A labeled polynucleotide is subjected to hybridization with oligonucleotide probes fixed respectively to regions on a supporting material. With a first restriction enzyme, a single-stranded moiety of the labeled polynucleotide having failed to form a double strand with each probe is separated from a double strand having been formed by the labeled polynucleotide and each probe. First detection data is obtained from each region. A mismatching moiety, at which the labeled polynucleotide and a certain probe have undergone a mismatch binding, is separated with a second restriction enzyme, and second detection data is obtained from each region. The region to which the mismatch polynucleotide was bound is specified in accordance with results of a comparison between the first detection data and the second detection data.

1. Field of the Invention
This invention relates to a method of detecting a mismatching region, in which a mismatch binding has occurred, on a biochemical analysis micro array used for detection, analysis, or the like, of a specific sequence contained in a polynucleotide, such as a DNA.
2. Description of the Related Art
DNA micro arrays are expected to be applied to a wide range of fields of life science, such as monitoring of genetic expression, determination of base sequences of genes, analysis of gene polymorphism (SNP), analysis of gene amplification or deletion at cancered parts, classification of diseases, such as cancers, prediction of drug response characteristics, and searching of disease genes.
Principles of assay techniques utilizing DNA micro arrays are based upon detection of nucleic acids through hybridization. Specifically, various different probe DNA's are arrayed at a high density at a plurality of regions of a surface of a supporting material, such as glass, silicon, or a membrane filter, and secured to the regions of the surface of the supporting material. Thereafter, a target DNA (i.e., a DNA having been labeled with a labeling substance) is subjected to hybridization with the probe DNA's having been fixed to the regions of the surface of the supporting material. Signals obtained from the regions (spots) are then detected in the manner described below.
For example, in cases where the target DNA has been labeled with a radioactive labeling substance, a stimulable phosphor layer of a stimulable phosphor sheet is exposed to radiation radiated out from the radioactive labeling substance, which is contained selectively in the regions of the supporting material. Thereafter, the stimulable phosphor layer is exposed to stimulating rays, which cause the stimulable phosphor layer to emit light in proportion to the amount of energy stored on the stimulable phosphor layer during the exposure of the stimulable phosphor layer to the radiation. The light emitted by the stimulable phosphor layer is detected photoelectrically. In this manner, the target DNA having been specifically bound to at least one of the probe DNA'S, which have been fixed to the regions of the surface of the supporting material, is detected.
In cases where the target DNA has been labeled with a fluorescent labeling substance, excitation light is irradiated to the regions of the supporting material, and the fluorescent labeling substance, which is contained selectively in the regions of the supporting material, is excited by the excitation light to produce fluorescence. The thus produced fluorescence is detected photoelectrically.
In cases where the target DNA has been labeled with a chemical luminescent labeling substance capable of producing the chemical luminescence when being brought into contact with a chemical luminescence substrate, the chemical luminescent labeling substance, which is contained selectively in the regions of the supporting material, is brought into contact with the chemical luminescence substrate. Also, the chemical luminescence produced by the chemical luminescent labeling substance is detected photoelectrically. (The aforesaid assay techniques are described in, for example, U.S. Patent Laid-Open No. 20020016009.)
The DNA micro arrays may be classified into two groups in accordance with the kinds of the DNA's, which are arrayed, and processes for producing the DNA micro arrays. One of the two groups is an oligonucleotide array produced with a process, wherein a light blocking plate referred to as a mask is overlaid on a silicon base plate, the silicon base plate is exposed to light via the mask by the utilization of photo-lithography, which is an exposure technique for semiconductors, the operation for exposing the silicon base plate to light via the mask is iterated, and DNA molecules are thereby superposed one by one on the base plate. (The oligonucleotide array produced with the process described above will hereinbelow be referred to simply as the oligonucleotide array.) The other group is a cDNA micro array produced with a process, wherein cDNA's having been subjected to PCR amplification previously are spotted onto slide glass by use of a thin pin, an ink jet technique, or the like.
Conditions (such as a temperature and a salt concentration) optimum for the hybridization may vary for the different kinds of the probe DNA's having been fixed respectively to the regions. However, it is not always possible to perform the reaction under the conditions optimum for each of the probe DNA's, which have been fixed respectively to the regions having been located at a high density. Therefore, ordinarily, the reaction is performed under the identical conditions with respect to all of the regions. Accordingly, it may often occur that a target DNA, which is not perfectly complementary to a certain probe DNA on the array and has a sequence similar to the perfectly complementary sequence, undergoes incorrect hybridization with the aforesaid certain probe DNA on the array. The incorrect hybridization described above is referred to as the mismatch binding. The target DNA, which has undergone the mismatch binding, causes noise to occur at the time of signal detection and adversely affects a detection accuracy.
In particular, the cDNA micro array is produced by directly subjecting the cDNA's, which have been isolated from cells of organisms, to the PCR amplification and fixed to the slide glass. The cDNA micro array is not produced by previously designing the probe DNA's so as not to undergo a mismatch bonding as in the cases of the oligonucleotide array. Therefore, the cDNA micro array has a high possibility that the probe DNA's on the cDNA micro array will under go the mismatch binding. Accordingly, in the cases of the cDNA micro array, the mismatch binding is suppressed through preparation of the probes located on the cDNA micro array. For such purposes, for example, sequences specific to a gene to be detected are selected, and oligo DNA's having been synthesized in accordance with the selected sequences are used as the probes.
However, in both the cases of the oligonucleotide array and the synthetic oligo array, it is not always possible to design such that the mismatch binding does not occur. Particularly, in cases where analysis is to be made with respect to a long gene, such as a cDNA, it is almost impossible to design such that the mismatch binding does not occur.
Attempts have been made to solve the problems with regard to the mismatch binding described above by, for example, finely adjusting the temperature or the pH value at the time of hybridization of a target DNA with probe DNA's having been fixed to an array. For example, a biochemical reaction detecting chip for the hybridization of a polynucleotide with oligonucleotide probes, with which biochemical reaction detecting chip the biochemical reaction is capable of being caused to occur at a temperature optimum for the hybridization at each of probe fixing surfaces, has been proposed in, for example, U.S. Patent Laid-Open No. 20020164778. The proposed biochemical reaction detecting chip is based upon characteristics concerning a melting out temperature (i.e., a Tm value) of a complementary strand binding of oligonucleotide probes. Specifically, under conditions of temperatures lower than the Tm value, background noise due to the mismatch binding increases. Also, under conditions of temperatures higher than the Tm value, it becomes difficult for the polynucleotide to undergo the binding with the probes. Therefore, with the proposed biochemical reaction detecting chip, a temperature, at which the polynucleotide is capable of undergoing the hybridization with a probe such that the mismatch binding does not occur, is adjusted at a value optimum for each of the probes.
Also, a technique for selectively separating and recovering a desired polynucleotide is proposed in, for example, U.S. Pat. No. 6,093,370. With the proposed technique for selectively separating and recovering a desired polynucleotide, oligonucleotide probes are fixed respectively to regions of a surface of a base plate, and polynucleotides are subjected to the hybridization with the oligonucleotide probes. Thereafter, only a specific region of the base plate is heated selectively, and only the polynucleotide, which has been complementarily bound to the probe, is separated from the probe.
However, with the technique for utilizing the biochemical reaction detecting chip proposed in U.S. Patent Laid-Open No. 20020164778, it is necessary that a plurality of islands are formed on the biochemical reaction detecting chip, and that the temperature adjustment is monitored finely for each of the islands. Also, in cases where the technique for selectively separating and recovering a desired polynucleotide, which is proposed in U.S. Pat No. 6,093,370, is utilized, it is necessary that only the specific region of the base plate is heated selectively. Further, the Tm values of the probes, which are contained in the islands or the regions described above, must be set previously at approximately identical values. As described above, with each of the technique for utilizing the biochemical reaction detecting chip proposed in U.S. Patent Laid-Open No. 20020164778 and the technique for selectively separating and recovering a desired polynucleotide, which is proposed in U.S. Pat. No. 6,093,370, complicated operations must be performed, and it is necessary for a specific apparatus to be utilized. Furthermore, in order for the mismatch binding to be suppressed, fine adjustment for raising or lowering the salt concentration in a liquid subjected to reaction must be made, and considerable labor and time are thus required. In cases where the hybridization is performed through adjustment of the pH value, the same problems as those described above arise.
As described above, in order for the mismatch binding during the hybridization to be suppressed, complicated adjustments must be performed, and considerable labor and time are required. Also, new problems occur in that, depending upon the conditions for the suppression of the mismatch binding, perfect match binding, i.e. perfect complementary binding, is weakened.
If a region, at which a target DNA having under gone the mismatch binding is located, is capable of being specified, for example, the thus specified region will be capable of being eliminated at the time of data analysis, and the detection accuracy will thus be capable of being kept high.


