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<SOH> SUMMARY OF THE INVENTION <EOH>The invention features an electrode array in which pairs of electrodes are geometrically arranged so that the broadest faces of the exposed electrodes are not directly opposed to each other. Rather, the broadest facing surfaces of the electrodes in the array are parallel, adjacent, or offset at an angle. The electrode geometry of an electrode array of the invention permits electrodes to be in close proximity, thereby lowering series resistance, while minimizing the possibility for short circuits that can cause electrical leakage. The electrode array of the present invention may function as an anode, cathode, or as a stand-alone capacitor or electrochemical cell combining cathodes and anodes in one sheet of material. Electrodes may be formed as patterns of lines, dots, or other shapes placed on, attached, or formed onto a substrate, including a conductive material, or, alternatively, a nonconductive sheet material. These lines, dots, etc., may be formed in very thin layers and may be spaced very close. An electrochemical cell, fuel cell, battery, capacitor, or flow-through capacitor is formed by the adjacent pairs or groups of lines and dots. Thus, one aspect the invention features an array of electrodes, whereby the broadest exposed electrode faces of the electrodes are adjacent to each other. Alternatively, the broadest faces of the exposed electrodes are coplanar or offset at an angle, but not directly opposite to each other, whether on the same sheet of a material containing both anodes and cathodes, or between separated anode sheets and cathode sheets. This minimizes the possibility for short circuits that cause leakage. The electric field comes up out of the anodes and curves back to the cathode. The electrodes of the array can be dots, shapes, or lines that can be recessed into the dielectric spacer as a further means of protecting against electrical leakage between them. The electrode array of the present invention uses a geometric arrangement of parallel, adjacent, or offset electrodes in order to create capacitors, electrochemical cells, or flow-through capacitors. A dielectric insulator serves as a spacer between adjacent electrodes; the spacer may be a porous, nonporous, ion-permeable, ion-selective, membrane, or other dielectric material. Optionally, current collectors are in electrical contact with the electrodes, either placed under, or embedded in, or sandwiched between, the electrodes. The adjacent geometry of the electrodes within the electrode array reduces series resistance and leakage sufficiently that the distance between the anode and the cathode can be less than 0.03 inches, or can preferably be reduced to 0.005 inches, or to less than 0.001 inches. Electrode materials in any shapes separated by insulating materials may be manufactured with a narrow distance between adjacent electrodes, reduced to 1 micron or less, by using manufacturing methods commonly used to print circuit boards in the semiconductor industry. Where it is preferred to use a wider space between the electrodes, for example, over 0.001 cm, manufacturing methods such as screen or other printing or coating methods will suffice to manufacture the electrode array of the present invention. The electrode array of the invention can be used in any type of electrochemical cell, such as capacitors, batteries, and fuel cells. An electrode array of the invention can be a used in a flow-through capacitor. Alternatively, an electrode array of the invention can be used in a lithium battery. In one embodiment, the electrode array is comprised of dots, shapes, or lines that may be recessed into the spacer as a further means of protecting against electrical leakage between them. The broadest faces of the electrodes are offset from each other. An additional advantage of the present invention is that the electrode arrays may be lines, dots, or any other shapes that can be arranged in patterns with small distances between shapes or nonfacing or adjacent surfaces of each pair of electrodes, for example, less than 3 millimeters, thereby allowing construction of an electrode array containing anode-cathode pairs within a single sheet of material. Therefore, the electrode array sheet may comprise anode-cathode pairs to act as an integrated capacitor, electrochemical cell, or flow-through capacitor. These electrode array sheets, containing one or more anode-cathode pairs per sheet, may be stacked together in any geometry known to prior art flow-through capacitors, electrochemical cells, water filters, batteries, or electronic capacitors. In one embodiment, the electrode pattern of the present invention forms a two-dimensional electrode array. The electrode array may be used as a double-layer capacitor, capacitor, flow-through capacitor, fuel cell, or any other electrochemical device. The thin electrodes, for example, less than 0.13 cm thick, and thin spacing, for example, less than 0.13 cm apart, of this array reduce leakage and ESR. Leakage is reduced because the broadest area of each electrode of an electrode pair is offset from the other electrode. Because the electrodes may be placed close together without generating electrical leakage, series resistance is reduced. Series resistance of less than 50 ohms/cm 2 of electrode array facing area and leakage resistance of more than 30 ohms/cm 2 of electrode array facing area can be achieved by the present invention. |
Method for analysing audio signals |
The present invention relates to a method for analyzing, separating and extracting audio signals. Due to the generation of a series of short-time spectra, a non-linear mapping into the pitch excitation layer, a non-linear mapping into the rhythm excitation layer, extraction of the coherent frequency streams, extraction of the coherent time events and the modeling of the residual signal, the audio signal can be decomposed into rhythm and frequency portions with which the signal can be further processed in a simple manner. The uses of said method are: data compression, manipulation of the time base, tune and formant structure, notation, track separation and identification of audio data. |
1. A method for analyzing audio signals by a) generating a series of short-time spectra, b) non-linear mapping of the short-time spectra into the pitch excitation layer (PEL), c) non-linear mapping of the short-time spectra into the rhythm excitation layer (REL), d) extraction of the coherent frequency streams from the audio signal, e) extraction of the coherent time events from the audio signal, f) modeling of the residual signal of the audio signal. 2. The method according to claim 1, wherein the short-time spectra are produced by means of short-time Fourier transform, by means of wavelet transform, or by means of a hybrid method consisting of wavelet transform and Fourier transform. 3. The method according to claim 1, wherein the mapping into the pitch excitation layer consists of the correlation of the logarithm of the spectral magnitude with a predetermined ideal harmonic spectrum, suppression of spectral echoes corresponding to the positions of possible harmonics, and of a subsequent separation of the frequency streams. 4. The method according to claim 3, wherein a lateral inhibition is performed according to at least one of the mappings logarithm, correlation and suppression of the echoes. 5. The method according to claim 4, wherein correlation, suppression of the echoes and lateral inhibition are linear mappings. 6. The method according to claim 3, wherein the separation of the frequency streams is carried out with a neuronal network. 7. The method according to claim 3, wherein the separation of the frequency streams is achieved by searching for time-coherent local maxima and calculation of the pitch data as a time series. 8. The method according to claim 1, wherein the mapping into the rhythm excitation layer consists of a linear mapping for frequency noise suppression and for time correlation, which is applied to the logarithm of the spectral magnitude. 9. The method according to claim 8, wherein the time correlation matrix is given by a differential correlation. 10. The method according to claim 1, wherein the extraction of a frequency stream from the audio signal is carried out with a filter having a variable center frequency. 11. The method according to claim 10, wherein the center frequency of the filter is controlled via frequency trajectories from the pitch excitation layer. 12. The method according to claim 10 wherein the extracted signal is multiplied by a complex-valued envelope to adapt the phase with an optimization method. 13. The method according to claim 12, wherein the complex-valued envelope is used for adapting the amplitude of the signal with an optimization method. 14. The method according to claim 1, wherein the frequency streams are calculated as a development according to the band signals of a filterbank, the coefficients being given by projections of a frequency evaluation onto the frequency responses of the filterbank. 15. The method according to claim 1, wherein the extraction of the time events consists of a frequency evaluation and a time domain evaluation. 16. The method according to claim 15, wherein the frequency evaluation is carried out with an FFT filter or an analysis filterbank. 17. The method according to claim 1, wherein the residual signal is statistically modeled. 18. The method according to claim 17, wherein several bands with frequency-localized noise are used for modeling, the bands being added according to a frequency analysis with a time-dependent weighting. 19. The method according to claim 17, wherein the residual signal is modeled by calculating a distribution function from the statistic moments at predetermined time intervals. 20. The method according to claim 19, wherein the interval windows are overlapping with 50% and are then added during resynthesis, evaluated with a triangle window. 21. A method for compressing audio signals by separating the audio signal according to claim 1, and subsequent compression of the PEL streams, REL events and the residual signal. 22. The method according to claim 21, wherein compression comprises the steps of: a) adaptive double-differential coding of the PEL streams, b) time-localized coding of the REL events, c) adaptive differential coding of the residual signal, d) statistic compression of the data from steps a), b) and c) by entropy maximization. 23. The method according to claim 22, wherein the events for REL coding are given as a linear combination of a finite amount of base vectors. 24. The method according to claim 22, wherein the final compression is carried out with LZW or Huffmann methods. 25. A method for manipulating the time base of signals which have been separated with the method according to claim 18, by a) determining the envelopes or trajectories of the PEL streams and the envelopes of the noise bands, b) adapting the time marks of the envelope or trajectory points, c) adapting the times of the events, d) adapting the envelope grid points of the noise bands. 26. A method for manipulating the time base of signals which have been separated with the method according to claim 19, by a) determining the envelopes or trajectories of the PEL streams, b) adapting the time marks of the envelope or trajectory points, c) adapting the times of the events, d) adapting the synthesis window lengths in moment coding. 27. A method for manipulating the tune of signals which have been separated with a method according to claim 1, by shifting the logarithmic frequency trajectories along the frequency axis. 28. A method for manipulating a formant structure of signals which have been separated according to the method according to claim 18, by a) determining the harmonic amplitudes of PEL streams, b) interpolating a frequency envelope from the harmonic amplitudes, c) shifting the frequency envelope, d) adapting the band frequencies in the noise band representation according to the formant shift. 29. A method for the notation of audio data into musical notes by a) separating the audio signal according to the method of claim 1, b) grouping the PEL streams according to their harmonic characteristics into at least one group by means of trainable vector quantizer, c) identifying the percussive instruments by comparing REL events with low-frequency PEL events or residual signal portions by means of a neuronal network, d) converting the frequency trajectories of each group and the percussion beats into notations. 30. A method for the track separation of audio data by a) separating the audio signal according to the method of claim 1, b) grouping the PEL streams according to their harmonic characteristics by means of a trainable vector quantizer, c) identifying PEL streams, REL events and residual signal portions pertaining to one group, by means of a neuronal network, d) resynthesis of the associated streams, events and residual signal portions into one track for each group. 31. A method for identifying an audio signal by separating the signal according to claim 1, and subsequent comparison of the relative positions and types of streams and events with a database. 32. A method for identifying an audio signal by separating the signal according to claim 1, and subsequent comparison of dominant structures with a database. 33. A method for identifying a voice in an audio signal by separating the signal according to claim 1, extrapolation of the formant position from the PEL streams and subsequent comparison with a database. 34. A method according to claim 31, wherein a hashing scheme is used for restricting the selection after separation of the signal and a checksum comparison is thus made with the database. |
<SOH> FIELD OF THE INVENTION <EOH>The present invention relates to a method for analyzing audio signals. By analogy with the function of the human brain, audio signals are analyzed in the present method with respect to frequency and time coherence. Data streams of the signals can be separated by extracting said coherences. |
Phase correction circuit |
There is provided a phase correction circuit capable of detecting a skew between a data signal and a clock signal without requiring a clock signal as pattern data upon initialization. The phase correction circuit is configured to include a variable delay device 10 to which a data signal in a DDR format is inputted, a first F/F 1 which fetches a delayed data signal in synchronization with the clock signal, a second F/F 2which fetches the delayed data signal in synchronization with a reverse clock signal, a third F/F 3 which fetches an output signal from the first F/F 1 in synchronization with the clock signal, and a fourth F/F 4 which fetches an output signal from the second F/F 2 in synchronization with the clock signal, and the phase correction circuit further includes a fifth F/F 5 which fetches a rate signal having the same cycle as that of the data signal in synchronization with the clock signal, a sixth F/F 6which fetches an output signal from the fifth F/F 5 in synchronization with the clock signal, and an AND circuit 8 to which an output signal from the third F/F 3 and an output signal from the sixth F/F 6 are inputted. |
1. A phase correction circuit comprising: a variable delay device to which a data signal in a DDR format is inputted; first fetch means to fetch a delayed data signal outputted from the variable delay device in synchronization with a clock signal; second fetch means to fetch the delayed data signal in synchronization with a reverse clock signal obtained by reversing the clock signal; third fetch means to fetch an output signal from the first fetch means in synchronization with the clock signal; and fourth fetch means to fetch an output signal from the second fetch means in synchronization with the clock signal, the phase correction circuit further comprising: fifth fetch means to fetch a rate signal having the same cycle as that of the data signal in synchronization with the clock signal; and an AND circuit to which an output signal from the third or fourth fetch means and an output signal from the fifth fetch means are inputted. 2. The phase correction circuit according to claim 1, further comprising: sixth fetch means to fetch an output signal from the AND circuit in synchronization with the clock signal; and a latch circuit which holds an output signal from the sixth fetch means. 3. The phase correction circuit according to claim 1 or 2, wherein all or some of the fetch means consist of flip-flops. 4. A phase correction method comprising: a variable delay step of receiving a data signal in a DDR format, and outputting it as a delayed data signal obtained by delaying the data signal by a predetermined time; a first fetch step to fetch the delayed data signal in synchronization with a clock signal; a second fetch step to fetch the delayed data signal in synchronization with a reverse clock signal obtained by reversing the clock signal; a third fetch step to fetch an output signal from the first fetch step in synchronization with the clock signal; a fourth fetch step to fetch an output signal from the second fetch step in synchronization with the clock signal; a fifth fetch step to fetch a rate signal having the same cycle as that of the data signal in synchronization with the clock signal; and a logical product step of receiving an output signal from the third or fourth fetch step and an output signal from the fifth fetch step, and obtaining their logical product. 5. The phase correction method according to claim 4, further comprising: a sixth fetch step to fetch an output signal from the logical product step in synchronization with the clock signal; and a latch step of holding an output signal from the sixth fetch step. |
<SOH> BACKGROUND ART <EOH>In recent years, a data signal is transferred in a DDR format between chips or between modules. In the DDR format, the data signal is processed with the both timing of a rise edge and a fall edge of a clock signal. Therefore, transferring the data signal in the DDR format can improve a transfer rate. Before explaining the present invention, a conventional example of a circuit (input circuit) receiving a data signal in a DDR format will be briefly described with reference to FIG. 4 . As shown in FIG. 4 , the input circuit for a data signal in a DDR format is generally constituted of a variable delay device (VD) 10 , a first flip-flop (first F/F) 1 , a second flip-flop (second F/F) 2 , a third flip-flop (third F/F) 3 and a fourth flip-flop (fourth F/F) 4 . A data signal in a DDR format is inputted to the variable delay device. Further, the first F/F 1 fetches a delayed data signal outputted from the variable delay device 10 in synchronization with a clock signal. That is, the data signal is fetched with a timing of a rising edge of the clock signal. Furthermore, the second F/F 2 fetches the delayed data signal in synchronization with a reverse clock signal obtained by reversing the clock signal. That is, the data signal is fetched with the timing of a fall edge of the clock signal. Therefore, a phase of the data signal outputted from the second F/F 2 is delayed from a phase of the data signal outputted from the first F/F 2 by a ½ cycle of the clock signal. Thus, the third F/F 3 fetches an output signal from the first F/F 1 in synchronization with the clock signal. Moreover, the fourth F/F 4 fetches an output signal from the second F/F 2 in synchronization with the clock signal. Since the third and fourth F/F 3 and 4 respectively fetch output signals in synchronization with the clock signal in this manner, the output signal of the first F/F 1 and the output signal of the second F/F 2 can be in phase. Meanwhile, the data signal in a DDR format is transferred at a high speed. Therefore, in the DDR format, an allowable range of a skew (signal phase shift) between the data signal and the clock signal is very narrow. Thus, in the input circuit, the skew is corrected by delaying the data signal by using the variable delay device 10 . Here, setting a delay time at the time of initialization can be performed by giving predetermined data and a clock as pattern data, gradually changing a delay time of the variable delay device 10 and reading output values of the first and second F/F 1 and 2 each time. As shown in FIG. 4 , the conventional input circuit includes a first AND circuit 11 to which the output signal from the first F/F 1 and a read mode signal are inputted and a second AND circuit 12 to which the output signal from the second F/F 2 and the read mode signal are inputted, thereby reading output values of the first and second F/F 1 and 2 . The output values are read in the following manner. First, the input circuit is operated for a predetermined clock number given from the pattern data, and predetermined data signals are outputted from the variable delay device 10 . The outputted data signals are held in the first and second F/F 1 and 2 . The clock signals whose number is equal to a predetermined pattern number are generated, and this generation is then terminated (stopped). The data signals held in the first and second F/F 1 and 2 are respectively read through the first and second AND circuits 11 and 12 based on the read mode signals. Then, the read values and the data signals are compared, and it is judged whether they have been correctly fetched into the first and second F/F 1 and 2 , respectively. If the data signals have not been correctly fetched, a delay time is increased in increments of a predetermined time, and the judgment is repeated. Then, a skew is corrected, and a delay time with which the data signals can be correctly fetched is set. As described above, in the conventional input circuit, since the first and second F/F 1 and 2 judge whether the data signals have been correctly fetched, the clock signals as pattern data must be generated every time the setting of the delay time is changed and judged. Therefore, besides the circuit configuration shown in FIG. 4 , a program, a circuit or the like which generates and controls the clock signals as the pattern data is required. In view of the above-described problems, it is, therefore, an object of the present invention to provide a phase correction circuit capable of easily detecting a skew between a data signal and a clock signal and correcting their phase difference without requiring a clock signal as pattern data upon initialization. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG. 1 is a circuit diagram illustrating a structure of a preferred embodiment of a phase correction circuit according to the present invention; FIG. 2 is a timing chart illustrating an operation of the preferred embodiment of the phase correction circuit according to the present invention; FIGS. 3 (A) and (B) are graphs illustrating an optimum delay time; and FIG. 4 is a circuit diagram of an input circuit according to a prior art. detailed-description description="Detailed Description" end="lead"? |
