An ATV receiver is provided with a 16-state trellis decoder to achieve improved performance in the presence of NTSC co-channel interference. In one embodiment, the ATV receiver comprises a tuner, a comb filter, and a trellis decoder. The tuner is configured to downmix a selected channel frequency signal to an intermediate frequency signal, where the comb filter is configurable to screen out most of the NTSC co-channel interference. The intermediate frequency receive signal is modified by the comb filter to resemble a partial response signal. The trellis decoder the demodulates the partial response signal in an improved fashion taking into account the state of the trellis encoder and the partial response channel. The trellis decoder may have a 16 state trellis comprised of four 4-state butterflies wherein each edge in the trellis is a single transition.

1. Field of the Invention
This invention relates to an apparatus for receiving and decoding a digital video broadcast signal. More specifically, this invention relates to a device with a new trellis code modulation decoder structure which demonstrates an improved performance in the presence of NTSC co-channel interference.
2. Description of the Related Art
A Digital Television System (DTS) standard was recently prepared by the Advanced Television Systems Committee (ATSC). The DTS standard outlines various system characteristics of the Advanced Television (ATV) system proposed for use in the U.S., and Annex D in particular specifies the Radio Frequency (RF) transmission subsystem for the DTS standard. The RF transmission subsystem performs amplitude and vestigial sideband modulation (VSB) in two modes: a terrestrial mode (8-level VSB, or simply 8 VSB)) and a high data rate mode (16-level VSB, or simply 16 VSB). Since no trellis code modulation is employed in the 16 VSB mode, the discussion hereafter will focus on the 8 VSB mode.
Turning now to the figures, FIG. 1 shows one embodiment of a ATV transmitter 100. The ATV transmitter comprises a data randomizer 102, a Reed-Solomon encoder 104, an interleaver 106, a trellis encoder 108, a sync word insertion module 110, a pilot insertion module 112, a VSB modulator 114, and a radio-frequency (RF) up-converter 116. The ATV transmitter 100 receives a digital audiovisual signal, e.g. an MPEG bitstream, in the form of 188-byte data packets. The first byte is a synchronization byte, and the remaining 187 bytes are payload data.
Data randomizer 102 drops the synchronization byte and changes each remaining byte value according to a known pattern of pseudo-random number generation in order to eliminate repetitious patterns and provide the data with a completely random noise-like character. Reed-Solomon encoder 104 encodes the 187 randomized bytes to add 20 redundancy bytes to enable future error correction of up to 10 byte errors. Interleaver 106 re-orders the encoded bytes to intermix the byes from different packets and thereby provide resistance to burst errors (since all the bytes for a given packet are no longer located in a short time interval). Trellis encoder 108 provides further encoding and modulation, and is discussed further below. At the trellis encoder output, 328symbols are produced for every 187 input bytes. Sync word insertion module 110 pre-pends a 4-symbol segment synchronization word to each group of 328 symbols to form a 332 symbol data segment, and further inserts a field synchronization segment for every set of 312 data segments to form a 313 segment data field. Pilot insertion module 112 provides a DC offset to all the symbols. The DC offset will appear as a carrier tone in the modulated signal. VSB modulator 114 modulates the symbols from module 112 onto an intermediate frequency (IF) carrier in 8-level amplitude-modulated vestigial-sideband form, and RF up-converter 116 moves the IF signal into the assigned frequency channel, and amplifies and filters the output signal before supplying it to a transmit antenna.
The trellis encoder 108 actually consists of 12 identical trellis encoders 202-224 as shown in FIG. 2. The bytes coming into the trellis encoder 108 are divided into two-bit symbols, and each trellis encoder 202-224 receives every 12th two-bit symbol, i.e. encoder 202 operates on the 0th, 12th, 24th, 36th, . . . two-bit symbols, encoder 204 operates on the 1st, 13th, 25th, 37th, . . . two-bit symbols, and so on. At the output of the encoders 202-224, a three-bit symbol is provided for each two-bit input symbol. During insertion of the segment synchronization byte, the output multiplexer of FIG. 2 is advanced by four symbols, although the state of the encoders is not advanced.
FIG. 3 shows one embodiment of trellis encoder 202. The most-significant bit (MSB) of the two-bit word is "precoded" using delay element 302 and XOR gate 304 to produce mapper input Z.sub.2. The least-significant bit (LSB) is convolutionally encoded using delay elements 306, 308 and XOR gate 310 to produce mapper inputs Z.sub.1, Z.sub.0. The three mapper inputs may be converted to a signal amplitude immediately by a mapper 312, but typically Z.sub.2, Z.sub.1, and Z.sub.0 are simply forwarded as a three bit symbol and converted later by VSB modulator 114.
FIG. 4 shows a second embodiment of trellis encoder 202. In this embodiment, the convolutional encoder formed by delay elements 406, 408 and XOR gate 410, is a moving-window encoder rather than a feedback encoder. The correspondence between input bits and output sequences is somewhat altered, but the overall code properties are the same. Either encoder may be employed.
During the transition period from the National Television Standards Committee (NTSC) standard to the DTS standard, many ATV transmissions will take place in channels which are shared by NTSC transmissions in neighboring broadcast regions. It is desirable to provide a system with increased immunity to co-channel interference from these NTSC transmissions.
FIG. 5 shows one embodiment of an ATV receiver 500 which includes provisions for screening out interfering NTSC transmissions. Receiver 500 comprises tuner 502, IF module 504, NTSC rejection filter 506, equalizer 508, phase tracker 510, trellis decoder 512, de-interleaver 514, Reed-Solomon decoder 516, and de-randomizer 518.
Tuner 502 receives all the signals in a designated frequency band from the receiver antenna, and downmixes a selected channel to IF (at 44 MHz). IF module 504 filters out undesired adjacent channels using a square-root raised-cosine bandpass filter, and locks on to the carrier tone using a narrowband frequency-and-phase locked loop (FPLL). NTSC rejection filter 506 is a "comb" filter with nulls near the standard NTSC luminance, color, and audio carrier frequencies to screen out interference from NTSC transmissions. Filter 506 may be switched in or out of the processing pipeline as needed.
Equalizer 508 is a moving-window adaptive equalizer that operates to remove any linear distortions (e.g. spectrum tilt, multi-path echo) from the received signal and thereby maximize the "eye openings" in the equalized signal. Phase tracker 510 is a wide-band first-order tracking loop that removes any remaining phase noise not tracked by the FPLL carrier recovery loop. The phase tracker 510 operates independently of the preceding modules.
Trellis decoder 512 operates according to the Viterbi algorithm to demodulate the data, in a manner discussed further below. De-interleaver 514 reverses the operation of interleaver 106 to gather the dispersed bytes from Reed-Solomon encoded packets back together, and Reed-Solomon decoder 516 decodes the packets, providing error correction as needed. The de-randomizer then reverses the operation of randomizer 102 to produce the original data packets.
FIG. 6 shows that trellis decoder 512 is actually comprised of 12 identical trellis decoders 602-624. The trellis decoders each process every 12th signal sample and form a two-bit symbol decision for each signal sample. To drop the synchronization bytes, the input multiplexer continues to advance during synchronization intervals, but the processing by the trellis decoders 602-624 is frozen. The two-bit symbol decisions are re-assembled into bytes by the output multiplexer before being forwarded to the de-interleaver 514.
FIG. 7 illustrates the modes of operation of the trellis decoder 602. Trellis decoder 602 implements a 4-state trellis decoder 702 when the NTSC rejection filter is not used. When the NTSC rejection filter is used, an 8-state trellis decoder 704 is implemented. The effect of the comb filter is indicated by the presence of the delay element 706 and sum element 708, which adds additional state dependence to valid received signal sequences. The delay element 706 and summer 708 are shown for explanatory purposes only and are not actually part of the trellis decoder 602. The point made here is that the 8-state trellis decoder must take into account the effect of the comb filter to operate properly. Although the comb filter reduces the NTSC interference, it also modifies the receive signal symbols. The eight-level symbols are converted to fifteen-level symbols by the "partial response" characteristic of the comb filter. This characteristic advantageously does not reduce the eye openings in the equalized data, but does amplify the white noise present in the receive signal. Consequently, a white-noise performance penalty is paid in return for removing the coherent NTSC interference, and to avoid unnecessary impairment, the comb filter is configured to be bypassed when no NTSC interference is present.
FIG. 8 illustrates an embodiment of the 4-state trellis employed by trellis decoder 702. The Viterbi algorithm tracks all the most likely received signal sequence evolutions through multiple trellis stages, and each stage is identical to that shown in FIG. 8. In one embodiment, the trellis decoder employs a Viterbi algorithm with a decoding depth of 18 stages. Each trellis stage consists of all possible code states at a given time t (802) and the transitions 804 to the possible code states at a subsequent time t+1 (806). Each of the possible code states 802 is assigned a value indicative of the probability of the most likely sequence that ends in that state. For each of the transitions 804, the receive signal is compared with an expected signal value to determine a likelihood that that transition was taken from the originating state, and this likelihood is combined with the value from the originating state to determine a potential value for the destination state. The destination state is then assigned a selected potential value from all incoming transitions which is indicative of the highest likelihood. By performing this iterative process and tracking the selected transitions, the Viterbi algorithm provides maximum-likelihood demodulation and decoding of the received signal.
The trellis code states 802, 806 are determined using the binary contents of the delay elements 306 (S.sub.0) and 308 (S.sub.1). The precoding delay element 302 does not provide for additional coding gain, and its inclusion here would require additional states, so it is ignored for the moment--the inverse precoding may be performed after the decisions made by the trellis decoder. The inputs to the trellis encoder are then I.sub.1 and Z.sub.2, and the corresponding expected receive signal value R is a function of the trellis state 802. For each state 802, ordered triplets (I.sub.1,Z.sub.2,R) are provided indicative of the expected values R for each pair of inputs I.sub.1 and Z.sub.2. Each ordered triplet corresponds to a transition from the state 802, and the ordering from left to right is selected to correspond with transitions in top-to-bottom order. It is noted that the four transitions leaving each state 802 exist in parallel pairs.
The embodiment of the decoder trellis in FIG. 8 corresponds to the feedback trellis encoder of FIG. 3. An embodiment of the decoder trellis for the moving window trellis encoder of FIG. 4 is shown in FIG. 9. Here the code states 902, 906 are determined by the binary contents of delay elements 406 (S.sub.0) and 408 (S.sub.1). As indicated by the triplets 908, the correspondence between expected results R and the transitions 904 is the same as that of FIG. 8, but the correspondence with the inputs I.sub.1 and Z.sub.2 is slightly different, and hence the decoded bits for selected transitions which are part of the most likely code sequence are different.
FIG. 10 illustrates an embodiment of the 8-state decoder trellis for trellis decoder 704. Trellis decoder 704 may have a decoding depth of 36 stages. The trellis states 1002, 1006 are determined by the binary contents of delay elements 306 (S.sub.0) and 308 (S.sub.1) and the eight-level contents of delay element 706 (D). Although this would allow for as many as 32 trellis states, the contents of D are not entirely independent of the contents of S.sub.0 and S.sub.1, which eliminates many of the possible states. Further, the remaining states may be combined in pairs to reduce the number of states without reducing the minimum distance properties of the trellis code. The increased number of states (relative to the 4-state trellis) does allow for the precoding of I.sub.2 to be accounted for in this trellis, so the triplets 1008 are now provided in (I.sub.1,I.sub.2,R') form, where R' is the expected value of the receive signal after the comb filter is taken into account.
The brackets in each state label indicate the combined values of delay element 706. The resulting trellis has six transitions 1004 leaving each state 1002 in parallel groups of three. Two transitions from each group share common information bits but have different expected signal values--the bracketed pairs in the ordered triplets 1008 indicate this sharing, but to determine the correspondence between triplets and transitions, these should be read as distinct, adjacent triplets.
The trellis in FIG. 10 is used when the transmitter uses the trellis code modulator of FIG. 3. When the transmitter uses the modulator of FIG. 4, the decoder 704 uses a trellis with the same structure but with a slightly different input bit correspondence, so that the decoded input bits for many of the transitions are different.
It is noted that the Viterbi algorithm assumes that the noise in the receive signal is uncorrelated, and it is further noted that this assumption is violated when the comb filter is employed. As a consequence, the performance (in terms of probability of decision error) of decoder 704 falls short of optimal. It is desirable to provide an ATV receiver which has improved performance in the presence of NTSC co-channel interference.

