rheumatism, pregnancy, hyperoxemia, and HIV infection (Committee for Creation of Guidelines for Prevention and Treatment of Osteoporosis, Guidelines for Prevention and Treatment of Osteoporosis 2006, Life Science Publishing, Co., Ltd., Japan, 2006).
Among the above-mentioned diseases, bone diseases resulting from osteoarthritis, articular rheumatism, malignant tumors, or renal diseases are specifically regarded as bone diseases that impose serious influences at the societal level, in addition to primary osteoporosis.
Osteoarthritis develops most often in locomotor regions. The number of patients afflicted therewith is said to be 10,000,000 in Japan, and it has been deduced that the number of patients will keep increasing as the aging of society advances. Advanced articular disorders are treated via artificial joint replacement; however, radical treatment of moderate or milder symptoms has not yet been reported (Nampei, A. & Hashimoto, J., The Bone, 22, 3, 109-113, 2008).
Articular rheumatism is a chronic and progressive inflammatory disease characterized mainly by multiple arthritis. Articular synovial proliferation gradually causes infiltration of cartilage or bones in the vicinity thereof, and articular rheumatism often leads to destruction and deformation of joints. It has been reported that treatment with the use of an antirheumatic drug (methotrexate) cannot sufficiently inhibit the progress of joint destruction, and a biological agent targeting a tumor necrosis factor (TNF) a produces significant effects of inhibiting joint destruction. Thus, it is considered to be a revolutionary agent. However, increased incidence, as a side effect, of opportunistic infection, tuberculosis (extrapulmonary tuberculosis), Pneumocystis pneumonia, or the like when using such agent is an issue of concern (Soen, S., The Bone, 22, 3, 103-107, 2008).
Major examples of bone diseases involved in malignant tumors include hypercalcemia and bone metastasis related to malignant tumors. Hypercalcemia causes loss of appetite and diuresis, and it causes dehydration and renal failure caused thereby. Bone metastasis is often observed in patients with breast cancer, prostate cancer, or lung cancer, in particular. While bone metastasis is hardly ever fatal by itself, it causes bone ache, pathologic fracture, neuroparalysis, or the like. It thus often significantly deteriorate patients' QOL, and bone metastasis control is a critical objective in clinical settings (Takahashi, S., The Bone, 22, 3, 115-120, 2008). These bone diseases related to malignant tumors are treated with the use of bisphosphonate preparations, although the problem of side effects has been pointed out.
Among bone diseases related to renal diseases, a pathological condition of bone damage caused by renal tissue damage is referred to as renal osteodystrophy. Bone disease experienced by kidney dialysis patients are mainly caused by secondary hyperparathyreosis. Because of the elevated PTH concentration caused by hyperparathyreosis and, for example, insufficient production of bone morphogenetic protein (BMP) 7, renal osteodystrophy advances. Dialysis patients often exhibit lowered reactivity of the bone with the parathyroid hormone (PTH). When the PTH concentration is chronically and significantly elevated, accordingly, fibrous ostitis (high bone turnover) develops. When the PTH concentration is maintained within a standard range, in contrast, bone aplasia (low bone turnover) develops.
When fibrous ostitis advances, collagen fibers are irregularly formed, such fibers are mineralized as non-crystalline calcium phosphate, and woven bone is then formed. This enhances bone formation, although the bone becomes easily fracturable. Basic treatment of fibrous ostitis involves inhibition of parathyroid hormone secretion, which mainly entails calcium ingestion and administration of active vitamin D. When a patient has a chronic kidney disease (CKD) and receives dialysis treatment, in particular, various regulations, such as restrictions on food or water intake, are necessary. When secondary hyperparathyreosis advances, hypercalcemia also becomes an issue of concern. When prescribing active vitamin D, extreme caution, such as via the monitoring of renal functions (i.e., serum creatinine level) and serum calcium level, is always required.
Bone aplasia develops because of prolonged use and excessive administration of active vitamin D preparations or suppression of parathyroid hormone after parathyroidectomy (PTX).
