A method encodes a video as video objects. For each candidate object, a quantizer parameter and a skip parameter that jointly minimizes an average total distortion in the video are determined while satisfying predetermined constraints. The average total distortion includes spatial distortion of coded objects and spatial and temporal distortion of uncoded objects. Then, the candidate objects is encoded as the coded objects with the quantizer parameter and the skip parameter, and the candidate objects is skipped as the uncoded objects with the skip parameter.

A number of video coding standards support variable frame rates, e.g., H.263 and MPEG-4. With variable frame-rates, any number of frames, or objects in the case of MPEG-4, can be skipped during the coding of the output video. That is, the skipped frames remain uncoded. With these video coding standards, the encoder may choose to skip frames of a video to either satisfy buffer constraints, or to optimize the video coding process. However, most encoders only skip frames to satisfy buffer constraints. Buffer constraints are usually due to bit-rate (bandwidth) limitations. The coder is forced to skip frames when insufficient bandwidth causes the buffer to fill up. Consequently, it is not possible to add any additional frames to the buffer, and these frames remain uncoded (skipped) until there is room in the buffer to store a new coded frame. This type of frame skipping can degrade the quality of the video because the content of the video is not considered. Note that skipping frames effectively reduces the frame-rate.
It is a problem to provide an optimal strategy for coding a video. Specifically, the video could be coded at a higher frame-rate having a lower spatial quality, or a lower frame-rate having a higher spatial quality. This trade-off between spatial and temporal quality is not a simple binary decision, but rather a decision over a finite set of coding parameters (constraints). Obviously, the best set of coding parameters will yield the optimal rate-distortion (R-D) curve that maximizes the frame-rate while minimizes the distortion. The two parameters of interest are the number of frames per second (fps or frame-rate) and a quantizer (Q) parameter. A higher quantizer parameter increases the spatial distortion. Lowering the frame rate, by skipping frames, reduces both the spatial and temporal distortion. In the known prior art, the distortion is measured only for coded frames, and is expressed as the mean-squared error (MSE) between pixels in the original video and the compressed video. That is, the prior art methods have two problems, only spatial distortion in coded frames is considered, and uncoded frames contributing to both the spatial and temporal distortion are not considered at all.
Generally, prior art optimized coding methods do not consider the temporal aspect of rate-distortion, see H. Sun, W. Kwok, M. Chien, and C. H. John Ju, "MPEG coding performance improvement by jointly optimizing coding mode decision and rate control," IEEE Trans. Circuits Syst. Video Technol., June 1997, T. Weigand, M. Lightstone, D. Mukherjee, T. G. Campbell, S. K. Mitra, "R-D optimized mode selection for very low bit-rate video coding and the emerging H.263 standard," IEEE Trans. Circuits Syst. Video Technol., and April 1996, J. Lee and B. W. Dickenson, "Rate-distortion optimized frame type selection for MPEG encoding," IEEE Trans. Circuits Syst. Video Technol., June 1997. Generally, it is assumed that the frame-rate is fixed.
These methods consider optimizations on the quantizer parameter, H. Sun, W. Kwok, M. Chien, and C. H. John Ju, "MPEG coding performance improvement by jointly optimizing coding mode decision and rate control," IEEE Trans. Circuits Syst. Video Technol., June 1997, mode decisions for motion and block coding, T. Weigand, M. Lightstone, D. Mukherjee, T. G. Campbell, S. K. Mitra, "R-D optimized mode selection for very low bit-rate video coding and the emerging H.263 standard," IEEE Trans. Circuits Syst. Video Technol., April 1996, and frame-type selection, J. Lee and B. W. Dickenson, "Rate-distortion optimized frame type selection for MPEG encoding," IEEE Trans. Circuits Syst. Video Technol., June 1997. Such methods can achieve an optimum coding when the frame-rate is fixed, and the bit-rate can be met for the given frame-rate. However, these methods are less than optimal for varying frame-rates.
It should be noted that the trade-off between spatial and temporal quality, while coding, has been described by F. C. Martins, W. Ding, and E. Feig, in "Joint control of spatial quantization and temporal sampling for very low bit-rate video," Proc. ICASSP, May 1996. However, in their method, the trade-off was achieved manually.
Therefore, it is desired to provide a method and system for encoding a video subject to a variable frame-rate, while minimizing the total average distortion.

Encoding a video with a variable frame-rate while minimizing total average distortion

Mitsubishi Electric Research Laboratories, Inc.


The present invention relates to an apparatus and a method for realizing all-optical NOR logic device using the gain saturation characteristics of a semiconductor optical amplifier(SOA). More particularly, the invention relates to a 10 Gbit/s all-optical NOR logic device among all-optical logic devices, in which a signal transmitted from a given point of an optical circuit such as an optical computing circuit is used as a pump signal and a probe signal.

The present invention relates to an apparatus and a method for realizing an all-optical NOR logic device using the gain saturation characteristics of a semiconductor optical amplifier SOA. More particularly, the invention relates to a 10 Gbit/s all-optical NOR logic device among all-optical logic devices, in which a signal transmitted from a given point of an optical circuit such as an optical computing circuit is used as a pump signal and a probe signal.
The recent trends show that the requirement for a high capacity and a high speed system is exponentially increasing.
Most of the present systems are based on silicon materials which operate on the basis of electric signals. For this reason, their future reliability is somewhat uncertain since they are expected to face a serious limitation in terms of their speed and the quantity of information that can be processed.
On the contrary, the systems utilizing the optical devices which are based on Indium Phosphide (InP) are expected to easily overcome all aspects of the aforementioned problems including the speed and quantity of information that can be processed by the systems.
When a system is being configured in general, the method which is based on integrating the single logic devices (AND, OR, XOR, NAND, NOR, NXOR) is utilized. This is also true in case of configuring an optical system.
The logic devices which have two stable states known as logic 0 and 1 are the basic building blocks of a digital computer.
Computers codify all the information using the two logic states. Accordingly, it is certain that logic devices will play a significant role in the development of all-optical logic devices and optoelectronic devices for the future information technology.
Up to the present, all-optical logic devices for ultra-high speed optical information processing utilize either the nonlinear characteristics or wavelength conversion characteristics of light.
Especially, implementations of all-optical NOR logic devices using the nonlinear gain of the SOA are listed chronologically as below;    (1) NOR based on a single-arm ultra-fast nonlinear interferometer (N. S. Patel, et al., Opt. Lett., 21, 1446 (1996))    (2) All-optical NOR implemented by two pump signals with the same wavelength (A. Sharaiha, H. W. L I, F. Matchese and J. Le Bihan, Electron. Lett., 33, 323(1997))    (3) All-optical NOR implemented by two pump signals with two different wavelengths (Young Tae Byun, Sang Hyuck Kim, Deok Ha Woo, Seok Lee, Dong Hwan Kim and Sun Ho Kim, New Physics, 40, 560(2000)), Apparatus and method for realizing All-optical NOR logic device (Young Tae Byun, Sang Hyuck Kim, Deok Ha Woo, Seok Lee, Dong Hwan Kim and Sun Ho Kim, U.S. Pat. No. 6,424,438 B1, Date of patent: Jul. 23, 2002)    (4) All-optical NOR implemented by connecting two semiconductor optical amplifiers (Ali Hamie, Ammar Sharaiha, Mikael Guegan and Benoit Pucel, IEEE Photon. Technol. Lett., 14, 1439 (2002)).
All-optical NOR logic devices that utilizes an ultra-fast nonlinear interferometer (UNI) as shown in (1) has an advantage of being able to operate at high speed.
However, it is unsuitable for an application in optical operating systems which require a high degree of integration. Also, their essential components consist of complicated optical fiber devices whose integration with other devices are also difficult.
On the contrary, all-optical logic devices which utilize the SOA are stable and small in size. They also have the advantages of being easily integrated with other optical devices and are independent of polarization and wavelength (T. Fjeld, D. Wolfson, A. kloch, B. Dagens, A. Coquelin, I. Guillemot, F. Gaborit, F. Poingt and M. Renaud, Electron. Lett., 36, 1863(2000)).
However, if only the nonlinear characteristic of single SOA is utilized without the optical fiber interferometer, then the structure of an all-optical logic NOR becomes simple and the integration with other devices is possible. However, its operating speed is less than 100 MHz.
In addition, an all-optical NOR logic device implemented by connecting two SOAs as listed in (4) has a better on/off ratio in a wide wavelength region in comparison to an all-optical NOR logic device that utilizes single SOA. However, its operating speed becomes below 62.5 MHz.
More specifically, in the conventional all-optical NOR logic devices as listed in (2) to (4), which do not use the optical fiber interferometer, a pump signal is None-Return to Zero (NRZ) pattern by using a square wave. In addition, a laser beam of continuous wave (CW) light is utilized as a probe signal.
In this case, the operating speed of the all-optical NOR logic devices is constrained to below 100 MHz due to the NRZ pattern of the pump signal and the CW light of the probe signal.
Accordingly, it is imperative to develop an all-optical NOR logic device which is not only possible to be integrated with other optical devices but also has a simple structure and an improved operating speed more than 10 GHz.

Apparatus and method for realizing all-optical NOR logic device using gain saturation characteristics of a semiconductor optical amplifier

Abstract: The method for realizing an all-optical NOR logic device using the gain saturation characteristics of the SOA according to the present invention comprises the steps of: utilizing A+B signal which couples together an input signal pattern A (1100) and an input signal pattern B (0110) as a pump signal (1110); utilizing a probe signal (1111) by generating a clock signal out of said input signal pattern A (1100); and obtaining a Boolean equation {overscore (A+B)} by making said probe signal and said pump signal incident upon the SOA simultaneously from the opposite direction.

Abstract: The all-optical logic device according to the present invention has a simple construction since it is realized through the XGM (Cross Gain Modulation) method which utilizes the gain saturation characteristics. Also, it is expected that the method employed in the present invention could be used for realizing other all-optical logic circuits and devices.

Korea Institute of Science and Technology


There is disclosed a transflective liquid crystal display having pixels each including a reflective region and a transmissive region. The aperture ratio of the pixels can be enhanced. End portions of two adjacent color filters are overlapped to form an optically shielding portion for each pixel. The optically shielding portion and the reflective region cooperate to form an optically shielding film acting as a black matrix for the transmissive region. The aperture ratio is enhanced by improving the positional relationship between the reflective regions and the color filters.