A new type of structure for mounting solar modules addresses the problem of solar access in several ways. Firstly, the structure can enclose a living plant and protect it and equipment. This allows improved acceptance and can provide a means for ground mounted solar to be classified as landscape by permitting authorities. This can make a large difference in the amount of area a land owner can use to mount solar modules. Also, the structure can make better use of recently developed solar technology like AC modules, microinverters and optimizers by allowing a solar designer to predefine a non-planar insolation surface that can be implemented using locally made materials. By allowing a custom non-planar design the solar plant support structure can more closely match loads, lower energy storage costs, energy transmission costs and make better use of limited solar access.

The present invention generally relates to the field of solar energy. More specifically, it relates to solar access, solar module mounting support and equipment protection. The invention also has secondary purposes that relate to fields associated with vegetative plant protection and support, shade providing structures and seating.
There are many options and methods for the mounting of solar modules yet the adoption of solar energy is still limited by many factors. These include electrical, geographical, economic, environmental, temporal as well as aesthetic limitations all of which could affect usefulness and adoption of solar energy equipment over conventional energy sources.
For photovoltaic modules specifically, when using a traditional string inverter it is a commonly accepted design practice to arrange solar modules within a plane or at the same azimuth and elevation angle since this optimizes the energy output. However, the proliferation of several technologies enables non-planar arrangement of a plurality of modules without a reduction in output associated with the electrical components of the solar equipment. This technology includes AC modules, DC modules with microinverters and also module level optimizers used with traditional string inverters. Most available solar support structure products are designed to meet the needs of traditional design practice and are limited in providing the design flexibility that microinverters, optimizers or AC modules allow.
The arrangement of solar modules particularly photovoltaic modules is also complicated by self shading. While humans may not be averse to shade in full sun, a small amount of shading or self shading of an array of solar modules can drastically reduce the output of modules or a series string of modules. A common traditional design is to mount strings of modules on planar frames at a specified azimuth and elevation that are arranged in rows and/or columns. Most mounting and or support products can be designed to support this arrangement. Unfortunately this leads to a classic design trade-off problem in using these products where the row spacing should be carefully selected to minimize shading that is caused by another row or column. There are currently limited available products or structural methods which can allow modules to be designed and arranged on a non-planar but continuous surface that can help minimize this self shading and make better use of microinverters, AC modules and optimizers. Implementing non-planar modules or modules with diversified azimuth angles has an additional advantage of possibly providing an energy production curve that more closely follows energy load. Energy load can vary throughout the day and year. By matching energy production to load, energy storage costs and/or transmission costs are minimized. This optimization can be done using existing technology but not without even greater challenges.
Solar module (Photovoltaic or other) height is also an important consideration in maximizing insolation and output. Adoption of solar energy often requires that sufficient insolation be available to the modules for years to come. Available locations are often affected by changing conditions such as the construction of new buildings or the growth of trees or other obstructions that are beyond the solar equipment owner or solar equipment operator's control. Positioning modules at sufficient height can often make a project more economically viability by increasing insolation over the life of the solar equipment. Height requirements further limit the range of products or structural methods that can address the above limitations.
While aesthetics do not directly affect utility, it is sometimes a key consideration in the adoption of solar. Often the economic viability of a solar equipment implementation is dependent on location. Specifically, if the solar equipment can be located near the end user an economic advantage is often achieved. Many end users can participate in net metering by an agreement with their energy provider or can store energy locally that can allow the end user to avoid cost at a retail energy rate. Additionally, some of an energy provider's transmission costs are avoided and the development of additional electrical distribution infrastructure can also be avoided. As an illustrative example of aesthetics playing an indirect role in utility let us consider a solar array for a residential property that is governed by a neighborhood association. A neighborhood association may be unlikely to approve a ground mount that occupies a large portion of a property owner's yard; however, it may be more inclined to approve a structure that integrates a plant that has inherent desirable aesthetic qualities or a structure that can provide additional value with shaded seating and integrated protection. Protection of solar equipment often requires a fence to protect sensitive and potentially dangerous equipment from animals or the general public. Also, many currently available options often sacrifice a large area of property to accommodate only the solar energy equipment and do not provide integrated shaded seating or the inherent aesthetics of a plant. By placing the solar energy equipment close to the end user as stated, transmission costs are avoided and the owner or operator may avoid some retail energy costs.
A variety of plant protection and support structures ranging from green houses and fences to trellises exist; however these are rarely designed to be integrated with a solar module support structure.
Shading and seating is obviously available but very often not specifically integrateable with solar energy equipment. Solar car parks and trees exist but are often limited by the arrangement of modules in a way that allows very little light to penetrate and are typically limited to the arrangement of modules in a single plane.
Finally, the durability of the support is also a major concern. For example, a designer may use a roof mounting solution on a roof that will only last a fraction of the expected lifetime of the solar components.
These and other limitations often present challenges for which few existing mounting solutions are ideal. Even fewer are readily integrated with the secondary purposes of the invention.


