This invention provides novel libraries of olfactory receptor odorant/ligand-binding domains and methods of making and using them. The invention also provides libraries of vectors and cells comprising these nucleic acid constructs. The compositions and methods of the invention are used to identify novel ligand-binding domains for olfactory neuron odorant receptors and their ligands. Thus, the compositions and methods of the invention can be used to generate novel odorants, to screen for toxic odorants, or to manipulate an animal""s olfactory response.

This invention generally pertains to the fields of cell biology and medicine. In particular, this invention provides novel libraries of nucleic acids encoding odorant/ligand-binding domains. Also provided are libraries of hybrid 7-transmembrane olfactory receptors comprising these odorant ligand-binding domains. The compositions and methods of the invention can be used to identify novel ligand-binding domains for olfactory neuron odorant receptors and their ligands. Thus, the compositions and methods of the invention can be used to generate novel odorants and to manipulate an animal""s olfactory response.
A better understanding of the vertebrate olfactory system would provide improved means to manipulate this process and possibly prevent disease or injury. For example, means to manipulate human olfactory neuron odorant receptors from healthy individuals and from individuals with neuro-psychiatric illnesses would offer systems for testing possible odorant/ligands for therapeutic and toxic effects. However, our ability to detect and discriminate between the thousands of beneficial or toxic odorants is complicated by the fact that odorant receptors belong to a multigene family with at least 500 to 1000 members. Furthermore, each olfactory receptor neuron may express only one, or at most a few, of these olfactory receptors. Any given olfactory neuron cell can respond to a small, arbitrary set of odorant-ligands. Odorant discrimination for a given neuron may depend on the ligand specificity of the one or few receptors it expresses. Thus, given this systems"" complexity, information about odorant/ligand-receptor recognition remains meager.
To analyze odorant/ligand-receptor interactions and their effects on cell physiology, it is first necessary to identify specific odorant/ligand(s) and the olfactory receptors to which they specifically bind. Such analysis requires isolation and expression of olfactory receptor polypeptides. However, despite the fact that many putative olfactory receptors have been cloned, only limited progress has been made in the functional expression of these receptors because present systems fail to efficiently translocate these 7-transmembrane proteins to the plasma membrane. This may be because olfactory receptors are a subclass of 7-transmembrane-domain receptors. For example, expression of one rat olfactory receptor in insect cells resulted in only a modest elevation in second messengers when exposed to a mixture of odorants; responses to single compounds were not seen (Raming (1993) Nature 361:353-356). The present invention addresses these and other needs.
The present invention provides novel compositions and methods to generate great numbers, or libraries, of odorant receptor ligand-binding regions. Also provided are novel chimeric olfactory receptors that incorporate these libraries of odorant binding domains. The present invention also provides novel compositions and methods to efficiently translocate polypeptides to the plasma membrane surface. Another aspect of the invention is based on the surprising discovery of a peptide domain that, when incorporated into a polypeptide, can with great efficiency xe2x80x9cchaperonexe2x80x9d or translocate the hybrid protein to the cell plasma membrane. Combining these two aspects of the invention also provides expression vectors and cells that efficiently express these recombinant proteins. Cells and transgenic animals efficiently expressing libraries of hybrid olfactory receptors can be used for screening potential beneficial and toxic odorant molecules.
The invention provides an amplification primer sequence pair for amplifying a nucleic acid encoding an olfactory receptor ligand-binding region comprising a first primer comprising a sequence 5xe2x80x2-GGGGTCCGGAG(A/G)(C/G)(A/G)TA(A/G/T)AT(A/G/P)A(A/G/P)(A/G/P)GG-3xe2x80x2 (SEQ ID NO:1) and a second primer comprising a sequence 5xe2x80x2-GGGGCTGCAGACACC(A/C/G/T)ATGTA(C/T)(C/T)T(A/C/G/T)TT(C/T)(C/T)T-3xe2x80x2 (SEQ ID NO:2). When used to amplify olfactory receptor nucleic acid sequences, it typically amplifies the receptor ligand-binding region comprising olfactory receptor transmembrane (TM) domains II through VII.
The invention also provides a method for generating nucleic acid sequence that encodes a ligand-binding region of an olfactory receptor, the method comprising amplification of a nucleic acid using the primer pair SEQ ID NO:1 and SEQ ID NO:2. In this method the amplified nucleic acid can be genomic DNA, mRNA or cDNA derived from olfactory neurons or olfactory epithelium. The amplification can be by polymerase chain reaction (PCR), wherein the PCR amplification comprises the following conditions and steps in the following order: about one cycle at about 94xc2x0 C. for about 2 min; and about 30 cycles of about 45xc2x0 C. to about 65xc2x0 C. for about 1 min, followed by about 72xc2x0 C. for about one min. followed by about 94xc2x0 C. for about 1 min. The PCR amplification protocol can further comprise the following conditions and steps in the following order: about one cycle of about 45xc2x0 C. to about 65xc2x0 C. for about 10 min; and about one cycle of about 72xc2x0 C. for about 10 min.
Also provides is a kit for amplification of olfactory receptor sequences comprising primer pairs that can amplify olfactory receptor transmembrane domain regions II through VII, II through VI, III through VII, or III through VI, e.g., SEQ ID NO:1 and SEQ ID NO:2 to amplify TM II through VII.
The invention also provides a library of olfactory receptor ligand-binding regions consisting essentially of olfactory receptor transmembrane domain regions II through VII, II through VI, III through VII, or III through VI, including partial domains, or a combination of domain sequences. The library of the olfactory receptor ligand-binding regions can be generated by PCR using degenerate primer pairs.
Also provided is a library of chimeric nucleic acid sequences comprising the following domains in 5xe2x80x2 to 3xe2x80x2 order: a nucleic acid encoding an amino terminal plasma membrane translocation domain; a nucleic acid encoding a first transmembrane domain; and a nucleic acid encoding an olfactory receptor ligand-binding region, wherein the chimeric nucleic acid sequence encodes a 7-transmembrane polypeptide that can transverse a plasma membrane seven times. The amino terminal plasma membrane translocation domain comprises an amino acid sequence as set forth in SEQ ID NO:3 (and encoded by a subsequence of SEQ ID NO:6):
In alternative embodiments, the nucleic acid encoding the first transmembrane domain can be just a polynucleotide sequence encoding SEQ ID NO:3, or, SEQ ID NO:6 (including 45 nucleotides upstream of the initiation codon) or a subsequence thereof.
The first transmembrane receptor of the sequences of the library can be a 7-transmembrane receptor region I domain, or subsequence thereof, e.g., the sequence between the Eco R1 and Pst 1 sites of the M4-chimeric olfactory receptor of the invention (SEQ ID NO:4), as schematically represented in FIG. 1A; the full length sequence of the hybrid receptor has an amino acid sequence as set forth in SEQ ID NO:55, a nucleic acid that can encode this protein is SEQ ID NO:54, described below.
The olfactory receptor ligand-binding regions of the library can comprise olfactory receptor transmembrane domain regions II through VII, II through VI, III through VII, or III through VI, or a combination thereof. These olfactory receptor ligand-binding regions can be generated by amplification, e.g., PCR, using degenerate primer pairs. The library""s nucleic acid sequence encoding transmembrane domain regions II through VII can generated by PCR amplification of nucleic acid using a first primer comprising a sequence 5xe2x80x2-GGGGTCCGGAG(A/G)(C/G)(A/G)TA(A/G/T)AT(A/G/P)A(A/G/P)(AG/P)GG-3xe2x80x2 (SEQ ID NO:1) and a second primer comprising a sequence 5xe2x80x2-GGGGCTGCAGACACC(A/C/G/T)ATGTA(C/T)(C/T)T(A/C/G/T)TT(C/T)(C/T)T-3xe2x80x2 (SEQ ID NO:2). The library can be generated from PCR-amplified nucleic acid isolated as or derived from genomic DNA, mRNA or cDNA derived from olfactory neurons or olfactory epithelium.
Exemplary ligand-binding regions comprising transmembrane domains II through VII ca be an amino acid sequence encoded by a nucleic acid selected from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45 and SEQ ID NO:47, or an amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48.
To generate the nucleic acids of the library, PCR amplification can comprise the following conditions and steps in the following order: about one cycle at about 94xc2x0 C. for about 2 min; about 30 cycles of about 55xc2x0 C. for about 1 min, followed by about 72xc2x0 C. for about one min. followed by about 94xc2x0 C. for about 1 min; about one cycle of about 55xc2x0 C. for about 10 min; and about one cycle of about 72xc2x0 C. for about 10 min.
The library can further comprise a carboxy terminal 7-transmembrane receptor transmembrane region VII domain or subsequence thereof, e.g., the sequence between the Bsp E1 and Xba 1 sites of the M4-chimeric olfactory receptor of the invention (SEQ ID NO:5), as schematically represented in FIG. 1A; the full length sequence of the hybrid receptor has an amino acid sequence as set forth in SEQ ID NO:55, a nucleic acid that can encode this protein is SEQ ID NO:54.
The library of nucleic acid sequences can also comprise the following domains in 5xe2x80x2 to 3xe2x80x2 order: a nucleic acid encoding an amino terminal plasma membrane translocation domain comprising a sequence as set forth in SEQ ID NO:3, a nucleic acid encoding a transmembrane region I domain comprising a sequence as set forth in SEQ ID NO:4, a nucleic acid sequence generated by polymerase chain reaction (PCR) amplification of mRNA or cDNA derived from olfactory epithelium using a first primer comprising a sequence 5xe2x80x2-GGGGTCCGGAG(A/G)(C/G)T(A/G)A(A/G/T)AT(A/G/P)A(A/G/P)(A/G/P)GG-3xe2x80x2 (SEQ ID NO:1) and a second primer comprising a sequence 5xe2x80x2-GGGGCTGCAGACACC(A/C/G/T)ATGTA(C/T)C/T)T(A/C/G/T)TT(C/T)C/T)T-3xe2x80x2 (SEQ ID NO:2), and a nucleic acid encoding a 7-transmembrane receptor transmembrane region VII domain comprising a sequence as set forth in SEQ ID NO:6.
Also provided are expression vectors (e.g., plasmids, viruses) comprising a nucleic acid sequence derived from the libraries of nucleic acid sequences of the invention. Transformed or isolated infected cells comprising a nucleic acid sequence derived from a library of nucleic acid sequences of the invention or an expression vector of the invention are also provided. Transgenic non-human animals comprising a nucleic acid sequence derived from a library of nucleic acid of the invention or an expression vector of the invention are also provided. In the transgenic animal, the expression vector can be a mammalian expression vector that can be expressed in olfactory epithelium or olfactory neurons.
The invention also provides a library of recombinant polypeptides translated or derived from the library of nucleic acids of the invention. Also provided are polypeptides isolated or derived from the library of polypeptides of the invention.
Also provided are methods of determining whether a test compound specifically binds to a mammalian olfactory receptor comprising the following steps: expressing a nucleic acid derived from a nucleic acid library of the invention under conditions permissive for translation of the nucleic acid to a receptor polypeptide; contacting the translated polypeptide with the test compound; and determining whether the test compound specifically binds to the polypeptide.
Also provided are methods of determining whether a test compound specifically binds to a mammalian olfactory transmembrane receptor comprising the following steps: contacting a cell stably or transiently transfected with a nucleic acid derived from a nucleic acid library of the invention; culturing the cell under conditions permissive for translation of the nucleic acid to a receptor polypeptide with the test compound; and determining whether the test compound specifically binds to the receptor polypeptide. In this method, the receptor polypeptide can be expressed as a transmembrane receptor with a ligand-binding site on the cell""s plasma membrane outer surface. The specific binding of the test compound to the polypeptide can be determined by measuring a change in the physiologic activity of the cell, wherein a change in the cell""s activity measured in the presence of the test compound compared to the cell""s activity in the absence of the test compound provides a determination that the test compound specifically binds to the polypeptide. The measured cell activity can be a change in the calcium ion (Ca2+) or cAMP concentration in the cell, which can be measured by loading the cell with a calcium ion-sensitive fluorescent dye before contacting the cell with the test compound. In this method any cell can be used, e.g., a human cell or a Xenopus oocyte.
Also provided are methods of determining whether a test compound specifically binds to a mammalian olfactory transmembrane receptor polypeptide in vivo comprising the following steps: contacting a non-human animal stably or transiently infected with a nucleic acid derived from the library of the invention or an expression vector of the invention with the test compound; and determining whether the animal reacts to the test compound by specifically binding to the receptor polypeptide, wherein the specific binding of the test compound to the polypeptide is determined by measuring a change in a physiologic activity of the animal, wherein a change in a receptor-encoding vector-infected animal""s activity measured in the presence of the test compound compared to a bare vector-infected animal""s activity in the presence of the test compound provides a determination that the test compound specifically binds to the mammalian olfactory transmembrane receptor polypeptide. In this method, the measured physiologic activity can be measured by an electroolfactogram. The vector can be a recombinant virus, e.g., an adenovirus expression vector.
The invention also provides a method of determining whether a test compound is neurotoxic to an olfactory neuron expressing an olfactory transmembrane receptor polypeptide comprising the following steps: contacting an olfactory neuron cell stably or transiently infected with a nucleic acid derived from a library as set forth in claim 8 or claim 10 or an expression vector as set forth in claim 23 with the test compound; and measuring the physiologic activity of the cell, wherein a change in the cell""s activity measured in the presence of the test compound compared to the cell""s activity in the absence of the test compound provides a determination that the test compound is toxic. In this method toxicity can be indicated by abnormal calcium ion, cAMP or plasma membrane homeostasis.
Also provided are peptide domains for the efficient translocation of a newly translated protein to a plasma membrane comprising an amino acid sequence as set forth in SEQ ID NO:3 or an amino acid sequence having conservative amino acid residue substitutions based on SEQ ID NO:3. Translocation domains within the scope of the invention include amino acid sequences functionally equivalent to the exemplary translocation domain of the invention SEQ ID NO:3. The peptide translocation domain can be at least about 20 amino acids in length, at least about 30 amino acids in length or at least about 40 amino acids in length. The peptide translocation domain can have a sequence as set forth in SEQ ID NO:3, or, be encoded by a nucleic acid comprising a sequence as set forth in SEQ ID NO:6. The newly translated protein can be a transmembrane protein, e.g., a 7-transmembrane protein receptor, e.g., an olfactory receptor.
The invention also provides a hybrid (chimeric) polypeptide comprising an amino terminal amino acid sequence comprising a peptide translocation domain of the invention and a second polypeptide sequence, wherein the second polypeptide sequence is not a rhodopsin polypeptide sequence. The second polypeptide sequence can be a transmembrane protein, e.g., a 7-transmembrane protein receptor, e.g., an olfactory receptor. Also provides are isolated or recombinant nucleic acid sequences encoding these hybrid polypeptides. For example, an exemplary chimeric polypeptide of the invention and a polynucleotide that encodes this hybrid, described in the Example below and schematically represented in FIG. 1A as the insert from BamH1 to XbaI, have the amino acid (SEQ ID NO:55) and nucleic acid (SEQ ID NO:54) sequence, respectively (restriction enzyme sites are also indicated):
A further understanding of the nature and advantages of the present invention is realized by reference to the remaining portions of the specification, the figures and claims.
All publications, GenBank deposited sequences, ATCC deposits, patents and patent applications cited herein are hereby expressly incorporated by reference for all purposes.