1. Field of the Invention
The present invention relates to a display device equipped with color filters and, more particularly, to a transflective liquid crystal display having a transmissive region and a reflective region within each one pixel.
2. Description of Related Art
In recent years, so-called transflective liquid crystal displays (LCDs) operating in both reflective and transmissive display modes have been often used as display portions of mobile electronic devices.
A transflective LCD has both transmissive and reflective regions within each one pixel. The transflective LCD operates either in a transmissive display mode in which light transmits through transmissive regions formed in pixels or in a reflective display mode in which light is reflected by reflective regions formed in the pixels.
When the transflective liquid crystal display is used outdoors, the display is employed under bright ambient conditions. In this state, the reflective mode is added to the transmissive mode, and external light is used for display.
The reflective LCD has the problem that the visibility deteriorates severely when the external light is weak. On the other hand, the transmissive LCD has the problem that in cases where the external light is very intense (e.g., under fine weather, outdoor conditions), the visibility deteriorates.
A transflective LCD uses a display mode in which the reflective mode and transmissive mode are combined to solve the above-described problems. This kind of device is proposed, for example, in U.S. Pat. No. 6,654,076.
Many transflective LCDs use an active matrix structure in which thin film transistors (TFTs) are used as switching elements for selectively supplying a video signal to pixel electrodes.
In an active-matrix liquid crystal display, a TFT substrate on which TFTs and pixel electrodes are formed is disposed opposite to a color filter substrate on which color filters are mounted to provide a color display. A liquid crystal material is sealed in between the two substrates, thus completing the display. Plural video signal lines and plural scanning lines intersect each other on the TFT substrate. Plural regions partitioned by the video signal lines and scanning lines are arranged in a matrix. TFTs and a pixel electrode are formed in each region, forming a pixel.
In a liquid crystal display, a counter electrode is disposed opposite to pixel electrodes. An electric field is produced between each pixel electrode and the counter electrode. The orientation of liquid crystal molecule is changed by applying a voltage between the pixel electrode and the counter electrode. Concomitantly, the characteristics of the liquid-crystal layer for light is change. By making use of this property, a display is provided.
Generally, VA (vertical alignment) displays in which a counter electrode is formed on a color filter substrate and IPS (in-plan switching) displays in which a counter electrode is formed on a TFT substrate are known.
Color filters are formed on a color filter substrate in a corresponding manner to pixels to provide a color display. Each color filter transmits certain wavelengths of light. Three pixels form a unit color element of a color display. Each pixel has color filters of red (R), green (G), and blue (B).


There is provided a multilayer film comprising a starch layer and at least one other layer. The multilayer film has excellent barrier properties. The starch layer comprises a modified starch having a degree of substitution less than 1.5. Suitable other layers include polyolefins. The multilayer film finds use in packaging, particularly in packaging foodstuffs.

Multilayer films find widespread use as a packaging material in the food industry. In such application not only does the packaging material have to contain the foodstuff, it must also preserve the product. Foods are by their nature perishable and the properties of the packaging material will greatly impact the useful shelf life of a given foodstuff. Chemical changes, such as oxidation, and microbiological growth can be accelerated in the presence of oxygen. Accordingly, controlling the oxygen content in a package or the rate of oxygen ingress into a package is often one of the most critical attributes of barrier packaging.
Multilayer barriers are usually constructed with a polar polymer as an internal layer which is covered with apolar polymer. The former acts as a gas barrier, and the latter as a hydrophobic skin having a low water vapour transmission rate to prevent fast water absorption in the internal layer. Polyolefin skin layers covering a polar polymer layer are commonly utilised, for example, a polyethylene skin covering a polyethylene-vinyl alcohol copolymer (PE-EVOH).
EVOH copolymers show good oxygen barrier properties at low humidity, typically in the range of 0 to 60%. However, their gas barrier property deteriorates dramatically under high humidity conditions when the humidity is in the range of 75 to 90%. In fact, due to the polar nature of EVOH, such films generally exhibit poor moisture barrier. Therefore, EVOH is typically laminated with polyolefins on both sides to provide barrier properties for practical packaging applications in order to protect the EVOH from humidity effects. However, over time sufficient moisture may permeate the polyolefin hydrophobic skin such that the oxygen barrier properties of the EVOH layer will be compromised.
EVOH materials also require the use of adhesion promoters and/or tie-layer resins in order for them to bond adequately to polyolefin substrates. Without such tie resins, EVOH materials tend to peel off easily from the polyolefin substrate resulting in loss of barrier properties and poor appearance.
A further disadvantage of EVOH is that it is relatively expensive. Additionally, from a renewable standpoint, EVOH is fully derived from fossil fuels.
It would be desirable to provide a multilayer film that has excellent barrier properties, particularly at high humidity and which utilises inexpensive, renewable materials.


A high-power fiber laser amplifier with a laser wavelength of about 1060 to 1100 nm and an average laser power of more than about 500 W and an almost diffraction-limited beam quality (M2<1.5) comprises a double-clad laser fiber that has a separate, laser-active fiber core with an outer diameter of at least 20 μm and a pump core that surrounds the fiber core and has an outer diameter of at least 50 μm, and ytterbium-doped solid state laser that emits pump light with a wavelength of approximately 1030 nm, a beam quality of M2< of approximately 30 and a laser power of at least approximately 600 watts, for pumping the double-clad laser fiber.

In accordance with the present state of the art, continuous laser radiation with an almost diffraction-limited beam quality can be obtained by individual laser diodes with small output powers (i.e., <1 W). Power sealing by coherent coupling of many diodes is possible for small output powers.
Solid state lasers that are pumped with laser diodes are currently used to produce higher fundamental mode powers with good electro-optical efficiency (i.e., 10-30%) in a spectral range around 1 μm. Pumping sources of reasonable brilliance can already realize an efficient fundamental mode operation by using laser transitions with small thermal occupation of the lower laser energy level (so-called four-level systems) and sufficient durability of the upper laser energy level and sufficiently large absorption and emission cross-sections. One decisive criterion in this connection is that only small thermo-optical effects (i.e., "thermal lens") occur in the laser-active medium. When laser-active media of rod-shaped or cuboid geometries are used, this is guaranteed only up to approximately 5 to 20 W output power per laser-active element. For fundamental mode powers of more than approximately 20 W per crystal, a rod laser can produce fundamental mode operation only with reduced efficiency and reduced dynamic stability range due to thermo-optical effects. With thoroughly homogenized pump light distribution, a side-pumped Nd:YAG rod achieves a maximum fundamental mode power of around 80 W with an optical-optical efficiency of approximately 20%.
It is possible to obtain a fundamental mode power of more than 100 W from a single laser-active medium having good efficiency (i.e., >40% optical-optical) by means of a laser-active medium of optimized geometry. Either a disc laser or a fiber laser are suitable for this purpose. The thermal-optical effects of the disc laser are considerably reduced due to its axial thermal flow, parallel to the resonator radiation, and its extremely short optical path, while the fiber laser is insensitive to thermal-optical effects due to wave guidance of the laser radiation. Both geometries require a relatively high pump radiation brilliance; a typical value for a Yb:YAG disc laser is approximately 5 kW/(mm·rad)2, whereas a unilaterally end-pumped fiber laser with 100 W output power, a typical pump core diameter of 0.4 mm and a numerical aperture of 0.45 requires approximately 20 kW/(mm·rad)2. The output power is scaled in the disc laser with approximately constant pump radiation brilliance, wherein the diffraction-limited beam quality can be maintained with good efficiency only up to approximately 200 W. The output power of the fiber laser can be doubled to approximately 200 W through pumping at both sides with the same pump light brilliance. The output power of the fiber laser can be further increased, and an increase in output power requires a proportional increase of the pump light brilliance if the diameter and the numerical aperture of the pump core are kept constant. Thus, high-power fiber laser with output powers of more than 500 W and an almost diffraction-limited beam quality put very high demands on the brilliance of the pump light source(s). The requirements with regards to brilliance can be reduced by increasing the numerical aperture of the pump core by increasing the pump core diameter.
Conventional high-power fiber lasers are end-pumped with laser diode arrays having complex beam shapes and a high brilliance. The wavelength of the pump light is usually about 980 or 915 nm. The required pump light brilliance increases with increasing output power, which also increases the expense for shaping the beams of the laser diodes. High-power laser diode arrays ("stacks") with good fast-axis collimation have a brilliance of approximately 10-20 kW/(mm·rad)2 such that further measures are used to increase the brilliance of a diode-pumped fiber laser with 1 kW output power by approximately five to ten times. For example, the further measures can include polarization coupling, slow-axis collimation, and wavelength multiplex. Due to the required multi-stage optical transformations, the electro-optical efficiency of these pump light sources with high brilliance is considerably smaller than that of a simple pump light source. At the same time, the required optical expense greatly increases the costs of the pump light sources.

High-power fiber laser amplifier and fiber laser oscillator


A locking device for opening or closing a door comprising a lock housing having a pair of holes; a slider slidable within the lock housing; a pair of clamps, the base end being pivotally mounted on the front end of the slider, and the other end having a spring force acting outward; a spring member for the clamp attached to the front end of the slider; rails each provided on the outside of the pair of clamps in the lengthwise direction; a pair of rollers each having a circular shape, which can be fitted with each of the rails, and are supported to be revolved in adjacent to the holes of the lock housing; a spring member for the slider which is intervened between the lock housing and the slider and which applies a spring force to the slider in the direction forward to the hole of the lock housing; a heart-shaped groove carved in one side of the slider, having a plurality of differences in depth, and a recess provided on the bottom portion thereof; a projecting member, one end being slidable in the groove only in one direction, and the other end being formed on the lock housing; a spring member for the projecting member; a resilient member which reinforces the resiliency of the spring member for the projecting member; and a key unit which can be clamped by the pair of clamps is provided.