To provide a developer supply roller including: a shaft core; and an elastic layer over the shaft core, wherein the elastic layer is composed of at least a first elastic layer and a second elastic layer, the first elastic layer and second elastic layer made of elastic bodies with different moduli of elasticity, the first layer being an outermost layer of the supply roller, and wherein the relationships 0%≦k1≦20% and 20%≦k2≦70% are satisfied, where k1 is the modulus of repulsion elasticity of the first elastic layer and k2 is the modulus of repulsion elasticity (%) of the second elastic layer.

1. Field of the Invention
The present invention relates to a developer supply roller (hereinafter, sometimes referred to as "toner supply roller" or "supply roller") in a developing device used for electrophotographic image formation process and to an image forming apparatus using a non-magnetic one component developer and the supply roller.
2. Description of the Related Art
Various types of elastic rubber rollers are used in electrophotographic copiers and printers. For example, a toner supply roller for supplying toner to a developing roller is used in a developing device in which a latent electrostatic image formed on a photoconductor is developed. A given amount of toner is supplied from the toner supply roller to the developing roller, and a latent electrostatic image is developed by the toner deposited on the surface of the developing roller. To achieve stable image formation, the toner supply roller plays an extremely important role; for example, poor supply of toner from the toner supply roller to the developing roller will result in the formation of blurred copies with reduced image density. Further, the supply roller not only functions to supply toner to the developing roller, but also to recover toner on the developing roller, which has not been subjected to development. If toner on the developing roller is not recovered but remains on the developing roller for a prolonged period of time, the toner particles are rubbed against each other many times at a developer regulation portion and, as a result, the toner shows an abnormal level of charge amount. Moreover, the toner is degraded due to stress caused by being rubbed and adversely affects image quality. Therefore, it is now strongly demanded to develop a toner supply roller, which can always supply a fixed quantity of toner and can stably recover the toner on a developing roller.
In recent years, smaller image forming apparatus and higher printing speed have been demanded. As the apparatus becomes smaller, there is a necessity for miniaturizing a developing unit, which in turn results in a necessity for manufacturing a developing roller and a supply roller smaller in terms of their diameter. Further, it is necessary to increase the rotational speed of each roller when increasing the printing speed. Accordingly, rollers with smaller diameters require increased rotational speed. In this case, toner charge amount and toner transfer efficiency decreases.
Further, in association with the miniaturization of apparatus and the necessity for greater printing speed, the supply roller is required to be made smaller in diameter and rotated at high speed. In order to retain toner chargeability and transfer efficiency and to attain a stable image output with such smaller rollers, it is contemplated to increase the penetrating amount (indentation amount) of the supply roller with respect to the developing roller. When the indentation amount the supply roller to the developing roller is increased, toner is more efficiently scraped by the developing roller. However, this results in increased frictional force between the rollers and the rotational torque of each roller increases. Moreover, the rollers with small diameters have low heat capacity and generate heat due to friction, whereby the toner is heated to elevated temperatures and is thereby firmly attached to the developing roller or regulation blade, resulting in unstable image formation. Furthermore, increasing the indentation amount of the supply roller into the developing roller gives undue stress to toner, leading to degradation of toner stored in the developing unit. Thus, there is a fear that image quality decreases when the toner is used for a prolonged period of time.
On the other hand, a method has been contemplated for decreasing the modulus of repulsion elasticity of the supply roller in order to reduce the rotational torque of the supply roller. A decrease in the modulus of repulsion elasticity of the supply roller makes it possible to reduce the rotational torque. Low repulsion foams are excellent in impact absorption and vibration absorption. Particularly, where such foams are employed as a material of a supply roller and a cleaning roller, they are in contact with a developing roller and photoconductor at a uniform contact pressure and thereby local stresses are relieved. Thus, excellent toner scraping property can be expected at low contact pressures. However, foams are less resilient after application of strain and thus it is difficult to stably form a nip between the rollers. Foams are particularly difficult to employ as a high-speed rotating body. A simple decrease in the modulus of repulsion elasticity of the supply roller will make a nip portion unstable, thereby adversely affecting the toner chargeability and transfer efficiency and cause image unevenness.
In an attempt to solve the foregoing problems Japanese Patent Application Laid-Open (JP-A) No. 2006-184602 has proposed a toner supply roller composed of a metal shaft, a first layer and a second layer, wherein the first layer (shaft side) is greater than the second layer (surface side) in average pore diameter. However, considering the fact that foams with larger average pore diameters have smaller modulus of elasticity, this proposal will result in the formation of unstable nip portion and unstable toner supply by a supply roller.
Further, JP-A No. 2006-154537 has proposed a supply roller in which an appropriate modulus of repulsion elasticity of the supply roller measured in accordance with JIS K6400 is in a range of 30% to 60%. With this proposal however, it is still difficult to make the contact pressure of the supply roller against the developing roller and photoconductor uniform for relieved local stresses, to keep the nip portion stable, and to provide stable toner chargeability and excellent toner efficiency at the same time.
Neither of the above Patent Literatures can fully solve the above problems, i.e., increased frictional resistance with an increase in the indentation amounts of the rollers, increased rotational torque, accelerated toner degradation, and heat generation from the rollers. On the other hand, where the elasticity moduli of the developing roller and supply roller are reduced, the rotational torque can be reduced. However, this raises another problem of unstable nip portion and unstable toner supply by the supply roller, leading to poor toner charge amount and poor toner transfer efficiency that may cause image unevenness.