Washing and chilling apparatus and method |
Sterically stabilized cationic liposomes (SSCL) encapsulating a K type oligodeoxynucleotide (ODN) including a CpG motif are disclosed. These SSCL encapuslating a K type ODN can be used to effectively deliver the ODN to a cell. A novel method is also disclosed for producing the SSCL encapsulating the K type ODN. Administration of the SSCL encapsulating a K type ODN and a chemotherapeutic agent, such as a chimeric molecule comprising a targeting molecule selected from the group consisting of an IL-13, and an anti-IL-13 receptor antibody; and an effector molecule selected from the group consisting of a Pseudomonas exotoxin, a Diphtheria toxin, and a radionuclide, can be used to dramatically reduce the growth of solid tumors. |
1. A sterically stabilized cationic liposome composition, comprising a cationic lipid, a co-lipid, a stabilizing agent and encapsulating an oligodeoxynucleotide of at least ten nucleotides in length comprising a cpg motif, wherein the oligodexoynucleotide comprising a CpG motif comprises a sequence represented by the formula 5′ N1N2N3Q-CpG-WN4N5N6 3′ (SEQ ID NO: 97), wherein Q is a T, G, or A, W is A or T, and N1, N2, N3, N4, N5, and N6 are any nucleotide, and wherein the lipid:co-lipid ratio is from about 3:7 to about 7:3, and wherein stabilizing agent comprises about 1 percent to about 5 percent of the co-lipid composition: 2. The sterically stabilized cationic liposome composition of claim 1, wherein the lipid is a cholesterol. 3. The sterically stabilized cationic liposome composition of claim 1, wherein the cholesterol is dimethyaminoethane-carbamol-cholesterol. 4. The sterically stabilized cationic liposome composition of claim 1, wherein the co-lipid is a phosphitidyl ethanolamine. 5. The sterically stabilized cationic liposome composition of claim 4, wherein the phosphitidyl ethanolamine is dioleyl phosphatidyl ethanolamine. 6. The sterically stabilized cationic liposome composition of claim 1, wherein the stabilizing agent is a polyethylene glycol. 7. The sterically stabilized cationic liposome composition of claim 6, wherein the polyethylene glycol is polyethylene glycol-phosphatidyl ethanolamine. 8. The sterically stabilized cationic liposome composition of claim 3, wherein the co-lipid is a dioleyl phosphitidyl ethanolamine, and the stabilizing agent is polyethylene glycol-phosphatidyl ethanolamine. 9. The sterically stabilized cationic liposome composition of claim 8, wherein the respective molar ratios of dimethyaminoethane-carbamol-cholesterol, dioleyl phosphitidyl choline are present as a molar ration of about 4:6. 10. The sterically stabilized cationic liposome composition of claim 8, wherein the respective molar ratios of dimethyaminoethane-carbamol-cholesterol, dioleyl phosphitidyl ethanolamine, and polyethyleneglycol-phosphatidyl ethanolamine are present as a molar ratio of about 4:6:0.06. 11. The sterically stabilized cationic liposome composition of claim 1, further comprising an antigen. 12. The sterically stabilized cationic liposome composition of claim 10, wherein the antigen is selected from the group consisting of a protein, a polypeptide, or a polysaccharide. 13. The composition according to claim 9, wherein the sterically stabilized cationic liposome further comprises a targeting molecule. 14. A pharmaceutical composition comprising a therapeutically effective amount of the sterically stabilized liposome composition of claim 1 in a pharmaceutically acceptable carrier. 15. A method for stimulating an immune response in a subject, comprising administering the subject a therapeutically effective amount of the sterically stabilized cationic liposome composition of claim 1, thereby stimulating the immune response. 16. The method of claim 15, wherein the immune response is expression of a cytokine. 17. The method of claim 15, further comprising administering an antigen to the subject, thereby stimulating an antigen-specific immune response. 18. The method of claim 15, wherein the subject has a tumor and wherein the immune response is an immunotherapeutic response against the tumor. 19. A method of stimulating an immune cell, comprising contacting the immune cell with the sterically stabilized cationic liposome composition of claim 1, thereby stimulating the immune cell. 20. A method of stimulating an immune cell, comprising contacting the immune cell with the sterically stabilized cationic liposome composition of claim 9, thereby stimulating the immune cell. 21. The method of claim 19, wherein the cytokine is interferon gamma. 22. The method of claim 19, wherein the immune cell is in vitro. 23. The method of claim 19, wherein the immune cell is in vivo. 24. A method for inducing an immune response in a subject, comprising contacting immune cells in vitro with a sterically stabilized cationic liposome encapsulating an oligodeoxynucleotide of at least ten nucleotides in length comprising a CpG motif, wherein the oligodexoynucleotide comprising a CpG motif comprises a sequence represented by the formula 5′ N1N2N3Q-CpG-WN4N5N6 3′ (SEQ ID NO: 97), wherein Q is a T, G, or A, W is A or T, and N1, N2, N3, N4, N5, and N6 are any nucleotide comprising a oligodeoxy; contacting the immune cells with an antigen for a time sufficient to generate antigen specific immune cells, and administering said antigen specific immune cells to the subject in an amount sufficient to induce an immune response. 25. The method of claim 24, further comprising administering an antigen to the subject. 26. A method of inducing an immune response against an infectious agent, comprising administering the oligonucleotide of claim 1 to a subject infected with the infectious agent, thereby inducing an immune response against the infectious agent. 27. The method of claim 25, wherein the infectious agent is a virus. 28. The method of claim 25, wherein the infectious agent is a fungus, bacteria, or a virus. 29. The method of claim 25, further comprising administering an anti-infectious agent. 30. The method of claim 25, wherein the anti-infectious agent is an antibiotic, an antiviral, or an anti-fungal agent. 31. A method for producing a sterically stabilized cationic liposome encapsulating an agent of interest, comprising contacting a unilamellar vesicle with an agent of interest; dehydrating the unilamellar vesicle and the agent of interest; rehydrating the unilamellar vesicle and the agent of interest; thereby producing the sterically stabilized cationic liposome encapsulating the agent of interest. 32. The method of claim 31, further comprising extruding the sterically stabilized cationic liposome encapsulating the agent of interest to produce a liposome of a specified size. 33. The method of claim 32, wherein the specified size is about 150 micrometers or less. 34. The method of claim 31, wherein the agent of interest is a oligodeoxynucleotide of at least ten nucleotides in length and comprises a CpG motif. 35. The method of claim 34, wherein the oligodexoynucleotide comprising a CpG motif comprises a sequence represented by the formula 5′ N1N2N3Q-CpG-WN4N5N6 3′ (SEQ ID NO: 97), wherein Q is a T, G, or A, W is A or T, and N1, N2, N3, N4, N5, and N6 are any nucleotide. 36. The method of claim 31, wherein the unilamellar vesicle comprises a cationic lipid, a co-lipid, a stabilizing agent. 37. The method of claim 36, wherein the lipid is a cholesterol. 38. The method of claim 37, wherein the cholesterol is dimethyaminoethane-carbamol-cholesterol. 39. The method of claim 36, wherein the co-lipid is a phosphitidyl ethanolamine. 40. The method of claim 39, wherein the phosphitidyl ethanolamine is dioleyl phosphatidyl ethanolamine. 41. The method of claim 36, wherein the stabilizing agent is a polyethylene glycol. 42. The method of claim 41, wherein the polyethylene glycol is polyethylene glycol-phosphatidyl ethanolamine. 43. The method of claim 38, wherein the co-lipid is a dioleyl phosphitidyl ethanolamine, and the stabilizing agent is polyethylene glycol-phosphatidyl ethanolamine. 44. The method of claim 43, wherein the respective molar ratios of dimethyaminoethane-carbamol-cholesterol, dioleyl phosphitidyl choline are present as a molar ratio of about 4:6. 45. The method of 43, wherein the respective molar ratios of dimethyaminoethane-carbamol-cholesterol, dioleyl phosphitidyl choline, and glycol-phosphatidyl ethanolamine are present as a molar ratio of about 4:6:0.06. 46. A method for impairing growth of a solid tumor cell bearing an IL-13 receptor in a subject, comprising administering to the subject a therapeutically effective amount of a chimeric molecule comprising a targeting molecule selected from the group consisting of an IL-13, and an anti-IL-13 receptor antibody; and an effector molecule selected from the group consisting of a Pseudomonas exotoxin, a Diphtheria toxin, and a radionuclide, wherein the effector molecule is covalently linked to the targeting molecule or is linked to the targeting molecule by a linker; and administering a therapeutically effective amount of the composition of claim 1, thereby impairing the growth of the solid tumor. 47. The method of claim 46, wherein the targeting molecule is IL-13. 48. The method of claim 46, wherein the effector molecule is a Pseudomonas exotoxin, a chemotherapeutic agent, or a Diphtheria toxin. 49. The method of claim 46, wherein chimeric molecule is a single-chain fusion protein. 50. The method of claim 46, wherein the effector molecule is a Pseudomonas exotoxin. 51. The method of claim 50, wherein said Pseudomonas exotoxin is PE38QQR, PE38, PE4E, or PE38KDEL. 52. The method of claim 46, wherein the tumor cell is a human tumor cell. 53. The method of claim 46, wherein the chimeric molecule is administered intravenously, into a body cavity, or into a lumen or an organ. |
<SOH> BACKGROUND <EOH>Bacterial DNA contains unmethylated “CpG motifs” that strongly stimulate the mammalian immune system. Synthetic oligodeoxynucleotides (ODN) expressing CpG motifs patterned after those found in bacteria trigger cells of the immune system to proliferate, mature, and produce cytokines, chemokines and immnunoglobulin (Ig) Krieg et al., Nature 374:546, 1995; Yamamoto et al., J. Immunol. 148:407, 1992; Klinman et al., Proc. Natl. Acad. Sci. USA 93:2879, 1996; Takeshita et al., Cell Immunology 206:101, 2000). These immunostimulatory activities are being harnessed therapeutically. CpG ODN show promise as immune adjuvants, significantly improving the immune response to co-administered antigens (Roman et al., Nature Medicine 3:849, 1997; Davis et al., J. Immunol. 160:870, 1998; Chu et al., J. Exp. Med. 186:1623, 1997; Klinman et al., Springer Semin Immunopathol 22:173, 2000; Tighe et al., J Allergy Clin Immunol 106:124, 2000). The strong Th1 response elicited by CpG ODN down-regulates Th2 mediated IgE and cytokine production, thereby interfering with allergic asthma (Sur et al., J. Immunol. 162:6284, 1999; Broide et al., J. Immunol. 161:7054, 1998; Kline et al., J. Immunol. 160:2555, 1998). Finally, by pre-activating the innate immune system, CpG ODN can protect naive animals against a variety of microbial and parasitic pathogens (Krieg et al., J. Immunol. 161:2428, 1998; Elkins et al., J. Immunol. 162:2291, 1999; Klinman, Antisense awed Nuc Acid Drug Dev 8:181, 1998; Klinman et al., Infect Immun 67:5658, 1999; Klinman et al., Immunity 11:123, 1999). Prolonging the bioavailability and duration of action of CpG ODN may improve their therapeutic efficacy. Unfortunately, phosphorothioate CpG ODN used in vivo are rapidly eliminated from the circulation due to adsorption onto serum proteins and degradation by serum nucleases (Litzinger et al., Biochim. Biophys. Acta 1281:139, 1996; Soni et al., Hepatology 28:1402, 1998; Gregoriadis, Pharm. Res 15:661, 1998). One potential method for protecting CpG ODN from degradation while increasing their uptake by cells of the immune system involves liposome encapsulation (MacDonald et al., Biochim. Biophys. Acta 1061:297, 1991; Takeshita et al., Eur. J. Immunol. 30:108, 2000). |
<SOH> SUMMARY <EOH>As disclosed herein, sterically stabilized cationic liposomes (SSCL) encapsulating a K type oligodeoxynucleotide (ODN) including a CpG motif can be used to effectively deliver the ODN to a cell. A novel method is disclosed for producing SSCL that encapsulate the K type ODN. When combined with other targeted biological agents, such as a chimeric molecule comprising a targeting molecule selected from the group consisting of an IL-13, and an anti-IL-13 receptor antibody; and an effector molecule selected from the group consisting of a Pseudomonas exotoxin, a Diphtheria toxin, a chemotherapeutic agent, and a radionucleotide, the SSCL encapsulating a K type ODN act synergistically to dramatically reduce the growth of solid tumors. A sterically stabilized cationic liposome (SSCL) composition is disclosed herein that includes a cationic lipid, a co-lipid, and a stabilizing agent. The SSCL encapsulates an oligodeoxynucleotide of at least ten nucleotides in length comprising a CpG motif, wherein the oligodexoynucleotide comprising a CpG motif comprises a sequence represented by the formula 5′ N 1 N 2 N 3 Q-CpG-WN 4 N 5 N 6 3′, wherein Q is a T, G, or A, W is A or T, and N 1 , N 2 , N 3 , N 4 , N 5 , and N 6 are any nucleotide, and wherein the lipid:co-lipid ratio is from about 3:7 to about 7:3, and wherein stabilizing agent comprises about 1 percent to about 5 percent of the co-lipid composition. Methods for using these SSCL encapsulating a K type ODN to stimulate a cell of the immune system are also disclosed. In one embodiment, a method is disclosed for inducing an immune response in a subject. The method includes contacting immune cells in vitro with a sterically stabilized cationic liposome encapsulating an oligodeoxynucleotide of at least ten nucleotides in length comprising a CpG motif, wherein the oligodexoynucleotide comprising a CpG motif comprises a sequence represented by the formula 5′ N 1 N 2 N 3 Q-CpG-WN 4 N 5 N 6 3′, wherein Q is a T, G, or A, W is A or T, and N 1 , N 2 , N 3 , N 4 , N 5 , and N 6 are any nucleotide; and contacting the immune cells with an antigen for a time sufficient to generate antigen specific immune cells. These antigen specific immune cells are administered to the subject in an amount sufficient to induce an immune response. In another embodiment, a method is disclosed for producing a SSCL encapsulating an agent of interest. The method includes contacting a unilamellar vesicle with an agent of interest, dehydrating the unilamellar vesicle and the agent of interest; and rehydrating the unilamellar vesicle and the agent of interest to produce the sterically stabilized cationic liposome encapsulating the agent of interest. In one embodiment, the agent of interest is a K type ODN. In a further embodiment, a method is disclosed for impairing growth of a solid tumor cell bearing an IL-13 receptor in a subject. The method includes administering to the subject a therapeutically effective amount of a recombinant chimeric molecule comprising a targeting molecule selected from the group consisting of an IL-13, and an anti-IL-13 receptor antibody; and an effector molecule selected from the group consisting of a Pseudomonas exotoxin, a Diphtheria toxin, a chemotherapeutic agent and a radionucleotide, wherein the effector molecule is genetically fused or covalently linked to the targeting molecule or is linked to the targeting molecule by a linker. The method also includes administering to the subject a therapeutically effective amount of an SSCL encapsulating a K type ODN. The foregoing and other features and advantages will become more apparent from the following detailed description of several embodiments, which proceeds with reference to the accompanying figures. |
Actuator unit for an electro-hydraulic brake system |
An actuator unit for an electro-hydraulic brake system of the ‘brake-by-wire’ type, the first and second pistons thereof being preloaded by a resetting spring each in opposition to the actuating direction. The first pressure chamber of the tandem master cylinder is connected to a hydraulic chamber limited by a simulator element. On the other hand, the simulator element defines a simulator chamber that accommodates a simulator spring and is connected to a pressure fluid reservoir. A valve device is provided to open or close the hydraulic connection between simulator chamber and pressure fluid reservoir. To increase the reliability in operation of a hydraulic fallback mode, at least one channel portion through the pressure chamber, closed at the edge at least in areas, is provided for the purposeful removal of pressure fluid contaminants by means of a compulsory pressure fluid scavenging operation. |
1-11. (canceled) 12. Actuator unit comprising: a first piston operable by means of an actuating pedal and preloaded by a first resetting spring, and a second piston preloaded by a second resetting spring, wherein said first and second pistons are arranged in tandem in a housing, one ore more limiting pressure chambers which are in connection to a non-pressurized pressure fluid reservoir by way of reservoir ports and are closably connected to brake circuits by way of outlets and valve devices associated with them, wherein the housing includes at least one channel portion that is closed at least in areas and extends through said one ore more limiting pressure chambers, for removing pressure fluid contaminants by means of a pressure fluid scavenging operation. 13. Actuator unit as claimed in claim 12, wherein the channel portion is arranged between the reservoir port of the first or the second pressure chamber and the respective outlet. 14. Actuator unit as claimed in claim 12, wherein a channel portion is formed by a piston and at least one other component added to the piston or abutting thereon. 15. Actuator unit as claimed in claim 14, wherein at least one of the first or second pistons includes a spring plate extending beyond a central valve and limiting along with piston said channel portion. 16. Actuator unit as claimed in claim 12, wherein said one or more limiting pressure chambers includes at least one axially extending channel portion and a substantially radially extending channel portion. 17. Actuator unit as claimed in claim 12, wherein the channel portion associated with the first pressure chamber there is comprised of a first tubular sleeve arranged centrically to the first piston and a second tubular sleeve that is arranged concentrically to the first sleeve and axially displaceable relative to the first sleeve. 18. Actuator unit as claimed in claim 17, wherein the first sleeve is fixed to the first piston and includes a stop for the second sleeve that is preloaded elastically to the first sleeve by means of resetting spring. 19. Actuator unit as claimed in claim 17, wherein the channel portion formed by the first and second sleeves is in alignment with a central valve that is arranged so that it is movably guided on a piston side. 20. Actuator unit as claimed in claim 17, wherein the second sleeve includes an end face for abutment on the second piston, and wherein the end face includes at least one groove pointing to the direction of the outlet for the pressure fluid scavenging operation. 21. Actuator unit as claimed in claim 20, wherein the direction the groove extends is essentially at a right angle relative to the channel portion formed by the first and second sleeves. 22. Actuator unit as claimed in claim 12, wherein a channel portion formed of telescopically movable sleeves associated with the pressure chamber, wherein said telescopically moveable sleeves are provided at the piston while sleeve is fixed to the housing and opens into said housing channel. |
<SOH> BACKGROUND OF THE INVENTION <EOH>DE 198 22 411 A1 discloses an actuator unit of this general type. Because a vehicle operator is decoupled from the physical generation of brake force in electro-hydraulic brake systems (EHB) and the braking demand is realized electrically ‘by wire’, a travel simulator is used reproducing the tactile pedal feedback which simulates the reaction force of a conventional brake system. In addition, the actuator unit includes means allowing a hydraulic emergency braking operation when the electronics is defective by means of direct application of the wheel brakes (so-called hydraulic fallback mode). Outside the hydraulic fallback mode a request for actuation is detected in the by-wire mode as a result of sensing e.g. an actuating travel of the brake pedal, and separating valves are closed to shut off the direct hydraulic throughgrip in the direction of the wheel brakes. While the braking pressure buildup is initiated in an electrohydraulic fashion, a reaction force is imparted to the vehicle operator due to a volume displacement into the travel simulator corresponding to the actuating movement. The hydraulic fallback mode is of utmost importance. Air that is drawn into the hydraulic circuit (due to leakages or the like) can impair or prevent the functioning of the hydraulic fallback mode because the pressure fluid when mixed with air) reacts compressibly. The minimum requirement of an emergency braking operation as mandated by law—without servo boosting—is not reached. Any air that enters the system will migrate to the highest points of the brake system. Preferred concentration points are the pressure chambers, what is due to a principally inclined mounting position of the actuator unit in the motor vehicle. The above-mentioned publication does not disclose measures that would allow eliminating or significantly reduce a malfunction that occurs in by-wire brake systems, which is caused by compressibility (or other contamination). Patent application DE 10147180.7, which is not prior-published, discloses an electronically controllable brake actuation system with means for the electronically controlled separation of contaminants contained and/or dissolved in the brake fluid. |