1. Field of the Invention
The present invention relates to a locking device for opening or closing a door which is used in kitchen furniture and the like.
2. Prior Art
In conventional, there is a locking device for opening or closing a door, for example, as described in Japanese examined Utility Model Publication 2-2857 (corresponding to U.S. Pat. No. 4,482,175). The locking device disclosed therein comprises a lock housing 36, having a case 38, formed in a cabinet A for kitchen furniture and a key unit 37 formed in a door B as shown in FIG. 4. The lock housing 36 and the key unit have the relation to be fitted with each other and to be released. The lock housing 36 of the cabinet A has a pair of holes formed in the surface thereof in which a slider which can be moved 40 is accommodated. At the front end of the slider 40, a pair of clamps 41 and 41 which can revolve are supported. A spring member (not shown) is provided so that it is applied to a spring force to open the front ends of the clamps 41 and 41 in a V-shape. At the same time, a heart-shaped groove 42 is provided on the side of the slider. A projecting member 43, one end of which slides within the slider only in one direction is provided, and the other end is formed on an appropriate position of the cabinet A, is applied to a spring force in the direction of groove 42 by a leaf spring 45.
Consequently, when the door B in the closed state is opened, the clamps 41 and 41, which are opened in the rear surface of the door in a V shape, are brought into contact with each other in the state where a spherical member 47 of the key unit is intervened between the clamps. When the door B is further pushed toward the direction of the cabinet A, the clamps 41 and 41 are guided by rollers 48 and 48 to be pushed inward. The clamps 41 and 41 then revolve in the direction where the open ends thereof are closed, and they clamp the spherical member 47 and move toward the direction of the cabinet A. At this time, the front end of the projecting member 43 residing within the heart-shaped groove is positioned at the recess via one side of the groove 42 to the top of the groove, and inhibits the slide of the clamps 41 and 41. The door B is locked on the cabinet A in this state.
The locking device as described above in which the leaf spring 45 is used to apply a spring force to the heart-shaped groove is disadvantageous in terms of the durability in that when the number of using the lock device is increased, the resilient of the spring force becomes poor.
Furthermore, when the door is further pushed, the front ends of the clamps 41 and 41 are opened in a V-shape by a spring force of the spring provided on the base ends of the clamps 41 and 41 whereby the slider 40 and the clamps 41 and 41 move forward to release the locking state in the locking device disclosed therein. However, since the movement is due to the spring force of the spring provided on the base ends of the clamps 41 and 41, it is difficult to smoothly move the slider 40 and the clamps 41 and 41 forward, and their actions tend to be slow.
Moreover, it is difficult to correctly fix the spherical member 47 of the key unit in the door side at the position opposite to the lock housing 36 and, thus, there is a tendency to have some gaps between the positions of the lock housing 36 and the key unit 37.

Tokyo Magnet Ohyo Seihin Kabushiki Kaisha


The subject matter disclosed herein relates to rofecoxib, also known as TRM-201 or RXB-201, its method of manufacture, and use. In certain aspects, the highly pure or substantially pure rofecoxib as provided herein has a favorable purity profile and is the active ingredient in a pharmaceutical composition that is administered to treat or prevent a number of conditions, including pain associated with a condition caused by a bleeding disorder.

Rofecoxib is a selective COX-2 inhibitor, nonsteroidal anti-inflammatory drug (NSAID), that was marketed under the brand name "VIOXX" until it was withdrawn from the market in 2004 over safety concerns. Before being withdrawn from the market, "VIOXX" was approved in the United States for the following indications: signs and symptoms of osteoarthritis (OA); signs and symptoms of rheumatoid arthritis (RA) in adults; signs and symptoms of pauciarticular or polyarticular course Juvenile Rheumatoid Arthritis; management of acute pain in adults; treatment of primary dysmenorrhea; and treatment of migraine attacks with or without aura in adults.
There is a long felt but unmet need in the art for new medications for the treatment of pain, fever, and inflammation. This is especially true with respect to subjects suffering from pain that is co-morbid or associated with diseases or conditions arising from bleeding disorders, including hemophilic arthropathy and von Willebrand's disease. Hemophilia is a bleeding disorder caused by inherited or spontaneous mutations in genes that code for clotting factor. The most common forms of hemophilia are the result of deficiencies in the coagulation factors VIII (hemophilia A) or IX (hemophilia B). The prevalence of subjects with hemophilia (PWH) in the United States is estimated at 20,000 subjects.
Blood-induced joint disease may follow acute joint injury and is associated with hemophilia, with intra-articular bleeding (hemarthrosis) accounting for more than 90% of all serious bleeding events in subjects with severe hemophilia (baseline factor (F)VII or FIX activity <1%). Over time, recurrent bleeding into the same articular joints results in progressive damage and the development of hemophilic arthropathy. Despite advances in treatment and the delivery of comprehensive care at dedicated centers, joint bleeding and arthropathy remain the single largest cause of morbidity in PWH.
Although the pathogenesis of the blood-induced joint injury of hemophilic arthropathy has not been fully elucidated, it appears to have similarities with the degenerative joint damage that occurs in mechanically-induced joint injury such as OA and the inflammatory processes associated with RA.
Arthritis as a result of hemophilia is the most common comorbidity in adult PWH, with 44-55% of subjects reporting suffering from hemophilic arthropathy. Pain/discomfort is the most frequently reported limitation for PWH, with 75% of PWH reporting this limitation. Eighty-nine percent of PWH reported that pain had interfered with their daily life in the past 4 weeks, and 50% reported suffering from constant pain.
In additional to its impact on quality of life, hemophilic arthropathy has a significant impact on the cost associated with the treatment of hemophilia. As acute pain associated with hemarthrosis and chronic pain associated with arthritis are largely indistinguishable for PWH, significant over-utilization of factor replacement to address pain symptoms have been reported and 58% of PWH report using factor replacement to treat chronic pain. In addition to being medically ineffective, this misutilization has significant economic implications, as factor replacement is one of the costliest known drug interventions.
Currently, management of the pain and inflammation associated with hemophilic arthropathy is difficult, due to a lack of approved treatments and limitations to existing off-label options. Acetaminophen is generally recommended as first-line therapy, however, it is of limited utility owing to its lack of anti-inflammatory effect and increased risk of hepatic adverse events in a subject population with a significant rate of comorbid chronic hepatitis C.
Opioids can be used to alleviate pain associated with hemophilic arthropathy, but also have little anti-inflammatory effect and long-term use of this mode of analgesic therapy may lead to tachyphylaxis, dependence, and the potential for abuse. Additionally, a 2010 observational study by Solomon et al. demonstrated that long-term chronic use of opioids to treat arthritis in the general population resulted in increased risk of (cardiovascular) CV events, fractures, hospitalization, and all-cause mortality relative to non-selective NSAIDs or a combined group of therapeutic and supra-therapeutic doses of COX-2 selective NSAIDs.
Due to their effect on platelet function, NSAIDs, particularly acetylsalicylic acid (ASA), are not recommended in PWH. In addition, even otherwise trivial NSAID-induced ulcerations might be expected to result in more severe or prolonged bleeding in the setting of an underlying coagulopathy, and it has been demonstrated that NSAIDs are associated with an increased risk of upper gastrointestinal (UGI) complications, including ulcers, bleeding and perforations, in PWH. Thus, there is a long felt but unmet need in the art for medications to treat pain in PWH.
Von Willebrand Disease (vWD) is a genetic disorder caused by defective von Willebrand factor, a clotting protein vWD is the most common form of bleeding disorder, affecting ˜1% of US population, though mild cases are often undiagnosed. Generally mild and characterized by heavy mucosal bleeding, more severe forms can manifest into significant broad-ranging internal bleeding. Evidence suggests that vWD subjects are more susceptible to migraine headaches. Due to risk of exacerbating hemorrhaging, NSAIDs are not recommended for pain management in subjects with vWD.
Juvenile idiopathic arthritis, including systemic juvenile idiopathic arthritis (SJIA) is one of several rheumatic diseases that affect children. SJIA affects the entire body, including the joints. SJIA typically occurs in flares with some subjects having healthy periods between flares. Diagnosis may be delayed by these ups and downs over the course of the disease. SJIA is generally easier to diagnose during a flare. The most common symptoms of SJIA are recurrent bouts of fever with high daily temperature spikes, skin rash, and painful, stiff joints.

This patent is subject to a terminal disclaimer.

Purified forms of rofecoxib, methods of manufacture and use


A laser imageable flexographic printing plate precursor comprises a thermoset elastomeric upper layer that is at least partially ablatable and comprises a radiation sensitive compound, and a non-ablatable elastomeric underlayer. This flexographic printing plate precursor can be imaged to provide a printing plate that is primarily useful for “high quality” printing because the resulting relief image is generally not greater than 600 μm and has an extremely even “floor”. The imaged flexographic printing plate also can have a high visual contrast between the imaged areas and the non-imaged background areas.