Olfactory receptor expression libraries and methods of making and using them


A method and apparatus in which a radio frequency transceiver is controlled via a plurality of connectors including providing control information for changing a mode of operation of the receiver. The modes can be transmit and receive modes, and the connectors can be configured to perform various functions depending on the mode of operation. Control signals, time critical control signals, and other types of signals may be received at one or more of the connectors in the different modes of operation. The functions may control a power amplifier of a transmitter portion of the transceiver, read/write data from/to registers of the transceiver, and/or other functions.

1. Field of the Invention
The present invention relates to an interface between base band circuitry and radio frequency transceiver circuitry, particularly circuitry operating in accordance with the Bluetooth Low Power Radio Frequency Specification. It additionally relates to devices having such an interface and either type of circuitry.
2. Description of the Prior Art
Low power radio frequency systems allow communication between devices over short distances typically ten's of meters. The devices must each be capable of receiving and transmitting according to the system's protocol.
One low power radio frequency system is the Bluetooth system. This system is designed to replace connecting wires and cables with wireless connectivity. For one device to communicate with another device, no wires connecting them will be necessary. Instead, each device will host a transceiver. A transceiver has a baseband part and an RF part. The host itself may have processing circuitry which is capable of doing the base band processing and that host will only require RF transceiver circuitry to be correctly connected to that processing circuitry.

This patent is subject to a terminal disclaimer.


Provided is a steam turbine overspeed protection system, includes a drive gear arranged to match a rotation speed of a rotor of a steam turbine; a rotating shaft parallel to an axis of the drive gear and capable of rotating at a critical rotation speed; a protective gear arranged on the rotating shaft and forming a lead screw nut mechanism with the rotating shaft, and arranged to be capable of engaging with the drive gear when the rotation speed of the drive gear exceeds the critical rotation speed; and an operating rod connected to the protective gear; wherein, when the drive gear engages with the protective gear, the protective gear can move in the axial direction of the rotating shaft and thereby drive the operating rod to move and produce an action that activates a protection device for preventing steam turbine overspeed.

Steam turbine overspeed may cause extremely high damages, and thus has drawn high attention from technicians. At present, steam turbine overspeed protection devices are implemented in two manners: electronic overspeed protection and conventional mechanical overspeed protection.
An electronic overspeed protection system is configured to measure the actual operating rotation speed of a steam turbine-generator set by means of an electronic speed measuring system, generate an analog signal and send it to a control system, in which a logical module operates and identifies the analog signal. When the rotation speed reaches a critical rotation speed (e.g., 3,300 rpm) or above, the control system will issue an automatic shutdown command to realize shutdown protection of the steam turbine. In such a case, if any problem occurs during measuring the rotation speed, protective action refused and protective action error accidents will happen very easily in the system.
In a conventional mechanical overspeed protection system, a fly-weight or fly-ring with certain mass is usually mounted at an end of the rotor. When the rotor operates at a rotation speed lower than the critical rotation speed (e.g., 3,300 rpm), the fly-weight or fly-ring will be restrained by a spring. When the rotation speed of the rotor reaches 3,300 rpm or above, a increased centrifugal force of the fly-weight or fly-ring will overcome the restraining force of the spring and make the fly-weight or fly-ring displace a certain distance, so that it may impact a protective stay hook. The protective stay hook works with other components in the mechanical overspeed protection system to release the pressure in the control oil system and accomplish a shutdown action. However, the reliability of the aforementioned system mainly depends on the quality of the spring, such as the manufacturing accuracy, coefficient of stiffness, and stability of the spring, which has a great influence on the system. In addition, periodical tests are required to verify the reliability of the system. The equipment may trip in the case of mal-function or mal-operation of any component in the online test process. Therefore, there is still a room for improvement to the reliability of conventional mechanical overspeed protection systems.
At present, there is no prior art providing a mechanical steam turbine overspeed protection system, in which the spring and fly-weight/fly-ring could be replaced by other components.