<SOH> BRIEF SUMMARY OF THE INVENTION <EOH>An object of the present invention is to disclose an actuator unit of the initially mentioned type that improves the availability of the hydraulic fallback mode. According to the invention, this object is achieved in that the actuator unit includes at least one channel portion that is closed at least in areas and extends through a pressure chamber, for the purpose of removing contaminants in the pressure fluid by means of a pressure fluid scavenging operation. Principally, the pressure chambers can comprise several indirectly or directly adjacent channel portions. The invention basically founds on the fundamental idea of providing a compulsory guide for the pressure fluid scavenging operation. The invention permits the purposeful removal of contaminants, in particular the entry of air or gas from the actuator unit by means of a pressure fluid scavenging operation by using small pressure fluid volume flows. Consequently, the scavenging operation at the wheel brakes brings about no or only a low impact pressure so that the friction linings are not pressed in the direction of the brake disc, or only to an insignificant extent. This feature allows executing a scavenging operation during driving and without unnecessary friction lining wear. Impairment of comfort due to the development of noise of great volume flows is avoided. Because an intense evacuation of the accumulator is prevented, the scavenging process can be executed with low energy consumption. According to a favorable embodiment of the invention, the channel portion is arranged between the reservoir port of the first or the second pressure chamber and the respective outlet. The result is that the pressure fluid flow is discharged specifically into the pressure fluid reservoir starting from the outlets on the brake circuit side through the pressure chambers and central valves associated with the pistons via the reservoir ports. Degassing/scavenging of the pressure fluid is executed there. The piston and another component added to the piston or abutting thereon can form a channel portion. For example, the channel portion can be defined by the second piston that is provided with a spring plate, extending beyond the central valve, for the second resetting spring so that the spring plate along with the second piston limits the channel portion. A change in direction is achieved when each pressure chamber includes an axially extending channel portion and a substantially radially extending channel portion according to a preferred embodiment of the invention. It is advantageous that for forming the channel portion associated with the first pressure chamber there is provision of a first tubular sleeve arranged centrically to the first piston and a second tubular sleeve (telescopic sleeve) that is arranged concentrically to the first sleeve and axially displaceable relative to the first sleeve. The sleeves allow simple and low-cost manufacture and permit a direct flow guide. When the first sleeve is fixed to the first piston and includes a stop for the second sleeve that is preloaded elastically to the first sleeve by means of the resetting spring, this achieves an elastically preloaded captivation of the first resetting spring on the first piston beside a directed flow guide. Preferably, the second sleeve includes an end face for abutment on the second piston, and the end face includes a groove pointing to the direction of the outlet for the pressure fluid scavenging operation. The channel portion is defined by groove surface and piston surface when these parts bear against one another in an initial position on account of the prevailing forces of the resetting springs. For rerouting of the flow, the groove is provided essentially at right angles relative to the channel portion formed by the sleeves. A further improved fluid inflow and penetration of central valves of the actuator unit is achieved when the channel portion formed by the sleeves is in alignment with a central valve, which is arranged so that it is movably guided on the piston side. In a favorable embodiment of the invention, a channel portion formed of telescopically movable sleeves is associated with the pressure chamber, one sleeve thereof being provided at the piston, while the other sleeve is arranged on the housing and opens into a housing channel. |
Resin composition, composition for solder resist, and cured article obtained therefrom |
A resin composition which is excellent in photosensitively and developability, gives a cured article excellent in flexibility, soldering heat resistance, resistance to thermal deterioration, and resistance to electroless gold plating, and is suitable especially for use in forming solder resists and interlayer dielectrics. The resin composition is characterized by containing an oligomer (A) obtained by reacting the following ingredients (a) to (d): an epoxy resin having at least two epoxy groups per molecule, a compound having two hydroxy groups and one carboxy group per molecule, a carboxy group per molecule, a carboxylated rubbery polymer, and a monobasic acid having an unsatuated group. |
1. A resin composition comprising: an oligomer (A) obtained by allowing the following components (a) to (d) to react with one another: (a) an epoxy resin having at least two epoxy groups per molecule; (b) a compound having at least two hydroxyl groups and one carboxyl group per molecule; (c) a carboxylated rubbery polymer; and (d) a monobasic acid having an unsaturated group. 2. A resin composition comprising: an oligomer (A) obtained by allowing the following compounds (a) to (e) to react with one another: (a) an epoxy resin having at least two epoxy groups per molecule; (b) a compound having at least two hydroxyl groups and one carboxyl group per molecule; (c) a carboxylated rubbery polymer; (d) a monobasic acid having an unsaturated group; and (e) a multibasic acid anhydride. 3. The resin composition according to claim 1 or claim 2, wherein the oligomer (A) has a weight-average molecular weight in a range of 1000 to 100,000. 4. The resin composition according to any one of claims 1 to 3, wherein the oligomer (A) has an acid value in a range of 1 to 300 mgKOH/g. 5. The resin composition according to any one of claims 1 to 4, wherein the epoxy resin (a) having at least two epoxy groups per molecule is represented by the following formula (1): in formula (1), X is —CH2— or —C(CH3)2—, m is an integer of 1 or more, and M is a hydrogen atom or a group represented by the following formula (G): (here, in the case when m is 1, M is represented by formula (G), and in the case when m is greater than 1, at least one of M is represented by formula (G) with the rest thereof being hydrogen atoms.) 6. The resin composition according to claim 5, wherein in formula (1), the rate of M being represented by formula (G) is in a range of 70 to 90 mole %. 7. The resin composition according to any one of claims 1 to 6, wherein the compound (b) having two hydroxyl groups and one carboxyl group per molecule is a dimethylolated compound of carboxylic acid having 2 to 6 carbon atoms. 8. The resin composition according to any one of claims 1 to 7, wherein the monobasic acid having an unsaturated group (d) is one kind or two or more kinds of compounds selected from the group consisting of (metha)acrylic acids and monobasic acids containing a maleimide group. 9. The resin composition according to claim 2, wherein the multibasic acid anhydride (e) is a phthalic anhydride derivative. 10. The resin composition according to any one of claims 1 to 9, further comprising a photo-polymerization initiator (C). 11. The resin composition according to any one of claims 1 to 10, further comprising a heat curing component (D). 12. The resin composition according to claim 11, wherein the heat curing component (D) is an epoxy resin. 13. The resin composition according to claim 12, further comprising a 1,4-dihydropyridine derivative as a curing accelerator agent for the epoxy resin. 14. The resin composition according to any one of claims 1 to 13, further comprising a diluent (B). 15. A composition, which contains the resin composition according to claim 14, and is used for a solder resist or an interlayer insulating layer for use in a printed circuit board. 16. A cured article comprising the composition according to any one of claims 1 to 15. 17. A product comprising a layer of the cured article according to claim 16. 18. The product according to claim 17, comprising a printed circuit board. |
<SOH> BACKGROUND ART <EOH>In a soldering process that is carried out so as to protect a wiring (circuit) pattern formed on a substrate through a screen printing method and the like from external environment and also to surface-assemble an electric part on a printed circuit board, conventionally, a protective layer, referred to as a cover coat or a solder mask, is applied on the printed circuit board so as to prevent the solder from adhering to unnecessary portions. Conventionally, with respect to solder resist ink to be used for such a purpose, a multi-functional epoxy resin-based ink has been mainly used; however, the problem with this ink is that the resulting cured film is inferior in its flexibility although it provides superior heat resistance. Therefore, the solder resist ink of this type is limited in its use to a rigid plate that does not require any flexibility (bending property) in the cured film, and it is difficult to apply this to flexible printed circuit boards (FPC) that have been widely used in recent years. Under these circumstances, recently, various resist inks having flexibility have been proposed. For example, Japanese Patent Application Laid-open No. 2-269166 has disclosed a heat-curing type solder resist ink made from polyparabanic acid, epoxy resin and a polar solvent, JP-A No. 6-41485 has disclosed a solder resist ink of a heat-drying type that uses polyparabanic acid and phenoxy resin as its essential components. However, these solder resists are formed as a resist pattern by using a screen printing process, with the result that there is a limitation in the line width and the like of the screen; thus, these fail to provide a fine image forming process that meets the current demands for high density. For this reason, in recent years, those of a photographic developing type, such as those disclosed in JP-A No. 2-172749, JP-A No. 2-173750 and JP-A No. 173751, have been proposed; however, these have not achieved sufficient flexibility. The objective of the present invention is to provide a resin composition and its cured article, and the resin composition, which is formed by using a specific oligomer (A), is superior in curability (photosensitivity), even when the used amount of a polymerization initiator is smaller than the conventional used amount, is excellent in developability even when the acid value (mgKOH/g) thereof is smaller than the conventional acid value, and provides a cured film, obtained by thermally curing the resin composition in a post curing process, which forms a coat film that is superior in flexibility, and excellent in soldering heat resistance, resistance to thermal deterioration, resistance to electroless gold plating, acid resistance and water resistance, and the resin composition is also suitably applied to an organic solvent or alkali developing type ink, in particular, a resist ink for use in flexible printed circuit boards. |
Delivery of broadcast information to a mobile station in a radio communication system |
A method (300) of delivering broadcast information to a mobile station (12) in a radio communication system (100) having a plurality of communication cells that are able to serve said mobile station (12) with a communication link. The method includes the steps of receiving a broadcast information trigger (310) caused by the mobile station; and transmitting (324), in response to receiving said trigger (310), broadcast information to the mobile station (112). |
1. A method of delivering broadcast information to a mobile station in a radio communication system having a plurality of communication cells that are able to serve said mobile station with a communication link, the method characterised by the steps of: receiving a broadcast information trigger caused by the mobile station; and transmitting, in response to receiving said trigger, broadcast information to the mobile station. 2. The method of delivering broadcast information according to claim 1, wherein the broadcast information includes serving cell and/or neighbour cell information. 3. The method of delivering broadcast information according to claim 1, the method further characterised by the step of: receiving a request from said mobile station for said broadcast information to be transmitted, wherein the request is the trigger. 4. The method of delivering broadcast information according to claim 3, wherein the mobile station requests transmission of the broadcast information in response to a deterioration in a signal condition. 5. The method of delivering broadcast information according to claim 3, wherein the mobile station transmits a request for said broadcast information after not receiving said information a pre-defined time period after moving into a cell. 6. The method of delivering broadcast information according to claim 5, wherein the pre-defined time period is variable dependent on whether said mobile station is in a call. 7. The method of delivering broadcast information according to claim 1, the method further comprising the step of: detecting when a mobile station arrives in a cell in order to initiate said trigger. 8. The method of delivering broadcast information according to claim 7, wherein the step of detecting includes the step of: detecting that said mobile station has performed a handover between a first cell and a second cell to initiate said trigger. 9. The method of delivering broadcast information according to claim 7, wherein the step of detecting includes at least one of the steps of: detecting an arrival of said mobile station on a cell; or detecting an exchange of signalling information with said mobile station, or detecting an uplink transmission from said mobile station. 10. The method of delivering broadcast information according to claim 7, wherein the step of detecting includes the step of: detecting a registration message of said mobile station. 11. The method of delivering broadcast information according to claim 2, the method further comprising the steps of: ranking neighbour cell information based on a likelihood of said mobile station selecting a particular cell to move to; and transmitting higher ranked neighbour cell information in preference to lower ranked neighbour cell information in response to the step of receiving a trigger. 12. The method of delivering broadcast information according to claim 11, the method further comprising the steps of: obtaining an indication of said mobile station's location, for example in a registration message or in determining a location from a transmission of said mobile station using a triangulation method; and ranking said neighbour cell information in response to said location. 13. The method of delivering broadcast information according to claim 1, wherein the transmission of broadcast information is only sent in response to a trigger. 14. A communication system having a plurality of communication cells that are able to serve a plurality of mobile stations with a communication link, the communication system comprising: a trigger response function for generating a broadcast information trigger, the trigger response function being responsive to at least one mobile station; and a transmitter, operably coupled to said trigger response function, to broadcast information to the at least one mobile station in response to said generated trigger. 15. The communication system according to claim 14, wherein the broadcast information includes serving cell and/or neighbour cell information. 16. The communication system according to claim 14, wherein the trigger response function is positioned in one or more of: an operations and management centre, a base transceiver station, a base station controller, a mobile switching centre. 17. A mobile station adapted to perform the method steps of claim 1. 18. A mobile station adapted to perform the method steps of claim 3. 19. The mobile station of claim 17 wherein the mobile station is a mobile phone, a portable or mobile PMR radio, a personal digital assistant, a lap-top computer or a wirelessly networked PC. 20. A storage medium storing processor-implementable instructions for controlling one or more processors to carry out the method of claim 1. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Wireless communication systems, for example cellular telephony or private mobile radio communication systems, typically provide for radio telecommunication links to be arranged between a plurality of base transceiver stations (BTSs) and a plurality of subscriber units, often termed mobile stations (MSs). The term mobile station generally includes both hand-portable and vehicular mounted ‘mobile’ radio units. The communication link from a BTS to an MS is generally referred to as a down-link communication channel. Conversely, the communication link from a MS to a BTS is generally referred to as an up-link communication channel. Wireless communication systems are distinguished over fixed communication systems, such as the public switched telephone networks (PSTN), principally in that mobile stations move between service providers (and/or different BTS) and in doing so encounter varying radio propagation environments. In a wireless communication system, each BTS has associated with it a particular geographical coverage area (or cell). The coverage area defines a particular range that the BTS can maintain acceptable communications with MSs operating in its serving cell. Often these cells combine to produce an expanded system coverage area, with the infrastructure supporting respective cells interconnected via centralised switching equipment. Furthermore, cells are often grouped into location areas for the purposes of tracking a MS within the coverage area, whilst minimising location-updating signalling. Mobile cellular and trunked radio systems typically allocate a broadcast information radio channel that is used to provide general system information to all mobile stations within coverage area of the system. Within such broadcast channels, each of the base transceiver stations broadcast information pertinent to the cell that they serve, as well as information about neighbouring cells. The serving and neighbour cell information is then used by mobile stations receiving the information to select the optimum cell for access to the system. Radio frequency (RF) resources, such as bandwidth, are generally limited. Transmission of broadcast information on a signalling channel, clearly takes up bandwidth that could be used for other control channel signalling and user traffic. As the broadcast channel is generally allocated a limited resource, it is desirable to optimise the use of that resource, i.e. bandwidth, available for broadcast information. It is known that neighbour cell information is sent periodically by each base transceiver station on the main control channel (MCCH) and may also be sent on the associated signalling channel (ACCH) on a trunked radio communication system. The MCCH is monitored by MSs whilst in idle mode, and the ACCH is monitored when MSs are communicating on assigned traffic channels in a call. Generally the base tranceiver station sends information about all of its neighbour cells in one or more downlink messages as a background low-priority task, i.e. when the BTS has little other information to transmit. Such a periodic broadcast transmission strategy can result in MSs not receiving information about its neighbour cells in an acceptable time period. This is particularly the case for those MSs involved in a call on the traffic channel where broadcast channel bandwidth is particularly scarce. Such delays have been known to result in dropped calls or link failure. Furthermore, such an unregulated broadcast transmission strategy can also result in transmitting broadcast information at times when no MSs actually need the information. Thus, this strategy wastes valuable bandwidth that could have been better used for control signalling or user traffic. In summary, the inventors of the present invention have recognised that the known prior art in the field of private mobile radio systems uses periodic delivery of neighbour cell information without regard to whether a particular MS is present or whether the MS wishes to receive the information. A need therefore exists for delivery of broadcast information, particularly broadcast neighbour cell information, to MSs in radio communication systems wherein the abovementioned disadvantages may be alleviated. Published US patent U.S. Pat. No. 5,289,525 describes a cellular radiotelephone network of the prior art. Hand-over of a call is described. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>Exemplary embodiments of the present invention will now be described, with reference to the accompanying drawings, in which: FIG. 1 shows a block diagram of a trunked radio communication system adapted to support the various inventive concepts of a preferred embodiment of the present invention; FIG. 2 shows a block diagram of a radio communication unit adapted to support the various inventive concepts of the preferred embodiment of the present invention; and FIG. 3 shows a flowchart depicting a delivery mechanism for delivering neighbour cell information in the communication system, in accordance with the preferred embodiment of the present invention. detailed-description description="Detailed Description" end="lead"? |
Novel g-protein coupled receptor and dna sequences thereof |
This invention relates to newly identified polypeptides and polynucleotides encoding such polypeptides, to their use in diagnosis and in identifying compounds that may be agonists, antagonists that are potentially useful in therapy, and to production of such polypeptides and polynucleotides, belonging to the class of G-protein coupled receptors. |