Flexography is a method of printing whereby a flexible plate with a relief image is wrapped around a cylinder and its relief image is inked up and the ink is then transferred to a suitable printable medium. The process has mainly been used in the packaging industry where the plates should be sufficiently flexible and the contact sufficiently gentle to print on uneven substrates such as corrugated cardboard as well as flexible materials such as polypropylene film. The quality of the printing in this manner is inferior to processes such as lithography and gravure, but nevertheless it is useful in certain markets. In order to accommodate the various types of printing media, the flexographic plates should have a rubbery or elastomeric nature whose precise properties can be adjusted for each particular printable medium.
In addition, where the flexographic printing plates are formed and/or imaged in a flat form, they should be flexible for bending around a cylinder for rotary printing. This can present more of a problem than with offset lithographic plates because the thickness of flexographic printing plates is generally several millimeters instead of fractions of a millimeter. Materials that are flexible as one or two μm films can be rigid and inflexible at one or more mm.
U.S. Pat. No. 4,323,636 (Chen) describes elements having thermoplastic elastomeric block copolymers (for example, those sold by Kraton Polymers under the trademark of KRATON) used in conjunction with an acrylate or methacrylate monomer and a photoinitiator. The upper surface may have on it a thin hard flexible solvent insoluble coating and on top of this a strippable thin film of e.g. polyethylene to protect the plate during storage. This would constitute a flexographic printing plate precursor that can be imaged by ultra-violet exposure through a negative mask, and the un-polymerized material washed away with solvent. Such elements usually have a thickness of one or more millimeters. Exposure from the front through an image bearing transparency is sufficient to polymerize both the image areas and the underlying layer. The block co-polymer materials formulated as printing blanks, whilst being able to be formed into solid blanks, retain a stickiness on their upper and lower surfaces that requires the use of protective films to prevent unused plates sticking together during storage.
U.S. Pat. No. 4,994,344 (Kurtz et al.) describes flexographic printing blanks prepared from ethylene-propylene-alkadiene terpolymers. It describes the process of initial back exposure to establish the "floor" of the plate before image exposure from the front of the plate through a negative mask. The image floor may be uneven due to differences in evenness of UV exposure and subsequent wash-out, but this would be of little consequence where image thickness (relief) is measured in millimeters. Because the plate requires back exposure, a polyester substrate is often used as the dimensionally stable backing material and must be transparent to UV light. In addition, the floor material of the plate is of the same material and formulation as the image areas. Thus, the finished plate generally has little or no visual contrast between the image areas and the floor and it is difficult for the user to make any visual assessment of the image because of this lack of contrast.
U.S. Pat. No. 5,719,009 (Fan) describes the use of a negative mask that is integral in the flexographic printing plate itself. The flexographic printing plate comprises photosensitive layers and an overcoat containing carbon black with a binder resin. The overcoat is ablated with an infrared laser in response to a digital signal generated by a computer. Digital imaging using a modulated laser source is an important part of the general technology that has become known as computer-to-plate (CTP) and is used for instance in the production of offset lithographic printing plates. The ablated areas in the overcoat permit a subsequent irradiation by UV light to expose the sensitive elastomeric layer and to harden it. The other unexposed areas situated under the non-ablated areas of the overcoat are washed away together with the remains of the carbon layer, leaving a relief image. In this case, there is good visual contrast between the masked areas that remain after ablation and the image areas that are exposed by ablation. However, after UV exposure and subsequent wash-out processing, any visual contrast disappears as the overcoat is washed away with the underlying unexposed background areas, so that the imaged flexographic printing plate has no visual contrast between the image and the background. The UV exposure and washing process still results in unevenness of the image floor.
It has long been recognized that the simplest way of making a flexographic printing plate would be by direct engraving using laser beam ablation, thereby eliminating all need for washing or drying the plate or multiple types of exposure.
U.S. Pat. No. 3,549,733 (Caddell) describes the formation of a laser engraved (or imaged) relief printing plate. However, the described plates do not have the elastomeric properties needed for flexographic printing but could be used in letterpress printing. Letterpress printing differs significantly from flexographic printing in that it is more like lithography in the complexity of the printing machine and the type of ink used. Letterpress inks must have high viscosity (paste-like), similar to offset inks and do not in general contain volatile solvents. If the letterpress printing is carried out using an offset blanket, the printing process is termed dry offset. As with offset printing, dry offset and letterpress require high pressure between the plate and blanket or printable media to achieve good ink transfer, whereas flexographic printing uses the minimum pressure possible. Thus a letterpress plate would be unsuitable for flexographic printing as it would not give good ink transfer under low pressure and similarly a flexographic plate would be unsuitable for letterpress as the high pressure would distort the softer plate and give very poor image quality with huge dot gain.
U.S. Pat. Nos. 5,798,202 and 5,804,353 (both Cushner et al.) describe the use of single or multiple layers of elastomers in flexographic printing plate precursors for direct laser engraving. The upper layer of the precursors is comprised of a thermoplastic elastomeric material.
Imaging sensitivity is limited by the use of large quantities of block polymers, such as those sold under the trade name of KRATON by Kraton Polymers. Poor melt edges are reported for flexographic engraving of layers containing such polymers in U.S. Pat. No. 6,627,385 (Hiller, in the Comparative Examples). The patent also describes the problem of using carbon black or opaque fillers in that the flexographic printing plate loses its transparency, which complicates mounting it with accurate register, since register crosses or similar marks, are no longer visible through the plate. Hiller suggests avoiding such layers.
U.S. Pat. No. 6,159,659 (Gelbart) describes a flexographic printing plate precursor having two ablatable layers, the upper layer comprising an elastomeric foam mounted on a thin non-ablatable backing layer where preferably ablation removes material right down to the backing layer. The method is intended to solve the problem in the prior art of small holes and nicks in the backing caused by exposure by the laser that reduce the life of the printing plate. However, no attempt is made to provide very high adhesion between the two layers that may be needed especially for small isolated image areas.
Problem to be Solved
Despite the limitations of carbon dioxide lasers, they are now being used commercially in flexographic engraving machines. They are known for slow and expensive imaging with limited resolution. However, the advantages of direct engraving are sufficient to ensure their commercial use in instances where fast imaging and high print quality are not required. It would be preferable to use infrared diodes that produce radiation in the near infrared and infrared (approximately 700 to 1200 nm) and have the advantages of high resolution and relatively low laser cost so that they can be used in large arrays. Until now, although the use of such lasers is described in many publications, they are not in industrial use because even when combined with the most sensitive imageable elements available, satisfactory engraving may not be achieved.
Infrared diode engraving (or ablative imaging) differs from that of carbon dioxide in that a compound absorbing suitable radiation (that is, IR radiation) is usually incorporated into the imaged coating. The use of an organic infrared radiation-sensitive dye (IR dye) may be prohibitive because it is costly and a large quantity of IR dye is needed throughout the imaging layer (which may be millimeters thick). The use of an opaque pigment such as carbon black reduces the possibility of visual contrast even further. Another problem experienced with high carbon and other fillers is the loss in layer resilience. Good resilience ensures the rapid elimination of any distortion of the plate during a printing cycle by permitting the plate to recover its original shape in time for the next cycle. Distortion may also occur from dirt entering the printing system and causing temporary indentations in the printing plate surface. Thus, good resilience is needed to provide fast recovery from distortion of any type.
The recent availability of high power (for example, 8 watts) IR-laser diodes opens up opportunity for the use of relatively low cost laser diode arrays capable of engraving flexographic blanks as described in WO 2005/84959 (Figov). Relief depth in the resulting image is an issue with laser engraving because the deeper the required relief, either more power is required or it takes longer to engrave or image the plate. Besides the segment of the flexographic market that involves deep relief, laser engraving is most suitable and competitive when applied to the high quality market segment using even substrates where low relief is a distinct advantage. In this market segment, it is of no advantage to work with high relief images as the image areas would have the possibility of more movement during printing and more dot gain and inaccuracy of printing. However, in order to achieve minimum image relief, the floor of the image should be extremely even, otherwise there is the danger that any slightly elevated floor area would give unwanted background printing.

Flexographic printing plate precursor and imaging method


A waterproof and water-vapor-permeable composite laminated material that has good snag resistance and puncture resistance is described. The material is suitable for making outdoor apparel for wear in rugged environments, for example, where brambles, briers or dense underbrush are likely to be encountered.

Waterproof fabrics for use in outdoor apparel suitable for wear in most rugged physical environments and weather conditions are well known. These fabrics are usually quite heavy and are typically made from yarns of high strength abrasion-resistant synthetic polymer fibers woven in tight smooth patterns having a high yarn count in both warp and fill directions. Such fabrics when coated with a continuous layer of polyvinyl chloride, polyurethane polymers, or other such compounds can be made into waterproof garments for protection in rainy or inclement weather. An example of such a coated fabric is a plain weave fabric of synthetic polymer yarn with a yarn count of 65.times.60 yarns per inch (210 denier, 24 filament yarn) that has been coated with a layer of polyvinyl chloride, polyurethane, or other such polymer. These continuous waterproof coating materials, however, do not permit passage of air or water vapor (from perspiration) through them and, consequently, the garments can become quite stuffy and uncomfortable to the wearer. Furthermore, when scratched by thorns, prickles, branches, and other sharp objects while traversing dense brush and brambles the coating may be punctured or worn off and a measure of the waterproofness lost.
Also known is weather-resistant waterproof and breathable outdoor apparel made from composite laminated fabric which consists of textile fabric laminated to a porous hydrophobic polymer film. The porous hydrophobic polymer film, which may also be formed as a composite in combustion with hydrophilic polymeric materials, resists the passage of liquid water but will readily transmit water vapor through it, i.e., is breathable, thus significantly increasing wearer comfort. Exemplary of materials and garments utilizing this method for weather resistance and waterproofness are those fashioned from GORE-TEX.RTM. membrane and GORE-TEX.RTM. fabric laminates prepared according to U.S. Pat. Nos. 4,194,041 (Gore, et al.), U.S. Pat. No. 4,187,390 (Gore), and U.S. Pat. No. 3,953,566 (Gore) which include films of porous expanded polytetrafluoroethylene, characterized by having a structure of nodes interconnected by fibrils, laminated to at least one layer of a textile fabric.
Despite the use of heavy densely woven outer fabrics in the apparel described above, when worn where dense brush or thickets of brambles, briers and other thorny bushes are routinely encountered, most are not entirely satisfactory in preventing damage to the garment or in protecting the wearer from injury or discomfort. Harm arising from sharp objects present in brambles, briers, and other dense brush can occur by immediate tearing or puncturing of the garment as the wearer passes through them or, more insidiously over a period of time, from thorns, prickles, and other sharp or needle-like objects that are picked up or become embedded in the outer fabric and, by movement of the yarn and weave of the fabric, are forced through to the wearer's side of the garment. In either case, physical damage to the garment, loss of waterproofness, and discomfort or injury to the wearer can result.


The present invention relates to a method of isolating stem cells from a mixed population of different cell types. This method involves selecting a promoter which functions only in said stem cells and not in the other cell types. The promoter can have a nucleotide sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8. A nucleic acid molecule encoding a marker protein under control of said promoter is introduced into all cell types of said mixed population and only the stem cells, but not the other cell types, within said mixed population are allowed to express said marker protein. The cells of said mixed population of cell types that are expressing the marker protein, which are restricted to the stem cells are identified and separated from the mixed population, where the separated cells are restricted to the stem cells. The promoter, nucleic acid constructs comprising it, and the resulting stem cells are also disclosed. The nucleic constructs can be incorporated in transgenic animals in order to monitor stem cell movement. A disorder mediated by stem cell proliferation can be carried out by administering both (1) a nucleic acid construct comprising the promoter of the present application and an enzyme capable of converting a prodrug to a cytotoxic drug and (2) the prodrug.