Steam turbine overspeed protection system, and steam turbine


An SS7 Network Preventative Maintenance System for detecting potential SS7 and switched network troubles, automatically analyzing the troubles, providing alarm and corrective action to avoid major network events. Real time monitors on SS7 links at the STP provide information on exceeded link load, exceeded Message Signaling Unit (MSU) frequency and network management status/error conditions in a Stage 1 Process. The Stage 1 Process provides alarm information to a Stage 2 Process which controls all Stage 1 associated monitors for an STP pair. Stage 2 reacts to Stage 1 signals to generate alarm and corrective action information which is passed on to a Stage 3 Process. The Stage 3 Process controls all Stage 2 Processes in the operating company. Stage 3 reacts to Stage 2 output to detect potential or real accompanying network trouble and generates alarm and corrective action information and displays in response thereto. Stage 3 also alerts a Stage 4 process which is connected to all Stage 3 Processes in a region. Stage 4 analyzes data from Stage 3 to determine if similar trouble could happen in another network where upon Stage 4 informs affected Stage 3 Processes regarding the same. Corrective action/trouble verification information is generated and passed on. An Interface to the network's surveillant system is provided.

One system for providing a Common Channel Signaling Network (CCSN) utilizes Signaling System 7 (SS7) protocol in a Packet Switched Data Network (PSDN) connecting Network Elements (NE) via packet switched 56 KB digital data circuits. In addition to providing call set signaling functions, the SS7 network also provides access to switching control points (SCP) used to permit line identification database (LIDB) queries for credit card verification and 800 database look-up for 800 services. Class services also use the SS7 network to provide custom call features. The latest services using the SS7 network comprise Advanced Intelligent Network (AIN) services. AIN services use the SS7 network to access an Integrated Switching Control Point (ISCP) where AIN service functions are performed.
This network currently employs various Network Traffic Management (NTM) and NE test and provisioning systems to maintain the NEs in the Public Switched Network. However, these systems can only report troubles and provide manual NE trouble resolution tools. The Bellcore SS7 Engineering and Administration System (SEAS) was developed to provide Network Management functions for the SS7 network, but lacks the ability to anticipate troubles and provide corrective action instructions to NE operations personnel. Also, both NTMs and SEAS use information supplied from the NEs or the SS7 switching transfer point (STP) hardware which, when in trouble, often cannot provide timely status information to maintenance personnel because of excessive NE and STP processor demands. SS7 STPs are especially susceptible to processor delays due to the extremely high volume of SS7 Message Signaling Units (MSUs) generated and processed during a major trouble condition. For these reasons an SS7 Real Time Monitoring System (RTMS) which is outside of the STP environment is needed to maintain the SS7 network and to ensure SS7 network reliability.
Current telecommunications and data networks using CCS generally utilize the protocol of Specification of Signaling System 7 (SS7) which is described in Section 6.5, LSSGR, Issue 2, July 1987, TR-TSY-000506, a module of TR-TSY-000064. Various methods and techniques for testing and analyzing the operation of such networks have been proposed including, by way of example, in co-pending application Ser. No. 07/953,173, filed Sep. 29, 1992, and commonly assigned with the present invention. Generally speaking, such prior arrangements for testing of SS7 networks have been responsive to malfunctions or else conducted on a routine basis at periodic intervals. Such techniques, while effective for their intended purposes, offer little, if any, assistance in detecting incipient developing problems so rapidly as to permit and provide prevention of network failure before the development of the problem is complete.
Accordingly it is an object of the present invention to provide a communications network preventative maintenance tool that detects potential CCS and switched network troubles, automatically analyzes such troubles and provides corrective action or instructions in time to avoid major breakdown.
It is another object of the invention to provide an SS7 Real Time Monitoring System (RTMS) which is outside of the STP environment and has the ability to anticipate troubles and provide corrective action instructions on a sufficiently rapid basis to detect incipient problems and provide corrective action before a serious network problem develops.

Common channeling signaling network maintenance and testing


Foot pads with foot straps are mounted to roller carriages which roll along a horizontal monorail with stationary middle supports and telescoping end supports. A rotatable shaft attached to one of the end supports is attached to an ergometric input-responsive variable resistance. Each of two cables wound around a spring-loaded retractable one-way clutch driver on the rotatable shaft connects to each of the foot pads. Stepper pedals hinged to the end support opposite the rotating shaft are each attached by a cable wound around the rotatable shaft. A long front rail with sliding hand grips and a short side rail are detachably mounted. A nordic double poling attachment may be mounted midway on the front rail with cables wound over pulleys and around the rotatable shaft. The platform is hinged in the center to fold in half for storage and transportation. A stationary foot pad is positioned adjacent to the hinges, and also serves as a stop for the moving foot pads. End foot pad stops may be positioned at any of a number of locations along the monorail. An electronic microprocesser performance monitor senses, interprets, and displays information about the output of the user on the invention. Any of a number of ergometric input-responsive resistance means can be used on the rotatable shaft including: a flywheel with a band brake, a vaned flywheel inside a variable opening case, a wind load, a water load, an eddy current load, a flywheel with a centrifugal braking device, and an electric motor and flywheel load.

1. Technical Field
The present invention relates to exercise machines, and in particular to a skating/skiing exercise machine with side-to-side foot pads rolling on a monorail with the pads pulling cables which are connected to ergometric variable input-responsive resistance.
2. Description of the Prior Art
Skating and skiing require practice and training for best performance, especially when engaged in competition. Actually performing the sports activities when possible (overcoming obstacles such as adverse weather and inadequate access to facilities) provides focused exercise but doesn't offer an adequate opportunity to vary the resistance involved in carrying out the activity to produce increased strength.
There are many exercise devices available on the market for providing cardiovascular and muscular development, but most of them fail to produce a true simulation of the actual sports activity for which the exerciser is training. Part of the difficulty lies in trying to shape exercise equipment to allow the full range of body movement in the same form as in the sport. Another difficulty lies in trying to create resistance in the exercise equipment which simulates actual resistance encountered in a sports activity while in motion in the sport having overcome inertial resistance.
Exercise equipment is often boring and uninvolving when the exerciser repeats the same action over and over again while remaining in a stationary position on the equipment. In most equipment, the exerciser does not experience the motion experienced in the actual sports activity.
Most prior art exercise devices for skating and skiing do not provide for side-to-side motion simulating the actual motion. Two U.S. Pat. Nos. (3,791,645 and 4,340,214) which do provide for side-to-side motion and two U.S. Pat. Nos. (4,781,372 and 4,915,373) which provide angled side/back motion do not provide ergometric variable resistance. U.S. Pat. No. 3,791,645 has a motor driving a belt which engages foot cradles to move the foot cradles to the side. U.S. Pat. No. 4,340,214 provides side sloping tracks. U.S. Pat. No. 4,781,372 uses variable weights lifted by cords connected to the foot cradles. U.S. Pat. No. 4,915,373 utilizes foot pedals in sliding tracks which have variable friction brake linings in the tracks. None provide the simulation of actual skating and skiing side-to-side motion when there is a gliding effect produced by inertia in motion after the initial inertia at rest is overcome.
Prior art exercise devices for skating and skiing generally do not provide for other exercise options.

Skating/skiing simulator with ergometric input-responsive resistance


A pair of shoes fastener for releasably coupling at least one pair of shoes together. The pair of shoes fastener includes a male portion and a female portion that are releasably coupleable together. The male portion has a neck member outwardly extending from the front face thereof and a bulbous end portion extending from the neck member. The female portion has an aperture extending through its top and bottom faces and a pair of axial breaks providing an opening through the front face of the female portion into the aperture. The aperture and the axial breaks define a groove of the female portion. The end and neck members of the male portion are slidably insertable in the groove of the female portion. The male portion is coupled to an inner side of a shoe. The female portion is coupled to an inner side of another shoe.