Developer supply roller and image forming apparatus


A method and apparatus for peeling protective adhesive tape 3 from work having the protective adhesive tape 3 applied to a surface of a semiconductor wafer W supported in a ring-shaped frame F by means of a supporting adhesive tape 1. Thin plates 4 with non-adhesive upper surfaces are applied to the supporting adhesive tape 1 adjacent a peel starting end 3a and a peel finishing end 3b of protective adhesive tape 3, respectively, to form regions for avoiding contact between a peeling tape 5 wound around an applying roller 8 and the like and the supporting adhesive tape 1. Subsequently, the peeling tape 5 is applied to the protective adhesive tape 3 at the peel starting end 3a with the applying roller 8, and a group of rollers 6-8 is moved horizontally on the work A to peel the peeling tape 5 and protective adhesive tape 3 together from the surface of semiconductor wafer W.

As processing steps executed after circuit patterns are formed on the surface of a semiconductor wafer, those shown in Japanese Patent Publication (Unexamined) H5-63077, for example, have been proposed heretofore.
The processing steps executed after the circuit patterns are formed include a back grinding step for cutting and grinding the back of the wafer to a required thickness, a dicing step for cutting and dividing the wafer into individual elements, and so on. Since the wafer undergoes mechanical stress in each of these steps, a separate step is provided for applying adhesive tape to the surface of the wafer to protect the circuit patterns on the surface of the wafer and fix the wafer in position.
The flow of these steps is approximately as follows. First, in the back grinding step, (1) a protective adhesive tape is applied to the surface of the wafer on which circuit patterns are formed. (2) After the wafer surface to which the protective adhesive tape has been applied is supported by a suction member and the wafer is fixed in position, its back is cut and ground. With the protective adhesive tape applied to the wafer surface, the next, adhesive tape applying and peeling steps are executed. (3) The protective adhesive tape on the surface of the wafer is peeled after a supporting adhesive tape having strong adhesion is applied to the back of the wafer and the wafer is mounted in a ring-shaped frame by means of this supporting adhesive tape. In the next dicing step, (4) the wafer is cut and divided into individual elements by a diamond cutter rotating at high speed.
Through the above flow of steps, the process after the circuit patterns are formed on the semiconductor wafer surface is performed. Details of the above step (3) for peeling the protective adhesive tape from the wafer surface will be described with reference FIGS. 25 through 27. First, as shown in FIG. 25, a wafer W mounted in a ring-shaped frame F by means of supporting adhesive tape 1 is suction-supported on a suction table 2. A draw roller 6 and a pinch roller 7 on which a peeling tape 5 drawn from a supply source not shown is wound, and an applying roller 8, are arranged slightly above a peel starting end 3a of protective adhesive tape 3 applied to the surface of wafer W. Next, as shown in FIG. 26, the applying roller 8 is lowered and biased to said peel starting end 3a to bond the peeling tape 5 and protective adhesive tape 3 together. Then, as shown in FIG. 27, the pinch roller 7 is lowered to the draw roller 6, and the three rollers 6, 7 and 8 are moved in the direction of a peel finishing end 3b ( afterward in FIG. 27). At this time, the applying roller 8 moves while pressing the peeling tape 5 on the protective adhesive tape 3, and the draw roller 6 peels the protective adhesive tape 3 by means of the peeling tape 5.
However, the above step of peeling the protective adhesive tape has the problem set out below. That is, when the applying roller 8 is lowered and biased to the peel starting end 3a to bond the peeling tape 5 and protective adhesive tape 3 together, since, as is well-known, the wafer W is very thin, with the thickness approximately 110-450 .mu.m, as shown in FIG. 28, for example, the supporting adhesive tape 1 exposed adjacent the peel starting end 3a of protective adhesive tape 3 and the peeling tape 5 lowered and biased by the applying roller 8 from above the peel starting end 3a have adhesive surfaces thereof stick to each other. When the three rollers 6, 7 and 8 are moved in this state to continue the peeling operation, the wafer W undergoes bending stress and the wafer W could crack.
To avoid the above inconvenience, as shown in FIG. 29, for example, the applying roller 8 may be lowered and biased slightly inwardly of the peel starting end 3a of protective adhesive tape 3 in the direction of the center of protective adhesive tape 3 to bond the peeling tape 5 and protective adhesive tape 3 together. Then, the peeling tape 5 is not applied to the peel starting end 3a of protective adhesive tape 3. The protective adhesive tape 3 cannot be peeled smoothly when the three rollers 6, 7 and 8 are moved in the direction of the peel finishing end 3b (leftward in FIG. 29).
This invention has been made having regard to the state of the art noted above, and its object is to provide a method and apparatus for peeling a protective adhesive tape from a semiconductor wafer reliably, without a peeling tape and a supporting adhesive tape applied to the back of the wafer sticking to each other.