Rapid revascularization of injured, ischemic and regenerating organs is essential to restore organ function. The angiogenic switch initiates the revascularization process and involves recruitment of endothelial cells that assemble into neovessels (Folkman et al., "Angiogenesis in Cancer, Vascular, Rheumatoid and Other Disease," Nat. Med. 1:27-31 (1995); Folkman et al., "Therapeutic Angiogenesis in Ischemic Limbs," Circulation 97:1108-1110 (1998); Hanahan et al., "Patterns and Emerging Mechanisms of the Angiogenic Switch During Tumorigenesis," Cell 86:353-364 (1996); Risau, W. "Mechanisms of Angiogenesis," Nature 386:671-674 (1997); Yancopoulos et al., "Vascular-specific Growth Factors and Blood Vessel Formation," Nature 407:242-248 (2000); Carmeliet et al., "Angiogenesis in Cancer and Other Diseases," Nature 407:249-257 (2000); Pepper et al., "Manipulating Angiogenesis. From Basic Science to the Bedside," Arterioscler. Thromb. Vasc. Biol. 17:605-619 (1997). Much effort has been focused on delivering aniogenic factors to accelerate tissue revascularization (Isner et al., "Myocardial Gene Therapy," Nature 415:234-239 (2002); Khurana et al., "Insights from Angiogenesis Trials Using Fibroblast Growth Factor for Advanced Arteriosclerotic Disease," Trends Cardiovasc. Med. 13:116-122 (2003); Cao et al., "Angiogenic Synergism, Vascular Stability and Improvement of Hind-Limb Ischemia by a Combination of PDGF-BB and FGF-2," Nat. Med. 9:604-613 (2003); and Carmeliet, P., "VEGF Gene Therapy: Stimulating Angiogenesis or Angioma-Genesis?" Nat. Med. 6:1102-1103 (2000). However, because tissue injury is associated with disruption of a permissive microenvironment necessary for recruiting pre-existing endothelial cells (ECs), exogenous introduction of vascular progenitors may facilitate restoration of organ revascularization. Accumulating evidence suggests that bone marrow contains vascular progenitor cells that can mobilize to ischemic sites and complement neo-angiogenesis afforded by pre-existing endothelium, thereby restoring rapid and timely organ revascularization.
Adult bone marrow is a rich reservoir of tissue-specific stem and progenitor cells. Among these, a scarce population of cells known as EPCs can be mobilized to the circulation and contribute to the neoangiogenic processes. Circulating EPCs (CEPs) have been detected in the circulation either after vascular injury or during tumor growth. CEPS primarily originate from EPCs within the bone marrow and differ from sloughed mature, circulating endothelial cells (CECs) that randomly enter the circulation as a result of blunt vascular injury. It is possible, however, that the parenchyma of the systemic vasculature or certain organs may harbor endogenous EPC-like cells. For example, distinct side population stem cells within the skeletal muscle can differentiate into ECs (Majka et al., "Distinct Progenitor Populations in Skeletal Muscle are Bone Marrow Derived and Exhibit Different Cell Fates During Vascular Regeneration," J. Clin. Invest. 111:71-79 (2003). Therefore, CEPs may originate either from bone marrow-derived EPCs or from resident EPCs embedded within organs and the systemic vasculature. Endothelial progenitor cells residing in the bone marrow will be referred to as EPCs, while endothelial progenitors detected in the circulation will be referred to as CEPs.
Emerging evidence suggests that angiogenic factors recruit subsets of proangiogenic hematopoietic cells, including hematopoietic stem cells (HSCs) and hematopoietic progenitor cells (HPCs). Corecruitment of HSCs and HPCs, along with EPCs and CEPs, may contribute to the initiation and sustenance of neoangiogenesis. The physiological role of corecruited HSCs and HPCs in formation of long-lasting functional neovessels remains to be determined.
Several studies have shown that bone marrow-derived cells functionally contribute to neoangiogenesis during wound healing and limb ischemia (Majka et al., "Distinct Progenitor Populations in Skeletal Muscle are Bone Marrow Derived and Exhibit Different Cell Fates During Vascular Regeneration," J. Clin. Invest. 111:71-79 (2003); Asahara et al., "Isolation of Putative Progenitor Endothelial Cells for Angiogenesis," Science 275:964-967 (1997); Asahara et al., "Bone Marrow Origin of Endothelial Progenitor Cells Responsible for Postnatal Vasculogenesis in Physiological and Pathological Neovascularization," Circ. Res. 85:221-228 (1999); (Asahara et al., VEGF Contributes to Postnatal Neovascularization by Mobilizng Bone Marrow-Derived Endothelial Progenitor Cells," EMBO J. 18:3964-3972 (1999); Iwaguro et al., "Endothelial Progenitor Cell Vascular Endothelial Growth Factor Gene Transfer for Vascular Regeneration," Circulation 105:732-738 (2002); Kalka et al., "Transplantation of ex vivo Expanded Endothelial Progenitor Cells for Therapeutic Neovascularization," Proc. Natl. Acad. Sci. USA 97:3422-3427 (2000); Schatteman et al., "Blood-Derived Angioblasts Accelerate Blood-Flow Restoration in Diabetic Mice," J. Clin. Invest. 106:571-578 (2000); Crosby et al., "Endothelial Cells of Hematopoietic Origin Make a Significant Contribution to Adult Blood Vessel Formation," Circ. Res. 87:728-730 (2000); Takahashi et al., "Ischemia- and Cytokine-Induced Mobilization of Bone Marrow-Derived Endothelial Progenitor Cells for Neovascularization," Nat. Med. 5:434-438 (1999); Luttun et al., "Vascular Progenitors: From Biology to Treatment," Trends Cardiovasc. Med., 12:88-96 (2002); Rafii et al., "Circulating Endothelial Precursors: Mystery, Reality, and Promise," J. Clin. Invest. 105:17-19 (2000), postmyocardial infarction (Orlic et al., "Bone Marrow Cells Regenerate Infarcted Myocardium," Nature 410:701-705 (2001); Orlic et al., "Mobilized Bone Marrow Cells Repair the Infarcted Heart, Improving Function and Survival," Proc. Natl. Acad. Sci. USA 98:10344-10349 (2001); Kocher et al., "Neovascularization of Ischemic Myocardium by Human Bone-Marrow-Derived Angioblasts Prevents Cardiomyocyte Apoptosis, Reduces Remodeling and Improves Cardiac Function," Nat. Med. 7:430-436 (2001); Jackson et al., "Regeneration of Ischemic Cardiac Muscle and Vascular Endothelium by Adult Stem Cells," J. Clin. Invest. 107:1395-1402 (2001); Edelberg et al., "Young Adult Bone Marrow-Derived Endothelial Precursor Cells Restore Aging-Impaired Cardiac Angiogenic Function," Circ. Res. 90:E89-E93 (2002), endothelialization of vascular grafts (Shi et al., "Evidence for Circulating Bone Marrow-Derived Endothelial Cells," Blood 92:362-367 (1998); Bhattacharya et al., "Enhanced Endothelialization and Microvessel Formation in Polyester Grafts Seeded with CD34+Bone Marrow Cells," Blood 95:581-585 (2000); Kaushal et al., "Functional Small-Diameter Neovessels Created Using Endothelial Progenitor Cells Expanded ex vivo," Nat. Med. 7:1035-1040 (2001); Noishiki et al., "Autocrine Angiogenic Vascular Prosthesis with Bone Marrow Transplantation," Nat. Med. 2:90-93 (1996), atherosclerosis (Sata et al., "Hematopoietic Stem Cells Differentiate into Vascular Cells that Participate in the Pathogenesis of Atherosclerosis," Nat. Med. 8:403-409 (2002), retinal and lymphoid organ neovascularization (Otani et al., "Bone Marrow Derived Stem Cells Target Retinal Astrocytes and Can Promote or Inhibit Retinal Angiogenesis," Nat. Med. 8:1004-1010 (2002); Grant et al., "Adult Hematopoietic Stem Cells Provide Functional Hemangioblast Activity During Retinal Neovascularization," Nat. Med. 8:607-612 (2002); Crisa et al., "Human Cord Blood Progenitors Sustain Thymic T-Cell Development and a Novel Form of Angiogenesis," Blood 94:3928-3940 (1999), vascularization during neonatal growth (Young et al., "VEGF Increases Engraftment of Bone Marrow-Derived Endothelial Progenitor Cells (EPCs) into Vasculature of Newborn Murine Recipients," Proc. Natl. Acad. Sci. USA 99:11951-11956 (2002) and tumor growth (Asahara et al., "Bone Marrow Origin of Endothelial Progenitor Cells Responsible for Postnatal Vasculogenesis in Physiological and Pathological Neovascularization," Circ. Res. 85:221-228 (1999); Lyden et al., "Impaired Recruitment of Bone-Marrow-Derived Endothelial and Hematopoietic Precursor Cells Blocks Tumor Angiogenesis and Growth," Nat. Med. 7:1194-1201 (2001); Reyes et al., "Origin of Endothelial Progenitors in Human Postnatal Bone Marrow," J. Clin. Invest. 109:337-346 (2002); Moore, M. A., "Putting the Neo into Neoangiogenesis," J. Clin. Invest. 109:313-315 (2002); Gehling et al., "In vitro Differentiation of Endothelial Cells from AC133-Positive Progenitor Cells," Blood 95:3106-3112 (2002); Marchetti et al., "Endothelial Cells Genetically Selected from Differentiating Mouse Embryonic Stem Cells Incorporate at Sites of Neovascularization in vivo," J. Cell. Sci. 115:2075-2085 (2002); Davidoff et al., "Bone Marrow-Derived Cells Contribute to Tumor Neovascular and, When Modified to Express an Angiogenesis Inhibitor, Can Restrict Tumor Growth in Mice," Clin. Cancer Res. 7:2870-2879 (2001)).
These studies have introduced the concept that vascular trauma and organ regeneration results in the release of chemokines that recruit EPCs and CEPs to the neoangiogenic site. Rapid incorporation of EPCs and CEPs accelerates vascular healing and prevents potential vascular complications secondary to thrombosis and hypoxia. Tissue ischemia results in upregulation of angiogenic factors, including vascular endothelial growth factor (VEGF)-A, which through interaction with its receptors VEGFR2 (KDR or Flk-1) and VEGFR1 (Flt-1) expressed on EPCs, CEPS, HSCs and HPCs, promotes migration of these cells to the site of the injury.
Despite the contribution of bone marrow-derived progenitors to tissue revascularization in animals models, the importance of these cells in restoring organ vascularization in a clinical setting remains unknown. Several recent clinical trials have challenged the potential of bone marrow-derived cells in restoring vascularization of ischemic tissues. The success of these strategies depends on defining the mechanisms by which stem and progenitor cells undergo appropriate molecular and differentiation, thereby permitting their functional incorporation into adult tissues.
The present invention is directed to overcoming the deficiencies in the art by providing a promoter which functions only in stem cells and not in other cell types to facilitate recovery and study of stem cells and to achieve therapeutic benefits.

Stem cell-specific promoters and their use


A seatbelt arrangement for restraining an occupant of a vehicle in a seat. The seatbelt arrangement has a latch plate and a buckle to which the latch plate can be locked. The buckle includes a housing having a plurality of sidewalls, a biasing member anchor provided on one of the plurality of sidewalls, and a mounting anchor. A mounting member is connected to the mounting anchor. The mounting member secures the buckle to the vehicle. A biasing member is attached to the biasing member anchor. The biasing member biases the buckle to a stowed position when the seatbelt arrangement is unfastened.