1. An isolated mGRR polypeptide selected from one of the groups consisting of: (a) an isolated mGRR polypeptide encoded by a polynucleotide comprising the sequence of SEQ ID NO: 3 or SEQ ID NO: 5; (b) an isolated mGRR polypeptide comprising a polypeptide sequence having at least 96% identity to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which shows properties in the ligand binding assay similar to those of mGRR; (c) an isolated mGRR1b or mGRR2 polypeptide comprising the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6; (d) an isolated mGRR polypeptide having at least 96% identity to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which shows properties in the ligand binding assay similar to those of mGRR; (e) the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6; (f) an isolated mGRR polypeptide having or comprising a polypeptide sequence that has an Identity Index of 0.96 compared to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which shows properties in the ligand binding assay similar to those of mGRR; or (g) a fragment or variant of such a polypeptide according to (a) to (f). 2. The isolated polypeptide as claimed in claim 1 which is the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6. 3. An isolated mGRR polynucleotide selected from one of the groups consisting of: (a) an isolated mGRR polynucleotide comprising a polynucleotide sequence having at least 80% identity to the polynucleotide sequence of SEQ ID NO: 3 or SEQ ID NO: 5 and which encodes for polypeptides which show properties in the ligand binding assay similar to mGRR; (b) an isolated polynucleotide comprising the polynucleotide of SEQ ID NO: 3 or SEQ ID NO: 5; (c) an isolated polynucleotide comprising at least 96% identity to the polynucleotide of SEQ ID NO: 3 or SEQ ID NO: 5 and which encodes for polypeptides which show properties in the ligand binding assay similar to mGRR; (d) the isolated polynucleotide of SEQ ID NO: 3 or SEQ ID NO: 5; (e) an isolated polynucleotide comprising a polynucleotide sequence encoding a polypeptide sequence having at least 96% identity to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which encodes for polypeptides which show properties in the ligand binding assay similar to mGRR; (f) an isolated polynucleotide comprising a polynucleotide sequence encoding the polypeptide of SEQ ID NO: 3 or SEQ ID NO: 5; (g) an isolated polynucleotide comprising a polynucleotide sequence encoding a polypeptide sequence having at least 96% identity to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which encodes for polypeptides which show properties in the ligand binding assay similar to mGRR; (h) an isolated polynucleotide encoding the polypeptide of SEQ ID NO: 4 or SEQ ID NO: 6; (i) an isolated polynucleotide comprising a polynucleotide sequence that has an Identity Index of 0.96 compared to the polynucleotide sequence of SEQ ID NO: 3 or SEQ ID NO: 5 and which encodes for polypeptides which show properties in the ligand binding assay similar to mGRR; or k) an isolated polynucleotide comprising a polynucleotide sequence encoding a polypeptide sequence that has an Identity Index of 0.96 compared to the polypeptide sequence of SEQ ID NO: 4 or SED ID NO: 6 and which encode for polypeptides which show properties in the ligand binding assay similar to mGRR; or polynucleotides that are fragments or variants of the above mentioned polynucleotides or that are complementary to above mentioned polynucleotides, over the entire length thereof. 4. An isolated polynucleotide as claimed in claim 3 selected from the group consisting of: (a) an isolated polynucleotide comprising the polynucleotide of SEQ ID NO: 3 or SEQ ID NO: 5; (b) the isolated polynucleotide of SEQ ID NO: 3 or SEQ ID NO: 5; (c) an isolated polynucleotide comprising a polynucleotide sequence encoding the polypeptide of SEQ ID NO: 4 or SEQ ID NO: 6; and (d) an isolated polynucleotide encoding the polypeptide of SEQ ID NO: 4 or SEQ ID NO: 6. 5. An expression system comprising a polynucleotide capable of producing a polypeptide of claim 1 when said expression vector is present in a compatible host cell. 6. A recombinant host cell comprising the expression vector of claim 5 or a membrane thereof expressing the polypeptide selected from one of the groups consisting of: (a) an isolated mGRR polypeptide encoded by a polynucleotide comprising the sequence of SEQ ID NO: 3 or SEQ ID NO: 5; (b) an isolated mGRR polypeptide comprising a polypeptide sequence having at least 96% identity to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which shows properties in the ligand binding assay similar to those of mGRR; (c) an isolated mGRR1b or mGRR2 polypeptide comprising the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6; (d) an isolated mGRR polypeptide having at least 96% identity to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which shows properties in the ligand binding assay similar to those of mGRR; (e) the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6; (f) an isolated mGRR polypeptide having or comprising a polypeptide sequence that has an Identity Index of 0.96 compared to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which shows properties in the ligand binding assay similar to those of mGRR; or (g) a fragment or variant of such a polypeptide according to (a) to (f). 7. A process for producing a polypeptide selected from one of the groups consisting of: (a) an isolated mGRR polypeptide encoded by a polynucleotide comprising the sequence of SEQ ID NO: 3 or SEQ ID NO: 5; (b) an isolated mGRR polypeptide comprising a polypeptide sequence having at least 96% identity to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which shows properties in the ligand binding assay similar to those of mGRR; (c) an isolated mGRR1b or mGRR2 polypeptide comprising the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6; (d) an isolated mGRR polypeptide having at least 96% identity to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which shows properties in the ligand binding assay similar to those of mGRR; (e) the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6; (f) an isolated mGRR polypeptide having or comprising a polypeptide sequence that has an Identity Index of 0.96 compared to the polypeptide sequence of SEQ ID NO: 4 or SEQ ID NO: 6 and which shows properties in the ligand binding assay similar to those of mGRR; or (g) a fragment or variant of such a polypeptide according to (a) to (f); comprising the step of culturing a host cell as defined in claim 6 under conditions sufficient for the production of said polypeptide and recovering the polypeptide from the culture medium. 8. A fusion protein consisting of the immunoglobulin Fc-region and any one polypeptide of claim 1. 9. An antibody immunospecific for the polypeptide of claim 1. 10. A method of screening to identify compounds that stimulate or inhibit the function or level of the polypeptide of claim 1, comprising a method selected from the group consisting of: (a) measuring or, detecting, quantitatively or qualitatively, the binding of a candidate compound to the polypeptide (or to the cells or membranes expressing the polypeptide) or a fusion protein thereof by means of a label directly or indirectly associated with the candidate compound; (b) measuring the competition of binding of a candidate compound to the polypeptide (or to the cells or membranes expressing the polypeptide) or a fusion protein thereof in the presence of a labelled competitor; (c) testing whether the candidate compound results in a signal generated by activation or inhibition of the polypeptide, using detection systems appropriate to the cells or cell membranes expressing the polypeptide; (d) mixing a candidate compound with a solution containing a polypeptide of claim 1, to form a mixture, measuring activity of the polypeptide in the mixture, and comparing the activity of the mixture to a control mixture which contains no candidate compound; or (e) detecting the effect of a candidate compound on the production of mRNA encoding said polypeptide or said polypeptide in cells, using for instance, an ELISA assay, and (f) producing said compound according to biotechnological or chemical standard techniques. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The drug discovery process is currently undergoing a fundamental revolution as it embraces “functional genomics”, that is, high throughput genome- or gene-based biology. This approach as a means to identify genes and gene products as therapeutic targets is rapidly superseding earlier approaches based on “positional cloning”. A phenotype, that is a biological function or genetic disease, would be identified and this would then be tracked back to the responsible gene, based on its genetic map position. Functional genomics relies heavily on high-throughput DNA sequencing technologies and the various tools of bioinformatics to identify gene sequences of potential interest from the many molecular biology databases now available. There is a continuing need to identify and characterise further genes and their related polypeptides/proteins, as targets for drug discovery. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention relates to purified m etabotropic g lutamate r eceptor r elated membrane receptor proteins of human origin referred to herein as human mGRR1a, mGRR1b and mGRR2 (all three are herein referred to as mGRR). The similarity of mGRR to metabotropic glutamate receptors suggests that these receptors also belong to family 3 of the GPCR. This receptor family comprises metabotropic glutamate receptors, GABA-B receptors, the Ca-sensing receptor, putative taste receptors and a family of olfactory vomeronasal receptors. The closest mammalian GPCR homologue found to the polypeptides of the invention in public domain databases is the metabotropic glutamate receptor type 3 (accession Q11923) with 23% identical and 43% similar amino acid residues to the polypeptide of mGRR1a described in SEQ ID NO: 2. Such polypeptides and polynucleotides are of interest in relation to methods of treatment of certain diseases, including, but not limited to the treatment of disorders associated with the central and peripheral nervous systems. In particular, mGRR receptor agonists or antagonists can e.g. be useful in treating neurological and/or psychiatric diseases, including but not limited to dementia, schizophrenia, depression, affective disorders, epilepsy, and motoric disorders, hereinafter referred to as “diseases of the invention”. In a further aspect, the invention relates to methods for identifying agonists and antagonists to mGRR (e.g., inhibitors) using the materials provided by the invention, and treating conditions associated with imbalance of such identified compounds. In still a further aspect, the invention relates to diagnostic assays for detecting diseases associated with inappropriate mGRR activity or levels. detailed-description description="Detailed Description" end="lead"? |
Processor system, task control method on computer system, computer program |
A mechanism for recording a timing in which a high urgency process is started is provided, and upon entry to a critical section in the middle of a low urgency process, by referencing the record, it is inspected whether a high urgency process will be started during execution of the critical section. If it will not be started, the critical section is entered, and if it will be started, control is exerted so that entry to the critical section is postponed until the high urgency process is completed. Exclusive access control in a critical section can be performed suitably under conditions where a plurality of task execution environments exist. |
1. (Cancelled) 2. (Cancelled) 3. A processing system in which a plurality of task execution environments, whose scheduling is performed according to distinct policies, exist, the processing system characterized in that it comprises: execution environment switching means for switching task execution environments; scheduled time managing means for managing a scheduled time at which a switching of task execution environments is to be performed next; and task execution managing means for managing execution of a task under a current task execution environment in accordance with said scheduled time. 4. The processing system according to claim 3 characterized in that said task execution managing means, upon entry, by a task under execution under the current task execution environment, into a critical section that cannot be referenced simultaneously by a plurality of tasks, determines whether or not to enter said critical section depending on whether or not there is room for the execution time of said critical section with respect to said scheduled time. 5. The processing system according to claim 4 characterized in that said task execution managing means allows entry to said critical section when there is room for the execution time of said critical section with respect to said scheduled time, but instructs switching of task execution environments to said execution environment switching means when there is no room for the execution time of said critical section. 6. The processing system according to claim 3 characterized by further comprising context saving means for saving, upon the switching of task execution environments by said execution environment switching means, a state of task execution at the time of switching. 7. A method of task control on a computer system in which a plurality of task execution environments, whose scheduling is performed according to distinct policies, exist, the method of task control on a computer system characterized in that it comprises: an execution environment switching step for switching task execution environments; a scheduled time managing step for managing a scheduled time at which a switching of task execution environments is to be performed next; and a task execution managing step for managing execution of a task under a current task execution environment in accordance with said scheduled time. 8. The method of task control on a computer system according to claim 7 characterized in that in said task execution managing step, upon entry, by a task under execution under the current task execution environment, into a critical section that cannot be referenced simultaneously by a plurality of tasks, it is determined whether or not to enter said critical section depending on whether or not there is room for the execution time of said critical section with respect to said scheduled time. 9. The method of task control on a computer system according to claim 8 characterized in that in said task execution managing step, entry to said critical section is allowed when there is room for the execution time of said critical section with respect to said scheduled time, but switching of task execution environments is instructed to said execution environment switching step when there is no room for the execution time of said critical section. 10. The method of task control on a computer system according to claim 7 characterized in that it further comprises a context saving step for saving, upon the switching of task execution environments by said execution environment switching step, a state of task execution at the time of switching. 11. (Cancelled) 12. A computer program in which a procedure of task control on a computer system, in which a plurality of task execution environments whose scheduling is performed according to distinct policies exist, is written in a computer readable format, the computer program characterized in that it comprises: an execution environment switching step for switching task execution environments; a scheduled time managing step for managing a scheduled time at which a switching of task execution environments is to be performed next; and a task execution managing step for managing execution of a task under a current task execution environment in accordance with said scheduled time. |
<SOH> BACKGROUND ART <EOH>Along with the innovative advances in modern LSI (Large Scale Integration) technology, various types of information processing devices and information communication devices have been developed and sold, and have come to permeate our daily lives. With these types of devices, a variety of processing services are provided by having predetermined program code executed by a CPU (Central Processing Unit) or some other processor under an execution environment provided by an operating system. However, in program design, it is sometimes useful to let a plurality of flows of control (sometimes referred to as “tasks”) to exist in a program. As used herein, the term a plurality of flows of control refers to the fact that, as shown in FIG. 6 , a plurality of “points currently under execution” exist in a program's process flow, or in other words in the flowchart. In the example shown in the same figure, at a certain point T 1 , step S 1 and step S 3 are executed in flow I and flow II, respectively. Then, as opposed thereto, at the next point T 2 after some time has passed, step S 2 and step S 3 are executed in flow I and flow II, respectively. In general, when a plurality of flows exist, and each of them manipulates data common to each, the consistency of data cannot be maintained unless they are synchronized. As used herein, common data includes task lists, conditional variables and the like. Conditional variable is a concept in which conditions a task has are made abstract, and is used as a means for communicating when a task should transition to a waiting state or when a task should return to an executable state. For example, in a case where two flows of control B and C exist, a case where each flow of control performs the following process will be considered. Procedure 1: Read the value of variable x Procedure 2: Substitute into variable x a value in which 1 is added to the read value When the two flows B and C each perform the process above once, the same process ends up being performed twice. Therefore, the value of variable x should increase by two. However, when flow B and flow C coincide as follows, the value of variable x only increases by 1. (1) Flow B executes procedure 1 (2) Flow C executes procedure 1 (3) Flow B executes procedure 2 (4) Flow C executes procedure 2 In order to prevent such an operational error, there is a need to prohibit the referencing and updating of data from other flows during a sequence of referencing and updating operations (transactions) performed in a certain flow. In the example above, because flow C referenced and updated variable x before the sequence of operations in flow B, namely procedure 1 and procedure 2, was completed, a problem in which the consistency of data was lost occurred. Such operations as procedures 1 to 2 described above could also be considered parts of a program which cannot be referenced simultaneously by a plurality of tasks, and will hereinafter be referred to as “critical sections.” Also, prohibiting the referencing and updating of data by other tasks in order to solve the problem of data consistency in critical sections can also be called “exclusive access control.” In other words, while a series of processes is performed with respect to some data in a flow of control, the act of operating on the same data by another flow of control is delayed, that is, operation of particular data is performed exclusively. The present inventors consider it preferable that an exclusive access control mechanism have the following features. (1) There is no possibility of having a high urgency process delayed by a low urgency process (of an occurrence of a reversal in priority). (2) Exclusive access control can be performed even between a plurality of task sets whose scheduling is performed in accordance with distinct policies. Of the features above, the reason (1) is necessary is obvious to those skilled in the art. Also, (2) is necessary in simultaneously running a plurality of operating systems on one computer system or in employing distinct scheduling methods for each of a plurality of task sets each having distinct characteristics. For example, for exclusive access control between tasks, “mutex mechanisms” and “semaphore mechanisms” are used. However, in these methods employing such exclusive access control lies the problem that there is a possibility of having high priority processes delayed by low priority processes, that is a possibility of an occurrence of a reversal in priority, and therefore feature (1) above is not satisfied. As a method for mitigating such a problem of reversal in priority, priority inheritance protocols (for example, see “Priority Inheritance Protocols: An Approach to Real-Time Synchronization” a paper by Lui Sha, Ragunathan Rajkumar and John P. Lehoczky, IEEE Transactions on Computers, Vol. 39, No. 9, pp. 1179-1185, September 1990) are proposed. Priority inheritance protocols refer to a method in which, in a case where a high priority task is trying to operate on the same data while a low priority task is executing a series of operations, the priority of the low priority task is temporarily raised to the same priority as the high priority task. The operation of priority inheritance protocols is illustrated in FIG. 7 . In this case, if high priority task B tries to start operating on some data while low priority task A is operating on the same data, a delay is inevitable. At this point, the priority of task A is temporarily raised to the same level as task B. Thereafter, even if task C that has a priority lower than task B but higher than task A tries to initiate execution, since the priority of task A is raised higher than task C, execution of task A is never interrupted. Then, after task A is finished, task B, while maintaining the consistency of the data, is able to initiate operation on the data without being interrupted by task C of a lower priority than itself. However, this priority inheritance protocol is predicated on the idea that the scheduling of all tasks is performed in accordance with a common criterion, namely priority. Therefore, it is difficult to apply it to a system in which a plurality of scheduling co-exists (especially, a system in which a task set that does not perform scheduling in accordance with priority exists) such as a task execution environment in which a plurality of operating systems operate simultaneously on a single computer system, for example. In other words, feature (2) above is not satisfied. As a method that does not entail these problems, the scheduler-conscious synchronization method may be given as an example (for example, see “Scheduler-conscious synchronization” a paper by Leonidas I. Kontothanassis, Robert W. Wisniewski, Michael L. Scott, (ACM Transactions on Computer Systems, Volume 15, Issue 1, 1997)). This method limits the effect a low urgency process has on a high urgency process by prohibiting other tasks from being dispatched during the execution of a critical section. Specifically, the delay time of a high urgency process is suppressed to below the maximum critical section execution time. Further, this method is predicated only on the presence of a mechanism for prohibiting dispatch. However, since this method is not one that takes into account application to a system in which a plurality of scheduling coexists, under such a task execution environment, there still remains the possibility that a high urgency process would be delayed by a low urgency process. In other words, it does not satisfy feature (2) above. In addition, as a method that satisfies both features (1) and (2) above, the non-blocking synchronization method may be given as an example (for example, see “Non-blocking Synchronization and System Design,” a paper by Michael Barry Greenwald (Ph.D. Thesis, Stanford University, 1999)). However, in order to apply it, a special hardware is required, and an increase in cost is incurred. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG. 1 is a schematic view of the hardware configuration of a processing system 10 used in the implementation of the present invention; FIG. 2 is a schematic view illustrating a plurality of operating systems running simultaneously; FIG. 3 is a schematic view of the configuration of a task execution environment related to a mode of the present invention; FIG. 4 is a flowchart showing a procedure for task set switching software to switch task sets; FIG. 5 is a flowchart showing a procedure carried out when a task that belongs to a task set enters a critical section; FIG. 6 is a diagram showing a flow of processes, in other words a flowchart, of a program in which a plurality of points currently under execution exist; and FIG. 7 is a diagram illustrating the operation of a priority inheritance protocol. detailed-description description="Detailed Description" end="lead"? |