The rate of fractures associated with bone aplasia is higher than that associated with fibrous ostitis, and it induces hypercalcemia or mineralization of blood vessels or other soft tissues. Thus, adequate treatment techniques have been desired. A pathological condition of bone aplasia is low bone turnover in which bone resorption and bone formation are inhibited, and there is no established treatment technique at present (Daugirdas, J. T., et al., Rinsho Toseki Handbook (Handbook of Dialysis), Fourth Edition, Medical Sciences International, Ltd., Japan, 2009).
Hyperphosphatemia or hypercalcemia caused by lowered capacity of the bone for phosphorus or calcium intake (low-turnover metabolic bone) or lowered storage capacity (high-turnover metabolic bone) is considered to be a cause of ectopic (vascular) mineralization. Cardiovascular complications account for 40% or more of the deaths of patients with chronic renal failures, and dialysis patients in particular, and arteriosclerosis involving vascular mineralization has drawn attention as a serious pathological condition. Treatment of mineralization of advanced lesions in patients with chronic renal failures remains difficult at present and the prognosis thereof is poor (Fujiu, A. et al., Rinsho Toseki (the Japanese Journal of Clinical Dialysis), 24, 43-50, Nihon Medical Center, Japan, 2008). In addition to agents for treating primary osteoporosis, accordingly, development of agents that more effectively act on bone diseases resulting from osteoarthritis, articular rheumatism, malignant tumors, or renal disease and vascular mineralization resulting from bone diseases with reduced side effects has been desired.
It is considered that bone metabolism is regulated by the balance between osteoblast functions and osteoclast functions, and osteoporosis develops when the bone-destroying activity exceeds bone-building activity (Cohen, M. M. Jr., American J. Med. Genetics, Part A, 140A, 2646-2706, 2006). In particular, secretion of the female hormone that assumes the role of protecting bones is lowered in postmenopausal women, a lowered capacity of osteoblasts for bone formation and the elevated bone resorption activity of osteoclasts are consequently observed, and it is highly likely that symptoms of osteoporosis would develop (Kousteni, S., et al., Cell, 104, 719-730, 2001; and Nakamura, T., et al., Cell, 130, 811-823, 2007). In order to overcome such problems, estrogen preparations have been used; however, application thereof has been restricted due to the increased risk of thrombosis and breast cancer caused by the use of such preparations. It is also reported that use of a selective estrogen receptor modulator would increase the risk of deep vein thrombosis (Wada, S., et al., Mebio, 25, 8, 89-95, 2008).
At present, calcitonin, bisphosphonate, and the like are used as agents that inhibit the bone resorption activity of osteoclasts. Calcitonin is known to bind to a calcitonin receptor expressed on the osteoclast surface to inactivate osteoclasts, and it is used for treatment of not only osteoporosis but also hypercalcemia, Paget's disease of bone, and the like in clinical settings. However, no effects thereof on bone fracture inhibition have yet been found, and calcitonin receptor expression is reported to be down-regulated by calcitonin administration (Wada, S., et al., Mebio, 25, 8, 89-95, 2008; and Wada, S. & Yasuda, S., Clin. Calcium, 11, 9, 1169-1175, 2001). Bisphosphonate exhibits potent bone resorption inhibitory activity, and amino-containing bisphosphonates, such as andronate and risedronate, are major therapeutic agents for osteoporosis in Japan. Such bisphosphonate preparations inhibit farnesyl diphosphate synthase, block lipid protein prenylation, and induce inhibition of bone-resorption functions and osteoclast apoptosis (Nakamura, T., The Bone, 22, 3, 147-151, 2008). However, the FDA warned of crises of severe skeletal, articular, or muscular pain in 2008 as problems of bisphosphonate preparations. In addition, side effects, such as jaw bone necrosis, caused by the prolonged use thereof (i.e., for 2 or 3 years or longer) after dental care have been reported (Sanna, G., et al., Ann. Oncol., 16, 1207-1208, 2005). An anti-RANKL antibody has been expected as a novel osteoclastic inhibitor other than those described above. Further, application of the anti-RANKL antibody as an inhibitor of articular destruction in the case of articular rheumatism or as a therapeutic agent for multiple myeloma has been expected, and clinical development thereof is in progress. Based on a report to the effect that the RANKL/RANK pathway is important for the survival and maintenance of dendritic cells (Theill, L. E., et al., Ann. Rev. Immunol., 20, 795-823, 2002) or a report to the effect that lymph node dysplasia is caused in