Trellis code modulation decoder structure for advanced digital television receiver


There is disclosed a replaceable inking cartridge for a printer. The inking cartridge includes an arrangement for indicating when the inking medium of the cartridge has been spent. The indicating arrangement is non-resettable so that the user is prevented from using a spent cartridge for additional printing. The indicating means is useful for signalling the user or the printer about the spent condition of the cartridge.

1. Field of the Invention
This invention relates to the art of inking cartridges.
2. Description of the Prior Art
Representative prior art inking cartridges are disclosed in U.S. Pat. Nos. 3,831,731 to Mack et al, granted Aug. 27, 1974, 3,904,015 to Boyden et al, granted Sept. 9, 1975, 3,941,231 to Matuck et al, granted Mar. 2, 1976 and 4,046,247 to La Spesa et al, granted Sept. 6, 1977. U.S. Pat. No. 3,143,963 to Schrempp, granted Aug. 11, 1964, discloses a device for locking out a printing device after a certain number of printing cycles.

Replaceable inking cartridge with depletion counter


The present invention includes well treatment fluids and methods utilizing nano-particles. An embodiment of a method of the present invention may comprise introducing a treatment fluid comprising nano-clay into a subterranean formation. The treatment fluid may be selected from the group consisting of a cement composition, a drilling fluid, a spacer fluid, and a lost circulation control composition. Another embodiment of the present invention may comprise a method of cementing. The method of cementing may comprise introducing a cement composition comprising a hydraulic cement, nano-clay, and water into a subterranean formation. The method further may comprise allowing the cement composition to set in the subterranean formation. Yet another embodiment of the present invention may comprise a treatment fluid, the treatment fluid comprising nano-clay. The treatment fluid may be selected from the group consisting of a cement composition, a drilling fluid, a spacer fluid, and a lost circulation control composition.

The present invention relates to well treatment fluids and methods utilizing nano-particles and, in certain embodiments, to well cement compositions and methods utilizing nano-clay.
In general, well treatments include a wide variety of methods that may be performed in oil, gas, geothermal and/or water wells, such as drilling, completion and workover methods. The drilling, completion and workover methods may include, but are not limited to, drilling, cementing, spacers, and lost circulation control methods. Many of these well treatments are designed to enhance and/or facilitate the recovery of desirable fluids (e.g., hydrocarbons) from a subterranean well.
In cementing methods, such as well construction and remedial cementing, well cement compositions are commonly utilized. For example, in subterranean well construction, a pipe string (e.g., casing and liners) may be run into a well bore and cemented in place using a cement composition. The process of cementing the pipe string in place is commonly referred to as "primary cementing." In a typical primary cementing method, a cement composition may be pumped into an annulus between the walls of the well bore and the exterior surface of the pipe string disposed therein. The cement composition sets in the annular space, thereby forming an annular sheath of hardened, substantially impermeable cement that supports and positions the pipe string in the well bore and bonds the exterior surface of the pipe string to the subterranean formation. Among other things, the annular sheath of set cement surrounding the pipe string functions to prevent the migration of fluids in the annulus, as well as protecting the pipe string from corrosion. Cement compositions also may be used in remedial cementing methods, such as squeeze cementing, repairing casing strings and the placement of cement plugs. In some instances, cement compositions may be used to change the direction of the well bore, for example, by drilling a pilot hole in a hardened mass of cement, commonly referred to as a "kickoff plug," placed in the well bore.
In operation, the annular sheath of cement formed between the well bore and the pipe string in primary cementing may suffer structural failure due to pipe movements which cause shear stresses to be exerted on the set cement. Such stress conditions are commonly the result of relatively high fluid pressures and/or temperatures inside the cemented pipe string during testing, perforating, fluid injection or fluid production. For example, such stress may occur in wells subjected to steam recovery or production of hot formation fluids from high-temperature formations. The high-internal pipe pressure and/or temperature can result in the expansion of the pipe string, both radially and longitudinally, which places stresses on the cement sheath causing the cement bond between the exterior surfaces of the pipe or the well bore walls, or both, to fail and thus allow leakage of formation fluids and so forth. Accordingly, it may be desirable for the cement composition utilized for cementing pipe strings in the well bores to develop high strength after setting and to have sufficient resiliency (e.g., elasticity and ductility) to resist loss of the cement bond between the exterior surfaces of the pipe or the well bore walls, or both. Also, it may be desirable for the cement composition to be able to resist cracking and/or shattering that may result from other forces on the cement sheath. For example, it may be desirable for the cement sheath to include structural characteristics that protect its structural integrity from forces associated with formation shifting, overburden pressure, subsidence, tectonic creep, pipe movements, impacts and shocks subsequently generated by drilling and other well operations.
Another problem that may be encountered in well cementing methods is the undesired gas migration from the subterranean formation into and through the cement composition. Problems with gas migration may be encountered during setting of the cement composition as it transitions from a hydraulic fluid to a solid mass. Gas migration may cause undesired flow channels to form in the cement composition that may remain in the cement composition after it has set into a hardened mass, potentially resulting in loss of zonal isolation.
Yet another problem that may be encountered in well cementing methods is associated with exposure to corrosive fluids. Examples of corrosive environments include exposure to acidic conditions either caused by actual placement of acid solutions for well treatment or in the presence of carbon dioxide (CO2). Carbon dioxide has been used for enhanced recovery methods by injecting CO2 into a permeable reservoir in order to displace oil and gas towards a producing well. Carbon dioxide sequestration activities involve placing CO2 into a reservoir for permanent storage. Upon exposure to water, the CO2 may yield carbonic acid. In addition, the carbon dioxide may also convert exposed cement surfaces to calcium carbonate, a process commonly referred to as carbonation. Calcium carbonate being acid soluble may then slowly be dissolved by the carbonic acid. Dissolution of the calcium carbonate by the carbonic acid may be more severe in a cement sheath with a higher permeability due to more flow paths for the carbonic acid into the cement sheath. To counteract problems associated with exposure to corrosive fluids, additives may often be added to a cement composition to reduce the permeability of the cement sheath. For example, latex additives have been added to reduce permeability. Reducing the water content by optimized particle packing also may reduce the permeability of the cement sheath. Reduction of the permeability of the cement sheath generally may reduce flow paths for the acid, thus reducing the exposure of the cement sheath to potentially damaging acid.

This patent is subject to a terminal disclaimer.

Cement compositions and methods utilizing nano-clay


A RAN server, a wireless communications system, and a terminal attach method are provided. The RAN server configured to centrally manage base stations of a plurality of wireless communications standards is introduced, and the RAN server is used as a central node to implement a unified interface to a core network. A base station that is of any wireless communications standard and that is managed by the RAN server may independently provide a data transmission service to a terminal, so that the wireless communications system provided in this application basically does not cause an additional latency. In addition, a TAI unrelated to a wireless communications standard of a base station is carried in an attach request message, an attach accept message, and an attach complete message that are exchanged in a terminal attach process.