Methods for preparing nucleic acid samples |
In one aspect the present invention provides methods of synthesizing a preparation of nucleic acid molecules, the methods comprising the steps of: (a) utilizing an RNA template to enzymatically synthesize a first DNA molecule that is complementary to at least 50 contiguous bases of the RNA template; (b) utilizing the first DNA molecule as a template to enzymatically synthesize a second DNA molecule, thereby forming a double-stranded DNA molecule wherein the first DNA molecule is hybridized to the second DNA molecule; (c) utilizing the first or second DNA molecule of the double-stranded DNA molecule as a template to enzymatically synthesize a first RNA molecule that is complementary to either the first DNA molecule or to the second DNA molecule; and (d) utilizing the first RNA molecule as a template to enzymatically synthesize a third DNA molecule that is complementary to the first RNA molecule. In another aspect, the present invention provides processed DNA samples prepared by a method of the invention for synthesizing a preparation of nucleic acid molecules. In another aspect, the present invention provides methods for hybridizing a processed DNA sample to a population of immobilized nucleic acid molecules. |
1. A method of synthesizing a preparation of nucleic acid molecules, the method comprising the steps of: (a) utilizing an RNA template to enzymatically synthesize a first DNA molecule that is complementary to at least 50 contiguous bases of said RNA template; (b) utilizing the first DNA molecule as a template to enzymatically synthesize a second DNA molecule, thereby forming a double-stranded DNA molecule wherein the first DNA molecule is hybridized to the second DNA molecule; (c) utilizing the first or second DNA molecule of the double-stranded DNA molecule as a template to enzymatically synthesize a first RNA molecule that is complementary to either the first DNA molecule or to the second DNA molecule; and (d) utilizing the first RNA molecule as a template to enzymatically synthesize a third DNA molecule that is complementary to the first RNA molecule. 2. The method of claim 2 wherein the double-stranded DNA molecule is enzymatically amplified before utilizing the first or second DNA molecule of the double-stranded DNA molecule as a template to enzymatically synthesize a first RNA molecule. 3. The method of claim 2, wherein the RNA template is messenger RNA. 4. The method of claim 2, wherein the first DNA molecule is synthesized using reverse transcriptase. 5. The method of claim 2, wherein the synthesis of the first DNA molecule is primed using a first primer mixture comprising a multiplicity of first primers, wherein each of the first primers comprises a random sequence portion, and a defined sequence portion. 6. The method of claim 5, wherein the defined sequence portion is located 5′ to the random sequence portion. 7. The method of claim 6, wherein the defined sequence portion comprises the nucleic acid sequence of an RNA polymerase promoter. 8. The method of claim 7, wherein the defined sequence portion comprises the nucleic acid sequence set forth in SEQ ID NO:1. 9. The method of claim 6, wherein each of the first primers consists of the nucleic acid sequence of primer ShT7N9 set forth in SEQ ID NO. 2. 10. The method of claim 5, wherein the random sequence portion consists of from 4 to 20 nucleic acid residues. 11. The method of claim 5, wherein the random sequence portion consists of from 4 to 15 nucleic acid residues. 12. The method of claim 5, wherein the random sequence portion consists of from 6 to 9 nucleic acid residues. 13. The method of claim 5, wherein the random sequence portion consists of 9 nucleic acid residues. 14. The method of claim 5 wherein each of the first primers consists of the nucleic acid sequence of primer ShDNP256 set forth in SEQ ID NO:3. 15. The method of claim 5, further comprising utilizing a poly-dT primer comprising a poly-dT portion and a defined sequence portion, wherein the poly-dT portion is located 5′ to the defined sequence portion. 16. The method of claim 15, wherein the sequence of the defined sequence portion of the poly-dT primer is identical to the sequence of the defined sequence portion of the primers of the first primer mixture. 17. The method of claim 15, wherein the poly-dT portion of the poly-dT primer consists of from 5 to 25 nucleic acid residues. 18. The method of claim 15, wherein the poly-dT portion of the poly-dT primer consists of from 15 to 25 nucleic acid residues. 19. The method of claim 15, wherein the poly-dT portion of the poly-dT primer consists of 18 nucleic acid residues. 20. The method of claim 5, further comprising the step of hydrolyzing the RNA template, and substantially removing the first primer mixture, after synthesizing the first DNA molecule and before synthesizing the second DNA molecule. 21. The method of claim 2, wherein the second DNA molecule is synthesized using the Klenow fragment of DNA polymerase I. 22. The method of claim 5, wherein the synthesis of the second DNA molecule is primed using a second primer mixture comprising a multiplicity of second primer molecules, wherein each second primer molecule comprises a random sequence portion and a defined sequence portion, wherein the sequence of the defined sequence portion of the second primer molecules is different from the sequence of the defined sequence portion of the first primer molecules. 23. The method of claim 22, wherein the defined sequence portion of the second primer molecules is located 5′ to the random sequence portion of the second primer molecules. 24. The method of claim 22, wherein the defined sequence portion of each second primer molecule comprises the nucleic acid sequence of an RNA polymerase promoter. 25. The method of claim 24 wherein the defined sequence portion of each first primer molecule does not comprise the nucleic acid sequence of an RNA polymerase promoter. 26. The method of claim 24, wherein the defined sequence portion of each second primer molecule comprises the nucleic acid sequence set forth in SEQ ID NO. 1. 27. The method of claim 24, wherein each second primer molecule consists of the nucleic acid sequence of primer ShT7N9 set forth in SEQ ID NO. 2. 28. The method of claim 22 wherein the defined sequence portion of each first primer molecule comprises the nucleic acid sequence of an RNA polymerase promoter, and the defined sequence portion of each second primer molecule does not comprise the nucleic acid sequence of an RNA polymerase promoter. 29. The method of claim 22 wherein the second primer molecule consists of the nucleic acid sequence of primer ShDNP256, set forth in SEQ ID NO:3. 30. The method of claim 22 wherein each first primer consists of the nucleic acid sequence of primer ShDNP256 set forth in SEQ ID NO:3, and each second primer consists of the nucleic acid sequence of primer ShT7N9, set forth in SEQ ID NO:2. 31. The method of claim 2, wherein the double-stranded DNA molecule is amplified using a polymerase chain reaction comprising from 1 to 25 amplification cycles. 32. The method of claim 31, wherein the number of amplification cycles is from 5 to 15. 33. The method of claim 31, wherein the number of amplification cycles is 10. 34. The method of claim 2, further comprising the step of purifying the amplified, double-stranded, DNA molecule before synthesizing the first RNA molecule. 35. The method of claim 2, wherein the first DNA molecule of the amplified, double-stranded, DNA molecule is utilized as a template to enzymatically synthesize the first RNA molecule. 36. The method of claim 35, wherein the first DNA molecule of the amplified, double-stranded, DNA molecule comprises a T7 RNA polymerase promoter that promotes synthesis of the first RNA molecule. 37. The method of claim 36, wherein the T7 RNA polymerase promoter comprises the nucleic acid sequence set forth in SEQ ID NO. 1. 38. The method of claim 36, wherein the T7 RNA polymerase promoter consists of the nucleic acid sequence set forth in SEQ ID NO. 1. 39. The method of claim 2, wherein the second DNA molecule of the amplified, double-stranded, DNA molecule is utilized as a template to synthesize the first RNA molecule. 40. The method of claim 39, wherein the second DNA molecule comprises a T7 RNA polymerase promoter which promotes the synthesis of the first RNA molecule. 41. The method of claim 40, wherein the T7 RNA polymerase promoter comprises the nucleic acid sequence set forth in SEQ ID NO. 1. 42. The method of claim 40, wherein the T7 RNA polymerase promoter consists of the nucleic acid sequence set forth in SEQ ID NO. 1. 43. The method of claim 2, further comprising the step of purifying the first RNA molecule before synthesizing the third DNA molecule, wherein the purification step removes substantially all nucleic acid molecules less than 100 bases long. 44. The method of claim 2 wherein the third DNA molecule is synthesized using reverse transcriptase, and the synthesis of the third DNA molecule is primed using a population of random primers. 45. The method of claim 44 wherein at least 99 percent of the random primers consist of nine nucleotides. 46. The method of claim 2 wherein the third DNA molecule is purified to remove substantially all nucleic acid molecules less than 100 bases long. 47. The method of claim 2 wherein dye molecules are joined to the third DNA molecule by aminoallyl linkages. 48. The method of claim 47 wherein the dye is a Cy dye. 49. A method of synthesizing a preparation of nucleic acid molecules, the method comprising the steps of: (a) utilizing an RNA template to enzymatically synthesize a first DNA molecule that is complementary to at least 50 contiguous bases of the RNA template, wherein: (i) the first DNA molecule is synthesized using reverse transcriptase; (ii) the synthesis of the first DNA molecule is primed using a first primer mixture comprising a multiplicity of first primers, wherein each of the first primers comprises a random sequence portion and a defined sequence portion located 5′ to the random sequence portion, wherein the defined sequence portion comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:3; (b) hydrolyzing the template RNA and removing substantially all of the first primer mixture after synthesis of the first DNA molecule; (c) utilizing the first DNA molecule as a template to enzymatically synthesize a second DNA molecule, thereby forming a double-stranded DNA molecule wherein the first DNA molecule is hybridized to the second DNA molecule, wherein: (i) the second DNA molecule is synthesized using the Klenow fragment of DNA polymerase I; (ii) the synthesis of the second DNA molecule is primed using a second primer mixture comprising a multiplicity of second primer molecules, wherein each second primer molecule comprises a random sequence portion and a defined sequence portion, wherein the sequence of the defined sequence portion of each second primer molecule is different from the sequence of the defined sequence portion of each first primer molecule, and wherein the second primer defined sequence portion comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:3; (d) removing substantially all of the second primer mixture after synthesis of the second DNA molecule; (e) amplifying the double-stranded DNA molecule using a polymerase chain reaction comprising from 5 to 15 amplification cycles, wherein the polymerase chain reaction is primed with a first PCR primer population and a second PCR primer population, wherein the first PCR primer population consists essentially of primer molecules consisting of the sequence set forth in SEQ ID NO:5, and the second PCR primer population consists essentially of primer molecules consisting of the sequence set forth in SEQ ID NO:6; (f) removing substantially all of the PCR primer mixture after amplification of the double-stranded DNA molecule; (g) utilizing the first or second DNA molecule of the amplified, double-stranded, DNA molecule as a template to synthesize, using an RNA polymerase molecule, a first RNA molecule that is complementary to either the first DNA molecule or to the second DNA molecule; (h) purifying the first RNA molecule to remove substantially all nucleic acid molecules less than 100 bases long; (i) utilizing the first RNA molecule as a template to enzymatically synthesize a third DNA molecule that is complementary to the first RNA molecule, wherein the third DNA molecule is synthesized using reverse transcriptase and the synthesis of the third DNA molecule is primed using a population of random primers wherein substantially all of the random primers consist of 9 bases; and (j) joining Cy dye molecules to the third DNA molecule by aminoallyl linkages. 50. A DNA sample prepared by a method comprising the steps of: (a) utilizing an RNA template to enzymatically synthesize a first DNA molecule that is complementary to at least 50 contiguous bases of said RNA template; (b) utilizing the first DNA molecule as a template to enzymatically synthesize a second DNA molecule thereby forming a double-stranded DNA molecule wherein the first DNA molecule is hybridized to the second DNA molecule; (c) utilizing the first or second DNA molecule of the double-stranded DNA molecule as a template to enzymatically synthesize a first RNA molecule that is complementary to either the first DNA molecule or to the second DNA molecule; and (d) utilizing the first RNA molecule as a template to enzymatically synthesize a third DNA molecule that is complementary to the first RNA molecule. 51. A method for hybridizing a processed DNA sample to a population of immobilized nucleic acid molecules, the method comprising the step of hybridizing a processed DNA sample to a population of immobilized nucleic acid molecules, wherein the processed DNA sample is prepared by a method comprising the steps of: (a) utilizing an RNA template to enzymatically synthesize a first DNA molecule that is complementary to at least 50 contiguous bases of said RNA template; (b) utilizing the first DNA molecule as a template to enzymatically synthesize a second DNA molecule, thereby forming a double-stranded DNA molecule wherein the first DNA molecule is hybridized to the second DNA molecule; (c) utilizing the first or second DNA molecule of the double-stranded DNA molecule as a template to enzymatically synthesize a first RNA molecule that is complementary to either the first DNA molecule or to the second DNA molecule; and (d) utilizing the first RNA molecule as a template to enzymatically synthesize a third DNA molecule that is complementary to the first RNA molecule. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The characterization of cellular gene expression finds application in a variety of disciplines, such as in the analysis of differential expression between different tissue types, different stages of cellular growth or between normal and diseased states. Recently, changes in gene expression have also been used to assess the activity of new drug candidates and to identify new targets for drug development. The latter objective is accomplished by correlating the expression of a gene or genes known to be affected by a particular drug with the expression profile of other genes of unknown function when exposed to that same drug. Genes of unknown function that exhibit the same pattern of regulation, or signature, in response to the drug are likely to represent novel targets for pharmaceutical development. DNA arrays are particularly useful in gene expression analysis at the level of transcription (see, e.g., Ramsay, Nature Biotechnol. 16:40-44, 1998; Marshall and Hodgson, Nature Biotechnol. 16:27-31, 1998; Lashkari et al., Proc. Natl. Acad. Sci . ( USA ) 94:130-157, 1997; DeRisi et al., Science 278:680-6, 1997). In such analysis, the identity and abundance of a selected nucleic acid sequence in a sample is determined by measuring the level of hybridization of the nucleic acid sequence to probes on the DNA array that comprise complementary sequences. The selected nucleic acid sequence in a sample can be an mRNA, or a nucleic acid molecule derived from an mRNA that has a nucleic acid sequence that is identical to, or complementary to, all, or a portion, of the mRNA. Using DNA array expression assays, complex mixtures of labeled nucleic acids (e.g., mRNAs, or nucleic acid molecules derived from mRNAs) can be analyzed. The nucleic acid molecules used to screen a DNA array should be representative of the mRNA population from which they are derived. All, or substantially all, of the sequences in the mRNA population should be represented in the nucleic acid molecule population used to screen the DNA array. For example, all portions of individual mRNA molecules should be equally represented in the nucleic acid molecule population used to screen the DNA array. In this regard, the use of oligo-dT primers, that hybridize to the polyA tail of mRNA molecules, to prime the enzymatic synthesis of complementary DNA molecules, results in the underrepresentation of the 3′ ends of long mRNA molecules in the population of complementary DNA molecules. A proposed solution to this problem is to use a population of oligonucleotides, having random nucleic acid sequences, to prime the enzymatic synthesis of DNA molecules complementary to the template mRNA molecules. It is statistically likely that at least one of the random oligonucleotides will hybridize to at least one portion of each mRNA molecule in a population, thereby yielding a population of complementary DNA molecules that represent all, or substantially all, portions of all, or substantially all, mRNA molecules in the template population. A drawback to this approach, however, is that there is little or no amplification of the sequences in the template mRNA population, thereby limiting the practical usefulness of the technique, for example to produce enough probe to screen numerous DNA arrays. Further, the nucleic acid molecules used to screen a DNA array should selectively hybridize to complementary nucleic acid molecules, and not hybridize, to a significant extent, to non-complementary nucleic acid molecules, immobilized on the DNA array. In this regard, the present inventors have observed that RNA molecules are typically more prone to hybridize to complementary nucleic acid molecules, immobilized on a DNA array, than are DNA molecules. Thus, there is a need for methods for synthesizing DNA molecules from mRNA template molecules, wherein: (a) the synthesized DNA molecules represent all, or substantially all, portions of all, or substantially all, template mRNA molecules; (b) the abundance of each template mRNA molecule, and each portion of each template mRNA molecule, is identical, or substantially identical, to the abundance of the identical, or complementary, DNA sequence in the population of synthesized DNA molecules; and (c) the synthetic method is capable of amplifying a small amount of template mRNA (e.g., 1 μg or less) to yield sufficient probe to screen numerous DNA microarrays. Preferably, the synthesized DNA molecules selectively hybridize to complementary nucleic acid molecules, and do not hybridize, to a significant extent, to non-complementary nucleic acid molecules immobilized on a DNA array. Moreover, it is desirable that the synthetic methods controllably yield a population of DNA molecules wherein all, or substantially all, of the DNA molecules are complementary to either the sequences of the template mRNA molecules, or to the complementary sequences of the template mRNA molecules. In one aspect, the present invention provides processed nucleic acid samples that meet the foregoing requirements, and methods for making such processed nucleic acid samples. |