an RANK- or RANKL-deficient mouse (Kong, Y. Y., et al., Nature, 397, 315-323, 1999; and Dougall, W. C., et al., Genes Dev., 13, 2412-2424, 1999), the influence of an anti-RANKL antibody preparation on the immune system has become an issue of concern. In 2008, AMGEN reported that an increased rate of development of some infectious diseases was found through a clincal test of the anti-RANKL antibody preparation (Denosumab). As a result of the clinical test of the anti-RANKL antibody conducted in 2009, development of jaw bone necrosis was found to be a side effect, as in the cases of the bisphosphonate preparations. Treatment via intermittent administration of PTH alone as an osteogenesis accelerator that activates osteoblasts has been conducted (Teriparatide, Eli Lilly; an unapproved drug in Japan), but such agent is not different from other therapeutic agents, such as bisphosphonate preparations, in that activity of increasing cortical bone thickness is not very high compared with activity of increasing cancellous bone mass. Accordingly, the effects thereof for bone fracture prevention are not considered to be very high. In relation to PTH, further, Asahi Kasei Pharma Corp. (Japan) has reported problems, such as side effects such as palpitation, tachycardia, and a lowering in blood pressure, and osteosarcoma observed in a long-term administration test to rats, unapproved continuous use thereof for 1.5 to 2 years or longer in Europe and the United States, and prohibited application thereof to cancer patients. Thus, it is impossible to use PTH for inhibition of cancer bone metastasis, treatment of cancer-induced hypercalcemia (paraneoplastic humoral hypercalcemia or local osteolytic hypercalcemia caused by the parathyroid-hormone-related peptide produced by tumor cells), or other purposes.
Accordingly, development of agents that more effectively work for osteoporosis caused by the lowered capacity of osteoblasts for bone formation or elevated bone resorption activity of osteoclasts in postmenopausal women, hypercalcemia, Paget's disease of bone, inhibition of bone metastasis inhibition of articular destruction associated with articular rheumatism, or multiple myeloma with reduced side effects has been awaited.
In addition thereto, osteohalisteresis and rachitis are known as bone diseases induced by selective inhibition of mineralization, unlike osteoporosis. A bone is formed by mineralization of a matrix layer comprising collagen or the like via hydroxyapatite deposition. Osteohalisteresis is a pathological condition in which such mineralization is blocked and osteoids increase, and it is referred to as rachitis if developed during childhood. Symptoms include bone and joint pains, such as chiropodalgia, arthralgia, lumbago, and backache, which lead to gait impairment and to a state in which bone is easily fractured. In the case of children, developmental disorders, limb deformities such as bow-legs, pigeon breast deformity, or other symptoms are observed. Such symptoms are generally treated with the use of vitamin D, calcium preparations, and phosphorus preparations, in addition to alimentary therapy. If the level of dysfunction caused by a deformity is high, however, surgical operation is the only possible symptomatic treatment. Therefore, development of agents that are more effective on osteohalisteresis or rachitis has been awaited.
As described above, bone is tissue that is always regulated by the balance between osteoblast functions and osteoclast functions and remodeled. In order to achieve tough bone that is more resistant to fracture, accordingly, a mere increase in bone mass may not be sufficient. In the case of hereditary diseases, such as osteopetrosis (Horiuchi A., CLINICIAN, 47, 401-404, 2000), Paget's disease of bone (Daroszewska, A., & Ralston, S. H., Nature Clinical Practice Rheumatology, 2, 270-277, 2006), or Camurati-engelmann's disease (CED) (Janssens, K., et al., Nature Genetics, 26, 273-275, 2000; and Tang, Y., et al., Nature Medicine, 15, 757-765, 2009), for example, it is known that the balance between bone formation and bone resorption becomes abnormal due to different causes, and bone strength is lowered even though bone mass is increased. Examples of factors that determine bone strength from the viewpoint of mechanisms of materials include form-related factors, such as connectivity of cancellous bones, thickness of cortical bones, porosity, and cross-sectional moment, and qualitative factors, such as mineralization or bone fatigue, in addition to quantitative factors represented by bone density (Mori S., CLINICIAN, 49, 621-626, 2002). Therefore, development of agents useful for improving bone strength, in addition to increasing bone mass, has been awaited for the purpose of treatment of primary osteoporosis and secondary osteoporosis.