An era of a new radio access (NR) technology is approaching, and the industry mainly proposes architectures of the following two wireless communications systems to address a problem of how to couple the NR technology to an existing wireless communications standard technology.
An architecture of a first wireless communications system is shown in FIG. 1, which is a scenario in which a Long Term Evolution (LTE) technology is tightly coupled to the NR technology. In FIG. 1, an eNB (evolved Node B) represents a base station in an LTE system, an EPC (Evolved Packet Core, evolved packet core) represents a core network of the LTE system, an NR node represents a base station in an NR system, and the eNB is connected to the NR node by using an existing X5 interface. The tight coupling means that the NR node has no independent core network, and the NR node needs to transmit data with the help of the eNB in the LTE system. In such a tightly-coupled architecture, user equipment (UE) cannot directly access the core network by using the NR node, and instead, the UE needs to access the core network by using the eNB, causing an additional latency.
An architecture of a second wireless communications system is shown in FIG. 2, which is a scenario in which the LTE technology is loosely coupled to the NR technology. A fifth generation mobile communications technology (5-Generation, 5G) CN (core network) in FIG. 2 represents a core network of an NR system. The loose coupling means that an eNB and an NR node each have an independent core network. In such a loosely-coupled architecture, UE needs to use signaling of an LTE wireless communications standard to access the core network by using the eNB, and the UE needs to use signaling of another new wireless communications standard to access the core network by using the NR node. As a result, signaling is not unified, increasing signaling complexity.
Therefore, a new wireless communications system is required to resolve a problem that the foregoing two wireless communications systems respectively cause an additional latency and increase signaling complexity.

RAN server, wireless communications system, and terminal attach method


The present invention relates to new DNA sequences, a method for producing new plants which contain a new DNA sequence, the coding sequence thereof being expressed after ozone induction. The invention also relates to said new plants and the use of DNA sequences to produce ozone-responsive gene expression in plants and plant cells. Moreover, it relates to a new promoter, the specificity thereof being increased by removal of the ozone response capacity thereof.