<SOH> SUMMARY OF THE INVENTION <EOH>In accordance with the foregoing, in one aspect the present invention provides methods of synthesizing a preparation of nucleic acid molecules, the methods comprising the steps of: (a) utilizing an RNA template to enzymatically synthesize a first DNA molecule that is complementary to at least 50 contiguous bases of the RNA template; (b) utilizing the first DNA molecule as a template to enzymatically synthesize a second DNA molecule, thereby forming a double-stranded DNA molecule wherein the first DNA molecule is hybridized to the second DNA molecule; (c) utilizing the first or second DNA molecule of the double-stranded DNA molecule as a template to enzymatically synthesize a first RNA molecule that is complementary to either the first DNA molecule or to the second DNA molecule; and (d) utilizing the first RNA molecule as a template to enzymatically synthesize a third DNA molecule that is complementary to the first RNA molecule. The double-stranded DNA molecule prepared in accordance with step (b) can optionally be enzymatically amplified before utilizing the first, or second, DNA molecule of the double-stranded DNA molecule as a template to enzymatically synthesize a first RNA molecule. The third DNA molecule prepared in accordance with the methods of this aspect of the invention can be labeled with a dye. In some embodiments of the methods of this aspect of the invention, the third DNA molecule is labeled via an aminoallyl linkage. The methods of this aspect of the invention yields populations of third DNA molecules that are representative of the population of RNA template molecules used to synthesize the third DNA molecules. In particular, substantially all of the RNA molecules in the population of template RNA molecules are represented in the population of third DNA molecules, and there is substantially no 5′ or 3′ bias within the population of third DNA molecules. The embodiments of the methods of this aspect of the invention that include amplification of the double-stranded DNA molecules typically yield an amount of third DNA molecules that is at least a thousand-fold greater than the amount of template RNA molecules. The methods of the invention for synthesizing a preparation of nucleic acid molecules are useful in any situation where it is desired to synthesize a preparation of nucleic acid molecules, such as DNA molecules, beginning with an RNA template. For example, the methods of this aspect of the invention are useful for synthesizing a population of third DNA molecules that is used to hybridize to a population of immobilized nucleic acid molecules, such as a population of DNA molecules immobilized on a DNA microarray. For example, third DNA molecules prepared in accordance with the methods of this aspect of the invention can be used to hybridize to a DNA chip in order to generate a gene expression profile. Gene expression profiling can be done, for example, for purposes of screening, diagnosis, staging a disease, and monitoring response to therapy, as well as for identifying genetic targets of drugs and of pathogens. In another aspect, the present invention provides processed DNA samples prepared by a method of the invention for synthesizing a preparation of nucleic acid molecules. The processed DNA samples of the invention can be utilized in any experiment, process or therapeutic treatment that requires DNA. For example, the processed DNA samples of the invention can be hybridized to a population of immobilized nucleic acid molecules, such as to a population of DNA molecules immobilized on a Southern blot, to a population of RNA molecules immobilized on a Northern blot, or to a population of DNA molecules immobilized on a DNA microarray (such as for the purpose of gene expression profiling). For example, the processed DNA samples of the invention can be used in the gene expression profiling method set forth in Hughes, T. R., et al., Nature Biotechnology 19:342-347 (2001), which publication is incorporated herein by reference. When used as probes to hybridize to a population of immobilized nucleic acid molecules, such as a population of nucleic acid molecules immobilized on a DNA array, processed DNA samples of the invention exhibit a high level of hybridization specificity and sensitivity. In another aspect, the present invention provides methods for hybridizing a processed DNA sample to a population of immobilized nucleic acid molecules, the methods each comprising the step of hybridizing a processed DNA sample to a population of immobilized nucleic acid molecules, wherein the processed DNA sample is prepared by a method of the invention for synthesizing a preparation of nucleic acid molecules. The methods of the invention for hybridizing a processed DNA sample to a population of immobilized nucleic acid molecules are useful in any hybridization experiment wherein DNA molecules are hybridized to a population of immobilized nucleic acid molecules, such as a population of DNA molecules immobilized on a DNA micro array. |
Applicator for tissue adhesive |
An adhesive applicator, for applying medical adhesives, and particularly cyanoacrylates, including a body (20) comprising an internal reservoir (22) and an opening between the internal reservoir (22) and the exterior of the body (20); a sponge (26) disposed over the opening; a slotted tip nozzle (30) disposed over the sponge (26); and a small orifice tip nozzle (40) disposed over the slotted tip nozzle.(30) |
1. An adhesive applicator, comprising: a body comprising an internal reservoir and an opening between the internal reservoir and the exterior of the body; a sponge disposed over the opening; a slotted tip nozzle disposed over the sponge; and a small orifice tip nozzle disposed over the slotted tip nozzle. 2. The adhesive applicator of claim 1, wherein the slotted tip nozzle is releasably attached to the body, thereby covering the sponge. 3. The adhesive applicator of claim 1, wherein the small orifice tip nozzle is releasably attached to the slotted tip nozzle, thereby covering the slotted tip of the slotted tip nozzle. 4. The adhesive applicator of claim 1, wherein the small orifice tip nozzle is releasably attached to the body, thereby covering the slotted tip of the slotted tip nozzle. 5. The adhesive applicator of claim 1, wherein the body is deformable such that squeezing on the body forces contents of the internal reservoir out through the opening in the applicator body. 6. The adhesive applicator of claim 1, further comprising: an adhesive in the internal reservoir. 7. The adhesive applicator of claim 6, wherein the adhesive is a medical adhesive. 8. The adhesive applicator of claim 7, wherein the medical adhesive is a cyanoacrylate ester, in monomeric form, represented by formula I: where R is selected from the group consisting of: alkyl of 1 to 10 carbon atoms, alkenyl of 2 to 10 carbon atoms, cycloalkyl groups of from 5 to 8 carbon atoms, phenyl, 2-ethoxyethyl, 3-methoxybutyl, and a substituent of the formula: wherein each R′ is independently selected from the group consisting of: hydrogen and methyl, and R″ is selected from the group consisting of: alkyl of from 1 to 6 carbon atoms, alkenyl of from 2 to 6 carbon atoms, alkynyl of from 2 to 6 carbon atoms, cycloalkyl of from 3 to 8 carbon atoms, aralkyl selected from the group consisting of benzyl, methylbenzyl and phenylethyl, phenyl, and phenyl substituted with 1 to 3 substituents selected from the group consisting of hydroxy, chloro, bromo, nitro, alkyl of 1 to 4 carbon atoms, and alkoxy of from 1 to 4 carbon atoms. 9. The adhesive applicator of claim 1, wherein the nozzle of the slotted tip nozzle has a width of between ⅓ and ⅔ of the width of the sponge. 10. A method of closing, sealing and dressing a wound with an adhesive applicator comprising a body with an internal reservoir, adhesive disposed in the internal reservoir and an opening between the internal reservoir and the exterior of the body; a sponge disposed over the opening; a slotted tip nozzle disposed over the sponge; and a small orifice tip nozzle disposed over the slotted tip nozzle; comprising: closing the wound by applying small drops of adhesive to the wound, the small drops being applied by forcing the adhesive from the internal reservoir through the sponge, through the slotted tip nozzle and then through the small orifice tip nozzle; removing the small orifice tip nozzle; sealing the wound by applying a line of adhesive with the slotted tip nozzle; removing the slotted tip nozzle; and dressing the wound by applying further adhesive with the sponge. 11. The method of claim 10, wherein forcing the adhesive from the internal reservoir through the sponge, through the slotted tip nozzle and then through the small orifice tip nozzle by squeezing on the body of the applicator. 12. The method of claim 10, further comprising: adjusting the width of the line of adhesive applied by the slotted tip nozzle by rotating the applicator, thereby rotating the slotted tip nozzle. 13. The method of claim 10, wherein the adhesive is a medical adhesive. 14. The adhesive applicator of claim 13, wherein the medical adhesive is a cyanoacrylate ester, in monomeric form, represented by formula I: where R is selected from the group consisting of: alkyl of 1 to 10 carbon atoms, alkenyl of 2 to 10 carbon atoms, cycloalkyl groups of from 5 to 8 carbon atoms, phenyl, 2-ethoxyethyl, 3-methoxybutyl, and a substituent of the formula: wherein each R′ is independently selected from the group consisting of: hydrogen and methyl, and R″ is selected from the group consisting of: alkyl of from 1 to 6 carbon atoms, alkenyl of from 2 to 6 carbon atoms, alkynyl of from 2 to 6 carbon atoms, cycloalkyl of from 3 to 8 carbon atoms, aralkyl selected from the group consisting of benzyl, methylbenzyl and phenylethyl, phenyl, and phenyl substituted with 1 to 3 substituents selected from the group consisting of hydroxy, chloro, bromo, nitro, alkyl of 1 to 4 carbon atoms, and alkoxy of from 1 to 4 carbon atoms. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Many different applicators for glues and adhesives have been developed. Unfortunately, some applicators are specifically designed to supply very small amounts of adhesive with pinpoint accuracy; whereas, other applicators have been specifically designed to supply large amounts of adhesives over wide surface areas. To date, applicators have not been developed with sufficient versatility to apply adhesives either in very small amounts with pinpoint accuracy, or over wide surface areas. Such applicators are specifically desirable in medical applications where the same adhesive compound may be used to close an open cut or wound, to seal the cut or wound, and then to dress the cut or wound. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention provides a versatile applicator which is ideally suited for controlled application of adhesives, especially medical adhesives. As such, the present invention is particularly useful in closing and sealing wounds and protecting tissue. In a preferred aspect, the present invention comprises an adhesive applicator, including a body having an internal reservoir and an opening between the internal reservoir and the exterior of the body; a sponge disposed over the opening; a slotted tip nozzle disposed over the sponge; and a small orifice tip nozzle disposed over the slotted tip nozzle. In various aspects, the slotted tip nozzle is releasable attached to the body of the applicator, and the small orifice tip nozzle is releasably attached either to the slotted tip nozzle, or directly to the body of the applicator. In accordance with the present invention, the small orifice tip nozzle may be removed first, followed by the removal of the slotted tip nozzle. In a preferred method of use, a cut or open wound can be sequentially closed, sealed and then dressed, as follows. First, the wound is closed by applying small drops of adhesive to the wound, the small drops being applied by forcing the adhesive from the internal reservoir through the sponge, through the slotted tip nozzle and then through the small orifice tip nozzle and into the wound. Then, after the wound is closed, the small orifice tip nozzle may be removed. Then, the wound may be sealed by applying a line of adhesive with the slotted tip nozzle. Then, after the wound has been sealed, the slotted tip nozzle may be removed. Lastly, the wound may be dressed by applying further adhesive and/or spreading the adhesive around the area of the wound with the sponge. Thus, advantages of the present invention include its ability to provide a versatile system which can be used to apply adhesives either as single small drops, in a thin line, in a wide line, or as a wide surface coating. In preferred aspects, the body is deformable such that squeezing on the body forces contents of the internal reservoir out through the opening in the applicator body. Also, in preferred aspects, the adhesive applied is a medical adhesive. Most preferably, the medical adhesive is a cyanoacrylate compound. |
Method for determining the load exerted on a vehicle tire |
A method for determining the load exerted on the tire of a motor vehicle or to monitor the tire pressure, the invention discloses a method wherein during operation of the vehicle the pressure in the individual wheels is detected and the rotational behavior of the individual wheels is monitored, and wherein load distribution characteristic quantities are determined by comparing the rotational behavior or changes in said rotational behavior of the individual wheels during given driving conditions taking onto account preset or learnt variables, and wherein finally the charge or load exerted on the tires or pressure loss is concluded from the tire pressure and the load distribution characteristic quantities. |
1-17. (canceled) 18. Method for determining the load exerted on the tire of a motor vehicle and for monitoring the tire pressure, comprising the steps of: determining the pressure in the individual tires during operation of the vehicle, monitoring the rotational behavior of the individual wheels, determining load distribution characteristic quantities by comparing the rotational behavior or the changes of the rotational behavior of the individual wheels in defined driving conditions in consideration of preset or learnt quantities, and deriving the load exerted on the tires and pressure loss from the tire pressure and one or more load distribution characteristic quantities. 19. Method as claimed in claim 18, wherein a pressure measuring system (TPMS) based on a pressure sensor system is used to determined the tire pressure, and wherein wheel speed sensors are used to monitor the wheel rotational behavior, and wherein a system based on the evaluation of wheel speed information is used to determine the load distribution characteristic quantities. 20. Method as claimed in claim 19, wherein the rotational speed of a front wheel is compared to the rotational speed of a rear wheel at the same vehicle speed or approximately identical vehicle speed and evaluated for producing a load distribution characteristic quantity, wherein the value or the change of the load distribution characteristic quantities in defined driving situations is taken into account for calculating the load exerted on the tires and the pressure loss. 21. Method as claimed in claim 20, wherein determining the load distribution characteristic quantities, includes using a quotient made up of the front-wheel and rear-wheel speeds of the wheels of the same vehicle side (VFL/VRL; VFR/VRR), and wherein the value or the variations of the load distribution characteristic quantities are evaluated at (approximately) equal vehicle speed or as a function of at least one driving parameter. 22. Method as claimed in claim 21, wherein the speeds of the wheels of one axle (VFL/VRL or VFR/VRR) are compared and evaluated for detecting relative load distribution changes vertically to the driving direction. 23. Method as claimed in claim 21, wherein said at least one driving parameter includes vehicle speed. 24. Method as claimed in claim 21, wherein said at least one driving parameter includes tire pressure. 25. Method as claimed in claim 21, wherein said at least one driving parameter includes wheel torque. 26. Method as claimed in claim 21, wherein the nominal values or normal values of the load distribution characteristic quantities applying to a defined load, are input manually, or are detected automatically as soon as defined conditions are satisfied. 27. Method as claimed in claim 26, wherein the values for the load distribution characteristic quantities that apply to the partial load operations are determined for the load distribution characteristic quantities in defined driving situations, or by taking into account the driving torque, by statistic analysis or by evaluation of criteria indicative of a partial load operation. 28. Method as claimed in claim 27, wherein the point of time where a normal condition prevails is fixed by manual or automatic release of a start signal. 29. Method as claimed in claim 28, wherein values for the load distribution characteristic quantities and the associated tire pressure values are learnt and memorized during predetermined driving conditions. 30. Method as claimed in claim 19, wherein the system based on evaluation of the wheel speed data learns the desired or normal condition in a learning phase. 31. Method as claimed in claim 30, wherein the quantities representative of the rolling circumference of the wheels are used as proportional factors of different wheel pairs. 32. Method as claimed in claim 31, wherein the system based on evaluating wheel speed data determines after the learning phase variations of the quantities representative of the rolling circumference of the wheels by comparison of current characteristic quantities representative of the rolling circumference of the wheels with learnt quantities indicating of the rolling circumference of the wheels. 33. Method as claimed in claim 32, wherein the difference between the learnt quantity and the current quantity is a standard of the wheel load. 34. Method as claimed in claim 19, wherein the system based on the evaluation of the wheel speed data is a complete DDS-system detecting tire pressure loss irrespective of the pressure measuring system (TPMS). 35. Method as claimed in claim 34, wherein a comparison is made and evaluated between the tire pressure loss determined by means of the pressure sensor system (TPMS) and the tire pressure loss detected on the basis of the DDS system. 36. Method as claimed in claim 35, wherein a compression triggered by increase of the wheel load is considered to prevail when the tire pressure loss found out on the basis of the wheel speed data is by a defined amount higher than the tire pressure loss detected on the basis of the pressure sensors (TPMS). |
<SOH> BACKGROUND OF THE INVENTION <EOH>It is disclosed in the art to determine in motor vehicles the tire pressure or a quantity representative of the tire pressure by means of pressure measuring systems (TPMS=Tire Pressure Measuring System) based on a pressure sensor or, without using pressure sensors, by means of systems based on the wheel speed (DDS=Deflation Detecting System). In the so-called TPMS system, the measured pressure along with the temperature is sensed by means of sensors arranged in the tire and transmitted in a wireless manner to a receiver installed in the vehicle. The pressure data is evaluated and the pressure prevailing in the individual tires is determined in a control device. For the conventionally used safety and motor vehicle control systems such as ABS, TCS, ESP, DDS, etc., a relatively exact knowledge of the rotational behavior of the individual vehicle wheels is necessary. Nowadays, vehicles are therefore equipped with passive or active wheel speed sensors, which supply the required input signals for the various safety and control systems. Thus, the tire pressure that cannot be derived directly from the wheel speeds is determined indirectly by way of the dynamic rolling circumference of the individual vehicle wheel e.g. by means of the DDS system (Deflation Detecting System). The load dependency of the dynamic rolling circumference rather must be considered a disturbance variable in pressure loss detection by means of DDS. A method for measuring the load exerted on the tire which is determined, among others, by the actual tire pressure and the wheel load or wheel load distribution during operation of the vehicle, meaning during driving, has not been known so far. It is not sufficient to monitor the tire pressure loss because the charge or load exerted on the tire, to a larger extent than the tire pressure, is responsible for the safety and durability of the tire and because different tire pressures are required for a comfortable driving behavior and ideal charge exerted on the tire depending on the wheel load or wheel load distribution. |