Method and apparatus for peeling protective adhesive tape from semiconductor wafer


In an optical waveguide reduction optical image sensor constituted by a polymeric optical waveguide array having a plurality of cores arranged into a one-dimensional array in a cladding substrate, and a photoelectric conversion element array for detecting light propagating through the polymeric optical waveguide array and converting the detected light into an electrical signal, a hollow thermal expansion suppressing member (jig) is mounted on the joint portion between the polymeric optical waveguide array and the photoelectric conversion element array, and the coefficient of linear thermal expansion of the thermal expansion suppressing member in the core array direction of the polymeric optical waveguide array is smaller than that of the polymeric optical waveguide array in the same direction.

(1) Field of the Invention
The present invention relates to an optical waveguide reduction optical image sensor using optical waveguides, which is used for a one-dimensional read optical system for producing hard copy and the like and is applied to a facsimile, a copying machine, an image scanner, and the like.
(2) Description of the Prior Art
With the recent increase in image readers such as facsimiles, image scanners, and digital copying machines, demands have arisen for a high-performance, compact one-dimensional image sensor for converting image information into an electrical signal. The conventional one-dimensional image sensors include a reduction optical image sensor for reading a focused image by using a reduction optical system such as lenses, and a contact type image sensor for reading a one-to-one focused image by using a one-to-one optical system.
The reduction optical image sensor is inexpensive and capable of high-speed read. However, since this sensor must perform a reduction imaging operation by using lenses, the apparatus size becomes large, resulting in difficulty in reducing the size. In addition, complicated adjustment is required for the optical system of each apparatus.
In contrast to this, the contact type image sensor does not require complicated adjustment for the optical system because the distance from an original to the photoelectric conversion element array is short. However, the size of the photoelectric conversion element array is large, and a complicated electronic circuit is required to drive the photoelectric conversion element array. For this reason, it is difficult to attain a reduction in cost.
Under the circumstances, a reduction image sensor using an optical waveguide array has recently been proposed. This sensor is constituted by a microlens array having a width almost equal to the width of an original surface, an optical waveguide array constituted by a plurality of waveguide cores used to guide light focused by the microlens array and formed in a cladding substrate, and a photoelectric conversion element array for receiving light and converting it into an electrical signal. According to this reduction image sensor using the optical waveguide array, reductions in the cost and size of the apparatus can be realized, and complicated adjustment for the optical system is not required.
Several methods of manufacturing the optical waveguides of such a reduction image sensor using a polymeric material for cores are known.
One method is a sandwich method. According to this method, a polymer precursor material for the cores of waveguides is filled in the groove portions of a pattern substrate made of a polymeric material such as PMMA and having groove patterns, and a flat substrate made of a polymer such as PMMA is brought into tight contact with the groove portions of the pattern substrate. Thereafter, photopolymerization is caused by ultraviolet radiation to form the cores of the optical waveguides made of the polymeric material.
Another method is a vacuum capillary method. According to this method, the pattern surface of a pattern substrate on which groove patterns serving as capillaries are formed is brought into tight contact with a flat substrate to form capillaries. Thereafter, a polymer precursor as a material for the cores of optical waveguides is filled in the capillaries by capillarity, and the polymer precursor is polymerized.
According to these methods, since there is no gap between the pattern substrate and the flat substrate, polymeric optical waveguides having excellent optical waveguide characteristics can be realized, which are free from crosstalk caused by light leaking from the gaps between the cores.
In the reduction optical image sensor using the polymeric optical waveguide array, an original surface is illuminated by an LED in an image read operation, and information on the original surface is guided to the photoelectric conversion elements through the core portions of the optical waveguides, thereby reading image information.
In the above conventional optical waveguide reduction optical image sensor using a polymeric material, if the coefficient of linear thermal expansion of a polymeric material such as PMMA for the polymeric optical waveguide array constituted by a plurality of cores differs from that of the photoelectric conversion element array, a pitch offset occurs between the polymeric optical waveguide array and the photoelectric conversion element array at the joint portion therebetween with a change in temperature. For this reason, a pixel offset occurs, and an output from each photoelectric conversion element deteriorates. An image cannot therefore be properly read.
When such polymeric optical waveguides are to be used, therefore, it is very important to prevent a decrease in image read precision due to a change in temperature.

Optical waveguide reduction optical image sensor


The image heating apparatus for heating an image formed on a recording material, comprises a flexible sleeve; a driving roller which contacts an outer peripheral surface of said sleeve and rotates said sleeve; a sliding member which contacts an inner peripheral surface of said sleeve and forms said driving roller together with a nip portion by nipping said sleeve between thereof; and an inner surface regulating member comprising an opposing area opposed to the inner peripheral surface of a generatrix direction end portion of said sleeve; wherein the recording material is nipped and conveyed by the nip portion; and wherein a outlineoutline of the opposing area of said inner surface regulating member is approximately similar to the outlineoutline of the end face of said sleeve when said sleeve is rotated by said driving roller in a state in which said inner surface regulating member is not attached to said apparatus. In this way, the durability of the flexible sleeve can be enhanced.