In recent years, factors associated with the Wnt/LRP signal control mechanism have drawn attention as targets for drug discovery regarding a bone formation accelerator. Wnt is a secreted glycoprotein that has been lipid-modified by palmitic acid having a molecular weight of about 40,000, and 19 types thereof are considered to be present in mammalian animals. As Wnt receptors, 10 types of seven-transmembrane receptors (i.e., Frizzled receptors) and two types of single-transmembrane receptors (i.e., LRP5/6 receptors) have been reported (Tamai, K., et al., Nature, 407, 530-535, 2000). A region referred to as a cysteine-rich domain (CRD) containing conserved 10 cysteine residues is present in an extracellular region of the Frizzled receptor family molecule to which Wnt is considered to bind. The region from the cysteine residue located closest to the N-terminus to the cysteine residue located closest to the C-terminus of such 10 cysteine residues may be exclusively designated as a CRD (Masiakowski, P. & Yancopoulos, G. D., Curr. Biol. 8, R407, 1998), or a region comprising such 10 cysteine residues and sequences each located closer to the C- or N-terminus may be designated as a CRD (R & D systems). CRDs were reported to have homodimer structures based on crystal structural analysis using a CRD of mouse Frizzled 8 (Dann, C. E., et al., Nature, 412, 86-90, 2001). At least three types of Wnt signaling pathways are considered to exist: a canonical-Wnt signaling pathway; a non-canonical Wnt signaling pathway, which is a PCP (planar cell polarity) pathway mediated by a small G-binding protein; and a Ca2+ pathway mediated by a trimeric G protein. Bone-metabolism-related research on the canonical-Wnt signaling pathway is the most advanced, and Wnt is considered to promote bone formation (Rawadi, G. & Roman-Roman, S., Expert Opin. Ther. Targets, 9, 5, 1063-1077, 2005). Therefore, regulation of functions of endogenous factors that inhibit this signaling pathway has been attempted in recent years for the purpose of application thereof to treatment of bone diseases.
Sclerostin was recognized as a BMP antagonist at first; however, it was reported to be a factor that would directly bind to LRP5/6 to inhibit the signaling pathway in research conducted later (Semenov, M., et al., J. B. C., 280, 29, 26770-26775, 2005). A significant increase was observed in bone density in a Sclerostin-knockout mouse (Li, X., et al., J. Bone Miner. Res., 23, 860-869, 2008). At present, a Sclerostin-neutralizing antibody is undergoing phase I trials in Europe and the United States of America (AMG785, Amgen & UCB), and the future development thereof has drawn attention. A DKK1 (Dickkopf-1)-neutralizing antibody that is known as another canonical-Wnt signal inhibitor was prepared, inhibition of lowered bone density was observed in an SCID mouse into which multiple myeloma (MM) cells had been transplanted (Yaccoby, S., Blood., 109, 2106-2111, 2007), and clinical trials using a neutralizing antibody (BHQ880, Novartis) have been conducted.
sFRP (soluble frizzled-related protein) that is considered to be a Wnt decoy receptor and has high amino acid sequence homology to the Frizzled extracellular domain is considered to negatively regulate Wnt signals (Nakanishi, R., et al., J. Bone Miner. Res., 21, 1713-1721, 2006), and an increase in the amount of cancellous bone in the femur of an sFRP1 knockout mouse has been reported (Trevant, B., et al., J. Cell. Physiol. 217, 113-126, 2008). Under such circumstances, research and development related to sFRP1 inhibitors have proceeded (Wyeth).