This invention relates to new DNA sequences, a method for producing new plants which contain a new DNA sequence, the coding sequence thereof being expressed after ozone induction. The invention also relates to said new plants and the use of DNA sequences to produce ozone-responsive gene expression in plants and plant cells. Moreover, it relates to a new promotor, the specificity of which is increased by removal of the ozone response capacity thereof.
The ozone concentrations in the lower troposphere above the continents of the northern hemisphere have steadily increased over the past one hundred years as a result of greater industrial activities (Volz and Kley (1988) Nature 332, 240-242). Meanwhile, ozone values reach intermittent peak concentrations of 100 nL/L up to nL/L in Europe and North America (Krupa et al. (1995) Environ. Pollut: 87, 119-126).
The phytotoxicity of the air pollutant ozone has been well tested and documented, e.g., in Heagle (1989) Annu. Rev. Phytopathol. 27, 397-423; Heath (1994) in: Alscher, Wellburn (ed) xe2x80x9cPlant responses to the gaseous environmentxe2x80x9d, pp. 121-145, Chapman and Hall, London. A decreased net photosynthesis and an increased early senescence are usually the result of such ozone impact which, consequently, results in diminished plant growth and a lower harvest yield.
Although ozone penetrates the plant cell through open stomata by means of diffusion, the ozone concentration in the intercellular spaces of the leaf is almost zero, irrespective of the environmental ozone concentration (Laisk et al. (1989) Plant Physiol. 90, 1163-1167). It is currently assumed that ozone reacts quickly with components of the cell walls and the plasmalemma and is converted into reactive oxygen species, such as peroxide-anions, hydroxyl radicals and hydrogen peroxide which were detected in ozone-treated plant material by using electron spin resonance spectroscopy (Mehlhorn et al. (1990) Physiologia Plantarum 79, 377-383). The so-called xe2x80x9coxidative burstxe2x80x9d, i.e. the fast development of a relatively high quantity of reactive oxygen species, can lead to a dramatic disturbance of the normal cell function due to alteration of the permeability of the plant membrane, inactivation of redox-sensitive proteins and increased lipid peroxidation.
Recent tests conducted on ozone-treated plants showed an increased biosynthesis of non-specific, defensive enzymes, the function of which is to protect live cells against damage due to oxidative stress (Kangasjxc3xa4rvi et al. (1994 Plant, Cell and Environment 17, 783-794). Yet the signal-transduction chain, which is responsible for the ozone-induced gene activation, which transmits to the cell core the relevant information about the formation of apoplastic, reactive oxygen species, has not been understood up to now. Various factors, such as the increase of calcium concentration in cytosol (Price et al. (1994) The Plant Cell 6, 1301-1310), the formation of salicylic acid (Klessig and Malamy (1994) Plant Mol. Biol. 26, 1439-1458) and the phytohormone jasmon acid (Farmer (1994) Plant Mol. Bio. 26 1423-1437) and ethylene (Ecker (1995) Science 268, 667-674) are currently being discussed as possible signal connections, caused by oxidative stress, which generally play a part in defence reactions of plants.
Tests conducted on ozone-gassed tobacco plants showed that ozone causes an increased expression of various disease-resistant genes, namely a few PR-(pathogenesis-related) proteins (Ernst et al. (1992) Plant Mol. Biol. 20, 673-682; Ernst et al. (1996) J. Plant Physiol. 148, 215-221; Eckey-Kaltenbach et al. (1994) Plant Physiol. 104, 67-74). These results indicate that oxidative stress, caused by ozone, influences the expression and regulation of defensive genes of plants in a similar way as that described regarding pathogenic attack. Only very limited information on cis-regulatory elements and transcription factors, which possibly play a part in the control of gene expression of non-specific defensive genes as a response to various environmental influences, is available at present (Lee et al. (1994) Eur. J. Biochem. 226, 109-114). However, based on previous results, it can be assumed that with respect to the genes coding for PR-proteins, separate or at least only partly overlapping ways of signal transduction exist (Somssich (1994) in: Nover (ed) xe2x80x9cPlant promoters and transcription factorsxe2x80x9d, pp. 163-179, Springer Publishing House, Berlin; Dolferus et al. (1994) Plant Physiol. 105, 1075-1087).
With respect to the activity of the stilbene synthase (STS), which takes part in the phytoalexin synthesis, it is known that in adult plants it is induced by environmental stress factors, such as, e.g., pathogenic attack (Langcake (1981) Physiol. Plant Pathol. 18, 213-226), ultraviolet light (Fritzemeier and Kindl (1981) Planta 151, 48-52) and ozone (Rosemann et al. (1991) Plant Physiol. 97, 1280-1286. Contrary to this a constitutive expression pattern was observed in embryos (Sparvoli et al. (1994) Plant Mol. Biol. 24, 743-755).
Stilbene synthase enzymes catalyze the synthesis of stilbenes such as resveratrol or pinosylvin from one molecule of p-cumaroyl-CoA or cinnamoyl-CoA and three units of malonyl-CoA. Resveratrol as well as pinosylvin have photoalexin properties and an antifungal activity, and perform, as phytoalexins in combination with other stilbenes derived from the phenylpropane metabolism, an important function in the defence against pathogens (Hart (1981) Annu. Rev. Phytopathol. 19, 437-458).
STS genes are found in some non-related plant species such as, e.g., peanut (Schrxc3x6der et al. (1988) Eur. J. Biochem. 172, 161-169), grapevine (Hain et al. (1993) Nature 361, 153-156) and pine (Fliegmann et al. (1992) Plant Mol. Biol. 18, 489-503) and are organized in larger gene families, comprising six or more genes (Lanz et al. (1990) Planta 181, 169-175; Wiese et al. (1994) Plant Mol. Biol. 26, 667-677).
Experiments with transgenic tobacco cells indicate that the expression of the stilbene synthase is regulated mainly at a transcription level, and that the stress-induced signal transduction chain has been preserved in various plant species during the course of evolution (Hain et al. (1990) Plant Mol. Biol.15, 325-335.
STS genes from peanut (Arachis hypogaea) and grapevine (Vitis vinifera) have already been isolated (Schrxc3x6der et al. (1988) supra or Hain et al. (1993) supra) and expressed in transgenic plants (Hain et al. (1990) supra or Hain et al. (1993) supra).
DNA sequences coding for stilbene synthase are known, e.g., from European Patent EP 0 309 862, German Patent Application DE-A-41 07 396, European Patent Application 0 464 461, as well as U.S. Pat. No. 5,500,367. These documents describe the isolation of stilbene-synthase genes and their use to produce transgenic plants. The resulting transgenic plants show greater resistance to various plant pests such as fungi, bacteria, insects, viruses and nematodes. Plasmids containing STS genes have been deposited with the German Collection of Microorganisms (DSM), Mascheroder Weg 1B, D-38124 Braunschweig. Also included in the deposition is the VstI-gene from grapevine in the pVstI plasmid, under deposit number DSM 6002 (DE-A-41 07 396, EP-A-0 464 461, U.S. Pat. No. 5,500,367).
While in the meantime also the use of STS coding sequences to produce male, sterile plants and altered blossom colour has been described (German Patent Application DE-A-44 40 200), a possible relation between STS gene expression and ozone induction has remained completely unexplored up to now.
Meanwhile there are various indications that in order to produce an effective as possible resistance to disease, based on the expression of STS genes in transgenic plants, it is advantageous if the expression of the heterologous STS gene (or the heterologous STS genes) in the plant is stimulated first of all by the attacking pathogen, i.e. if it is stimulated first of all by the interaction of plant and pathogen (Fischer and Hain (1994) Current Opinion in Biotechnology 5, 125-130; Fischer (1994) xe2x80x9cOptimization of the heterological Expression of Stilbene-synthase Genes for the Protection of Plantsxe2x80x9d, Hohenheim University). This is endorsed particularly by the observation that the pathogen-induced STS gene expression is locally limited to the place of infection and is of a transient nature, which means that the STS expression rises relatively fast to a maximum and declines again in less than 48 hours (Hain et al. (1993) supra). Also experiments conducted on transgenic tobacco plants, in which STS genes were expressed under the constitutive 35S RNA promotor of cauliflower mosaic virus, showed that the resistance to disease achieved in said plants is lower than in plants which expressed the identical genes under the control of the pathogen-inducible homologous STS promotor after fungus infection (Fischer and Hain (1994) supra; Fischer (1994) supra). Anyway, it is desirable that the STS expression in transgenic, cultivated plants which, due to the inserted STS genes show greater resistance to disease, are activated and controlled solely (and not until) by the attack of pathogens and not additionally (or already before) by undesirable environmental stress factors such as ozone.
Thus, it is an important task of biotechnological research of plant protection to realize a more specific expression of defensive genes in plants, in order to be able to materialize molecular biological strategies for producing plants of greater resistance in an efficient and controllable manner. An important aspect in doing so is to eliminate undesirable, non-specific environmental stimuli such as, e.g., the induction of certain defensive genes through ozone, ultraviolet light, heavy metals, extreme temperatures and other abiotic stimuli.
Thus, it is an important object of this invention to make available new DNA sequences which play a direct part in the ozone-induced expression of resistance genes.
Another object of the invention is to show possibilities for removing the ozone induction, i.e. to eliminate undesirable stimulation of the gene expression through ozone.
Furthermore, an important object of this invention is to provide a DNA sequence with the help of which stilbene-synthase genes can be expressed in transgenic plants only after contact of the plant with the pathogen and not through ozone stimulation.
As mentioned already at the outset, a steady increase in ozone impact can be observed which also has drastic effects on vegetation. The observation and determination of ozone concentrations in the air constitute already today a focal point of chemical, physical and biological, environmental research. An important instrument in this context are the so-called biomonitors, with the help of which ozone impact and the consequences thereof, particularly phytotoxic effects can be easily determined both qualitatively and quantitatively.
Thus, a further object of this invention is to provide DNA sequences that can be used to produce targeted ozone-inducible promoters. With the help of such promotors it is possible for so-called reporter genes, the expression of which can be proven by simple, enzymatic tests, and which are well known in biotechnical research, to be used as biomonitors.
Another object of the invention is to provide a system with the help of which certain genes,xe2x80x94whose gene products are able to detoxify oxygen species in cellsxe2x80x94can be xe2x80x9cturned onxe2x80x9d, if necessary, as for instance in case of great ozone impact. In other words, by providing DNA sequences, which are responsible for ozone-responsive gene regulation, an ozone-inducible expression of said genes such as, e.g., catalase and/or superoxide-dismutase genes, shall be rendered possible. Thus, it is an object of the invention to make available DNA sequences which can be used for producing an ozone-inducible, cellular xe2x80x9cozone protection systemxe2x80x9d. Further objects of the invention will become apparent as the following description proceeds.
These problems are solved by the subject-matter of the independent claims, based particularly on the provision of the DNA sequences, according to the invention, which are directly involved in the ozone-induced gene expression in plants.
We found, to our surprise, that a certain plant nucleic acid sequence is directly involved in the ozone-responsive STS expression. While experiments with transgenic tobacco embryos and plantsxe2x80x94which express the customary reporter gene uidA from E. coli that codes for a xcex2-glucuronidase under the control of variously long 5xe2x80x2 deletions of the VstI-promotor from grapevinexe2x80x94indicate that at least a few cis-elements, responsible for the fungus induction, are within the range of the promotor which comprises base pairs xe2x88x92140 to xe2x88x92280 (calculated from the start of transcription) (Fischer (1994) supra), the range of the VstI sequence, which comprises base pairs xe2x88x92280 to xe2x88x92430, is essential for a strong activation of the gene expression through ozone. Based on our experiments, it was possible to show that a VstI promoter, which is left with only base pairs up to and including xe2x88x92280 (and which thus is lacking the VstI-promoter sequences, located further upstream) is no longer ozone inducible. As mentioned above, said shortened promotor is nevertheless still able to indicate pathogen-induced gene expression of the coding sequence controlled by same (see Fischer (1994) supra).
Our analyses also lead us to suspect a relation between the treatment of plants with ozone and an increased biosynthesis or release of ethylene in plant cells. Therefore, an involvement of ethylene-responsive elements in the ozone-induced gene expression cannot be excluded. Thus, by taking into consideration familiar cis-elements, which are currently being discussed in connection with ethylene-response capacity (Sessa et al. (1995) Plant Mol. Biol. 28, 145-153; Shinshi et al. (1995) Plant Mol. 27, 923-932), an involvement of the sequence range of the VstI promotor, which comprises base pairs xe2x88x92283 to xe2x88x92273, cannot be excluded in an ozone-induced STS-gene expression.
Accordingly, the ozone-responsive DNA sequence range, which is described here for the first time, comprises base pairs xe2x88x92270 to xe2x88x92430 of the VstI promotor from grapevine.
Thus, the present invention relates to the DNA sequence (SEQ ID NO:1), as defined in Claim 1:
ACTTTTCGAG CCCCTTGAAC TGGAAATTAA TACATTTTCC ACTTGACTTT TGAAAAGGAG GCAATCCCAC GGGAGGGAAG CTGCTACCAA CCTTCGTAAT GTTAATGAAA TCAAAGTCAC TCAATGTCCG AATTTCAAAC CTCANCAACC CAATAGCCAA T, which is essential for the ozone-induced gene expression in plants. A preferred version of the DNA sequence, according to the invention, deals with a DNA sequence, which originates from grapevine, and especially preferred from the stilbene synthase gene VstI from grapevine (base pairs xe2x88x92270 to xe2x88x92430).
Furthermore, the invention relates to a promotor region of the VstI-gene which lacks at least the DNA sequence that comprises base pairs xe2x88x92270 to xe2x88x92430 of the VstI-gene. A preferred version concerns a promotor region of the VstI gene, which only comprises the sequence range from the start of the translation to base pair xe2x88x92270 of the VstI gene. It is particularly preferred that the promoter region, which lacks the sequence range xe2x88x92270 to xe2x88x92430 of the VstI gene, is able to convey a pathogen-induced gene expression in plants.
The invention also relates to chimeric nucleic acid molecules, into which the DNA sequence of base pairs xe2x88x92270 to xe2x88x92430 of the VstI-gene or at least a fragment of this sequence range has been inserted. It is especially preferred that the chimeric nucleic acid molecules, according to the invention, render possible, due to the presence of the DNA sequence of base pairs xe2x88x92270 to xe2x88x92430 of the VstI gene or at least a fragment thereof, an ozone-inducible expression of the coding regions in plants contained therein.
The nucleic acid molecules can be any nucleic acid molecules, especially DNA or RNA molecules, e.g., cDNA, genomic DNA, mRNA, etc. They can be naturally occurring molecules or molecules produced by gene technology or by chemical synthetic processes.
By making available, according to the invention, DNA sequences, promotor regions, nucleic acid molecules or vectors, it is now possible to mutate plant cells by means of gene technology methods in such a way that they show ozone-inducible characteristics. Furthermore, it is now possible to mutate plant cells by means of gene technology methods in such a way that they characterize one or more genesxe2x80x94which are naturally ozone inducible, due to the presence of the DNA sequence set out in Claim No. 1 or a DNA sequence derivable therefrom or one that is homologous with said DNA sequencexe2x80x94as being no longer inducible by ozone but preferably inducible mainly by pathogens.
A special advantage of the invention is the fact that the ozone induction of naturally ozone-inducible genes in plants and plant cells can be eliminated by deleting the DNA sequence, as set out in Claim No. 1, or at least a fragment thereof, in the genes which naturally contain this DNA sequence or a DNA sequence which can be derived therefrom or one that is homologous with said DNA sequence.
Another advantage of the invention is that genes which cannot or cannot substantially be naturally induced through ozone, can be characterized as being ozone inducible by using the invented nucleic acid sequences in plant and plant cells. In a preferred version the nucleic acid sequence, which is responsible for the ozone-inducible expression or at least a fragment thereof, controls the expression of genes, the gene products of which are able to detoxify reactive oxygen species that can develop among other things, as a consequence of ozone in plant cells. In a particularly preferred version, the nucleic acid sequence controls the expression of catalase and/or superoxide-dismutase genes.
In an alternative version, the DNA sequence that is responsible for the ozone-inducible gene expression controls the expression of reporter genes which are measured in order to determine ozone concentrations quantitatively and/or qualitatively and to evaluate the effects of ozone. Such reporter genes can be, e.g., the uidA gene from E. coli, which codes for the enzyme xcex2-glucuronidase (GUS), luciferase genes or other genes, customary in plant biotechnology. Every expert in biotechnology, biochemistry or molecular biology is familiar with appropriate reporter genes.
Furthermore, the invention relates to vectors which contain the above-mentioned DNA sequences or promoter regions or fragments thereof. Thus, this invention relates also to vectors, particularly plasmids, cosmids, viruses, bacteriophages and other vectors, common in gene technology, which contain the above-mentioned nucleic acid molecules, according to the invention and which, if required, can be used for transferring said nucleic acid molecules to plants or plant cells.
The invention also relates to transformed microorganisms, such as bacteria, viruses and fungi which contain the nucleic acid sequences, according to the invention.
It is also an object of the invention to provide plants and plant cells which are characterized by the absence of the ozone-inducible expression of genes that are naturally induced in plants through ozone.
This problem is solved by providing the DNA sequence responsible for the ozone induction and by making available promoters which lack said sequence and which for this reason render possible a no longer ozone-inducible gene expression of genes in plant and plant cells controlled by the promotors.
The problem of rendering possible an ozone-responsive gene expression of genes which are not naturally inducible through ozone is likewise solved by providing the DNA sequence, according to the invention. Thus, plants and plant cells are made available which in the presence of ozone specifically define certain characteristics.