1. Field of the Invention
The present invention relates to an image heating apparatus suitable if used as a fixing apparatus mounted on a copying machine or a printer, and in particular, it relates to an image heating apparatus using a flexible sleeve.
2. Related Background Art
A description will be made with a fixing apparatus in an image forming apparatus such as an electrophotographic copying machine and a printer as an example. In general, in the image forming apparatus, as the fixing apparatus (a fixer), which heats and fixes an unfixed toner image on a recording material surface as a permanent fixed image indirectly (transferred) or directly formed and borne on a recording material (paper) by an adequate image forming process means such as the electrophotographic process, a heating apparatus of a heat roller system has been conventionally in wide use.
In recent years, an apparatus of a film heating system has been put into practical use in view of a quick start and energy saving. Further, the heating apparatus of an electromagnetic induction heating system, which allows a film composed of a metal to heat itself, has been proposed.
a) Fixing Apparatus of Film Heating System
The fixing apparatus of the film heating system, for example, has been proposed in Japanese Patent Application Laid-Open Nos. S63-313182, H02-157878, H04-44075, H04-204980, and the like. That is, a pressure welding nip portion (hereinafter referred to as fixing nip portion) is formed by nipping a heat resisting film (hereinafter referred to as fixing film) between a ceramic heater as a heating body and a pressure roller as a pressure member, and a recording material forming and bearing an unfixed toner image is introduced between the fixing film of the fixing nip portion and the pressure roller, and is nipped and conveyed together with the fixing film, so that the unfixed toner image is fixed on the surface of the recording material by an applied pressure of the fixing nip portion, while giving a heat of the ceramic heater through the fixing film.
This fixing apparatus of the film heating system can constitute an on-demand type apparatus by using a member of low heat capacity for the ceramic heater and the film, and may generate heat of a predetermined fixing temperature by turning on the ceramic heater of a heat source only when an image formation is performed, and has advantages in that a waiting time from turning on the power source of the image forming apparatus until reaching an image formation executable state is short (quick startability), and a power consumption during a standby time period is sharply reduced (power saving) and the like.
Further, in the publications of among the above described Japanese Patent applications, those described in Japanese Patent Application Laid-Open Nos. H04-44075 and H04-204980, the film (flexible sleeve) by driving the pressure roller (pressure roller driving type), and comparing to the type which allows the flexible sleeve to rotate by providing a driving roller and a tension roller in the interior of the flexible sleeve, have an advantage of being simple in the structure.
b) Fixing Device of Electromagnetic Induction Heating System
For example, in the Japanese Patent Application Laid-Open No. H07-114276, there has been disclosed an induction heat fixing device, in which an eddy current is induced in a metallic layer (heat generating layer) of a fixing film by magnetic flux, and heat is generated by its joule heat. This device can make the fixing film to directly generate heat by utilizing the generation of the induced current, and achieves a fixing process of higher efficiency than the fixing device of the heat roller system which has a halogen lamp as its heat source.
In FIG. 14 is shown a schematic structure of an example of the fixing apparatus of the electromagnetic induction heating system which has improved its efficiency by concentrating the alternating magnetic flux of an excitation coil into a fixing nip portion. This fixing apparatus comprises a fixing film (flexible sleeve) 10 having a metallic layer, a film guide member 16C disposed in the inner face of this film, a magnetic field generating means 15 having an excitation coil 18 and a magnetic core 17, and a pressure roller 30 allowing the film 10 to rotate. In this example, a nip portion N conveying a recording paper P is formed by the film guide member 16C and the pressure roller 30. When the excitation coil 18 is energized, an eddy current is generated in the metallic layer of the fixing film 10, and the fixing film generates heat. By this heat, a toner image t on the recording paper P is heated and fixed.
As described above, both of the fixing device of the system which heats the toner image by a heater through the flexible sleeve and the fixing device of the system which heats the toner image by heating the flexible sleeve itself use the flexible sleeve.
In such a fixing device using the flexible sleeve, particularly in the device using a flexible sleeve made of a metal, it is necessary to take into consideration a breakage of sleeve due to metal fatigue. Further, from among the devices using the above described flexible sleeve, the device of a pressure roller drive type is simple in structure, and moreover, a load given to the flexible sleeve can be made small. However, particularly in case the flexible sleeve made of a metal is used, even if it is the device of the pressure roller drive type, it is necessary to take into consideration a breakage of sleeve due to metal fatigue. For example, when the sleeve 10 is driven by the pressure roller 30, the sleeve 10 approaches one side in a generatrix direction, and in case its approaching force is strong, a buckling stress is generated in the end portion in the generatrix direction, thereby accelerating a fatigue phenomenon. Alternatively, since a large frictional force acts on the end portion by its approaching force, the sleeve 10 is unable to aptly slide on the surface against which the sleeve 10 abuts, and the sleeve 10 is locally over deformed, thereby accelerating the fatigue phenomenon.
Thus, the flexible sleeve made of a metal is prone to break, comparing to the flexible sleeve made of resin such as polyamide, and consequently, various designs have been made to consider and control the metal fatigue of the flexible sleeve.
Incidentally, the present inventors have found a new cause which invites the metal fatigue of the flexible sleeve.
This cause is such that, when the sleeve 10 is driven by the pressure roller 30, though the sleeve is deformed into a shape as natural as it desires, it is forced to be partially reformed into another shape, and as a result, the stress partially acting on the sleeve becomes large, so that the fatigue phenomenon is accelerated.

Image heating apparatus having flexible sleeve


We have generated novel molecularly marked FMDV A24LL3DYR and A24LL3BPVKV3DYR vaccine candidates. The mutant viruses contain a deletion of the leader coding region (LL) rendering the virus attenuated in vivo and negative antigenic markers introduced in one or both of the viral non-structural 3Dpol and 3B proteins. The vaccine platform includes unique restriction endonuclease sites for easy swapping of capsid proteins for different FMDV subtypes and serotypes. The mutant viruses produced no signs of FMD and no shedding of virulent virus in cattle. No clinical signs of disease or fever were observed and no transmission to in-contact animals was detected in pigs inoculated with live A24LL3DYR. Cattle immunized with chemically inactivated vaccine candidates showed an efficacy comparable to a polyvalent commercial FMDV vaccine. These vaccine candidates used in conjunction with a cELISA provide a suitable target for DIVA companion tests.