Frizzled 7 has been identified as a receptor that binds to a Wnt ligand and transmits signals thereof (Wang, Y., et al., J. B. C., 271, 8, 4468-4476, 1996; and Huang, H-C., & Klein, P. S., Genome Biology, 5, 234, 1-7, 2004). The amino acid sequence of the human Frizzled 7 extracellular cysteine-rich domain (when a region from the cysteine residue located closest to the N-terminus to the cysteine residue located closest to the C-terminus of such 10 conserved cysteine residues is exclusively designated as a CRD) is completely identical to that of the mouse Frizzled 7 extracellular cysteine-rich domain (i.e., there is no difference between species). Involvement thereof with generation and differentiation of individual organisms (Wheeler, G. N., Current Biology, 10, 849-852, 2000) and involvement thereof with liver cell multiplication (Matsumoto, K., et al., Dev. Biol., 319, 2, 234-247, 2008) have been reported.
Expression patterns of such molecules have been reported: an expression pattern localized in the crypt base of the mouse small intestine or large intestine (Gregorieff, A., et al., Gastroenterology, 129, 626-638, 2005); elevated expression levels in various cancer cells (Katoh, M. & Katoh, M., Int. J. Mol. Med., 19, 529-533, 2007); expression in various tissues (the brain, eyeball, heart, kidney, liver, lung, or spermary) other than those of the spleen via expression analysis of adult mouse-derived tissues of (Wang, Y., et al., J. B. C., 271, 8, 4468-4476, 1996); and expression in tissue (the lung or kidney) other than those of the brain and the liver via expression analysis of human fetal tissue and potent expression in the skeletal muscle and relatively potent expression in the heart, weak expression in the brain, the placenta, and the kidney; and no expression in the lung, the liver, the pancreas, the spleen, the thymic gland, the prostate, the testicle, the ovary, the small intestine, or the large intestine via expression analysis of adult human-derived tissue (Sagara, N., et al., B. B. R. C., 252, 117-122, 1998).
An extracellular cysteine-rich domain that is a soluble receptor of the Frizzled receptor is considered to bind to Wnt and inhibit functions thereof. It is reported by an in vitro experimentation system that a fusion product of the Frizzled 7 extracellular cysteine-rich domain (comprising a region from the cysteine residue located closest to the N-terminus to the cysteine residue located closest to the C-terminus of the conserved 10 cysteine residues and sequences each located closer to the C- or N-terminus) and Fc (R & D Systems) inhibits stabilization of cytoplasmic β-catenin by Wnt3a (Kemp, C. R., et al., Dev. Dynanics, 236, 2011-2019, 2007). Since the expression level of Frizzled 7 is elevated in cancer cells, it has drawn attention as a target molecule for tumor treatment (WO 2008/031009; and Merle, P., et al., J. Hepatol., 43, 5, 854-862, 2005). Regarding colon cancer cells into which a vector that expresses a Frizzled 7 extracellular domain has been introduced, for example, growth thereof was inhibited to a greater extent in a xenograft tumor cell transplantation model compared with colon cancer cells into which a control vector had been introduced (Vincan, E., et al., Differentiation, 73, 142-153, 2005). This suggests the possibility that Frizzled 7 would be a target of drug discovery for tumor treatment.
As described above, 10 types of Frizzled family molecules have been reported, and Frizzled 1 and Frizzled 2 have been reported as molecules having particularly high primary sequence homology with Frizzled 7 in the extracellular cysteine-rich domain (when a region from the cysteine residue located closest to the N-terminus to the cysteine residue located closest to the C-terminus of the conserved 10 cysteine residues is exclusively designated as a CRD, Daroszewska, A., & Ralston, S. H., Nature Clinical Practice Rheumatology, 2, 270-277, 2006). The amino acid homologies of Frizzled 7 in the cysteine rich domain (when a region from the cysteine residue located closest to the N-terminus to the cysteine residue located closest to the C-terminus of the conserved 10 cysteine residues is exclusively designated as a CRD) of such molecule to Frizzled 1 and Frizzled 2 are 91% and 93% respectively in humans and mice. That is, such amino acid sequence homology is very high. As with the case of Frizzled 7, Frizzled 1 and Frizzled 2 do not show differences between mouse-derived and human-derived amino acid sequences in the cysteine rich domain (when a region from the cysteine residue located closest to the N-terminus to the cysteine residue located closest to the C-terminus of the conserved 10 cysteine residues is exclusively designated as a CRD); i.e., such sequences are 100% consistent with each other.