A preferred version concerns transformed plants and plant cells in which genes are ozone-inducibly expressed, and the gene products thereof are able to detoxify reactive oxygen species in plant cells. Particular preference is given in this connection to catalase and/or superoxide-dismutase genes. As an alternative, it is possible to produce plants and plant cells (including protoplasts) which characterize so-called reporter genes after induction through ozone and which, if required, can be used as biomonitors.
Thus, the subject-matter of the invention is transgenic plants which contain, integrated in the plant genome, the recombined nucleic acid molecules, as described above. Such plants can, in principle, be any plants. It concerns preferably a monocotyle or dicotyle useful plant. Examples of monocotyle plants are plants which belong to the genuses of Avena (oat), Triticum (wheat), Secale (rye), Hordeum (barley), Oryza (rice), Panicum, Pennisetum, Setaria, Sorghum (millet), Zea (corn). Dicotyle, useful plants are, e.g., cotton, leguminous plants such as legumes, and especially alfalfa, soya bean, rape, tomato, sugar beet, potato, ornamental plants, trees. Other useful plants can be fruit-bearing plants (particularly apples, pears, cherries, grapes, citrus fruits, pineapples and bananas), oil palms, tea and cacao shrubs, tobacco, sisal, as well as medicinal plants, such as rauwolfia and digitalis. Special preference is given to grain, such as wheat, rye, oat, barley, rice, corn and millet, sugar beet, rape, soya, tomato, potato and tobacco.
Furthermore, the subject-matter of the invention is the propagation material of plants, according to the invention, such as, e.g., seeds, fruits, cuttings, tubers, rootstock, etc., as well as constituents of such plants, such as plant cells, protoplasts and calli.
The plant cells include differentiated and non-differentiated plant cells (including protoplasts), as well as plant cells (including protoplasts) in which nucleic acid molecules are integrated in the plant genome or are present as autonomous molecules (including transient transformation).
In another version, the invention relates to host cells, particularly prokaryontic or eukaryontic cells, which have been transformed or inoculated with a recombined nucleic acid molecule or a vector, as described above, and cells which originate from said host cells and which contain the described nucleic acid molecules or vectors. The host cells can be bacteria or fungus cells, as well as plant or animal cells.
The object of this invention is also to show methods for the production of plant and plant cells which are characterized by the lack of an ozone-inducible expression of a gene, the expression thereof in plants and plant cells is naturally stimulated by ozone.
This problem is solved through processes by means of which the production of new plants and plant cells, which do not have this naturally occurring ozone-induced gene expression, is made possible.
As already mentioned above, it is also the object of this invention to provide methods for the production of plants and plant cells which express such genesxe2x80x94the expression of which is naturally not, or not substantially activated by ozonexe2x80x94after ozone stimulation. This problem is solved by methods with the help of which plants and plant cells can be produced which, after the invented DNA sequences or at least a fragment thereof has been inserted into naturally not ozone-inducible genes, or genes which are ozone-inducible only to a minor degree express such genes after ozone stimulation.
There are various methods by which such new plants or plant cells can be produced. For one thing, plants or plant cells can be mutated by conventional gene-technological transformation methods in such a way that the new nucleic acid molecules are being integrated in the plant genome, which means that stable transformants are produced.
According to the invention, plants or plant cells which, due to the absence of the invented nucleic acid sequence or at least a fragment thereof, no longer show an ozone induction of the gene(s) which naturally contains/contain said sequence, are produced by a method which includes the following steps:
a) Deletion of the DNA sequencexe2x80x94as defined in Claim No. 1, or a sequence which can be derived from said sequence, or which is homologous with said sequence, or at least a fragment of such sequencexe2x80x94from a gene which after the deletion of the invented DNA sequence, includes regulatory elements essential for the possibly regulated transcription and translation in plant cells, and has at least one coding sequence, as well as possibly a termination signal for the termination of the transcription and the addition of a poly-A-tail to the respective transcript.
b) Transformation of plant cells with the gene or nucleic acid molecule, produced in step a), and
c) possibly the regeneration of transgenic plants and possibly the propagation of the plants.
As an alternative it is possible in step a) that instead of deleting the sequence responsible for the ozone-induction, said sequence or at least a fragment thereof can be inactivated or blocked, e.g., through mutagenesis, and thus remain in the gene in an inactivated form. Irrespective of the manner in which the ozone-responsive gene range is deleted, all manipulative measures can be carried out by means of conventional methods and aids of recombined gene technology (see, e.g., Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbour Laboratory Press, Cold Spring Harbour, N.Y.).
In a particularly preferred version the nucleic acid molecule, which in step b) is transferred to plants or plant cells, contains regulatory elements which allow, e.g., a pathogen-induced gene expression of the coding sequence.
According to the invention, plants or plant cells which, due to the presence of the invented nucleic acid sequence that is essential or co-responsible for an ozone-induced expression of the genes containing same, or at least a fragment of said sequence, are produced by a method which includes the following steps:
a). Insertion of at least one DNA sequence, according to the invention, which in plants can produce an ozone-induced gene expression, or a sequence which is derivable from said sequence, or which is homologous with said sequence, or at least a fragment of said sequence, in a gene which is not naturally or not substantially expressed as ozone-inducible.
b) Transformation of plant cells by way of the gene or nucleic acid molecule, produced in step a), which has all elements that are naturally required for the expression in plant cells, and
c) possibly the regeneration of transgenic plants and perhaps the propagation of plants.
In a preferred version the gene concerned is a catalase dismutase, superoxide-dismutase or a common reporter gene.
Another object of the invention is to show advantageous usage of the invented nucleic acid sequences.
The invention includes, therefore, uses of the new DNA molecules to produce the aforementioned plants and plant cells, according to the invention, which are characterized either by the absence of a certain phenotypical distinguishing mark that is normally influenced by ozone, or which precisely due to the presence of the invented DNA sequence distinguish themselves from non-transgenic plants and plant cells by ozone-induced characteristics.
Furthermore, the invention includes the use of the invented nucleic acid molecules to produce plants which are characterized by an increased pathogen-induced but not an ozone-induced resistance to disease.
The invention also relates to the use of the invented nucleic acid sequences or fragments thereof for detecting and identifying ozone-responsive nucleic acid elements.
The expert can identify such ozone-responsive nucleic acid elements by applying customary molecular biological methods, e.g., hybridizing experiments or DNA protein-binding studies. For example, as a first step, the poly (A)+ RNA is isolated from a tissue which was treated with ozone. Then a cDNA-bank is set up. In a second step and with the help of cDNA-clones, which are based on poly(A)+ RNA molecules from a non-treated tissue, those clones from the first bank are identified by way of hybridizing whose corresponding poly(A)+ RNA molecules are induced strictly in the ozone treatment. With the help of cDNAs identified in this manner, promoters which have ozone-responsive elements are subsequently isolated. The nucleic acid sequences and molecules can be useful instruments when examining and characterizing these isolated promoters.
The subject-matter of the invention includes also nucleic acid molecules or fragments thereof which hybridize with one of the above-described nucleic acid molecules or one of the above-mentioned DNA sequences of the invention. Within the scope of this invention the term xe2x80x9chybridizingxe2x80x9d means hybridizing under conventional hybridizing conditions, preferably under stringent conditions, such as are described, for example, in Sambrook et al (1989) Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Laboratory Press, Cold Spring Harbor, N.Y.
Nucleic acid molecules which hybridize with the molecules, according to the invention, can be isolated, e.g., from genomic or cDNA banks.
The identification and isolation of such nucleic acid molecules can be accomplished by using the nucleic acid molecules, according to the invention, or fragments of said molecules or the reverse complements of such molecules, e.g., by way of hybridizing according to standard procedure (see, e.g., Sambrook et al, supra).
Thus, the invention relates also to the use of an invented DNA sequence or fragments thereof to identify and isolate homologous sequences from plants or other organisms.
For example, nucleic acid molecules which have exactly or substantially the invented nucleotide sequences or fragments of such sequences can be used as a hybridizing probe. The fragments used as a hybridizing probe can also be synthetic fragments which were produced with the help of customary synthesis techniques and the sequence of which basically corresponds with that of a nucleic acid molecule, according to the invention. Once genes that hybridize with the invented nucleic acid sequences have been identified and isolated, it is necessary to determine the sequence and analyze the properties. To do so, a number of molecular biological, biochemical and biotechnological standard methods are available to the expert.
The molecules that hybridize with the nucleic acid molecules, according to the invention, include also fragments, derivatives and allelic variants of the above-described DNA molecules that contain an ozone-responsive sequence in an active or inactivated form, or which are characterized by the fact that they no longer have such sequence. The term xe2x80x9cderivativexe2x80x9d means in this context that the sequences of these molecules distinguish themselves from the sequences of the above-described nucleic acid molecules in one or several positions and are to a great extent homologous with said sequences. Homology in this connection means a sequence identity of at least 40%, especially an identity of at least 60%, preferably above 80%, and especially preferable above 90%. The deviations from the above-described nucleic acid molecules can have been caused by deletion, addition, substitution, insertion or recombination.
With respect to the nucleic acid molecules which are homologous with the above-described molecules and which constitute derivatives of such molecules, it concerns usually variants of such molecules that constitute modifications which perform the same biological function. These may concern naturally occurring variations, e.g., sequences from other organisms, or mutations in which these modifications can have occurred naturally or were introduced through specific mutagenesis. Furthermore, the variations may concern synthetically produced sequences. With respect to the allelic variants, these may occur naturally as well as be synthetically produced variants, or variants produced by recombined DNA methods.
To prepare the insertion of foreign genes into higher plants or into the cells thereof, a large number of cloning vectors are available which contain a replication signal for E. coli and a marker gene to select transformed bacteria cells. Examples of such vectors are pBR322, pUC-series, M13mp-series, pACYC184, etc. The desired sequence can be inserted into the vector at a suitable restrictive cut. The plasmid obtained is used for the transformation of E. coli cells. Transformed E. coli cells are cultured in a suitable medium and subsequently harvested and lysed. The plasmid is recovered. Usually restrictive analyses, gel electrophoreses and other biochemical, molecular biological methods are applied as an analyzing method to characterize gained plasmid DNA. After each manipulation the plasmid DNA can be split and the gained DNA fragments can be linked with other DNA sequences. Each plasmid-DNA sequence can be cloned in identical or other plasmids.
Many well-known methods are available to introduce DNA into a plant host cell. The expert can determine, without difficulty, the respectively suitable method. These techniques include the following: the transformation of plant cells with T-DNA by using Agrobacterium tumefaciens or Agrobacterium rhizogenes as transformation medium, the fusion of protoplasts, the direct gene transfer of isolated DNA in protoplasts, the electroporation of DNA, the introduction of DNA by means of the biolistic method, as well as other possibilities. In doing so, stable, as well as transient tranformants, can be produced.
When injecting and electroporating DNA into plant cells no specific demands per se is made on the plasmids used. The same applies to direct gene transfer. Simple plasmids as, e.g., pUC derivatives, can be used. If, however, whole plants are to be regenerated from cells transformed in this way, the presence of a selectable marker gene is required. The expert is familiar with the usual selection markers, and it does not pose a problem for him to select an appropriate marker. Selection markers in common use are those that make the transformed plant cells resistant to a biocide or an antibiotic, such as kanamycin, G418, bleomycin, hygromycin, methotrexate, glyphosate, streptomycin, sulfonyl-urea, gentamycin or phosphinotricin, etc.
Depending on the method selected for introducing genes into the plant cell, additional DNA sequences may be required. If, for example, the Ti or Ri-plasmid is used for the transformation of the plant cell, at least the right boundary, but frequently, however, the right and left boundary of the T-DNA, contained in the Ti and Ri-plasmid must, as flank region, be linked with the genes to be introduced.
If agrobacteria is used for the transformation, the DNA to be introduced must be cloned in special plasmids, i.e. either in an intermediary or in a binary vector. Due to sequences which are homologous with sequences in the T-DNA, the intermediary vectors can be integrated in the Ti- or Ri-plasmid of the agrobacteria through homologous recombination. In addition, the latter includes the vir-region required for the transfer of the T-DNA. Intermediary vectors cannot replicate in agrobacteria. The intermediary vector can be transferred to Agrobacterium tumefaciens (conjugation) by means of a helper plasmid. Binary vectors can replicate in E. coli as well as in agrobacteria. They contain a selection-marker gene and a linker or a polylinker which are framed in by the right and left T-DNA boundary region. They can be directly transformed into the agrobacteria (Holster et al (1978) Molecular and General Genetics 163, 181-187). The agrobacterium which serves as host cell shall contain a plasmid that has a vir-region. The vir-region is necessary for transferring the T-DNA to the plant cell. Additional T-DNA can be present. The agrobacterium, transformed in the manner described, is used for the transformation of plant cells.
The use of T-DNA for the transformation of plant cells has been thoroughly studied and is adequately described in EP 120 515; Hoekema in: The Binary Plant Vector System, Offsetdrokkerij Kanters B. V., Alblasserdam (1985) Chapter V; Fraley et al (1993) Crit. Rev. Plant. Sci., 4, 1-46 and An et al (1985) EMBO J. 4, 277-287.
For the transfer of the DNA to the plant cell, plant explantates can be appropriately cultivated with Agrobacterium tumefaciens or Agrobacterium rhizogenes. Out of the infected plant material (leaf fragments, stem segments, roots, but also protoplasts or suspension-cultivated plant cells) whole plants can be regenerated in a suitable medium which can contain antibiotics or biocides for the selection of transformed cells. The regeneration of plants is carried out according to customary regeneration methods by using familiar culture media. Plants or plant cells obtained in the above-described manner can then be examined for the presence of the introduced DNA. Other possibilities for introducing foreign DNA by applying the biolistic method or through protoplast transformation are known (see, e.g., Willmitzer L. (1993) Transgenic Plants, in: Biotechnology, A Multi-Volume Comprehensive Treatise (H. J. Rehm, G. Reed, A. Pxc3xchler, P. Stadler, eds.) Vol. 2, 627-659, V. C. H. Weinheimxe2x80x94New Yorkxe2x80x94Baselxe2x80x94Cambridge).
Although the transformation of dicotyl plants or their cells via Ti-plasmid vector systems and with the help of Agrobacterium tumefaciens is well established, new studies indicate that also monocotyl plants or their cells are very receptive to transformation by means of Agrobacterium-based vectors (Chan et al (1993) Plant Mol. Biol. 22, 491-506; Hiei et al (1994) Plant J. 6, 271-282; Deng et al (1990) Science in China 33, 28-34; Wilmink et al (1992) Plant Cell Reports 11, 76-80; May et al (1995) Bio/Technology 13, 486-492; Conner and Domiss (1992) Int. J. Plant Sci. 153, 550-555; Ritchie et al (1993) Transgenic Res. 2, 252-265).
Alternative systems for transforming monocotyl plants or their cells are transformations by means of the biolistic setup (Wan and Lemaux (1994) Plant Physiol. 104, 37-48; Vasil et al (1993) (Bio/Technology 11, 1553-1558; Ritala et al (1994) Plant Mol. Biol. 24, 317-325; Spencer et al (1990) Theor. Appl. Genet. 79, 625-631), the protoplast transformation, the electroporation of partially permeabilized cells and the introduction of DNA by means of glass fibres.
The transformed cells grow within the plant in the usual manner (see also McCormick et al (1986) Plant Cell Reports 5, 81-84). The resulting plants can be cultivated normally and grafted with plants which have the same, transformed genetic trait or other genetic traits. The resulting hybrid individual plants have the respective phenotypical properties.
Two or more generations should be cultivated in order to ensure that the phenotypical characteristic is firmly maintained and propagated. Seeds should also be harvested in order to ensure that the respective phenotype or other characteristics are maintained.
By applying the usual methods, transgenic lines can be determined, which are homozygous for the new nucleic acid molecules and, furthermore, their phenotypical behaviour can be examined for an existing or non-existing ozone-response capacity and compared to that of hemizygous lines.
Naturally, plant cells which contain the nucleic acid molecules, according to the invention, can also be further cultivated as plant cells, (including protoplasts, calli, suspension cultures, etc.).
Another object of this invention is the use of the nucleic acid molecules, according to the invention, or fragments of such molecules, or the reverse complements of such molecules to identify and isolate homologous molecules which include ozone-responsive elements from plants or other organisms. As to the definition of the term xe2x80x9chomologyxe2x80x9d, please refer to the definition given earlier in the text.
The following examples serve the purpose of explaining the invention.