1. Field of the Invention
This invention relates to a rationally designed engineered attenuated antigenic marker vaccine-virus production platform comprising a deletion of the Lpro coding sequence resulting in complete attenuation and mutations (negative markers) introduced in two non-structural viral proteins resulting in the elimination of two antigenic epitopes recognized by specific antibodies, one located in 3B and the other in 3D, thus providing a target for DIVA (Differentiation of naturally Infected from Vaccinated Animals) serological tests. The attenuated marker vaccine production virus also comprises unique restriction endonuclease sites flanking the capsid-coding region to facilitate the replacement of the capsid region making possible the exchange of cassettes representing relevant capsid coding regions of different serotypes and subtypes of FMDV field isolates for the design of custom vaccines.
2. Description of the Relevant Art
Foot and mouth disease (FMD) is an extremely contagious viral disease of cloven-hoofed ungulates which include domestic animals (cattle, pigs, sheep, goats, and others) and a variety of wild animals. The most prominent disease symptoms in FMDV-infected cattle include vesicular lesions of the epithelium of the mouth, tongue, teats and feet. Although some countries, among them United States, Canada, Mexico, Australia and most of Europe, are considered to be free of FMD, the disease is distributed worldwide and has a great economic impact on the export industry. Indeed, several economically devastating outbreaks have occurred over the past decade on almost every continent.
Control methods to eradicate FMD depend upon the prevalence of the disease in particular geographical regions/states and often include mass annual prophylactic vaccination campaigns and the application of stringent zooprophylactic measures following outbreaks. A chemically inactivated whole virus vaccine has been used to contain the disease, but it is slow acting and does not always permit distinction between infected and vaccinated animals. Indeed, in recent years the differentiation of infected animals from those that have been vaccinated is of paramount importance as a protective activity following emergency vaccination. Historically, the use of non-structural viral protein as serological indicators of viral replication has been widely applied. Among these proteins, the highly conserved FMDV 3D polymerase (3Dpol) of 52-KDa has been identified as the main determinant of infection and has been called the FMD-Virus Infection-Associated Antigen (FMD-VIAA; Berger et al. 1990. Vaccine 8:213-216; Bergmann et al. 1993. Am. J. Vet. Res. 54:825-831; Cowan and Graves. 1966. Virology 30:528-540; McVicar and Sutmoller. 1970. Am. J. Epidemiol. 92:273-278; Sorensen et al. 1998. Arch. Virol. 143:1461-1476). Studies by Newman and Brown (1997. J. Virol. 71: 7657-7662; Newman et al. 1994. Proc. Natl. Acad. Sci. USA 91:733-737) suggested that purified 140S FMDV preparations contain small quantities of 3Dpol and therefore, could account for seroconversion to 3Dpol in animals that have received inactivated FMDV vaccines.
Previous strategies to select highly attenuated vaccines for FMDV have relied on the selection of less-pathogenic variants produced by serial passages of the virus in non-natural hosts such as embryonated chicken eggs and rabbits (Giraudo et al. 1990. Virology 177:780-783; Xin et al. 2009. Vet. Microbiol.). Those empirical strategies failed when tested in susceptible species due to reversion to virulence by the mutant viruses harboring point-mutations and therefore, were not pursued for being too risky (Sutmoller, P. 2001. Rev. Sci. Tech. 20:715-722; Sutmoller et al. 2003. Virus Res. 91:101-144). Modern approaches to produce genetically engineered FMDV with altered virulence have relied on the deletion of the cell-receptor binding site (Mason et al. 1994. Proc. Natl. Acad. Sci. USA 91:1932-1936; McKenna et al. 1995. J. Virol. 69:5787-5790; Rieder et al. 1996. Proc. Natl. Acad. Sci. USA 93:10428-33), the viral leader coding sequence (Lpro, Piccone et al. 1995. J. Virol. 69:5376-82) or elements within the non-translated region (NTR) (Rodriguez et al. 2009. J. Virol. 83:3475-3485). The FMDV Lpro together with the 3Cpro and 2A proteinases play an important role in processing of the viral polyprotein. In addition to cleaving itself from the nascent polyprotein, Lpro cleaves the eukaryotic initiation factor 4G (eIF4G) causing inhibition of the cellular translation machinery. Lpro is also known to relocate to the nucleus in the FMDV-infected cells and to induce degradation of nuclear factor kappa B (NF-κB) with the consequent inhibition of the innate immune response (de Los Santos et al. 2006. J. Virol. 80:1906-1914). FMDVs of serotype A lacking Lpro have been shown to be infectious, to grow more slowly in BHK-21 cells (A12-LLV2, (Piccone et al., supra) and be attenuated for pigs (Brown et al. 1996. J. Virol. 70:5638-5641; Chinsangaram et al. 1998. Vaccine 16:1516-1522; Mason et al. 1997. Virology 227:96-102).
Currently killed-antigen FMDV vaccines are produced in expensive biological containment facilities, by growing large volumes (thousands of liters) of virulent FMDV that has been adapted to grow in cells, which can be sometimes difficult. This process has resulted in escape of virulent virus from the manufacturing facility causing costly outbreaks in livestock (see Cottam et al. 2008. PLoS Pathogen 4:1-8). After growth, virus is then inactivated using chemicals and antigen concentrates are prepared, followed by purification steps required to remove contaminant proteins, making it difficult to differentiate infected from vaccinated animals (DIVA) through serological diagnostic tests. There is little to no cross protection across serotypes and subtypes requiring the appropriate matching between vaccine and circulating field strains to achieve protection. Despite these shortcomings of the vaccines, billions of doses are manufactured every year around the world. Their use has been the basis for eradicating FMDV from Europe and for controlling the disease in many parts of the world through mass vaccination campaigns. Thus, there is an urgent need for the development of effective marker FMDV vaccine candidates with DIVA capabilities.

Development of a marker foot and mouth disease virus vaccine candidate that is attenuated in the natural host

The United States of America, as represented by The Secretary of Agriculture


A disposable bag making apparatus that comprises a container for holding an elongate flexible plastic tube. The container includes a first wall and an aperture in the first wall through which a free end of the compacted elongate tube can project. A cutter is formed on the container whereby any desired length of the tube can be pulled through the aperture and brought into contact with the cutter and severed to form various sized disposable bags.

Various shapes and designs have been utilized to construct a container capable of dispensing disposable refuse or storage bags. Conventional containers of this type may be manufactured from rigid plastic materials that are neither flexible nor compact, and must be large enough to house an established number of plastic bags for dispensing. Such containers are normally too large to carry on one's person and generally are manufactured from expensive plastics involving complex and costly manufacturing techniques. In fact, some containers and bags are environmentally hazardous, requiring special disposal.
Furthermore, conventional containers typically dispense flexible bags of a predetermined size and are incapable of dispensing tubular lengths of flexible polymeric material that may be formed into disposable bags of various selected sizes. For example, although a number of conventional bags used for garbage, groceries and the like may be disposable, all are dispensed in predetermined sizes regardless of the consumer's preference for various or different sized disposable bags. Thus, a consumer is required to purchase more disposable bags than may be necessary for the consumer's particular needs at the time, resulting in unnecessary expense to the consumer. Therefore, the prior art is replete with various containers, dispensers and the like, that are limited to dispensing disposable bags of predetermined sizes from containers that are rigid, bulky, costly and not easily disposed of.
Accordingly, the disadvantages of the prior art are overcome by the present invention which provides a disposable bag making apparatus that is capable of dispensing various select lengths of an elongate flexible plastic tube that may be subsequently formed into disposable bags and is compact, lightweight and environmentally safe for disposal after use.