1. Field of the Invention
The present invention relates to footwear attachments and more particularly pertains to a new pair of shoes fastener for releasably coupling at least one pair of shoes together.
2. Description of the Prior Art
The use of footwear attachments is known in the prior art. More specifically, footwear attachments heretofore devised and utilized are known to consist basically of familiar, expected and obvious structural configurations, notwithstanding the myriad designs encompassed by the crowded prior art which have been developed for the fulfillment of countless objectives and requirements.
Known prior art includes U.S. Pat. No. 3,041,743; U.S. Pat. No. 2,761,223; U.S. Pat. No. 2,266,281; U.S. Pat. No. 5,269,690; U.S. Pat. No. 3,000,067; and U.S. Pat. No. Des. 273,659.
While these devices fulfill their respective, particular objectives and requirements, the aforementioned patents do not disclose a new pair of shoes fastener. The inventive device includes a male portion and a female portion that are releasably coupleable together. The male portion has a neck member outwardly extending from the front face thereof and a bulbous end portion extending from the neck member. The female portion has an aperture extending through its top and bottom faces and a pair of axial breaks providing an opening through the front face of the female portion into the aperture. The aperture and the axial breaks define a groove of the female portion. The end and neck members of the male portion are slidably insertable in the groove of the female portion. The male portion is coupled to an inner side of a shoe. The female portion is coupled to an inner side of another shoe.
In these respects, the pair of shoes fastener according to the present invention substantially departs from the conventional concepts and designs of the prior art, and in so doing provides an apparatus primarily developed for the purpose of releasably coupling at least one pair of shoes together.


A method of forming a film layer and a substrate comprising the film layer to reduce occurrence of defects and improve the quality of the film layer are described. The method of forming a film layer comprises forming a plurality of sub-film layers of a same material overlapped with each other on a substrate by multiple steps to constitute the film layer, wherein each time a sub-film layer is formed, the newly-formed sub-film layer is cleaned immediately. The substrate comprises a film layer formed by the above method.

Indium tin oxide film (ITO film), as an important kind of transparent conductive film due to its excellent conductivity and visible-light transmittance, is widely used in photoelectric devices. At present, the ITO film is fabricated generally by using magnetron sputtering technique. In the magnetron sputtering technique, the plasma discharging is implemented under vacuum conditions, the movements of charged particles are controlled through electromagnetic field so that a target is bombarded by the moving particles, and atoms of the target are knocked off and attached to the substrate, thereby a desired thin film is formed on the substrate.
Nowadays, it is necessary for a Twist Nematic (TN) type liquid crystal display panel to have an ITO film formed on the color film substrate as a common electrode. However, the ITO film formed by using the magnetron sputtering technique has defects on film surface, for example, pin holes are likely to appear on the film surface. The cause and process of producing the pin holes are illustrated in FIGS. 1 and 2. During film coating, if there is a foreign matter 21 (e.g. dust, foreign particle and the like) on the color film substrate 1, a bump will be formed on the ITO film surface after film coating. In subsequent cleaning process, the bump is removed by washing, and a pin hole 22 is produced passing through the ITO film 2. The pin hole 22 will be displayed as an abnormal spot when the cell-forming process of the display panel is finished, which will negatively impact on the qualified rate of the liquid crystal display panel. Obviously, it is necessary to avoid forming the pin hole 22. An existing method for reducing the occurrence rate of the pin holes 22 is that a pre-cleaning process is performed before the film coating process, and the devices are all sealed and cleaned periodically before the film coating process. However, it is obvious that such method can only reduce the foreign matters on the substrate before the film coating process, and can hardly eliminate the foreign matters completely. Furthermore, in the liquid crystal display panels, a voltage should be applied to the common electrode so as to control the switch between dark and light for the pixel units. Thus, in order to ensure that the common electrode can withstand a desired voltage, the ITO film acting as the common electrode should have a certain thickness. Presently, the thickness of the ITO film on the color substrate in the TN type liquid crystal display panel is generally 150 nm. When fabricating the ITO film 2 with a thickness of 150 nm, the power of the film-coating equipment is 14.6 kW, which corresponds to a production time of 35 second per product. However, in actual production, when the film-coating power is higher than 8 to 9 kW, the probability that knobbles (protuberances) are generated on the surface of the target and on the ITO film 2 on the color substrate increases dramatically. The knobbles on the ITO film 2 are apt to form pin holes. Furthermore, the knobbles on the surface of the target are apt to cause point discharge so that the surface of the ITO film 2 formed by film coating protrudes to form a protruding part which cannot be removed by washing and should be removed by polishing during repairing process, thus deteriorating the quality of film coating, and resulting in undesirable values of film-surface resistance and transmittance. Further, due to these effects, the production line has to be suspended so as to polish and wash the surface of the target to remove the knobbles periodically. Consequently, the maintenance time for the equipments increases greatly, and the utilization, which means the percentage of time spent on creating value by the equipment in a planned time, is reduced.
When the film layer is formed by using other processes (e.g., the evaporating process, the sputtering process, the chemical vapor deposition process, the spraying process, and so on), the phenomenon of defective film surface caused by the foreign matters also occurs, and thereby the performance of subsequent products is affected.

Method for forming film layer and substrate including the film layer

Beijing BOE Display Technology Co., Ltd.


A fuel cell system includes a fuel cell stack, a fuel processor, a heater and a controller. The fuel cell stack provides an output power that includes a first power that is consumed by a first load. The fuel processor furnishes a fuel flow to the fuel cell stack, and the heater furnishes heat to the fuel processor when enabled. The controller is coupled to the fuel cell stack to monitor the output power and is coupled to the heater to selectively enable the heater to provide heat to the fuel processor when the first power decreases below a predetermined threshold.

The invention relates to operating a fuel cell system during low power demand.
A fuel cell is an electrochemical device that converts chemical energy that is produced by a reaction directly into electrical energy. For example, one type of fuel cell includes a proton exchange membrane (PEM), often called a polymer electrolyte membrane, that permits only protons to pass between an anode and a cathode of the fuel cell. At the anode, diatomic hydrogen (a fuel) is reacted to produce hydrogen protons that pass through the PEM. The electrons produced by this reaction travel through circuitry that is external to the fuel cell to form an electrical current. At the cathode, oxygen is reduced and reacts with the hydrogen protons to form water. The anodic and cathodic reactions are described by the following equations:
H2xe2x86x922H++2exe2x88x92 at the anode of the cell, and
O2+4H++4exe2x88x92xe2x86x922H2O at the cathode of the cell.
Because a single fuel cell typically produces a relatively small voltage (around 1 volt, for example), several fuel cells may be formed out of a stack of fuel cells in series to produce a higher voltage. The fuel cells may include plates (graphite composite or metal plates, as examples) that are stacked one on top of the other, and each plate may be associated with more than one fuel cell of the stack. The plates may include various channels and orifices to, as examples, route the reactants and products through the fuel cell stack. PEMs (each one being associated with a particular fuel cell) may be located throughout the stack between the anodes and cathodes of the different fuel cells.
As an example, a residential fuel cell system may include a fuel processor to convert a hydrocarbon (a natural gas or propane, as examples) into a hydrogen-rich reformat that is consumed by the fuel cell stack pursuant to the above-described reactions. As an example, the fuel system may be used to deliver power to a house, a load that typically varies over the course of each day. In this manner, during the early morning hours, the house typically demands far less power than the house requires during other parts of the day. This variance may affect operation of the fuel processor. As an example, the efficiency of the fuel processor may be optimized to perform over a predefined power range, such as a reformat output sufficient to correspond to a system power output in the range from 2 kilowatts (kW) to 7 kW. When the power demand drops below the 2 kW level, the fuel processor may have difficulty in maintaining a desired quality of the reformat, or the efficiency of the fuel processor may decline.
Thus, there is a continuing need for an arrangement that addresses one or more of the problems that are stated above.
In an embodiment of the invention, a fuel cell system includes a fuel cell stack, a fuel processor, a heater and a controller. The fuel cell stack provides an output power that includes a first power that is consumed by a first load. The fuel processor furnishes a fuel flow to the fuel cell stack, and the heater furnishes heat to the fuel processor when enabled. The controller is coupled to the fuel cell stack to monitor the output power and is coupled to the heater to selectively enable the heater to provide heat to the fuel processor when the first power decreases below a predetermined threshold.
Advantages and other features of the invention will become apparent from the following description, from the drawings and from the claims.

Operating a fuel cell system during low power demand


The present invention includes method and apparatus for grading a poultry carcass intended for human consumption. An ultrasound machine is provided which includes a probe for transmitting and receiving ultrasound signals such that the machine is capable of producing a image of a breast muscle of the carcass. The probe is positioned on the carcass parallel to the keel such that the image of the breast muscle of the carcass is detected by the ultrasound machine. A breast muscle size indicator of the breast muscle is determined from the detected image. The calculation of a grade for the carcass is based upon the ratio of the indicator to the weight of the carcass. The determination of the breast muscle size indicator can be performed by an image analyzer and pattern recognition equipment modified to recognize the breast muscle image. The probe can be positioned automatically by probe location equipment automatically.

Many different types of evaluations are carried out on meat products before they are approved for commercial sale. Some of these procedures involve a system of grading depending upon fat and muscle content and distribution, while others involve condemnation decisions.
Various types of ultrasonic apparatus for examining the internal structure of humans and animals are, of course, well known in the prior art. The use of ultrasonic techniques for inspecting live animals for fat thickness has long been proposed, particularly in percentage of fat or lean, so that the fat indication provides a reasonable indication of carcass composition.
The advantage of using ultrasonic apparatus to inspect and grade livestock are numerous. Primarily, an animal's value can be accurately assessed without causing any damage to the carcass. In meat animals a grading determination is based upon the relative quantity of fat or muscle within a normal set of parameters. The normal set of parameters are the same for each animal within a particular species (e.g. the number of fat or muscle layers) whereas the proportion each type of tissue within these parameters differs for each individual animal. Ultrasound techniques have been used extensively to determine grading characteristics of livestock.
Currently poultry carcasses are graded A, B, or C subjectively based primarily upon general appearance, fleshiness and freedom from defects, torn skin and other blemishes. However there has been no practical way of accurately predicting the relative ratio of muscle to bone and the overall muscle composition of poultry carcass without dissection. As the breast is usually considerably the most valuable portion of poultry, it would be extremely valuable to provide a meaningful grading system based upon the size of the breast muscle to the overall weight of the carcass.
The industry is in need of a method of grading for poultry that provides a meaningful, accurate information on the size of the breast muscle to the weight of the carcass.