It is known to provide a vehicle with a seatbelt arrangement to restrain an occupant in a vehicle seat in the event of collision. A typical 3-point seatbelt arrangement includes a lap belt section and a shoulder belt section formed from a continuous length of webbing. When an occupant is seated in the vehicle seat the lap belt section is configured to extend across the occupant's pelvis and the shoulder belt section is configured to extend across the occupant's upper torso. A latch plate attached to the webbing is received by a buckle to allow the seatbelt arrangement to be fastened for enabling restraint and unfastened to allow entrance and egress from the vehicle.
The vehicle seat can have a seat back that is moveable relative to a seat cushion. This relative movement can be for facilitating access to a desired part of a vehicle or for allowing reconfiguration of an interior space of the vehicle to carry oversized cargo. In such arrangements it can be desirable to prevent the buckle from becoming trapped between the seat back and the seat cushion in order to prevent cosmetic and/or functional damage to the buckle. Furthermore, even in instances where the seat back is fixed in position relative to the seat cushion, it can be desirable to stow the buckle when the seatbelt arrangement is unfastened in order to maintain a neat and orderly appearance of the vehicle interior as well as to prevent the buckle from contacting other vehicle components and potentially generating noise.


Agonists of A.sub.2a adenosine receptors are effective for the treatment of inflammatory diseases.

1. Field of the Invention:
The present invention relates to methods and compositions for treating inflammatory diseases.
2. Discussion of the Background:
The release of inflammatory cytokines such as tumor necrosis factor-alpha (TNF.alpha.) by leukocytes is a means by which the immune system combats pathogenic invasions, including infections. Cytokines stimulate neutrophils to enhance oxidative (e.g. superoxide and secondary products) and nonoxidative (e.g. myeloperoxidase and other enzymes) inflammatory activity. Inappropriate and over-release of cytokines can produce counterproductive exaggerated pathogenic effects through the release of tissue damaging oxidative and nonoxidative products (Tracey, K. G., et al, J. Exp. Med., vol. 167, pp. 1211-1227 (1988); and Mannel, D. N., et al, Rev. Infect. Dis., vol. 9 (suppl 5), pp. S602-S606 (1987)).
For example, inflammatory cytokines have been shown to be pathogenic in: arthritis (Dinarello, C. A., Semin. Immunol., vol. 4, pp. 133-45 (1992)); ischemia (Seekamp, A., et al, Agents-Actions-Suppl., vol. 41, pp. 137-52 (1993)); septic shock (Mannel, D. N., et al, Rev. Infect. Dis., vol. 9, (suppl 5), pp. S602-S606 (1987)); asthma (Cembrzynska Nowak M., et al, Am. Rev. Respir. Dis., vol. 147, pp. 291-5 (1993)); organ transplant rejection (Imagawa, D. K., et al, Transplantation, vol. 51, pp. 57-62 (1991)); multiple sclerosis (Hartung, H.-P., Ann. Neurol., vol. 33, pp. 591-6 (1993)); and AIDS (Matsuyama, T., et al, AIDS, vol. 5, pp. 1405-1417 (1991)). In addition, superoxide formation in leukocytes has been implicated in promoting replication of the human immunodeficiency virus (HIV) (Legrand-Poels, S., et al, AIDS Res. Hum. Retroviruses, vol. 6, pp. 1389-1397 (1990)).
It is well known that adenosine and some relatively non-specific analogs of adenosine decrease neutrophil production of inflammatory oxidative products (Cronstein, B. N., et al, Ann. N.Y. Acad. Sci., vol. 451, pp. 291-314 (1985); Roberts, P. A., et al, Biochem. J., vol.227, pp. 669-674 (1985); Schrier, D. J., et al, J. Immunol., vol. 137, pp. 3284-3289 (1986); Cronstein, B. N., et al, Clinical Immunol. and Immunopath., vol. 42, pp. 76-85 (1987); lannone, M. A., et al, in Topics and Perspectives in Adenosine Research, E. Gerlach et al, eds., Springer-Verlag, Berlin, pp. 286-298 (1987); McGarrity, S. T., et al, J. Leukocyte Biol., vol. 44, pp. 411-421 (1988); De La Harpe, J., et al, J. Immunol., vol. 143, pp. 596-602 (1989); McGarrity, S. T., et al, J. Immunol., vol. 142, pp. 1986-1994 (1989); and Nielson, C. P., et al, Br. J. Pharmacol., vol. 97, pp. 882-888 (1989)). For example, adenosine has been shown to inhibit superoxide release from neutrophils stimulated by chemoattractants such as the synthetic mimic of bacterial peptides, f-met-leu-phe (fMLP), and the complement component C.sub.5 a (Cronstein, B. N., et al, J. Immunol., vol. 135, pp. 1366-1371 (1985)). Adenosine can decrease the greatly enhanced oxidative burst of PMN first primed with TNF.alpha. and then stimulated by a second stimulus such as f-met-leu-phe (Sullivan, G. W., et al, Clin. Res., vol. 41, p. 172A (1993)). There is evidence that in vivo adenosine has anti-inflammatory activity (Firestein, G. S., et al, Clin. Res., vol. 41, p. 170A (1993); and Cronstein, B. N., et al, Clin. Res., vol. 41, p. 244A (1993)).
It has been suggested that there is more than one subtype of adenosine receptor on neutrophils that have opposite effects on superoxide release (Cronstein, B. N., et al, J. Clin. Invest., vol. 85, pp. 1150-1157 (1990)). The existence of A.sub.2a receptor on neutrophils was originally demonstrated by Van Calker et al (Van Calker, D., et al, Eur. J. Pharmacology, vol. 206, pp. 285-290 (1991)).
There has been progressive development of compounds that are more and more potent and selective as agonists of A.sub.2a adenosine receptors based on radioligand binding assays and physiological responses. Initially, compounds with little or no selectivity for A.sub.2a receptors were used, such as adenosine itself or 5'-carboxamides of adenosine, such as 5'-N-ethylcarboxamidoadenosine (NECA) (Cronstein, B. N., et al, J. Immunol., vol. 135, pp. 1366-1371 (1985)). Later, it was shown that addition of 2-alkylamino substituents increased potency and selectivity, e.g. CV1808 and CGS21680 (Jarvis, M. F., et al, J. Pharmacol. Exp. Ther., vol. 251, pp. 888-893 (1989)). Additionally, it has been reported that adenosine can decrease the rate of HIV replication in a T-cell line (Sipka, S., et al, Acta. Biochim. Biopys. Hung., vol. 23, pp. 75-82 (1988)). 2-Alkoxy-substituted adenosine derivatives such as WRC0090 are even more potent and selective as agonists on the coronary artery A.sub.2a receptor (Ukena, M., et al, J. Med. Chem., vol. 34, pp. 1334-1339 (1991)). The 2-alkylhydrazino adenosine derivatives, e.g. SHA 211 (also called WRC-0470) have also been evaluated as agonists at the coronary artery A.sub.2a receptor (Niiya, K., et al, J. Med. Chem., vol. 35, pp. 4557-4561 (1992)).
There is one report of the combination of relatively non-specific adenosine analogs, D-phenylisopropyladenosine (D-PIA) and 2-chloroadenosine (Cl-Ado) with a phosphodiesterase inhibitor resulting in a lowering of neutrophil oxidative activity (Iannone, M. A., et al, in Topics and Perspectives in Adenosine Research, E. Gerlach et al, Eds., Springer-Verlag, Berlin, pp. 286-298 (1987)). However, such non-specific analogs are actually more potent activators of A.sub.1 adenosine receptors than of A.sub.2a adenosine receptors and, thus, are likely to cause side effects due to activation of A.sub.1 receptors on other tissues causing effects such as "heart block".
Thus, there remains a need for a method for treating inflammation. There also remains a need for pharmaceutical compositions useful for treating inflammation.

Method for treating inflammatory diseases with A.sub.2a adenosine receptor agonists

University of Virginia Patent Foundation


A residual chlorine meter detects a pre-calibration residual chlorine reaction amount of sample water in a detection section, inputs a calibration standard residual chlorine standard concentration of the sample water in an input section, determines calibration factors from data for computing the calibration factors based on the amount and the concentration stored in a calibration factor computation data storage section in a calibration factor computation section, and stores the calibration factors in a calibration factor storage section. Then, the meter detects an on-measurement residual chlorine reaction amount of the sample water in the detection section and determines a post-calibration residual chlorine concentration of the sample water from data for computing the post-calibration residual chlorine concentration based on the on-measurement residual chlorine reaction amount and the calibration factor stored in a residual chlorine concentration computation data storage section in a residual chlorine concentration computation section.

(i) Field of the Invention
The present invention relates to a residual chlorine meter which can measure a residual chlorine concentration in water such as tap water, water for a pool or industrial water easily.
(ii) Description of the Related Art
Water such as tap water, water for a pool or industrial water is chlorinated for the sake of sanitation. To keep a residual chlorine concentration resulting from the chlorination at an appropriate level, means for measuring the residual chlorine concentration has been disclosed.
For instance, a residual chlorine concentration measuring device disclosed in Patent Publication 1 comprises pH measuring means (pH meter) and electric conductivity measuring means (electric conductivity meter) and determines calibration factors corresponding to the measured pH and electric conductivity of sample water based on calibration curves associated with the pH dependency and electric conductivity dependency of an apparent residual chlorine concentration resulting from normalization of a true value of a residual chlorine concentration and calibrates the apparent residual chlorine concentration of the sample water based on the calibration factors so as to determine a true residual chlorine concentration. The device is means for measuring a residual chlorine concentration with excellent measurement accuracy.
Meanwhile, a residual chlorine concentration measuring method disclosed in Patent Publication 2 is a method using a phosphate buffer having a more alkaline pH than 6.5 together with a liquid DPD reagent. The method is capable of measuring a residual chlorine concentration without any problem.    Japanese Patent Laid-Open Publication No. 9-329577    Japanese Patent Laid-Open Publication No. 2003-215119    However, the residual chlorine concentration measuring device disclosed in Patent Publication 1 has a problem that since the device has generally expensive measuring means including the pH meter and the electric conductivity meter, the device as a whole becomes expensive accordingly. Meanwhile, the residual chlorine concentration measuring method disclosed in Patent Publication 2 has a problem that it requires a high running cost and is troublesome to use since it uses a liquid DPD reagent for every measurement.
Thus, in view of such conventional problems, an object of the present invention is to provide a residual chlorine meter capable of measuring a residual chlorine concentration inexpensively and easily.