As with Frizzled 7, it is reported that both Frizzled 1 and Frizzled 2 interact with Wnt and Frizzled 1 interacts with Wnt3a to protect the hippocampal neuron from being destroyed by amyloid β peptide (Chacon, M. A., et al., J. Cell Physiol., 217, 215-227, 2008). In addition, regarding Frizzled 1 expression patterns, potent expression in the heart, the placenta, the lung, the kidney, the pancreas, the prostate, and the ovary observed via expression analysis of adult human-derived tissue and potent expression in the lung and the kidney observed via expression analysis of fetus-derived tissue have been reported (Sagara, N., et al., B. B. R. C., 252, 117-122, 1998). Since the expression levels of both Frizzled 1 and Frizzled 2 are elevated in the case of colon cancer or breast cancer, the correlation thereof with canceration is suggested, and they have drawn attention as target molecules for tumor treatment (WO 2008/061013; Holcombe, R. F., et al., Mol. Pathol., 55, 220-226, 2002; and Milovanovic, T., et al., Int. J. Oncology, 25, 1337-1342, 2004). Further, it was reported that Frizzled 1 would not cause any changes in the phenotype of the Frizzled 1 gene-disrupted mouse (Deltagen, Inc., "NIH initiative supporting placement of Deltagen, Inc. mice into public repositories" MGI Direct Data Submission 2005 (www.informatics.jax.org/javawi2/servlet/WIFetch?page=alleleDetail&key=40116)). When a protein comprising an extracellular cysteine-rich domain derived from the Frizzled 1, Frizzled 2, or Frizzled 7 receptor is expressed in vivo at high levels or when a protein comprising an extracellular cysteine-rich domain derived from Frizzled 1, Frizzled 2, or Frizzled 7 is administered in vivo, accordingly, it has been very difficult to deduce that such protein would promotively and specifically function so as to increase bone mass and to enhance bone strength.

This patent is subject to a terminal disclaimer.

Pharmaceutical composition for treating bone diseases which comprises protein comprising Frizzled 1, Frizzled 2 or Frizzled 7 extracellular cysteine-rich domain


Device for coupling an ignitive reaction or percussive shock wave to a relatively movable body member such as an airbag mounted on the steering wheel of an automobile. The device includes first and second relatively movable body members with a transmission channel for ignitive reactions in one of the body members. Pyrotechnic transmission lines are connected to the body members with the gas channels of the transmission lines positioned for communication with the transmission channel notwithstanding relative movement of the body members.

This invention pertains generally to pyrotechnic systems and more particularly to systems in which an ignitive reaction or shock wave is transmitted to a relatively movable body such as an airbag mounted on an automotive steering wheel.
Pyrotechnically fired airbags have recently been developed as an alternative to earlier systems in which inflation of an airbag was initiated by the firing of an electrically actuated squib. Instead of electrical impact sensors and electrical conductors interconnecting the sensors and squib, a pryotechnic system utilizes a mechanical sensor which fires a percussion primer and a shock tube or pyrotechnic transmission line for transmitting the ignitive reaction or percussive wave from the primer to the gas generator associated with the airbag. Pyrotechnic systems have certain advantages over electrical airbag systems in that they operate independently of electrical power sources and cannot be fired inadvertently by electrical disturbance or electromagnetic radiation. Moreover, a pryotechnic system provides long term reliability, requires very little maintenance and is relatively safe to manufacture and install since there is no electric squib to be fired inadvertently.
One problem with a pryotechnic system such as an airbag system is the difficulty of coupling the ignitive reaction or shock wave between a relatively stationary portion of the system and a relatively movable portion such as an airbag and gas generator mounted on the steering wheel. One prior art system uses a length of shock-tube cord or transmission line wrapped about the steering wheel hub and a detonator mounted on the steering column for igniting the cord.