Grading of poultry carcasses with ultrasound


Fibers comprising 10 to 100% of bicomponent fibers, composed of a core polymer covered by a sheath polymer, are heated to a temperature between the softening point and the melting point of the sheath polymer for 5 seconds to 5 minutes at a pressure of 0 to 2000 psi until the fibers are bonded together. The products are used for drive belts and seals, nib felts for marking pens, filter cloths for plate and frame filters, filtration cartridges, stamp pad ink reservoirs, and battery separators.

This invention relates to the field of non-woven textile materials and articles made therefrom.
Current non-woven textile technology consists of dry and wet processing varied to control the properties of the end product. Wool felts cannot be made by dry processes if the felting properties of wool are to be used. Needled felts are limited in their hardness, rigidity and other properties, unless chemical and/or wet processes are added, for example through the use of impregnants. The continued development of new types of synthetic fibers has increased the range of properties of synthetic fiber needled felts, resulting in the displacement of wool felts from many areas of use. Among these areas are: marking pen nibs or tips, resin bonded filters, plate and frame filter sheets, marking stamp pad reservoirs and battery separator media.
Prior art for the modification of these and other types of non-woven textile materials has included the post-manufacture treatment of the textile with a variety of chemical materials, such as lattices, resins, plastics and the like, and has included also laminations, coatings and mechanical finishes. Marking pen nib felts are normally treated with formulations of melamine-formaldehyde to enhance rigidity and wear resistance, while retaining the ability to allow ink to flow freely through the fiber interstices. Phenolic complex resins continue to be used to produce rigid fibrous filter cartridges capable of withstanding pressures exceeding 100 psi. Plate and frame cloths have been coated with various resins (viz., acrylic, urethane, etc.) to provide a clean cake release from the surface. Stamp pad felts have been treated with wetting agents to promote ink leveling, and have been laminated to inert materials to provide rigidity and reduce cost. Battery separators have been produced which accommodate the special chemical requirements associated with advanced battery technology.
In general, these modifications have required wet processing and the use of chemical materials which are increasingly becoming categorized as hazardous or unsafe. Therefore, increased environmental concerns have vastly reduced the range of materials which can safely be used to modify the basic fibrous structures of non-woven textiles.
Within the last decade, certain specialty synthetic fibers identified generically as "bicomponent" or "bico" fibers have been produced. In general, these consist of, but are not limited to, two varieties of a single polymer type. Structurally, they consist of a core polymer and a sheath polymer. Creation of these new fibers has permitted the redevelopment of some products formerly requiring the post-treatment of basic non-woven structures, through a re-alignment of customary manufacturing techniques, adapted to the use of these fibers. Because the core and sheath polymers are often varieties of the same polymer, they retain their polymeric identity, but have different melting points. This feature permits their use as bonding agents. While bondable fibers have existed for a considerable period of time, their melting points rendered them unusable for many purposes, or their chemical properties excluded them from important uses.
A non-woven melded fabric, known under the trademark "Cambrelle" is composed of bico fibers. Upon heating to the melting point of the sheath polymer, the fibers soften and unite to form a fabric, which is used for carpet backing, road reinforcement, upholstery, interlinings, tablecloths and other household applications.

Non-woven textile articles comprising bicomponent fibers and method of manufacture


A communication device is described along with various methods and systems for controlling the same. Methods are provided that monitor audio from a local and remote audio stream when determining whether to provide a mute notification on a muted call. In general, when a mute function of a communication device is active and a user attempts to speak, the system can analyze the user's local audio stream and the call's remote audio stream to determine whether characteristics exist to provide a mute notification to the user. The system provides mechanisms for controlling false positive determinations of speaking through mute by utilizing a combination of recorded characteristics and initiated timers that can be adjusted based on determinations made.

When making calls between entities, a user may find it convenient, or even necessary, to mute the microphone of the user's communication device. In other words, a user may initiate a mute function associated with a communication device to prevent the microphone conveying sound to one or more other users participating in the call. Among other things, muting the microphone can provide a user with a certain level of privacy by preventing one or more other users from hearing what the user is doing or saying while the mute function is active. Some of the benefits associated with muting the communication device may include the ability for a user to hold a private conversation, cough, sneeze, perform other tasks, use another communication device, type on a keyboard, prevent distraction in a noisy environment, and the like.
Although it is common practice to mute a telephone while on a call, it is even more common when a user is on a conference call. It is a frequent occurrence, however, that one or more of the users participating in a conference call may forget to deactivate, or unmute, the mute function of their device. In this instance, a user may begin to speak to other conference participants without realizing that the mute function is still active, and as a result, the user cannot be heard by the other participants. Complicating the issue, the user may not be aware that the mute is still active until no other participant in the call acknowledges or responds to their dialogue. As can be appreciated, this scenario does not result in a desirable user experience. Not only is it frustrating for a user to speak and later realize that nothing spoken was heard, but it can also be embarrassing for the user and others in the call. Moreover, any delay caused between what the user thought was heard during an active mute and what the user must repeat when the mute is deactivated wastes the time of the teleconference participants and can disrupt the flow of a discussion.
Some have attempted to solve the problem of notifying users that a mute function has been engaged by providing an alert to a user. The alert may be provided in the form of a periodic tone, or "beep," that is output to the speaker of a user's telephone device. In some cases, the alert may be provided in the form of a flashing light, or text, to try and alert a user when the mute function of a telephone device is activated. In any event, it is expected that a user should hear, or see, the alert and respond by deactivating the mute function before attempting to participate in a conversation.
Unfortunately, these alerts do not adequately solve the problem associated with mute notification in communications across a network. For example, the periodic tone can be distracting to a user and may even mask the call content with each tone output. In other words, as the periodic tone is output to the user's device, any coincident audio output by the one or more other participants in the call may be cutoff, misunderstood, or result in unintelligible sounds. Furthermore, if the periodic tone is output to sound at longer intervals, a user may speak during the tone intervals only to realize later (e.g., at a subsequent tone output) that the mute function was active. The flashing light, or text, alert may be distracting in some implementations and unnoticeable in other implementations. For example, a bright light, or one having a quick illumination pulse, may be found annoying by a user. In contrast, if the light alert is dim, or displays at long intervals, the alert may go unnoticed. Furthermore, flashing lights suffer from context in their application. For instance, without knowing that the flashing light indicates that mute function of the device is active a user may associate the flashing light with some other meaning (e.g., a call is on hold, incoming call on another line, a message has been left on the device, etc.).

System and method for providing intelligent and automatic mute notification


The present invention generally relates to an apparatus for structural mapping of a macromolecule comprising an amount of persulfide effective to generate hydroxyl radicals upon contact with an aqueous solution. The present invention further relates to methods for structural mapping a macromolecule in an aqueous solution and methods for structural mapping a plurality of macromolecules in parallel, wherein each macromolecule is in a separate aqueous solution.