<SOH> SUMMARY OF THE INVENTION <EOH>In view of the above, an object of the present invention is to provide a method that allows detecting the charge or load exerted on a tire under different conditions, in particular in the event of different wheel load or wheel load distribution. A method of this type renders it possible to exclude overload on the tires, safeguard durability of the tires, and increase the general safety of vehicle and driver. The method of the present invention determines the pressure in the individual tires during operation of the vehicle and monitors the rotational behavior of the individual wheels and determines the load distribution characteristic quantities of the tires by comparing the rotational behavior or the changes of the rotational behavior of the individual wheels in defined driving conditions in consideration of preset or learnt quantities, and deriving the charge or load exerted on the tires and pressure loss from the tire pressure and the load distribution characteristic quantities. The invention is based on the knowledge that the variable, i.e. the charge exerted on the tire, influencing the safety and lifetime of a tire to a great degree can be determined by way of measuring the pressure and monitoring the wheel rotational behavior. The dependence of the dynamic rolling circumference on the tire pressure and the wheel load is e.g. known from the development of wheel-speed based pressure monitoring systems (DDS). By comparing the wheel speeds of the various vehicle wheels and evaluating the deviations it is possible to find out parameters representative of the charge exerted on the tires when knowing the tire pressure—e.g. measured by TPMS. According to an embodiment of the invention, a pressure measuring system (such as TPMS=Tire Pressure Measuring System) based on a pressure sensor system is used to determine the tire pressure, wheel speed sensors are used to monitor the wheel rotational behavior, and a system based on the evaluation of wheel speed information is used to determine the load distribution characteristic quantities, the latter system corresponding in its mode of functioning to a system (DDS) for detecting the conditions of the dynamic rolling circumferences of the individual tires. Thus, systems known in the art are used to realize the detection of the exerted charge. According to another favorable embodiment of the invention, the rotational speed of a front wheel is compared to the rotational speed of a rear wheel at the same vehicle speed or approximately identical vehicle speed (e.g. vehicle reference speed), is evaluated for producing a load distribution characteristic quantity, and the value or the change of the load distribution characteristic quantities in defined driving situations is taken into account for calculating the charge or load exerted on the tires or the pressure loss. It has proven especially suitable for determining the load distribution characteristic quantities to produce in each case a quotient made up of the front-wheel and rear-wheel speeds of the wheels (or the corresponding rotational speeds) of the same vehicle side and to evaluate the value or the variations of the load distribution characteristic quantities at (approximately) equal vehicle speed or in consideration of the vehicle speed. The quotients F FL /F RL or F FR /F RR , which can be linked to each other additionally to enhance the ‘safety’ of detection and the accuracy, are appropriate quantities to determine the load distribution. Other types of combinations, e.g. the diagonal, or corresponding other types of functional relationships, such as especially quotients of speed sums, are also possible. This feature depends on the respective concept of the vehicles and the monitoring systems. Further, it is arranged to respectively compare and evaluate the speeds of the wheels of an axle for detecting relative load distribution variations vertically to the driving direction, for example as a result of boarding of a passenger. To increase the measuring and evaluation accuracy it is also possible or even preferred to make the rolling times of the wheel instead of the wheel revolutions or wheel speeds the basis for the calculations. The nominal values or normal values of the load distribution characteristic quantities applying to a defined charge or load, e.g. partial load or full load, can be input manually, or the system can be designed so that the corresponding nominal or normal values are detected automatically as soon as defined conditions are satisfied. Further, it has proven expedient to determine the values for the load distribution characteristic quantities that apply to partial load operations in defined driving situations, for example, with (at least approximately) freely rolling wheels, at constant straight driving, etc., or by taking into account the driving torque into the calculations by statistic analysis or by evaluation of criteria indicative of a partial load operation. Although characteristic quantities obtained in driving situations with freely rolling wheels are generally more precise and, therefore, can be evaluated in a preferred fashion, it is frequently additionally suitable, for reasons of a number of data limited in practice, to equally include data from driving situations where the wheels do not run freely. In the last-mentioned cases, the data obtained must be cleared from the influence of the respective driving parameter on the characteristic quantity. The necessary information for determining the driving situation is preferably obtained from a per se known ABS or ESP system. The point of time where a normal condition, meaning a defined condition or desired condition, prevails can be fixed according to the invention by manual or independent release of a start signal, e.g. after adjustment of a predetermined inflation pressure and load condition. After a changing of tires or after the first assembly one will not be able in many cases to avoid manually triggering a reset action or a start signal. According to another embodiment of the method of the invention, values for the load distribution characteristic quantities and the associated tire pressure values are learnt and memorized during predetermined driving conditions. These learnt characteristic quantities can be established in particular in dependence on a driving parameter, such as in dependence on the speed. In the last-mentioned case it is particularly suitable when successive intervals for the driving parameter are produced and a learning value is determined for each interval. Learning processes are preferably used also for the system based on the evaluation of wheel speed data. In particular, the desired or normal condition is learnt in a learning phase, especially by determining and memorizing quantities representative of the rolling circumference of the wheels. The quantities representative of the rolling circumference of the wheels may also be proportional factors of different wheel pairs. Depending on the case of application, the proportional factors may of course also be determined by different calculations or a different type of comparison of the individual wheels. According to another preferred embodiment of the invention, the system based on evaluating wheel speed data determines after the learning phase variations of the quantities representative of the rolling circumference of the wheels by comparison of current characteristic quantities representative of the rolling circumference of the wheels with learnt quantities indicating of the rolling circumference of the wheels. The difference between the learnt quantity and the current quantity is a standard of the wheel load in this case. The system based on the evaluation of the wheel speed data may be extended with little effort to become a complete DDS, i.e. a system capable of detecting tire pressure loss irrespective of the pressure measuring system (TPMS). In this system, a comparison can be made and evaluated between the tire pressure loss determined by the pressure sensor system (TPMS) and the tire pressure loss detected on the basis of the DDS system. When the tire pressure loss found out on the basis of the wheel speed data is by a defined amount higher than the tire pressure loss detected on the basis of the pressure sensor system, a compression triggered by increase of the wheel load can be concluded from that fact. |
Method for the production of nucleic acids consisting of stochastically combined parts of source nucleic acids |
The present invention relates to a method for the production of nucleic acids consisting of stochastically combined parts of source nucleic acids as well as to a kit for carrying out said method. |
1. A method for the production of polynucleotide molecules with modified proper-ties, comprising the following steps: (1) providing a population of source nucleic acid molecules, the individual nucleic acid molecules of said population having homologous and heterologous segments and having at least one marker nucleotide incorporated within its nucleic acid sequence; (2) forming double-stranded polynucleotide molecules of the population of source nucleic acid molecules provided according to step (1) comprising double strands with heterologous segments (heteroduplices); (3) producing single-stranded breaks at the incorporated marker nucleo-tides of the double-stranded heteroduplices produced according to step (2); and (4) performing template-directed single-strand synthesis, with or without incorporation of marker nucleotides starting from single-stranded breaks produced according to step (3). 2. The method of claim 1, wherein (i) more than one cycle, preferably at least two cycles, more preferably at least ten and most preferably at least twenty cycles, comprising the aforementioned steps (2) to (4) are performed; and/or (ii) in all cycles but the last, step (4) is carried out with the incorporation of new marker nucleotides ; and/or (iii) steps (3) and (4) are carried out subsequently or contemporaneously. 3. The method of claim 1, wherein (i) homologous segments have a length of at least 5, preferably of at least 10 and more preferably of at least 20 nucleotides and/or are not longer than 5,000 nucleotides, preferably not longer than 2,000 nucleotides, more preferably not longer than 1,000 nucleotides; and/or (ii) the homologous segments are flanked by heterologous segments. 4. The method of claim 1, wherein (i) the incorporation of marker nucleotides into the nucleic acid molecules according to step (1) is achieved by using a template-directed polymerase reaction or by chemical synthesis of oligonucleotides; and/or (ii) the production of double-stranded heteroduplex polynucleotides according to step (2) is achieved by hybridization of the homologous segments of complementary polynucleotides; and/or (iii) the single-stranded breaks at the positions of the incorporated marker nucleotides of step (3) are nicks or gaps which are achieved by using enzymatic reactions; and/or (iv) the template-directed single-strand synthesis of step (4) utilizes a polymerase. 5. The method of claim 1, wherein more than one cycle comprising steps (2) to (4) is performed and the average distance between the starting points of the template-directed synthesis according to step (4) in each of two consecutive cycles is controlled by adjusting the probability of incorporating marker nucleotides in step (4) of the first of the two consecutive cycles. 6. The method according to claim 5, wherein the probability of incorporating marker nucleotides is controlled by adjusting the ratio of concentrations of marker nucleotides to standard nucleotides; and/or wherein the probability of incorporating marker nucleotides is preferably lower than one and higher than the reciprocal of the source nucleic acid length in base pairs; and/or wherein the probability of incorporating marker nucleotides is altered from cycle to cycle. 7. The method of claim 4, wherein the nucleic acid molecules are DNA molecules and in the template-directed polymerase reaction deoxyuridine triphosphate (dUPT) is utilized as a marker nucleotide in combination with the four standard deoxynucleoside triphosphates; and/or the uracil base of the incorporated marker uridine residues is separated from the ribose using an uracil-DNA glycosylase. 8. The method of claim 4, wherein the nucleic acid molecules are DNA molecules and in the template directed polymerase reaction 8-oxo-doxyguanosine triphosphate (8-oxo-dGTP) is utilized as a marker nucleotide in combination with the four standard deoxynucleoside triphosphates; and/or the 8-oxo-guanine base of the incorporated 8-oxo-GMP residues is separated from the ribose using formamidopyrimidine-DNA glycosylases. 9. The method of claim 4, wherein the nucleic acid molecules are DNA molecules and in the template directed polymerase reaction marker nucleotides with the following modified bases are used in combination with the four standard dNTPs: 3-methyladenine, 7-methyladenine, 3-methylguanine, 7-methylguanine,7-hydroxyethylguanine, 7-chloroethylguanine, O2-alkylthymine, O2-alkylcytosine, 5-fluorouracil, 2,5-amino-5-formamidopyrimidine, 4,6-diamino-5-formamidopyrimidine, 2,6-diamino-4-hydroxy-5-formamidopyrimidine, 5-hydroxycytosine, 5,6-dihydrothymine, 5-hydroxy-5,6-dihydrothymine, thymine glycol, uracil glycol, isodialuric acid, alloxan, 5, 6-dihydrouracil, 5-hydroxy-5,6-dihydrouracil, 5-hydroxyuracil, 5-formyluracil, 5-hydroxymethyluracil, hypoxanthine, 1, N6-ethenoadenine or 3,N4-ethenocytosine; and/or a DNA N-glycosylase which detects one of the aforementioned modified base, preferably E. coli endonuclease III or alkylbase DNA glycosylase, is utilized. 10. The method of claim 4, wherein the nucleic acid molecules are DNA molecules and in the template directed polymerase reaction one, two, three or all four ribonucleoside triphosphates (rNTPs) are utilized as marker nucleotides in combination with the four standard dNTPs in the template directed polymerase reaction; and/or, the rNMP residues incorporated in the DNA polynucleotide are recognized by a specific ribonuclease H, preferably by human RNase HI. 11. The method of claim 4, wherein the nucleic acid molecules are DNA molecules, any or all of the four ribonucleoside monophosphates (rNMPs) are used as marker nucleotides, and the marker strand is cleaved by alkaline hydrolysis at the rNMP residues, and/or the 2′- or 3′-rNMP at the 3′-end of a nick resulting from the alkaline hydrolysis is removed by a class II AP endonuclease, preferably by Exonuclease III or Endonuclease IV. 12. The method of claim 4 wherein in step (4) the 3′OH-group at a nick or gap resulting from the enzymatic reactions is extended with a template directed polymerase reaction with or without the incorporation of additional marker nucleotides, preferably (i) strands containing 5′-dRp group resulting from the action of a class II AP endonuclease are bound with a surplus of the corresponding template strands and the 3′-group is extended with a template directed polymerase ; and/or (ii) the 3′OH-group of the nick is template directed extended with a polymerase showing strong strand displacement properties; and/or (iii) the 3′OH-group of the nick or gap is extended by a template directed polymerase showing a 5′3′-exonuclease activity or with other template directed polymerases in combination with an additional 5′3′-exonuclease. 13. The method of claim 1, wherein the template strands in step (4) at which the template-directed single-strand synthesis takes place are RNA molecules, whereby an RNA-dependent DNA polymerase, preferably AMV reverse transcriptase from the avian myeloblastosis virus, HIV reverse transcriptase from the human immunodeficiency virus or MMLV reverse transcriptase from the Moloney murine leukemia virus are used for the template-directed single-strand synthesis. 14. A kit for carrying out the method as defined in claim 1, preferably said kit containing of the following components: (i) marker nucleotides for incorporation in the polynucleotide molecules; (ii) agents permitting the single-stranded breaks at the incorporated marker nucleotides; and (iii) buffers for carrying out the incorporation of the marker nucleotides and producing the single-stranded breaks at these sites. |
<SOH> SUMMARY OF THE INVENTION <EOH>The technical problem has been solved by providing the embodiments characterized in the claims. The present invention thus provides (A) a method for the production of polynucleotide molecules with modified properties, comprising the following steps: (1) providing a population of source nucleic acid molecules, the individual nucleic acid molecules of said population having homologous and heterologous segments and having at least one marker nucleotide incorporated within its nucleic acid sequence; (2) forming double-stranded polynucleotide molecules of the population of source nucleic acid molecules provided according to step (1) comprising double strands with heterologous segments (heteroduplices); (3) producing single-stranded breaks at the incorporated marker nucleotides of the double-stranded heteroduplices produced according to step (2); and (4) performing template-directed single-strand synthesis, with or without incorporation of marker nucleotides starting from single-stranded breaks produced according to step (3); and (B) a kit for carrying out the method as defined in (A) above, preferably said kit containing at least one of the following components: (i) marker nucleotides for incorporation in the polynucleotide molecules; (ii) agents permitting the single-stranded breaks at the incorporated marker nucleotides; and (iii) buffers for carrying out the incorporation of the marker nucleotides and producing the single-stranded breaks at these sites. In the method of embodiment (A) of the invention, in step (1)—if the source nucleic acid molecule is double stranded—the strands may be complementary or partially complementary. Moreover, steps (3)-(4) may be carried out subsequently or contemporaneously. The following figures further explain the embodiments of the invention. The figures are, however, not to be construed to limit the invention. |
Electronic apparatus, page display method, program, and storage medium |
An electronic apparatus capable of displaying linked pages in turn by an easy operation for users is provided. In a <body> element in an SMIL document, the attribute ??dur=t is added to URI description for specifying a media element. The resulting notation implies that a file name placed before ?? indicates a designated page and a link destination of the designated page is displayed. A numerical value following dur= indicates the duration for which a given link contained in the designated page can be activated. An SMIL player interprets an SMIL document having such a description, so as to allow auto-linking to the link destinations of the designated page with the defined link intervals. |
1. An electronic apparatus comprising: a display unit having a display screen; means for reading a program including at least allocation information for defining a display region to be allocated in the display screen, page specification information for specifying a desired page containing one or more links, and information regarding linking of the specified page to linked pages; means for configuring the display region in the display screen based on the allocation information of the read program; means for obtaining the specified page based on the page specification information of the read program; and linking means for sequentially obtaining the linked pages of the specified page based on the linking information of the read program and displaying the obtained pages in the display region configured in the display screen. 2. An electronic apparatus according to claim 1, wherein the linking information includes information for defining a link time interval, and the linking means allows the linked pages displayed in the display region to be changed based on the link time interval. 3. An electronic apparatus according to claim 2, wherein the linking information further includes information for defining a link range, and the linking means allows the linked pages displayed in the display region to be changed within the link range. 4. An electronic apparatus according to claim 3, wherein the link range includes a hierarchical link depth. 5. An electronic apparatus according to claim 2, wherein the linking information further includes information for defining the number of linked pages to be concurrently displayed, and the linking means allows the defined number of linked pages to be concurrently displayed in different display regions which are configured in the display screen. 6. A page display method in which a display unit having a display screen, a control unit, and a storage unit are provided, wherein the control unit causes: a program to be read and stored in the storage unit, the program including at least allocation information for defining a display region to be allocated in the display screen, page specification information for specifying a desired page containing one or more links, and information regarding linking of the specified page to linked pages; the display region to be configured in the display screen based on the allocation information of the program stored in the storage unit; the specified page to be obtained based on the page specification information of the program stored in the storage unit; and the linked pages of the specified page to be sequentially obtained based on the linking information of the program stored in the storage unit and to be displayed in the display region configured in the display screen. 7. A page display method according to claim 6, wherein the linking information includes information for defining a link time interval, and the control unit causes the linked pages displayed in the display region to be changed at the link time interval. 8. A page display method according to claim 7, wherein the linking information further includes information for defining a link range, and the control unit causes the linked pages displayed in the display region to be changed within the link range. 9. A page display method according to claim 8, wherein the link range includes a hierarchical link depth. 10. A page display method according to claim 7, wherein the linking information further includes information for defining the number of linked pages to be concurrently displayed, and the control unit causes the defined number of linked pages to be concurrently displayed in different display regions which are configured in the display screen. 11. A program including: allocation information for defining a display region to be allocated in a display screen; page specification information for specifying a desired page containing one or more links; and information regarding linking of the specified page to linked pages. 12. A program according to claim 11, wherein the linking information includes information for defining a link time interval. 13. A program according to claim 12, wherein the linking information further includes information for defining a link range. 14. A program according to claim 13, wherein the linking range includes a hierarchical link depth. 15. A program according claim 12, wherein the linking information further includes information for defining the number of linked pages to be concurrently displayed. 16. A program according to claim 11, wherein the program is described in an extensible markup language. 17. A storage medium having a program stored therein, the program including: allocation information for defining a display region to be allocated in a display screen; page specification information for specifying a desired page containing one or more links; and information regarding linking of the specified page to a linked page. |