A hard disk drive that utilizes a third order curve fit equation to generate a skew table for a head of a hard disk drive. The skew table is used to compensate for offset between a write element and a read element of the head at different radial positions of a disk. A linear algebraic matrix equation can be used to solve the third order equation using offset values measured at different radial positions of the disk.

1. Field of the Invention
The present invention relates to a process to generate a skew table for a head of a hard disk drive.
2. Background Information
Hard disk drives contain a plurality of magnetic heads that are coupled to rotating disks. The heads write and read information by magnetizing and sensing the magnetic fields of the disk surfaces. Each head is attached to a flexure arm to create a subassembly commonly referred to as a head gimbal assembly ("HGA"). The HGA's are suspended from an actuator arm. The actuator arm has a voice coil motor that can move the heads across the surfaces of the disks.
Information is typically stored in radial tracks that extend across the surface of each disk. Each track is typically divided into a number of segments or sectors. The voice coil motor and actuator arm can move the heads to different tracks of the disks between the inner and outer diameters of the disks.
The tracks have servo information that is used to center the heads relative to the tracks. The heads typically have a write element for writing information and a separate read element for reading information on the disks. The read element is typically offset from the write element relative to the center of a track. Consequently, centering the read element will not necessarily center the write element and vice versa. The offset will vary for different radial positions of the disks.
Some disk drives have a skew table that contains offset values for various radial disk positions. The offset values in the skew table are used to adjust the position of the heads to center the read and write elements. The skew table is generated by measuring the offset at different radial positions. Offset values for non-measured radial positions are interpolated from the measured values. As track densities increase more measured offset values are required to generate an accurate skew table. Taking more measurements increases the time required to generate the skew table and the overall time to manufacture disk drives. An increase in process time decreases production efficiency for mass producing hard disk drives. It would be desirable to minimize the time required to generate a skew table.

Third order curve fit for head skew in a hard disk drive


A device, system and method for modifying workflows based on call profile inconsistencies is provided. A device monitors a call, received from a caller reporting an incident. The device performs one or more sentiment analysis and semantic analysis on one or more of: video of the caller on call; audio of the caller on call; and transcribed text of the audio of the call. The device determines a profile for the call, from a plurality of predetermined profiles stored at a memory accessible to the device, the plurality of predetermined profiles previously generated from historical data. In response to determining an inconsistency between the profile for the call and one or more of the sentiment analysis and the semantic analysis, the device determines a modified workflow for handling the call. The device provides, at a notification device, the modified workflow for handling the call reporting the incident.

In public safety and/or private security operations, call takers and/or dispatchers may need to make split-second decisions to ensure that the certain workflows are followed such that a correct responder gets to an incident scene.
Skilled artisans will appreciate that elements in the figures are illustrated for simplicity and clarity and have not necessarily been drawn to scale. For example, the dimensions of some of the elements in the figures may be exaggerated relative to other elements to help to improve understanding of embodiments of the present invention.
The apparatus and method components have been represented where appropriate by conventional symbols in the drawings, showing only those specific details that are pertinent to understanding the embodiments of the present invention so as not to obscure the disclosure with details that will be readily apparent to those of ordinary skill in the art having the benefit of the description herein.

Device, system and method for modifying workflows based on call profile inconsistencies


The invention illustrates a system and method of displaying a base image and an overlay image comprising: capturing a base image of a real event; receiving an instrumentation data based on the real event; identifying a visual segment within the base image based on the instrumentation data; and rendering an overlay image within the visual segment.

Typical television sport event coverage includes many video cameras covering different parts of the event. Some televised football games have as many as 20 video cameras covering the football field and are capable of providing a viewpoint from many different directions.
In many televised events, it is desirable to include overlays in portions of the televised broadcast. These overlays may include graphical or captured images such as scoreboards, games statistics, advertisements, logos, and play-by-play graphics. To produce a television program of a live event with overlays such as a football game, a large amount of manual input is typically required to create a television program displaying scenes of the football game in conjunction with the overlays placed in an appropriate position. For the overlays to be effective, they should not block an important portion of the live programming. However, positioning the overlays is typically a manually intensive operation.

System and method for data assisted chroma-keying


A disposable pant-type absorbent article includes a chassis and an absorbent body. The chassis has a front portion and a back portion and each of the front and back portions has a waist edge, a pair of leg edges and a pair of side edges. The front and back portions are joined to each other at opposite side edges to at least partly define a waist-opening and a pair of leg-openings. The absorbent body is located mainly in a crotch portion of the article. An elastic leg feature is fastened to the chassis at least partly along leg edges. The elastic leg feature extends from the associated side edge towards a centre line in the longitudinal direction of the article. The disclosure also relates to a corresponding method of manufacturing a pant-type absorbent article, as well as an array of gender-specific, disposable, absorbent articles.

There is general desire in the field of disposable pant-type absorbent articles to provide absorbent articles with increased comfort and fit and that provides the user with confidence with respect to its leakage protection. Document US 2005/0137563 describes a solution for reducing waste of materials while providing undergarments with elastic elements that fit the curve of the leg opening to provide additional reinforcement during use.
While the known disposable absorbent garment is satisfactory for its intended use, such a disposable absorbent garment is nonetheless susceptible to improvement.

Disposable pant-type absorbent article with a leg cuff


In order to provide a compact and low power consumption optical repeater capable of amplifying a plurality of wavelength ranges, the optical repeater is provided with: an excitation means which generates excitation light in a single wavelength range; a first light amplification means which is excited by the excitation light and the amplification band of which is a first wavelength range; and a second light amplification means which is excited by the excitation light and the amplification band of which is a second wavelength range different from the first wavelength range.

An optical repeater used in an optical submarine cable system is laid on the seabed, and thus, is required to have a small size and low power consumption. Meanwhile, in order to make a capacity of the optical submarine cable system large, an optical repeater capable of amplifying L-band signal light as well as widely used C-band signal light has been studied.
FIG. 6 is a block diagram illustrating a configuration example of an optical repeater 900 related to the present invention. The optical repeater 900 is used in an optical submarine cable system. The optical repeater 900 includes a C-band optical amplifier 91 and an L-band optical amplifier 92. In the present description, the C-band designates a wavelength band of approximately 1530 nm to 1565 nm, and the L-band designates a wavelength band of approximately 1570 nm to 1610 nm.
The C-band optical amplifier 91 includes a control circuit 911, an excitation light source 912, and optical amplification media 913 and 923. The optical amplification media 913 and 923 amplify C-band signal light, with excitation light generated by the excitation light source 912. In order to amplify C-band signal light, four laser diodes (LDs) of a wavelength band of 980 nm are used in the excitation light source 912.
The L-band optical amplifier 92 includes a control circuit 921, an excitation light source 922, and optical amplification media 914 and 924. The optical amplification media 914 and 924 amplify L-band signal light, with excitation light generated by the excitation light source 922. In order to amplify L-band signal light, four lasers of a wavelength band of 1480 nm are used in the excitation light sources 922. Excitation light output from the LDs included in the excitation light sources 912 and 922 excites the optical amplification media 913 and 914 through couplers and multiplexer/demultiplexers.
Signal light of a C-band and an L-band is wavelength-multiplexed and input from an up input (UP IN) in FIG. 6. The input signal light is separated into the C-band and the L-band. The signal light of the C-band is amplified by the optical amplification medium 913, and the signal light of the L-band is amplified by the optical amplification medium 914. The amplified C-band and L-band signal light is wavelength-multiplexed and output from an up output (UP OUT).
Similarly, signal light of a C-band and an L-band input from a down input (DOWN IN) in FIG. 6 is amplified by the optical amplification media 923 and 924. The signal light amplified by the optical amplification media 923 and 924 is output from a down output (DOWN OUT).
Note that in a relation to the present invention, PTLs 1 to 3 describe an optical amplifier including a plurality of optical amplification media.

Optical repeater and control method for optical repeater


A body of a car stereo has a first substrate and a detachable grille has a second substrate. The second substrate has an operating device, a display and a control unit where an identification data is stored. The first substrate has a grille attachment detecting device a power supply means and a control unit having an identification data discriminating device which discriminates the identification data in the control unit on the second substrate of the grille. When the identification data discriminating device determines that an appropriate grille is attached, the power is fed to the grille through the power supply device.

Increase in use of sophisticated car stereos in recent years has caused increase in the theft of the stereos. In order to prevent the theft of the stereo, there has been proposed measures for the stereos. In accordance with the one of the measures, a grille having a plurality of push button for a stereo is provided to be separated from the body of the stereo, and is detachably attached to the front portion of the stereo.
FIG. 4 shows the above described prior art. The car stereo 1 comprises a car stereo body 4 and a detachable grille 2 attached to the body 4. The body 4 has recessed panel 3 mounted at the front thereof. The panel 3 has a cassette opening 5 having a lid 6. A series of terminals 9 which are connected to terminals (not shown) provided at the back of the grille 2 when the grille is attached, are formed on the panel 3. The grille 2 has a cassette opening 7 which coincides with the cassette opening 5 of the body 4 and various operation keys 8.
In operation, a cassette tape is inserted into the car stereo body 4 through the cassette openings 7 and 5. A loading mechanisms provided in the body 4 is operated to carry the cassette to a playback position where the cassette is played.
In order to detach the grille 2 from the panel 3, the grille 2 is depressed at one side. A push-push mechanism (not shown) provided in the body 4 is operated to project the grille 2. The grille 2 is then pulled off the panel 3 of the body 4.
When the grille 2 is taken out, the stereo body loses the appearance as a car stereo. Hence, if the driver takes the grille 2 with him when leaving the vehicle, the theft of the car stereo is prevented.
When the grille 2 is attached on the panel 3 of the car stereo body 4, the terminals on the back of the grille 2 is connected to the terminals 9 of the body 4. A control unit provided in the car stereo body 4 detects that the grille 2 is attached when predetermined corresponding pairs of terminals of the grille 2 and the body 4 are connected with one another. The control unit is then operated to supply power to the terminals of the grille 2 from a battery provided in the body 4.
However, for convenience of manufacturing various stereos, it is required that the same parts may be used in common in various types of car stereo. As a result, the dimensions of grilles of various types are determined at the same value. Thus, a grille can be fit in a car stereo body which do not correspond to the grille. If a grille is erroneously attached to an unsuitable car stereo body, current inappropriately flows in electronic equipment in the car stereo body and the grille, which possibly damages them. In some case, since the terminals are arranged differently in each type of the grille, the car stereo may be erroneously operated because of erroneous connection of the terminals.