Throughout this application various publications are referred to by Arabic numerals in brackets. Full citations for these references may be found at the end of the specification immediately preceding the claims. The disclosures of these publications are hereby incorporated by reference in their entireties into the subject application to more fully describe the art to which the subject application pertains.
Structural mapping or 'footprinting' refers to methods in which the surface of nucleic acids and proteins accessible to the solvent is mapped with as fine as single residue resolution. Footprinting assays can be viewed as a collection of 'molecular cameras' that snap pictures of the position of each residue. Footprinting assays examine structure, ligand binding and/or conformational changes by determining the accessibility of the backbone or residues of macromolecules through their sensitivity to chemical or enzymatic modification or cleavage (reviewed by [3, 4]). The key characteristics of a footprinting assay are that i) the reaction of the footprinting probe with the polymer is limited such that each position along the chain is sampled with equal probability, and ii) cleavage or modification products are uniquely identified (FIG. 1).
The ability of proteins to undergo association and folding reactions has long been known to be a fundamental feature of cellular function across all the kingdoms of life. Macromolecular binding and folding reactions critical to unique biological functions are generally referred to as 'reversible associations'. Among the techniques that have been developed for the study of reversible associations, footprinting occupies a unique niche. Unlike structural methods such as x-ray crystallography and nuclear magnetic resonance (NMR), footprinting achieves site-specific resolution without extensive infrastructure, makes parsimonious use of biological samples and can be performed at dilute concentrations of macromolecules. Footprinting can map static structures as well as equilibrium and time-dependent transitions. The single residue resolution of footprinting can be used to develop detailed models of macromolecular structure, map ligand binding sites and follow conformational changes (FIG. 1). Computational tools are being developed that utilize the ensemble of individual measurements of the residue solvent accessibility to generate structural models of proteins and nucleic acids [6-10].
Cleavage of RNA and DNA by hydroxyl radicals is relatively insensitive to base sequence and whether a nucleic acid is single or double stranded [11, 12]. That hydroxyl radical cleavage of nucleic acids is quantitatively correlated with the solvent accessibility of the phosphodiester backbone has been demonstrated through comparisons of hydroxyl radical footprints with solvent accessibility calculations from crystal structures for protein-DNA complexes [13, 14] and RNA tertiary structures [15-19]. Backbone cleavage of DNA by hydroxyl radical is correlates with the accessible surface of the hydrogen atoms of the nucleotide sugar [20]. Hydroxyl radical footprinting yields a robust and readily interpretable measure of the structure and interactions of nucleic acids (FIG. 1). The availability of modern analysis tools such as CAFA [1] strengthens the feasibility of this method.
Hydroxyl radical footprinting was first extended to proteins by monitoring cleavage of the peptide backbone by gel electrophoresis [21, 22]. However, peptide bond cleavage is inefficient [23]. Thus, further development of protein hydroxyl radical footprinting has focused on the oxidation of amino acid side chains (reviewed in [24] and [25, 26]). Mass spectrometric analysis of proteolytic fragments is used to quantitate the oxidation rate of individual or groups of amino acid side chains. The differential reactivity of the amino acid side chains to oxidation is addressed in thermodynamic and kinetic analyses by quantitating the relative change in residue reactivity [24, 27]. A relationship between hydroxyl radical reactivity and solvent accessibility is emerging for proteins (reviewed in [24] & [25, 26]).
An important virtue of footprinting is that it can provide solution structural information with single residue resolution coupled to thermodynamic and kinetic transitions. Quantitative protocols have been extensively used to determine thermodynamic [3] and kinetic [31-34] constants describing protein-DNA interactions (reviewed in [3]). These protocols have been successfully extended to multiple implementations of quantitative hydroxyl radical footprinting [35-37] (see below). Protocols for thermodynamic protein hydroxyl radical footprinting have been published [38]. The individual-site isotherms [39, 40] and kinetic progress curves [31, 32] determined from thermodynamic and kinetic footprinting studies, respectively, provide an ensemble of local measures of macromolecular transitions from which detailed energetic and mechanistic portraits can be painted [41-44].
Footprinting assays for DNA, RNA and proteins have been developed using a wide range of reagents including the hydroxyl radical (.OH). The hydroxyl radical is among the most reactive and promiscuous of chemical oxidants [45]. Hydroxyl radical can be generated in solution by the Fenton-Haber-Weiss reaction according to the reactionFe(II)-EDTA+H2O2→Fe(III)-EDTA+.OH+OH−  (1).Tullius and co-workers showed that a convenient implementation of this chemistry for footprinting is to reductively cycle Fe(III) back to Fe(II) by the addition of ascorbate [13, 46, 47],
allowing low concentrations (μM) of the iron catalyst in the reaction mixture. This method is widely applied and inexpensive to perform. The reagent concentrations typically used in static and equilibrium .OH footprinting studies are μM in Fe(II)-EDTA and mM in H2O2 and ascorbate with reaction times of several to tens of minutes. Obviously, long reaction times are incompatible with high-throughput implementations.
A method using equation 1 where Fe(II) is stoichiometrically consumed by reaction with H2O2 to produce hydroxyl radicals on the millisecond timescale was recently developed [48-50]. While the reaction time is fast, impediments to high-throughput implementation of equation 1 include the need to precisely add high concentrations of the two reactants and auto oxidation of Fe(II).
Peroxonitrite has been used to hydroxyl radical footprint macromolecules [35, 51]. This reagent has not gained wide acceptance due to limitations on the solution conditions under which the reagent produces significant quantities of hydroxyl radical.
A recently developed method photolyzes H2O2 with UV radiation [25, 26, 52]. The use of UV radiation precludes this approach for DNA and RNA; nucleic acids are damaged by even limited exposure to short wavelength UV light. Thus, a disadvantage of H2O2 photolysis is that it is not general to both proteins and nucleic acids.
Radiolysis of water by ionizing radiation produced either from low flux gamma sources and high flux synchrotron beams has been effectively used to footprint DNA, RNA and proteins [16, 37, 53-56]. The advantage of radiolysis compared to the above described methods is that 'nothing but light' is added to the solutions containing the macromolecules to be footprinted. The disadvantage of low flux gamma sources is the need for an expensive gamma source, the long exposure required and cumbersome sample loading and unloading. While high flux synchrotron beams allow very short exposure times, substantial heat may be generated that is intolerable to biological samples. More importantly, the use of a synchrotron requires an application for beamtime and the transport of samples to a remote facility. Therefore, synchrotron footprinting is incompatible with laboratory-based high-throughput implementation.
The past decade has seen the advent of high-throughput structure determination initiatives focused on proteins, DNA and RNA. These initiatives are typically grouped under the rubric 'structural genomics'. High-throughput structure determination may be particularly valuable in screening studies where large numbers of structures and/or complexes need to be interrogated. Biological function may be revealed only by understanding the acquisition of structure as a function of time or the binding of a ligand requiring the determination of multiple structures for a single reaction.
High throughput structural initiatives have had limited to moderate success despite the infusion of many millions of dollars. While atomic resolution structures are the gold standard for structure determination, biological function can often be gleaned from lower resolution structures. In many cases, the complexity or size of macromolecules and complexes precludes determination of an atomic resolution structure. RNA molecules of even moderate size are notoriously refractory to structure determination.

Solid state synthesis hydroxyl radicals for high throughput structure determination of proteins and nucleic acids by oxidative footprinting

Albert Einstein College of Medicine, Inc.


A method and apparatus for estimating a parameter of interest of an earth formation ahead of a carrier within borehole. The method including estimating one or more functions based on received signals that may reduce the sum of the signals received by a first receiver, and estimating the parameter of interest based on signals received by one or more additional receivers by applying the one or more functions and, if necessary, a defocusing factor, such that reception of information from downhole of a selected downhole location is enhanced relative to reception of information uphole from the selected location. The apparatus including two or more transmitters, two or more receivers, and a processor to estimate one or more functions, and, based on these functions, the parameter of interest.

1. Field of the Disclosure
The present disclosure relates to estimating one or more parameters of interest relating to an earth formation. In one aspect, the present disclosure relates to a method of determining one or more parameters relating to the earth formation ahead of a selected location in a borehole.
2. Background of the Art
Exploration and production of hydrocarbons generally requires drilling a borehole into the earth. The borehole can be used to gain access to depths of the earth for performing measurements related to the exploration and production.
Well logging is a technique used to perform the measurements in the borehole. In well logging, a logging tool is conveyed through the borehole. For "logging-while-drilling" (LWD), a logging tool is coupled to a drill string. Thus, the measurements can be performed while the borehole is being drilled, during interruptions in drilling, or as the drill string is conveyed along the borehole. In post drilling applications, a logging tool may be conveyed via a non-rigid carrier such as a wireline in order to log the earthen formation. Generally speaking, logging tools may transmit signals into the earthen formation and then detect reflected signals and/or the response of the earthen formation to the transmitted signals.
In some situations, it may be desirable to have a tool that is predominantly sensitive to the one or more formation features downhole of the selected downhole location and, at the same time, is minimally affected by the formation properties uphole the selected downhole location.

TDEM forward focusing system for downhole use


An acoustic viscous damper for a compressor including a porous liner located within the discharge duct, the porous liner having a plurality of holes for passage of a purge working fluid therethrough into the discharge duct.

The present invention relates generally to compressors and more particularly, but not exclusively, to centrifugal gas compressors.
Centrifugal gas compressors are known to have problems with noise believed to originate from the airfoils of the impellers. Many such compressors will have at least one rotating radial impeller mounted within a rigid casing and will further include an inlet pipe and inlet plenum and discharge pipe and a discharge plenum. The rapid rotation of the blades of the impeller causes them to set up some harmful pressure pulsations inside the working fluid. These pulsations can be transmitted through the discharge pipe, or other components of the compressor, resulting in potential mechanical damage or environmental noise.
It is believed that the space-varying pressure field surrounding each of the airfoils of the impeller is acting as a source of noise. This noise is due to the rapid rotation of the airfoils inside the pressurized casing of a typical gas compressor. The pressure field of each airfoil may interact with other components, such as diffuser vanes. Similarly, the pressure field of each airfoil may interact with the geometry of the collector plenum itself. In both such scenarios, pressure pulsations are developing inside the collector plenum at approximately the so-called blade passing frequency. The blade passing frequency is approximately equal to rotation speed of the impeller in rotations per second times the number of impeller blades. These pulsations are potentially of a high enough level to cause mechanical or environmental concerns. Consequently, significant efforts have focused on developing mechanisms to minimize pressure pulsations within compressors.
Heretofore, there has been a need for compressors having improved noise characteristics when considered in view of other characteristics such as unit cost, ease of design, and other competing tensions.

Acoustic viscous damper for centrifugal gas compressor


The present invention is directed to a chimeric crystal protein comprising CryIF and CryIAc. Bacillus thuringiensis isolates comprising said protein and a method of controlling lepidopterans are also included in the present invention.