A knitted fabric comprises fibers. At least part of these are metal fibers. The fabric has 90 or more stitches per square centimeter. The fabric is used as a separation cloth (12) between mould (11) and glass (14). The increased number of stitchers per square centimeter reduces the risks for markings on the glass.

The invention relates to a knitted fabric comprising fibres (fibers), at least part of these fibres being metal fibres and the use of such fabric as a separation cloth for moulds in glass bending processes.
Such knitted fabrics comprising metal fibres are described in the PCT-patent applications WO97/04152, WO94/01372 and WO94/01373 and are utilised in various fields of application.
The use of a textile fabric as separation cloth between mould and glass, to form side-lites and back-lites for automotive business is known. During this contact, temperatures of 650 to 700xc2x0 C. are used. It is of great importance that no marks are left on the glass surface after the contact of the glass and the textile fabric.
The use of textile fabrics out of 100% glass fibres is known. The disadvantage of these glass fibre cloths is that it doesn""t resist the mechanical action during the glass shaping process. Also the use of textile fabrics, partially or fully consisting out of metal fibres is known. Using these fabrics as mould coverings, the mechanical action of the bending process is withstand better, but there is still the risk of marking the glass, by transferring the woven or knitted pattern into the glass surface which has contacted the textile fabric.
Further, it is known that the use of knitted structures is more suitable to cover moulds, since knitted surfaces can be draped better on moulds and less or no folds will be created when bending the knitted fabric, especially on three-dimensionally shaped surfaces or moulds.
The risk of having marks, caused by use of textile fabrics as the separation cloth for moulds in glass bending processes, is influenced by several parameters, such as glass temperature and pressure used to bend the glass. Since for example the automotive industry requires more complex glass surfaces, that is glasses which show a deeper bend, the glass has to be heated to a higher temperature and the pressure to bend the glass, is increased as well. These two adjustments to the production parameters of the bending process, makes the glass more sensitive to markings since higher temperature makes the glass softer, and creates a more obvious transfer of the textile structure, either woven or knitted, on the glass surface because of the higher pressure.
Another parameter that influences the risk of marking, is the wear of the textile fabric, used as a separation cloth between moulds and glass, due to the repetitive contacts with glass sheets, and the temperature. This temperature makes the fibres become more sensitive to breaking forces, and the mechanical action of the glass sheets against the fabric makes the fabric wear out little by little. Since the fibres which are standing out on the yarn surface, will suffer most on this mechanical action, and so will disappear after several contacts with glass, the stitches out of which the knitted fabric is made or the weaving pattern, used to provide the woven fabric, will be transferred more obviously to the glass surface.
Separation cloths should preferably meet next requirements:
1. The cloth should resist the bending temperature. Typically, these temperatures raise up to 700xc2x0 C. when the bending takes place in the heated part of the furnace. When the glass bending takes place out of the furnace, this temperature will be less.
2. The cloth should be able to follow the mould shape as close as possible.
3. The separation material should show enough air permeability. It is taken as a limit that separation cloths should at least have an air permeability of 2400 l/10 cm2*h, and preferably more than 4500 l/10 cm2*h.
4. The weight of the separation cloth is preferably between 600 g/m2 and 2000 g/m2. Fabrics with less weight usually wear out too fast, where too heavy fabrics tend to elongate too much under its own weight, so causing obstruction in the furnace for the glasses to pass in the neighbourhood of the fabric before or after the bending action.
5. The thickness for the separation cloth is preferably more than 0.8 mm and even better more than 1 mm. Too thin fabrics show a lack of elasticity in the direction perpendicular to the fabric surface.
6. And as already mentioned, the risk for remaining marks on the glass surface should be reduced to a minimum.
The higher the number of requirements met, the better the performance of the separation cloth between mould and glass in the glass bending process will be.
It is an object of the present invention to provide a fabric with a reduced risk for markings on the glass surface.
It is also an object of the present invention to provide a fabric which meets all of the above-mentioned minimum requirements.
The invention relates to a knitted fabric, which comprises fibres, at least part of these fibres being metal fibres, and which has more than 90 stitches per square centimetre.
Preferably the knitted fabric has more than 95, most preferably more than 100, e.g. more than 105 or even more than 110 stitches per square centimetre.
The first requirement, being the resistance to the temperatures used for the bending of the glass, is met by the use of metal fibres, usually stainless steel fibres.
Possibly, other high temperature fibres, such as glass fibres, ceramic fibres, TWARON(copyright), NOMEX(copyright), meta-aramid fibres, para-aramid fibres, carbon fibres, preox-fibres and other high temperature resistant man-made fibres can be used, next to the metal fibres. The fibres, of which at least one are metal fibres, can be intimately blended and possibly plied to a two or more plied yarn or the yarn can be a two- or more plied yarn, where some or all of the single yarns are made out of one fibre type.
By plying yarns, it is meant that two or more yarns are given a torsion round the direction of the axis""s of the yarns.
To meet the second requirement, being the drapeability, usually knitted structures are used.
The other characteristics, air permeability, thickness, weight and number of stitches, are largely influenced by the gauge of the knitting machine, the metrical number of the used yarns, the knitting structure and the settings of the knitting machine during the knitting action. The higher the number of stitches per square centimetre, the heavier and thicker the fabric and the lower the air permeability. The inventors, however, have discovered that the risk for glass markings can be substantially reduced, if not avoided, if the fabric has a higher number of stitches per surface unit and that this higher number of stitches can be reached with values of air permeability, thickness and weight which still fall within the above-mentioned ranges.
The reduced risk for markings on the glass surface can be explained as follows:
To reduce the risk on creating marks on the bent glass surface, it is important to use a fabric with as much yarn surface as possible on the fabric side which contacts the glass during the bending operation. This for 2 reasons:
1. By having more yarn surface on this contact side, the force to bend the glass is distributed over more contact surface. The depth to which extend the fabric might be pressed into the softened glass largely depends on this force per surface, so less force per unit decreases the risk on having a too large impression of the fabric in the glass, and so creating marks on the glass surface.
2. Because this less force per contact surface unit, the wear due to the repetitive mechanical action on the fabric surface will be reduced. This makes the time to have too much yarn pronunciation longer and the risk to have marks will be decreased in time.
The yarns which can be used to realise fabrics, as subject of the invention are made out of metal fibres, usually stainless steel fibres, possibly blended with glass fibres or ceramic fibres, other high temperature fibres, such as TWARON(copyright), NOMEX(copyright), meta-aramid fibres, para-aramid fibres, carbon fibres, preox-fibres and other high temperature resistant man-made fibres. The fibres, of which at least a part being metal fibres, can be intimately blended and possibly plied to a two or more plied yarn or the yarn can be a two- or more plied yarn, where some or all of the single yarns are made out of one fibre type.
At least partially, the yarns will contain metal fibres. Usually, but not necessarily, stainless steel fibres are used. Alloys such as AISI 316 or AISI 316L, AISI 347, or other alloys out of the AISI 300 type are used. Also alloys out of the AISI-400 type or Aluchrome-type alloys can be used. These fibres can be bundle drawn, as described in patent U.S. Pat. No. 3,379,000, be made by shaving them from a coil, as described in patent U.S. Pat. No. 4,930,199 or melt extracted. Also metal fibres produced as described in U.S. Pat. No. 4,220,112 can be used.
These metal fibres have an equivalent diameter usually between 1 and 100 xcexcm, and more typically between 6 and 25. The equivalent diameter is the diameter of the circle, which has the same surface as the fibre section when cut perpendicularly to the axis of the fibres.
Typically, the fabric which is subject of the invention, has an air permeability of more than 2400 l/10 cm2*h and preferably more than 4500l/10 cm2*h. The weight of the fabric will be more than 600 g/m2 and less than 2000 g/m2. The thickness of the fabric will be not less than 0.8 mm and preferably more than 1 mm.
Different knitting structures can be used to provide the fabric as subject of the invention. It was found that knitting structures single jersey xc2xd, single jersey ⅓ and single jersey {fraction (1/4 )} can be used to provide knitted fabrics, comprising metal fibres with more than 90 stitches per square centimetre. Other single jersey structures, with more floating yarns such as single jersey ⅕, single jersey ⅙ or more, can be used.
By single jersey structures is meant a knitting structure, obtainable by using one needle bed, providing one stitch for every needle in the needle bed per row of stitches.
Different gauges can be used to provide the fabric as subject of the invention. The gauge are the number of needles per inch on the needle bed or beds of the knitting machine. Typically gauges from 10 to 32 can be used. However it is shown that to obtain more than 90 stitches per cm2, gauge 16 or more should be used. Best fabrics were provided using gauge 20 or more, such as gauge 22 or more.
Different yarns with different metrical numbers can be used to provide the fabric as subject of the invention. The metrical number (Nm) of a yarn, as mentioned in the list, is an expression for the fineness of the yarn. It gives you the length of yarn that has a weight of 1 gram. For reason of comparison, all metrical numbers were recalculated as if all fibres were metal fibres of type AISI 316L. To obtain a fabric as subject of the invention, yarns with metrical number Nm 5.5 can be used. Finer yarns such as Nm 7.5 or Nm 10 could also be used to reach 90 or more stitches per cm2.
Reinforcement multifilament weft yarns with a titre of less than 180 tex, such as e.g. metal yarn or glass fiber yarns, can be incorporated, as described in the international application number PCT/BE98/0010.
A fabric as subject of the present invention, with two surfaces having a different fibre content can be provided by using the plating technique as described in Belgian patent application number 9800212.
According to another aspect of the present invention, there is provided a use of a fabric according to any one of the preceding fabrics for covering moulds and tempering or press-on rings which are utilised in the process of forming glass plates, or for covering the means of transport by which glass plates are moved during the forming process.
Still according to the present invention, there is provided a method for reducing the risk for marking the glass surface during bending.