System for supplying power to a detachable grille of a car stereo


The invention is directed to methods of culturing rickettsiae in Ixodes scapularis cell lines. The methods of the invention provide for culture of microorganisms such as Anaplasma marginale, Ehrlichia canis, and Rickettsia rickettsii. A method of the invention involves incubating a rickettsia with an Ixodes scapularis tick cell culture in a culture medium under reduced oxygen and increased CO.sub.2 at a sufficient temperature until growth of the rickettsia is detected. The culture medium comprises a medium suitable for the growth of invertebrate cells supplemented with an organic buffer. The cell culture method can be used in large scale production of rickettsia containing products useful in diagnostic assays and vaccine preparations.

Rickettsiae with the exception of Rochalimaea are obligate intracellular parasites. As such, these microorganisms are often difficult to grow and the diseases they cause are difficult to diagnose. Because of the difficulty of growing these microorganisms, large scale preparation of vaccines and antigens has been costly and sometimes impossible. Many of the rickettsiae are the pathogenic agents of vector-transmitted diseases of human and animals. They are transmitted by invertebrate vectors such as ticks. Within rickettsiae, the microorganisms of the Anaplasma, Ehrlichia and Rickettsia genera are the causative agents of vector transmitted diseases and are difficult to grow, especially on a large scale.
Anaplasmosis is the only major tick-borne disease of cattle endemic in the United States. The causative agent, Anaplasma marginale, invades and multiplies in erythrocytes of cattle causing mild to severe anemia. Annual mortality and morbidity due to anaplasmosis among U.S. beef cattle has been estimated at 50,000 to 100,000 head and a cost of $300 million. The average weight loss during acute infections is 86 kg, with increased veterinary costs estimated at $52/head (Palmer in Veterinary Protozoan and Hemoparasite Vaccines, J. G. Wright (Ed.) CRC Press Inc., Boca Raton, Fla. 1989). The resultant loss of cattle for the export market has been estimated at $45 million annually (McCallon 1973).
Anaplasmosis can be transmitted biologically by several species of hard ticks (Dikmans Amer. J. Vet. Res. 38:5 (1950); Ewing, S. Transmission of Anaplasma marginale in Arthropods in Proc: 7th National Anaplasmosis Conference, Mississippi State MS p 395, 1981) or mechanically by blood-contaminated mouthparts of biting flies or fomites. In some geographical areas, transmission has been thought to occur primarily by biting flies and thus, the disease has been called "fly fever". However, ticks most likely play a major role in transmission of this disease. Male ticks are believed to contribute markedly to transmission of A. marginale. (Coan and Stiller, Abst. 70th Am. Pacific Branch Mtg. of Entomological Soc. Amer., Sand Diego, Calif. p.66 (1986); Coan et al., Ann. Western Conf. of Food Ani. Vet. Med., Boise, ID p.34 (1987); Coan et al., Proc. 8th National Vet. Hemoparasit. Dis. Conference, St. Louis, Mo. pp. 1161 (1989); Kocan et al., Am. J. Vet. Res. 53:499 (1992a); Kocan et al., J. Med. Entomol. 29:657 (1992b)). Male ticks can acquire infection as adults by feeding for short periods on infected cattle. Because male ticks are intermittent feeders, they readily detach and can reattach to other cattle during their life. Interhost transfer of male Dermacentor ticks among cattle has been clearly demonstrated. These male ticks have been shown to be persistently infected with A. marginale and are able to transmit the rickettsia repeatedly to susceptible cattle (Kocan et al. 1992b cited supra).
The widening distribution of anaplasmosis is probably contributed to by increased movement of cattle and the lack of sensitivity of the complement-fixation test. The complement-fixation test for anaplasmosis has been shown to lack sensitivity, especially in calves, carrier cattle and in cattle that have been treated with tetracyclines which lowers antibody titers often to undetectable levels (Goff et al., Vet. Microbiol. 24:381 (1990)). Thus, cattle may be shipped as serologically negative when they actually are active carriers of the organism. These carrier cattle are then likely to serve as reservoirs of infection for mechanical transmission by blood-contaminated fomites, biting flies or biologically by ticks.
Serologic diagnosis of anaplasmosis is predominantly accomplished by the complement-fixation test (USDA, A microtiter technique for complement fixation text for anaplasmosis, Manual pp. 1-16, USDA Animal and Plant Health Inspection Service, Beltsville, Md. (1968)). Many other tests have been developed to measure Anaplasma-specific antibodies, including the capillary-tube agglutination test (Ristic, J. Am. Vet. Assoc. 141:588 (1962)), card test (Amerault & Roby, J. Am. Vet. Assoc. 153:1828 (1968)), latex agglutination (Montenegro-James et al., Vet. Parasitology 8:241 (1981)), indirect fluorescent antibody test (IFAT) (Goff et al. 1990, cited supra), conventional ELISA (Shkap et al., Vet. Microb. 25:45 (1990)) and radioimmunoassay (Schunter & Leatch, Am. J. Vet. Res. 49:504 (1988)). Growth of A. marginale in cell culture may provide an inexpensive and more consistent antigen for use in serologic tests.
Control strategies for anaplasmosis in Oklahoma and elsewhere varies owing to the complexity of the disease and mode of transmission, and may include vector control, administration of tetracyclines, vaccination and maintenance of uninfected cattle. Current vaccines marketed in the U.S. are killed vaccines made from infected bovine blood. These vaccines reduce clinical symptoms but do not prevent infection. Vaccinated cattle that are challenge-exposed thereafter become carriers and their blood can serve as an infective source for ticks and/or mechanical transmission via biting flies and blood-contaminated instruments. Subunit vaccines which have been described provide only partial protection against challenge (Vidotto et al., Inf. and Imm. 62:2940 (1994)). In addition, many vaccine preparations can be contaminated with bovine red blood cell antigens as well as with other bovine pathogens.
Numerous attempts to grow A. marginale in a variety of mammalian and invertebrate cells have failed. Attempts included cell lines from D. variabilis ticks but none resulted in a continuous culture system capable of producing cattle-infective rickettsiae. A. marginale apparently does not invade nucleated cells in its vertebrate host, only erythrocytes. In contrast, the parasite develops in several types of nucleated tick cells (in midgut, gut muscle and salivary glands).
Erlichieae are rickettsial pathogens closely related to Anaplasma. Those species for which a biological vector is known are transmitted by ticks. While Anaplasma is confined to red cells in its mammalian host, the tick-borne Ehrlichieae infect white blood cells. Typically, Anaplasma and Ehrlichia are contained within membrane-bound vacuoles of their respective host cells.
The monocytic Ehrlichia species include Ehrlichia canis of dogs and Ehrlichia chaffeensis, E. risticii and E. sennetsu. These microorganisms are genetically related and cross-react serologically. The first Ehrlichia species recognized was E. canis. It occurs in all areas of the world where the vector tick, Rhipicephalus sanguineus (the brown dog tick), lives. The disease it causes is sometimes called canine tropical pancytopenia. It is a problem especially in all warm areas of the world, e.g., the southern U.S., Central and South America, the Mediterranean, South Asia (Ristic, M. and Huxsoll, D. L., 1984); Tribe II. Ehrlichieae Philip 1957. In: Bergey's manual of Systematic Bacteriology; Vol. 1, section 9). Ehrlichia canis can be cultivated in the dog cell line DH82 as well as human-dog hybrid cell lines (See: Rikihisa, Y. 1991. Clinical Microbiology Reviews, 4:286). Ehrlichia chaffeensis has been recognized only recently, and is associated with human ehrlichiosis. Maeda, K. et al., 1987, N. Eng. J. Med. 316:853; Dawson, J. E. et al., 1991, J. Clin. Microbiol. 29:2741). E. chaffeensis can also be cultured in DH82 cells. E. risticii is the causative agent of Potomac horse fever. This disease is known to occur in North America, France and India. E. risticii can be grown in macrophage-monocyte cell lines such as P388D.sub.1, T-84 and U937. E. sennetsu is the causative agent of Sennetsu erlichiosis of humans. E. sennetsu grows in mice and cell lines such P388D.sub.1 cells, L929 cells, and Hela cells.
Infections with monocytic Ehrlichiae can be diagnosed by direct microscopic examination and/or serodiagnosis. Treatment with antibiotics is effective. Two vaccines prepared from inactivated cell cultured E. risticii are commercially available for Potomac Horse Fever but none have been developed for large scale prevention of other ehrlichial diseases.
Rickettsia rickettsii is the causative agent of Rocky Mountain spotted fever. In the western U.S., the main vector of this human pathogen is the Rocky Mountain wood tick, Dermacentor andersoni. A close relative, Dermacentor variabilis also known as the American dog tick or common wood tick, is responsible for transmission in the eastern parts of the country. The disease is characterized by an acute febrile phase, typically with a disseminated rash. Unlike Anaplasma and Ehrlichia, which are passed only transstadially in ticks (from larvae to nymphs and/or nymphs to adults), R. rickettsii is transmitted transovarially and transstadially in the vector. After entry into the host cell, spotted fever rickettsiae do not remain in the phagosome, but quickly escape into the host cytoplasm, and commonly invade the nucleus. Their ability to move quickly from cell to cell by making use of the host cell's cytoskeletal elements is a characteristic feature leading to early dissemination to multiple tissues.
Vaccines against Rocky Mountain spotted fever were produced from inactivated organisms grown in embryonated hen's eggs, and more recently, in mammalian cell cultures. These vaccines are capable of preventing clinical illness, but not necessarily infection. Broad spectrum antibiotics, particularly chloramphenicol and tetracyclines, are highly effective when used promptly.
Thus there is a need to develop a culture system for growing rickettsiae in tick cell culture for use in diagnostics, and preparation of antigens and vaccines. There is a need to develop vaccines and antigen preparations that effectively protect against infection with Anaplasma marginale but that are not contaminated with bovine red blood cells or bovine pathogens. There is a need to develop a large scale culture system for preparation of large amounts of antigen from rickettsiae such as Anaplasma marginale and Ehrlichia canis for use in diagnostics and vaccines.

Method of growing rickettsiae in Ixodes scapularis tick cell culture and preparing antigens and vaccines of rickettsiae