<SOH> BACKGROUND ART <EOH>In pages created in a markup language such as HTML (Hyper Text Markup Language), hyperlinks allow users to jump from a currently viewed page to pages associated with desired objects in this page. The hyperlinks are generally triggered by an active operation of a reader, such as by the reader clicking a mouse on a desired object. When a reader wants to successively view a plurality of pages linked to a given page, the reader must specify link destinations on the link source page in turn. A known technique for displaying linked pages without being specified by a reader is described in, for example, Japanese Unexamined Patent Application Publication No. 10-40062. In this technique, a page is parsed to find out page addresses linked from this page, and the page addresses are used to obtain and concurrently display one or a plurality of linked pages. However, this technique does not allow the linked pages to be displayed in order over time. The pages cannot be displayed with predetermined time intervals, either. Another problem with the active operation of readers required for obtaining linked pages is that the latest information of a page whose content is frequently updated, such as news, can be missing if the page content is updated on a server. In view of such situations, it is an object of the present invention to provide an electronic apparatus capable of displaying linked pages in turn by an easy operation for users, a page display method, a program, and a storage medium. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIGS. 1A and 1B are illustrations of a television set 100 according to an embodiment of the present invention. FIG. 2 is a block diagram showing the structure of the television set shown in FIG. 1 . FIG. 3 is a diagram showing the structure of an SMIL document processing unit (SMIL player). FIG. 4 is a view showing an example of extended SMIL notation. FIG. 5 is a view showing an example of linking between a plurality of pages. FIG. 6 is a view showing an example of page presentation of the SMIL notation shown in FIG. 4 . FIG. 7 is a view showing an example of page presentation including a designated page and a plurality of pages linked from the designated page. FIG. 8 is a view showing an example of page presentation including a designated page and a plurality of pages hierarchically linked from the designated page. FIG. 9 is a view showing an example of SMIL notation which realizes the presentation shown in FIG. 7 . FIG. 10 is a view showing an example of SMIL notation which realizes the presentation shown in FIG. 8 . FIG. 11 is a view showing another example of SMIL notation which realizes the presentation shown in FIG. 8 . FIG. 12 is a flowchart of a procedure for creating an extended SMIL document. FIG. 13 is a flowchart showing how an SMIL player processes an SMIL document. FIG. 14 is a flowchart showing how an extended SMIL document is parsed. FIG. 15 is a flowchart of a timer interrupt. FIG. 16 is a view showing a URL example shown in a status bar. FIG. 17 is a flowchart of an interrupt induced by an event where a file name or URL shown in the status bar changes. FIG. 18 is a flowchart of an interrupt induced by completely reading received data. detailed-description description="Detailed Description" end="lead"? |
Method of screening pigs for resistance to salmonellosis |
The present invention relates to methods for assessing an animal's susceptibility to bacterial infection. Methods for selecting suitable breeding animals are also provided. |
1. a method for assessing an animal's susceptibility to bacterial infection, which comprises measuring at least one immune function of the animal: 2. A method as claimed in claim 1, wherein the immune function is measured prior to infection. 3. A method as claimed in claim 2 wherein the method comprises determining the intrinsic activity of non-stimulated polymorphonuclear neutrophils (PMNs). 4. A method as claimed in claim 2 wherein the method comprises measuring lymphocyte proliferation following appropriate stimulation with a mitogen. 5. A method as claimed in claim 3 wherein the intrinsic activity is measured by means of measuring oxidative burst response. 6. A method as claimed in claim 4 wherein lymphocyte proliferation is measured following stimulation by Concavalin A (Con A). 7. A method as claimed in claim 1, wherein the immune function is measured post infection. 8. A method as claimed in claim 7, which comprises determining the number of neutrophils in a sample from the animal. 9. A method as claimed in claim 7, wherein the method comprises measuring the response to anti-LPS antibody. 10. A method as claimed in claim 7, wherein the method comprises monitoring the pyrexic response. 11. A method as claimed in any one of claims 1 to 10 wherein the assessment is carried out in respect of Salmonella infection. 12. A method as claimed in claim 1 wherein the assessment is carried out on a blood sample obtained from the animal. 13. A method of selecting animals for suitability as breeding animals which comprises carrying out a method as defined in claim 1. 14. A method as claimed in any one of claim 1 wherein the animal is a mammal. 15. A method as claimed in claim 1 wherein the animal is a pig. |
Oscillating-rotary engine |
This invention refers to an oscillating-rotary engine with internal combustion in four strokes and it is made from a block-engine with a single cylinder 1, tightened by to lids 2 and 3, the frontal lid 2 has two orifices 4 and 5 for the inlet and outlet galleries and a spark-plug 6; inside the cylinder, in two bearings 7 and 8 there is an oscillating-rotor unit consisting of a special central hub 9 and two pairs of blades (A-B, C-D), which produces mechanical work under the form of an oscillating rotation; this mechanical work is changed into a continuous rotation by a special gear-group that is placed in case 12; the rotation is transmitted to a shaft-engine 13. |
1. The oscillating-rotary internal combustion engine is characterized by the fact that for the scope to obtain a simple construction and high efficiency it is made up of a stator unit consisting of a tightened cylinder (1) closed by two lids (2, 3), of a front lid (3) with two orifices (4, 5) for the inlet and outlet gallery (4, 5) and a spark-plug (6): inside the cylinder (1), sustained by two bearings (7, 8) set in the center of the lids, there is an oscillating-rotor unit, balanced from centrifugal power point of view, made from two pairs of blades (A-B, C-D), on which edges are set segments (S) which assure the tightness, the blades are set on a central hub (9), its rotation axle is the same with the geometrical point (0) of the two lids 12, 3), from this hub there are going out two concentric shafts (10, 11) and that are thus designed that when the shaft (10) towards the other shaft (11) a pair of blades (A-B) are allowed to rotate independently towards the other pair of blades (C-D), in this way there are created four distinct rooms inside the cylinder (1), between the lids (2, 3) and the surface of the blades (A-B, C-D); during the working of the engine in the inside the rooms there is developed, through the alternative rotation and an 180° anti-phase of the blades (A-B, C-D) a shifting with a volume variation of all rooms thus, inside every room, at a shifting of 360° of the two pair of blades (A-B, C-D), which have also the role of permitting the communication of a certain room through the two windows (4, 5) with a certain inlet gallery or outlet gallery at pre established moments and by the positioning of the windows on the front lid (2) and thus the four strokes are realized (admission, compression, combustion and evacuation) of an internal combustion engine and due to the fact there are four rooms, every moment there will exist an active phase (combustion), which through the pressure caused by the burnt fuel-mixture lighted by the spark-plug (6) set on the front lid (2), will drive the pairs of blades (A-B, C-D) with a power-couple with the help of a gear-group (coupling group) built in two variants: the model “reversible lever” and the model “variable driving” from a protector case (12); the model “reversible lever” works on the principle of levers with variable positions of the points where both the active force and the resistance force act with a variable support point, which turns reversible from saving-movement levers into saving-power levers and produce a one-way rotation movement and will turn the oscillation movement of the two concentric shafts (10, 11) into a uniform rotation movement which is maintained inertial by the whole system and having also a role of a fly-weel: it is made from a shaft-engine (13) which is set in the center of the case (12) with the help of a bearings group (14) and on which there is set a rotating arm (15) which has a counter-weight (16) on one edge and on the other edge a short shaft (17) which through the agency of a bearing (18) sustains a rack wheel (19) which has a number “n” of teeth and rotates inside the case on a toothed-crown (20) which has a number of teeth “2n” set inside the case (12), on the rack-wheel (19) there is set a disk (21) having in two opposite points two bolts (22) which are driven by the help of two pieces named “skates” (23) by two arms with counter-weights (24, 25) which arms have a groove where the pieces named “skates” (23) are sliding fixed to the two concentric shafts. 2. The oscillating-rotary engine, according to claim 1, characterized by the fact that the blades (A-S, C-D), which modify the volume of the four rooms and determine the volumetric proportion, can be made in such a size and shape, that a big volumetric proportion will result and this way the engine will function by ignition compresion (Diesel). 3. The oscillating rotary engine according to claims 1 and 2, characterized by the fact that it has on the contact surface of the blades (A-B, C-D) segments for tightness, lubrication is made by oil, which is poured inside the blades and if there is a circulation of oil inside the blades and an external radiator a good cooling of the entire oscillating-rotor unit will exist. 4. The oscillating-rotary engine, as claim 3 says, is characterized by the fact that oscillating-rotor unit does not touch the cylinder or the lids of the stator; only the rectangular segments touch and are pressed by flexible bands with spring effect whose length doesn't change while functioning; segments are made from porous steel or other self-lubricants, being wide enough so that they needn't be replaced repeatedly, the engine can function a long time without modifying its characteristics or the level of the polluting gases. 5. As claim 1 says, the stator unit is made of double walls; and in the empty space between them the cooling liquid will circulate achieving the cooling of the engine. 6. According to claim 1 the lubrication system of the whole gear-group (coupling-group) is made by introducing oil to an established level in the protector case (12) and by stirring up the oil through the agency of the rack wheel (19) and the two arms (14, 25). 7. According to claim 1, the orifices of the inlet and outlet galleries (4, 5) and the spark-plug (6) are set on the front lid (2); at the high revolution engines, the orifices will be replaced by slots or more orifices placed on the length of the cylinder and the spark-plug will be replaced ‘by a row’ of spark-plugs placed at the same-distance from each other and on the whole length of the cylinder to achieve a complete burning of the air-fuel mixture, in different places assuring this way a complete burning; there will no longer be unburnt gases in the evacuated gases. 8. On the cylinder (1), for cooling the stator unit, there will be set: a few small wings with thermal radiation effect; the air-flow from: a blower will be controlled by an electronical device set on the cylinder. 9. The oscillating-rotary engine acording to claim 1, is characterized by the fact that for obtaining a simple construction, high efficiency and a long liability, the coupling-group unit model “variable driving”—which works on the principle of transmission without slide of a couple of forces having the transmission ratio of periodic variable, is built from a protective oval case (26) which sustains by the help of a bearings-group (27) the engine-shaft (28) in which exist two jointly fixed rack wheels (29 and 30) with form of elliptical cylinder having the correspondents radii in perpedincular position; these rack wheels have a role of fly-wheel and are put to work by two other rack wheels (31 and 32) fixed independently in the two concentric shafts (10 and 11) of oscillating-rotor unit; these rack wheels have the same geometrical form as the rack wheels (29 and 30) fixed an the engine-shaft (28) but are made of light materials or they are hollow and their resistance is ensured by spokes in the interior with a view to keep inertia, as low as possible, the whole gear-group being attached with the stator unit through an adapting plate (33). 10. This type of oscillating-rotary engine with internal combustion in four strokes may to be used to equip flying and nautical devices (ships, boats) transport vehicles (lorries, cars, buses, motorcycles), agricultural machines (all types of tractors and combines) and for all types of machines which work by mechanical driving, depending on the construction mode: sparkling or compresion. 11. The four stroke rotary-oscillating engine with internal combustion is, according to claim 1, characterized through the fact that—taking into account its working principle—it may be adapted constructively to fulfill the role of a compressor or a volumetric pump. |
Shaping method and drum for manufacturing a tyre for vehicle wheels |
A method for building a tyre for a vehicle wheel includes positioning a casing sleeve on a shaping drum and securing the sleeve on the drum. During at least one of positioning and securing the sleeve, the sleeve is kept in an essentially centered position on the drum by radial forces directed against a radially inner surface of the sleeve. A shaping drum for making a tyre for a vehicle wheel includes a rotation shaft and two coaxial half-drums integral with each other with respect to rotation. At least one of the half-drums is axially movable in both directions with respect to the other half-drum. Each half-drum includes a support device, a gripping device, and a centering device. A first equatorial plane defined the centering devices is axially movable with respect to a second equatorial plane defined by the gripping devices. A plant for building a tyre is also disclosed. |
1-21. (canceled) 22: A method for building a tyre for a vehicle wheel, comprising: positioning a casing sleeve on a shaping drum; securing the casing sleeve on the shaping drum; toroidally shaping the casing sleeve; applying a pair of sidewalls to lateral surfaces of the shaped casing sleeve; and applying an outer sleeve to a radially outer surface of the casing sleeve; wherein during at least one of toroidally shaping the casing sleeve, applying the pair of sidewalls, and applying the outer sleeve, the casing sleeve is kept in an essentially centered position on the shaping drum by axial forces directed against axially inner surfaces of the casing sleeve, and wherein during at least one of positioning the casing sleeve and securing the casing sleeve, the casing sleeve is kept in an essentially centered position on the shaping drum by radial forces directed against a radially inner surface of the casing sleeve. 23: The method of claim 22, further comprising: positioning the sidewalls on the shaping drum; wherein the sidewalls are positioned with axially inner edges of the sidewalls in axially inner positions with respect to beads of the casing sleeve. 24: The method of claim 23, further comprising: turning the axially inner edges of the sidewalls onto axially inner surfaces of the beads. 25: The method of claim 24, wherein turning the axially inner edges takes place before toroidally shaping the casing sleeve. 26: The method of claim 24, wherein turning the axially inner edges takes place after toroidally shaping the casing sleeve. 27: The method of claim 24, wherein turning the axially inner edges is carried out by inflating at least one inflatable chamber. 28: The method of claim 24, wherein turning the axially inner edges is carried out using turning devices made to act against radially inner surfaces of the axially inner edges. 29: The method of claim 28, wherein the turning devices are used as elements for maintaining the essentially centered position of the casing sleeve during applying the outer sleeve. 30: The method of claim 23, wherein positioning the sidewalls takes place before positioning the casing sleeve. 31: The method of claim 23, wherein positioning the sidewalls takes place after positioning the casing sleeve. 32: The method of claim 23, wherein for each sidewall, positioning the sidewalls comprises: securing one end of a sidewall strip on a surface of the shaping drum before the strip is wound around the shaping drum. 33: A shaping drum for making a tyre for a vehicle wheel, comprising: a rotation shaft; and two coaxial half-drums; wherein the rotation shaft is connected for operation to the half-drums, wherein the half-drums are integral with each other with respect to rotation, wherein at least one of the half-drums is axially movable in both directions with respect to the other half-drum, wherein each half-drum comprises: a support device; a gripping device; and a centering device; wherein the support device positions and applies a sidewall on a casing sleeve, wherein the gripping device secures one axially opposed bead of the casing sleeve on the half-drum, wherein the centering device positions one of the axially opposed beads on the half-drum, and wherein a first equatorial plane defined by the centering devices is axially movable with respect to a second equatorial plane defined by the gripping devices. 34: The shaping drum of claim 33, wherein the gripping devices comprise first and second guides that delimit a fixed path for a moving element. 35: The shaping drum of claim 34, wherein the centering devices comprise a mechanism that, when operated, forces the moving element to move along the fixed path, thus varying the axial centering position of the casing sleeve with respect to the equatorial plane of the shaping drum. 36: The shaping drum of claim 33, further comprising: turning devices for assembling the axially inner edges of the sidewalls onto corresponding surface portions of the casing sleeve. 37: The shaping drum of claim 33, further comprising securing devices for securing the sidewalls to the shaping drum during winding of sidewall strips onto the shaping drum. 38: A method for building a tyre for a vehicle wheel, comprising: positioning a casing sleeve on a shaping drum; and securing the casing sleeve on the shaping drum; wherein during at least one of positioning the casing sleeve and securing the casing sleeve, the casing sleeve is kept in an essentially centered position on the shaping drum by radial forces directed against a radially inner surface of the casing sleeve. 39: A method for building a tyre for a vehicle wheel, comprising: positioning a casing sleeve on a shaping drum; securing the casing sleeve on the shaping drum; positioning a pair of sidewalls on the shaping drum; and turning axially inner edges of the sidewalls onto axially inner surfaces of beads of the casing sleeve; wherein the sidewalls are positioned with the axially inner edges of the sidewalls in axially inner positions with respect to the beads of the casing sleeve. 40: A method for building a tyre for a vehicle wheel, comprising: positioning a casing sleeve on a shaping drum; and securing the casing sleeve on the shaping drum; wherein the shaping drum comprises centering devices for the casing sleeve, wherein the shaping drum comprises gripping devices for the casing sleeve, wherein the casing sleeve is secured on the shaping drum using the gripping devices, wherein the casing sleeve is secured on the shaping drum with an equatorial plane of the casing sleeve being different from an equatorial plane of the shaping drum, wherein the equatorial plane of the casing sleeve is defined by a position of the centering devices, and wherein the equatorial plane of the shaping drum is defined by the gripping devices. 41: A plant for building a tyre for a vehicle wheel, comprising: a first-step building drum; a device for transferring a first-step sleeve; at least one shaping drum; an auxiliary drum for building an outer sleeve; a device for transferring the outer sleeve; and a feed device provided next to the at least one shaping drum; wherein the at least one shaping drum is supported by a motor unit, wherein the at least one shaping drum projects from one side of the motor unit, wherein the outer sleeve comprises: a belt structure; and a tread; wherein the feed device is coaxial with the building drum, wherein the at least one shaping drum comprises: a rotation shaft; and two coaxial half-drums; wherein the rotation shaft is connected for operation to the half-drums, wherein the half-drums are integral with each other with respect to rotation, wherein at least one of the half-drums is axially movable in both directions with respect to the other half-drum, wherein each half-drum comprises: a support device; a gripping device; and a centering device; wherein the support device positions and applies a sidewall on the first-step sleeve, wherein the gripping device secures one axially opposed bead of the first-step sleeve on the half-drum, wherein the centering device positions one of the axially opposed beads on the half-drum, and wherein a first equatorial plane defined by the centering devices is axially movable with respect to a second equatorial plane defined by the gripping devices. 42: The plant of claim 41, further comprising: a pair of shaping drums; wherein the pair of shaping drums are supported by the motor unit, wherein the pair of shaping drums project from opposite sides of the motor unit, and wherein the motor unit is mounted on a rotatable turret designed to change positioning of the pair of shaping drums with respect to each other. |
Cool oxygen chemical gas generator |
The present invention is directed to a chemical oxygen generator to produce cool oxygen gas comprising: a. a charge housing, b. a solid but porous charge contained in the said housing, the charge being made of a chemical mixture that generates oxygen upon decomposition and that will undergo a self-sustained exothermal decomposition after initiation, the said charge containing at most 3.0 wt. % of binder material, the said porous charge allows the generated oxygen to pass through the charge without damaging the virgin material and without creating volumetric burning, the said charge is mounted in the housing in such a way that the generated oxygen passes through the charge and under the pressure difference flows from the moving decomposition front towards the vent, c. an ignition device mounted at one end of the cartridge in such a way that it is capable to initiate a self-sustained decomposition of the charge at the charge surface adjacent to the initiator, d. one or more vents mounted in such a way that the generated oxygen that has passed through the generating porous charge leave the gas generator through the said vents. |
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