Knitted fabric of steel fibers with increased number of stitches


A silver halide photographic light-sensitive material includes a support having thereon at least one silver halide light-sensitive emulsion layer, characterized in that at least one of a plurality of emulsions contained in the emulsion layer contains tabular grains having a thickness of less than 0.5 .mu.m, a diameter of at least 0.3 .mu.m, and an average ratio of grain diameter to grain thickness of at least 2, the tabular grains occupy at least 50% of the entire projected area of all the grains contained in the emulsion, the surfaces of the grains are chemically sensitized, and a ratio of surface sensitivity to internal sensitivity of the grains is 0.5 to 2.

1. Field of the Invention
The present invention relates to a silver halide photographic light-sensitive material having improved image sharpness and resistance to stress.
2. Description of the Related Art
Normally, in a photographic light-sensitive material comprising a silver halide emulsion layer, light scattering caused by silver halide grains tends to reduce the sharpness of the emulsion layer.
U.S. Pat. Nos. 4,434,226, 4,439,520, and 4,433,048 describe that image sharpness and the like can be improved by use of a tabular silver halide emulsion and that this technique can be applied to a photographic light-sensitive material.
However, tabular silver halide grains have poor resistance to stress, and form many stress marks and therefore results in difficulties in designing photographic light-sensitive material, as will be explained below.
More precisely, a photographic light-sensitive material on which is coated a silver halide emulsion is subjected to a variety of stresses when in the form of a conventional photographic negative film; being rolled up inside a cartridge, bent when it is inserted into a camera, and pulled when it is wound and re-wound.
On the other hand, a sheet-like film such as a light-sensitive print material or a roentgen light-sensitive material for medical and direct X-ray often becomes bent or folded on account of it being handled directly by an operator.
In addition to the above, all light-sensitive materials are subjected to high stress when cut and during manufacture.
When a photographic light-sensitive material is subjected to such stress, silver halide grains are stressed through gelatin which is a binder of the silver halide grains, or through a plastic film which acts as a support therefor. When the silver halide grains undergo such stress, the photographic properties of the photographic light-sensitive material are changed, as is described in detail in K.B. Mather, J. Opt. Soc. Am., 38, 1054 (1948), P. Faelens and P. de Smet, Sci. et Ind. Phot., 25, No. 5. 178 (1954) P. Faelens. J. Phot. Sci, 2, 105 (1954).
Consequently, there is widespread demand for a photographic light-sensitive material whose photographic properties are not adversely affected by any type of stress.
In order to improve resistance to stress, a plasticizer such as a polymer or emulsion is added, or the ratio of silver halide to gelatin in the silver halide emulsion is reduced so that no stress reaches the grains.
For example, British Patents Nos. 738,618, 738,637, and 738,639 disclose methods of using a heterocyclic compound, alkylphthlate, and alkylester, respectively; U.S. Pat. Nos. 2,960,404 and 3,121,060 disclose methods of using polyhydric alcohol and carboxyalkylcellulose, respectively; JP-A-49-5017 discloses a method of using paraffin and a carbonic acid salt; and JP-B-53-28086 discloses a method of using alkylacrylate and an organic acid. Hereinafter, the symbol "JP-A-" will be used to denote a Japanese Patent Disclosure, and the symbol "JP-B-" will be used to designate a Japanese Patent Publication. Further, the symbol "JP-A-(examined)" will be used to specify a published Japanese patent application without having been laid open which was filed before Jan. 1, 1971 when the system of laying open any patent application came into existence.
However, since mechanical strength of an emulsion layer is reduced when a plasticizer is added, the amount of the plasticizer which can be used limited. In addition, when the amount of gelatin is increased, sharpness is degraded or a developing speed is reduced. Therefore, neither of these methods can achieve a sufficient effect.
As a method of improving resistance to stress of the tabular silver halide grain itself, JP-A-59-99433 discloses a method of forming an iodide rich annular region inside a grain.
Although resistance to stress can be improved by this method, further improvements are desired.