Bacillus thuringiensis ("B.t.") is a gram-positive bacterium that typically produces proteinaceous crystalline inclusions during sporulation. These B.t. crystal proteins are highly toxic to certain insects. Crystal proteins from various B.t. strain isolates have been identified as having insecticidal activity against insect larvae from the insect orders Lepidoptera (caterpillars), Coleoptera (beetles), Diptera (mosquitoes, flies), and Homoptera (aphids). The insecticidal crystal proteins (ICPs) of B. thuringiensis were originally classified as CryI, CryII, CryIII, and CryIV proteins based on their insecticidal activities (Hofte H. and H. R. Whiteley, Microbiol. Rev. 53:242-255 (1989)). The Hofte and Whiteley (1989) review provides an overview of the B.t. insecticidal crystal protein genes. CryI proteins, which encompass crystal proteins of approximately 130-140 kilodaltons (kDa) in molecular mass, display lepidopteran toxicity. CryII proteins are approximately 71 kDa in mass and may display both lepidopteran and dipteran toxicity. CryIII proteins are approximately 73-74 kDa in mass and display coleopteran toxicity. The CryIV proteins represent a diverse group of proteins that exhibit dipteran toxicity. For the purpose of the present invention, the Hofte and Whiteley nomenclature will be used.
Commercial B.t. bioinsecticide products currently being marketed for lepidopteran insect control are based on either naturally occurring ("native") strains or transconjugants strains. Transconjugant strains are created by transferring a crystal protein-encoding plasmid from a donor strain to a recipient strain via a conjugation-like process, resulting in a new B.t. strain. Plasmids may also be transferred from one strain to another by phage transduction. Native and transconjugant strains are fermented in a broth medium, the spores and crystals harvested, either by spray-drying or by centrifugation, and subsequently formulated for spray application.
The insecticidal activity of conventional B.t. bioinsecticides results from insect larvae feeding on the crystal protein, typically in sprayed-on deposits of the bioinsecticide on leaves or other plant surfaces. General details of the mode of action of the insecticidal crystal proteins (ICPs) are apparent. The ICPs contained within the proteinaceous crystals are released into the insect midgut after ingestion and solubilization of the crystals. In many instances, the full-length proteins or protoxins are processed by midgut proteases to a fully active state. The 130-140 kDa CryI protoxins, for instance, are processed to a core toxin moiety of 60-65 kDa, derived from the amino terminal half of the full-length protoxin. This processed core toxin is regarded as the active toxin within the insect midgut. The discarded carboxyl domain of the protoxin, though not essential for toxicity, is apparently indispensable for CryI crystal formation in B.t. For purposes of the present invention, the carboxyl half of the CryI protein not contained within the active or core toxin will be referred to as the tail domain. As depicted in FIG. 1, the processed, activated toxin, derived from the chimeric CryIF-IAc crystal protein of the present invention, binds to the brush border membranes (BBMs) of the insect midgut epithelium, a step that frequently requires the presence of fortuitous "receptor" proteins. This binding is followed by an apparent intercalation event in which the active toxin moiety, or a portion of it, contributes to the formation of ion channels as well as aggregates to form larger pores within the BBM, leading to osmotic imbalance, cellular swelling and lysis. Intoxicated insect larvae stop feeding within minutes and eventually die.
For many lepidopteran insect pests, such as armyworms, the B.t. spore present in the bioinsecticide formulation also contributes substantially to toxicity. The synergistic effect of spores has been reported for a number of important lepidopteran insect pests, including Spodoptera exigua (Moar, W. J., et al., Appl. Environ. Microbiol. 61:2086-2092 (1995)), Lymantria dispar (DuBois, N. and D. H. Dean., Biological Control 24:1741-1747 (1995)), and Plutella xylostella (Tang, J. D., et al., Appl. Environ. Microbiol. 62:564-569 (1996)). This spore effect on the insecticidal activity of B.t. is apparently due to septicemia: the ability of the spore to germinate within the insect midgut, to penetrate the disrupted midgut epithelium, and to enter and proliferate within the hoemcoel. For many lepidopteran insect pests, it is therefore desirable that the B.t. bioinsecticide formulation contain a mixture of spores and crystals to achieve maximal efficacy.
Among the lepidopteran insect pests, armyworms are particularly difficult to control, regardless of the insecticide used. Thus, there is a need for bioinsecticide products for armyworm control that are both efficacious and cost-effective. To satisfy this need, the amount of crystal protein produced by B.t. in fermentation should be maximized as much as possible in order to provide for its economic and efficient utilization in the field. Increased concentration of crystal protein in the formulated bioinsecticide promotes use of reduced amounts of bioinsecticide per unit area of treated crop, without reducing the actual amount of crystal protein applied per unit area, thereby allowing for more cost-effective use of the bioinsecticide product. Alternatively, increased fermentation yields of crystal protein, resulting in more concentrated formulations, may be used to increase the amount of crystal protein applied per unit area, thereby enhancing the performance of the bioinsecticide product.
Previous efforts to create mutants or variants of B.t. strains that show enhanced production of crystal proteins have related primarily to the production of coleopteran- or dipteran- toxic crystal proteins, not CryI lepidopteran-toxic crystal proteins. Also, most of these examples describe oligosporogenous or asporogenous (produce few, if any, spores) variants of B.t. that show increased crystal protein production. As noted above, the full production of spores is a desirable feature for a lepidopteran-active B.t. strain used for the production of a comnmercial bioinsecticide.
U.S. Pat. No. 5,006,336, issued to Payne, describes a native B.t. isolate (PS122D3), active against coleopteran insects, which produces more coleopteran-toxic protein (CryIIIA) than does an unrelated coleopteran-toxic B.t. strain, B.t. san diego. Strain PS122D3 is not a variant of B.t. strain san diego.
U.S. Pat. No. 4,996,156, issued to Zaehner et al., describes a dipteran-active B.t. israelensis mutant strain which produces crystal proteins but is asporogenous.
Published European Patent Application Publication No. O 099 30 of Fitz-James, describes mutants of B.t. israelensis, obtained using a chemical mutagen, that produces up to 1.5 times the amount of dipteran-toxic crystal protein as does the progenitor strain.
Published European Patent Application Publication No. O 228 228, of Mycogen Corporation, describes asporogenous Bacillus thuringiensis mutants obtained by treatment of the progenitor strains with ethidium bromide. Such B.t. mutants are described as being more efficient at producing coleopteran-toxic (CryIIIA) crystal protein.
Published PCT International Patent Application Publication No. WO 91/07481, of Novo Nordisk A/S, describes a mutant of Bacillus thuringiensis tenebrionis, which was obtained by gamma irradiation and which produces two times the amount of coleopteran-toxic crystal protein (CryIIIA) obtained from the progenitor strain.
U.S. Pat. No. 4,990,332, issued to Payne et al., describes a lepidopteran-toxic B.t. kurstaki mutant strain (PS85al-168) that produces crystal protein in amounts "equal to or higher than the wild type" but is asporogenous.
The efficacy and cost effectiveness of a B. thuringiensis-based bioinsecticide product may also be improved by manipulation of the crystal protein composition in the bioinsecticide product, engineering of crystal proteins for improved insecticidal activity or stability, and by improvements in the formulation that promote longer shelf-life and longer persistence upon field application.
With respect to engineered crystal proteins, Geiser and Moser, in published Canadian Patent Application No. 2035199, disclose a chirneric CryIAb-CryIAc protoxin that shows improved temperature stability when compared to the native CryIAb protoxin. The native CryIAb protoxin does not form crystal protein inclusions in B. thuringiensis when grown at temperatures of 30-35.degree. C., the preferred temperature for cultivation of B. thuringiensis, and therefore cannot be produced efficiently in B. thuringiensis under these conditions. The chimeric CryIAb-CryIAc-protein, in which most of the carboxyl half or tail domain of the CryIAb protein has been replaced with that of the CryIAc protein, forms crystal protein inclusions in B. thuringiensis at 30.degree. C. and thus can be produced efficiently. This chimeric protoxin was reported to have the same toxicity profile for H. virescens and Trichoplusia ni larvae as the parental CryIA(b) protoxin (Aronson, A. 1993. Insecticidal toxins in Bacillus subtilis and other gram-positive bacteria. Sonensheim, A. L., Hoch, J. A., and Losick, R. (eds.). Am. Soc. Microbiol., Washington D. C., pp. 953-963). Furthermore, the reciprocal chimeric protein, in which most of the carboxyl half or tail domain of the CryIAc protein has been replaced with that of CryIAb, does not form stable inclusions in B.t. at the preferred cultivation temperature thus mimicing the temperature sensitive crystal forming phenotype of CryIAb.
Thompson et al., in PCT International Publication No. WO 95/30752, disclose a chimeric CryIC-CryIAb protoxin that shows improved insecticidal activity when produced in Pseudomonas fluorescens. This protoxin alone may not be produced efficiently in B. thutingiensis at the preferred cultivation temperature because it contains the tail domain of CryIAb that confers the temperature-sensitive crystal-forming phenotype of CryIAb described by Geiser and Moser.
A cryIF lepidopteran-toxic crystal protein gene is described in U.S. Pat. No. 5,188,960 issued to Payne et al., and in PCT International Publication No. WO 91/16434 of Ecogen Inc. Chambers et al. (1991) J. Bacteriol. 173:3966-3976, discloses a cryIF crystal protein gene from B.t. strain EG6346 subsp. aizawai and the insecticidal activity of its encoded CryIF crystal protein. A recombinant B.t. strain expressing this cryIF gene and designated strain EG7826 is disclosed by Baum in PCT International Publication No. WO 95/02058. Thompson and Schwab, in PCT International Publication No. WO 95/30753, disclose a chimeric CryIF-CryIAb protoxin that can be produced more efficiently in Pseudomonas fluorescens than the native CryIF protoxin. By analogy to the work of Geiser and Moser, the production of this chimeric CryIF-CryIAb protoxin in B. thuringiensis , at the preferred cultivation temperature, would be expected to be less efficient than that of the native CryIF protein. Thus, the requirements for efficient expression in B. thuringiensis and Pseudomonas appear to differ.
U.S. Pat. No. 5,508,264, issued to Bradfisch et al.discloses a synergistic effect as a result of the combination of the chimeric CryIF crystal protein and a chimeric CryIAc crystal protein produced in Pseudomonas fluorescens. The disclosed chimeric CryIF proteins includes the chimeric CryIF-CryIAb protoxin or a chimeric CryIF-CryIAc/CryIAb protoxin. The protoxin domain of the latter protein is comprised of both CryIAb and CryIAc sequences.
There is a need to improve the efficacy and cost-effectiveness of lepidopteran-toxic insecticides, particularly those used for the control of armyworms. This may be achieved, in part, by improving the amount of CryI crystal protein obtained in fermentation, thereby allowing for more economic use of the crystal protein in bioinsecticide formulations. The toxicity of the bioinsecticide product towards armyworms may be further improved by the use of a Bacillus thuringiensis strain that is proficient in the production of spores as well as in the production of CryI crystal protein. As evidenced by the prior art, standard methods for optimizing the production of CryI crystal proteins in a spore-forming entomopathogenic bacterium such as Bacillus thuringiensis remain elusive.