Silver halide photographic light-sensitive material


An assembly for the sorted conveyance of substantially plate-like products, the assembly includes a stroke limiting shoulder, downstream of at least one supply line, a first pusher having a lateral surface that faces and is proximate to the stroke limiting shoulder, and at least one second pusher, which is adjacent to the other pusher. The pushers are substantially perpendicular to the advancement direction of the products and are independently movable from a first configuration, in which the respective end face faces and is proximate to the edge of the at least one supply line, to a second configuration, in which the end face faces and is proximate to the edge of the exit channel, surmounting the respective supply line. The exit channel includes consecutive receptacles for accommodating clusters of products.

A packaging apparatus for chewing gum in sticks is constituted by a series of consecutive stations: a wrapping machine for the individual wrapping, with adapted sheets, of each stick, a supply assembly for arranging the wrapped sticks in consecutive clusters, a stacker assembly for creating the bundle to be packaged and a cartoning machine for inserting the bundle into an adapted package.
The wrapping machine transfers the wrapped sticks to the supply assembly on one or more separate channels; the supply assembly, which is also provided with corresponding channels, has the task of transferring the products to conveyance chutes that lead to the stacker assembly.
Downstream of the stacker, the sticks sorted in bundles are in turn inserted into the package by the cartoning machine.
The format of the bundle to be inserted in the package comprises a plurality of sticks: particularly there is particular interest in bundles made up of seven sticks that are arranged in series on three superimposed layers.
The possibility is not excluded of having to make up bundles that are constituted by a different number (more or fewer) of sticks with respect to the number given in the previous paragraph by way of example.
Unfortunately however the sticks have very considerable size tolerances which negatively influence their correct arrangement in the conveyance chutes.
The size tolerance of sticks of chewing gum is of the order of magnitude of approximately ±4% (and cases are known in which it is even higher), and this means that products with maximum size differences of 8%, i.e. particularly pronounced, could end up being arranged side by side.
So if we are operating on sticks with a nominal width of 11 mm, there may be sticks that are 10.61 mm wide and sticks that are 11.54 mm wide.
In these cases, if we are working on a bundle made up of seven sticks, the maximum tolerance on the length of the bundle, which is made up of seven laterally adjacent sticks, is 6.51 mm (this value is calculated as the difference between the maximum length deriving from seven sticks that are 11.54 mm wide and arranged side by side, i.e. 80.78 mm, and the minimum length of seven sticks that are 10.61 mm wide and arranged side by side).
It should be noted that these sticks generally have a scant thickness, in the region of 5 mm (comprised between 4.74 and 5.08 mm).
When we proceed with the stacking of several bundles of sticks, we find ourselves in an extremely difficult position, in that the risk arises that a stick could rotate and "fall" into the space between two adjacent sticks of the lower layer: in fact it could happen that two adjacent sticks end up at a mutual distance of 6.51 mm (the maximum tolerance allowed on the length of the bundle), which can thus accommodate a stick that is arranged transversely with respect to the others.
If this condition of incorrect positioning arises, the apparatus will not be able to effectively proceed with the cartoning, since this will be made impossible by the transverse arrangement of the stick that "fell" into the channel between two contiguous sticks of the lower layer.
This problem is more evident the bigger the size tolerance is for each individual stick, and the bigger the bundle to be stratified (i.e. constituted by a high number of sticks) is.
In order to guard against these problems, the stacking could be carried out very slowly and by holding each individual stick: this would result in a considerable reduction in efficiency of the plant and in the necessity of having dedicated grip means for each individual type of stick, with a consequent increase in costs and in complexity if a change of format is necessary.

Assembly for the sorted conveyance of substantially plate-like products


The present application provides a communication method for vehicle remote diagnosis, which is applied to a WEB server; the method comprises: establishing a first socket connection with the WEB front end when receiving a landing request of the WEB front end; receiving a vehicle remote diagnosis request of the WEB front end through the first socket connection and obtaining an identification of a target vehicle diagnosis equipment carried in the vehicle remote diagnosis request; establishing a second socket connection with the target vehicle diagnosis equipment according to the identification of the target vehicle diagnosis equipment; performing interaction of the vehicle diagnosis data between the WEB front end and the target vehicle diagnosis equipment through the first socket connection and the second socket connection. The application also provides a communication device and system for vehicle remote diagnosis. The application can improve the real-time capability and convenience of vehicle remote diagnosis.

With the rapid development of the automobile industry, electronic control technology has entered the various components of an automobile, and with the increase of the complexity of the structure of the automobile, fault diagnosis and elimination for vehicles have become important technical requirements and security guarantees. The vehicle remote diagnosis can achieve the remote diagnosis and consultation to fault vehicles of inexperienced technicians or non-professional staff, such that the advantage of professional automobile technicians can be brought into full play.
In the existing remote diagnosis technology of vehicles, the professionals obtain vehicle diagnosis data collected by remote diagnosis equipment in the sites of the vehicle through near-end diagnosis equipment, so as to realize the diagnosis for the vehicle fault. This method has the following limitations:
On one hand, the near-end diagnosis equipment is generally professional diagnosis equipment (for example, a professional computer with a special operating system), and the equipment must be stalled with a relevant diagnosis APP or other diagnosis program; in the event of the automobile fault, since ordinary users cannot use these equipment to run the program anytime and anywhere, the convenience is very low, and it is inconvenient for the promotion of the remote diagnosis technology; on the other hand, due to the restriction of network transmission condition, only some simple data interchanges, such as unidirectional document viewing and data reception, and sending some simple commands, etc., can be achieved between near-end diagnosis equipment and remote diagnosis equipment, which cannot be applied to the occasions where the requirement for the real-time performance and complexity of vehicle diagnosis is higher.

Communication method, device, and system for vehicle remote diagnosis


An ultrasound image processing apparatus (10) is disclosed comprising a processor arrangement (16) adapted to map a model (1) of an anatomical feature of interest onto an ultrasound image showing at least a section of said anatomical feature of interest and to segment said ultrasound image in accordance with the mapped model; and a touchscreen display (18, 19) adapted to display said ultrasound image including the mapped anatomical model. The processor arrangement is responsive to the touchscreen display and adapted to recognize a type of a user touch motion (3) provided through the touchscreen display (18, 19), each type of user touch motion being associated with a particular type of alteration of said mapping and alter said mapping in accordance with the recognized type of user touch motion. Also disclosed are an ultrasound imaging system, a computer-implemented method and a computer program product.

Ultrasound plays an essential role in many diagnostic imaging techniques including but not limited to cardiovascular imaging. In this context, diagnosis and treatment planning typically rely on a clear delineation of relevant anatomical sites such as the ventricles, the atria or surrounding vessels in the cardiac ultrasound images in order to facilitate meaningful evaluation of the heart under investigation. Historically, measurements (of the volume) of cardiac chambers, such as the left ventricle, have been generated by a user hand-tracing the endocardial boundary of the chamber. Such tracings are subject to significant variability due to differences in the criteria different users utilize in determining where to locate the tracing.
In order to address such unwanted variability, this evaluation process has been automated by using anatomically intelligent model-based segmentation, where generic, shape-constrained heart models are fitted to the imaging data. For example, such a heart model is disclosed in WO 2016/142204 A1. The heart model may be subsequently used as an aid to subject the mapped cardiac structure in the ultrasound imagery to segmentation algorithms in order to obtain the dimensions of interest of the cardiac anatomical sites under investigation, e.g. by mapping such a model onto a volumetric image. A suitable segmentation algorithm is disclosed by O. Ecabert et al. in "Automatic Model-Based Segmentation of the Heart in CT Images", IEEE Transactions on Medical Imaging, 2008 (27), pages 1189-1201.
The success of the segmentation algorithm requires an accurate placement of the heart model within the imaging data, e.g. a volumetric image. While this placement generally includes basic coordinate transformations such as rotation, translation and scaling, further processing also involves affine transformations of single model components (e.g. ventricles or atria) or the selection from a set of mean shapes representing different heart pathologies (e.g. dilated or normal shape). In some cases, parts of this automatic placement process fail, for example, due to poor ultrasound image quality or a limited field-of-view (FoV) within the ultrasound imagery.
Two typical examples are shown in FIG. 1 and FIG. 2. In FIG. 1, due to the imaging data showing a heart of rather small size, the automatic placement of the heart model 1 including left ventricle (LV), right ventricle (RV), left atrium (LA) and right atrium (RA) anatomical site models failed by misjudgment of the overall model scale, causing the entire heart to be wrapped by the left ventricle model of the heart model 1. In FIG. 2, the ultrasound imaging data exhibits a limited field of view in combination with rather poor imaging contrast, particularly at the apex. Consequently, the translation parameter was misjudged by the automatic process of the placement of the heart model 1 within the imaging data, such that the actual right ventricle is covered by the left side of the heart model 1.
In case of such placement failures of the heart model 1, the analysis pipeline cannot be performed unless the user manually places the heart model 1 in the correct position within the ultrasound image, which can be tedious and time-consuming. This for example is disclosed in WO 2005/078666 A1. As will be readily understood by the skilled person, such model placement problems are not limited to heart models but equally apply to placement of models of other anatomical features of interest, e.g. organ models, fetal models and so on.

Ultrasound evaluation of anatomical features