Chimeric lepidopteran-toxic crystal proteins


Proposed is a method for the finishing treatment of various kinds of fabric materials with an aminoalkyl-modified dimethylpolysiloxane to impart the fabric material with excellent softness and smoothness with less disadvantages of yellowing of the treated fabric material than in the prior art using similar aminoalkyl-modified dimethylpolysiloxanes. In the inventive method, the aminoalkyl-modified dimethylpolysiloxane is represented by the general formula EQU A--SiR.sub.2 --O--(--SiR.sub.2 --O--).sub.p --(--SiRQ--O--).sub.q --SiR.sub.2 --A,. in which R is a monovalent hydrocarbon group having 1 to 20 carbon atoms, Q is an aminoalkyl group expressed by the general formula EQU --R.sup.1 --(--NH--CH.sub.2 --CH.sub.2 --).sub.n --NH.sub.2,. R.sup.1 being a divalent hydrocarbon group having 1 to 8 carbon atoms and the subscript n being 2 or 3, A is a hydroxyl group, R, OR or Q, the subscript p is an average number in the range from 5 to 2000 and the subscript q is 0 or an average number not exceeding 100 with the proviso that q is not 0 when the groups denoted by A in a molecule are each a hydroxyl group, R or OR, and having an amine equivalent in the range from 5000 to 100,000 g/mole.

The present invention relates to a method for the finishing treatment of a fabric material or, more particularly, to a method for the finishing treatment of a fabric material by using a silicone-containing fabric finishing agent capable of imparting the treated fabric material with excellent touch feeling of softness and smoothness as accompanied by less significant troubles of yellowing of the treated fabric material than with conventional silicone-containing fabric finishing agents.
It is a conventional prior art method widely under practice that various kinds of fabric materials are imparted with improved touch feelings of softness and smoothness by a finishing treatment using a silicone-containing fabric-finishing agent. The silicones as the principal ingredient in the prior art fabric-finishing agents include dimethylpolysiloxanes, epoxy-modified organopolysiloxanes and aminoalkyl-containing organopolysiloxanes, of which aminoalkyl-containing organopolysiloxanes are most widely employed in respect of the particularly excellent softness imparted to the finished fabric material as well as versatility thereof relative to the kinds of the fibers forming the fabric material. While the aminoalkyl groups contained in the aminoalkyl-containing organopolysiloxanes are not particularly limitative, the aminoalkyl groups of the most preferred types include 3-aminopropyl group of the formula --C.sub.3 H.sub.6 NH.sub.2 and 3-N-(2-aminoethyl)aminopropyl group of the formula --C.sub.3 H.sub.6 NHCH.sub.2 CH.sub.2 NH.sub.2 because a fabric-finishing agent containing an organopolysiloxane having these aminoalkyl groups imparts the treated fabric material with very excellent touch feeling of softness as is taught in Japanese Patent Publications 48-1480, 54-43614 and 57-43673 and Japanese Patent Kokai 60-185679, 60-185880 and 64-61576.
Though excellent in respect of the touch feeling of softness of the treated fabric materials, the fabric-finishing agents containing an organopolysiloxane having the aminoalkyl groups of the above mentioned types are not quite satisfactory due to a very serious problem that the fabric materials finished therewith sometimes cause yellowing when subjected to a heat treatment or dried under sunlight or in the lapse of time due to degradation of the amino groups presumably by the influences of heat or ultraviolet light eventually resulting in the loss of the softness. This problem is particularly detrimental against white-colored or light-colored fabric materials.
Various attempts and proposals have been made heretofore with an object to solve the above mentioned problem due to yellowing of the fabric material finished with an aminoalkyl silicone-based fabric-finishing agent of the above mentioned types, in which the aminoalkyl-containing organopolysiloxane is modified by the reaction with an anhydride or chloride of an organic acid disclosed in Japanese Patent Kokai 57-101076, with an epoxy compound disclosed in Japanese Patent Kokai 59-179884, with a higher fatty acid disclosed in Japanese Patent Kokai 1-306683 and with a carbonate disclosed in Japanese Patent Kokai 2-47371. Modification of the aminoalkyl groups is of course not ineffective, though insufficient, to solve the problem of yellowing of the treated fabric materials but is accompanied by a decrease in the effectiveness of imparting softness and smoothness to the finished fabric materials as compared with a silicone-based fabric finishing agent containing an organopolysiloxane having unmodified aminoalkyl groups.
Besides the types of the aminoalkyl groups, another important factor which influences the usefulness of an aminoalkyl-containing organopolysiloxane as the principal ingredient in the silicone-based fabric-finishing agent is the content of the aminoalkyl groups in the organopolysiloxane. For example, the aminoalkyl-containing organopolysiloxane having 3-N-(2-aminoethyl)amino propyl groups most widely employed in he prior art fabric finishing agents has an amine equivalent in the range from 1500 to 2000 g/mole because of the highest softness imparted to the treated fabric materials. The amine equivalent here implied is the amount of the organopolysiloxane in grams giving 1 mole of the amino groups --NH.sub.2 and imino groups --NH-- as a total. When the amine equivalent of the aminoalkyl-containing organopolysiloxane is increased, for example, in the range from 3000 to 4000 g/mole, the trouble due to yellowing of the treated fabric materials can be dissolved to some extent though at a sacrifice of the softness imparted to the treated fabric materials.

Method for finishing treatment of a fabric material


A tool system for working on a bone plate is described. The tool system includes a first tool and a second tool, used at the same time as the first tool to shorten or bend the bone plate. Each of the two tools includes one or more introduction openings with which in turn one or more blades may be associated.

In surgical operations such as reconstructions in the region of the lower jaw, implant systems including bone plates are frequently used. In the context of a surgical operation of this kind, it is frequently necessary to adapt the bone plate to certain anatomical conditions. Adaptations of this kind include, for example, bending and shortening the bone plate.
Bending a bone plate may be performed by means of two clamping pincers. In this case, the clamping pincers are placed at two mutually spaced points on the bone plate and then moved relative to one another. To cut a bone plate, a pincer with cutting jaws is conventionally used.
The object of the invention is to specify an easily handled tool system for working on a bone plate.


In a digital camera 1, in an automatic shooting stand-by state, when judged at Step S26 that an automatic shooting condition has been satisfied (N at Step S26), whether or not to issue an AE re-processing command is judged. In other words, whether or not the current shooting condition is similar to a condition where an automatic shooting condition is satisfied is judged. When judged to issue the AE re-processing command, exposure tracking in an aperture-priority mode is performed (S28), and shooting exposure conditions excluding the aperture value for still-image shooting in the aperture-priority mode are determined. Then, the storage is updated (Step 29), and the process returns to Step S26. When judged at Step S26 that an automatic shooting condition has been satisfied (Y at Step S26), record processing for still-image shooting is performed with the updated shooting exposure conditions without changing the aperture value (Step S30).

1. Field of the Invention
The present invention relates to an imaging apparatus and a program thereof. In particular, the present invention relates to an imaging apparatus providing an automatic recording function and a program thereof.
2. Description of the Related Art
A technology is known in which automatic recording is performed when the subject enters a predetermined condition. To actualize a technology such as this, there is a method in which imaging conditions (such as aperture value, shutter speed, and gain value) used during automatic shooting are calculated upon entering an automatic shooting stand-by state, and automatic shooting and recording is performed using the calculated imaging conditions to prevent time lag in automatic shooting timing caused by the calculation of imaging conditions used during automatic shooting and mechanical aperture operations (Japanese Patent Application Laid-Open (Kokai) Publication No. 2003-092701).
However, according to the above conventional technology (Japanese Patent Application Laid-Open (Kokai) Publication No. 2003-092701), when a prolonged time is taken to perform automatic recording after entering the automatic shooting stand-by state, the luminance of the subject may change. As a result, image data captured with correct exposure cannot be obtained.