A fan impeller system has a rotor element (10), a multiplicity of fan impeller blades (30) and a multiplicity of adapter elements (20). The rotor element (10) has a multiplicity of receptacles (11) that are evenly distributed in a circumferential direction (U) with an adapter element (20). The adapter element (20) is arranged between the respective fan impeller blade (30) and the respective receptacle (11). The adapter elements (20) may differ at least in their shape and/or dimensions. The fan impeller blades (30) may differ at least in their shape and/or dimensions.

A large number of different variants of fan impeller has been known for a long time in the art.
Mostly, however, a provision is made to provide a specific one-piece fan impeller or a specific multi-part fan impeller for one application or for a specific application. Thus, different fan impellers are provided for different applications and a large number of tools must be provided for their production. This results in high production costs and logistical challenges or problems, since the fan impellers, the parts, and the tools have to be stored and managed, which is costly.
The disclosure is therefore based on the object that the aforementioned drawbacks can be overcome and a wide variety of fan impellers can be provided in a simple manner for a wide variety of applications.

Fan impeller system for configuring and constructing a fan impeller


A method of operating a substrate processing chamber that includes, prior to a substrate processing operation, flowing a seasoning gas comprising silane and oxygen into said chamber at a flow ratio of greater than or equal to about 1.6:1 oxygen to silane to deposit a silicon oxide film over at least one aluminum nitride nozzle exposed to an interior portion of the chamber. Also, a substrate processing system that includes a housing, a gas delivery system for introducing a seasoning gas into a vacuum chamber, where the gas delivery system comprises one or more aluminum nitride nozzles exposed to the vacuum chamber, a controller and a memory having a program having instructions for controlling the gas delivery system to flow a seasoning gas that has an oxygen to silane ratio greater than or equal to about 1.6:1 to deposit a silicon oxide film on the aluminum nitride nozzles.

During chemical vapor deposition (CVD) of silicon oxide and other layers onto the surface of a substrate, the deposition gases released inside the processing chamber may cause unwanted deposition on areas such as the walls of the processing chamber. Unless removed, this unwanted deposition is a source of particles that may interfere with subsequent processing steps and adversely affect wafer yield.
To avoid such problems, the inside surface of the chamber is regularly cleaned to remove the unwanted deposition material from the chamber walls and similar areas of the processing chamber. This procedure may be performed as a standard chamber dry clean operation where an etchant gas, such as nitrogen trifluoride (NF3), is used to remove (etch) the deposited material from the chamber wall and other areas. During the dry clean operation, the chamber interior is exposed to a plasma from the etchant gas so that the etchant gas reacts with and removes the deposited material from the chamber walls. Such cleaning procedures are commonly performed between deposition steps for every wafer or every n wafers.
The clean step can, in itself, be a source of particle accumulation however. Fluorine from the clean plasma can be adsorbed and/or trapped in the chamber walls and in other areas of the chamber such as areas that include ceramic lining or other insulation materials. The trapped fluorine can be released during subsequent processing steps (e.g., by reacting with constituents from the plasma in a high density plasma CVD (HDP-CVD) step) and can be adsorbed in subsequently deposited silicon oxide layers.
To prevent such fluorine adsorption and to provide protection against other contaminants within the chamber walls (e.g., the diffusion of metal fluorides) a CVD chamber is often "seasoned" after the dry clean operation. Such seasoning includes depositing a thin silicon oxide layer over the chamber walls before a substrate is introduced into the chamber for processing. The deposited silicon oxide layer covers the chamber walls, reducing the likelihood that contaminants will interfere with subsequent processing steps. After deposition of the seasoning layer is complete, the chamber is used for 1 to n substrate deposition steps before being cleaned by another clean operation as described above and then reseasoned.
The dielectric properties of the seasoning layer also provide an insulating barrier that prevents electrical arcing between the plasma and the walls of the chambers. The chambers typically include conductive materials (e.g., metals), and when the electric potential of the plasma exceeds a threshold level it begins to ground itself by discharging through the exposed conductive surfaces of the chamber. This discharging can damage the chamber by ablating away parts of the conductive surface. Coating these exposed conductive surfaces with the seasoning film reduces (or prevents) plasma arcing.
A conventional method used to season deposition chambers includes the steps of forming a plasma from a process gas of silane (SiH4), oxygen (O2), and argon (Ar). In one particular implementation of this previously known method, the preferred flow ratio of oxygen to silane used in the method is 1.375:1. O2 is introduced at 110 sccm, SiH4 is introduced at 80 sccm and Ar is introduced at 20 sccm. This seasoning process has been used to protect subsequent processing steps form contaminants in the HDP-CVD chamber.
Semiconductor device geometries have dramatically decreased in size since such devices were first introduced decades ago. Today's wafer fabrication plants are routinely producing integrated circuits having 0.13 micron feature sizes and smaller.
As device sizes become smaller and integration density increases, issues that were not previously considered important by the industry are becoming a concern. Also, improved control over criteria such as particle generation and contaminant control is necessary to ensure that deposited layers meet stringent manufacturers specifications. In order to meet processing demands created by such small scale geometry devices, new technology for substrate processing equipment is constantly being developed.
Another trend in semiconductor fabrication is for a larger numbers of copies of an integrated circuit design to be formed on a single substrate. For example, the current industry standard is to form integrated circuits on circular silicon wafers having a 200 mm diameter. Current trends in the industry, however, favor forming ICs on larger, 300 mm diameter wafers. The larger wafers allow more copies of an IC to be fabricated in a single process than could be accommodated on the smaller 200 mm wafers.
However, the larger wafers can place extra demands on the fabrication chamber, including the need to operate the chamber at higher power to generate an adequate sized plasma for depositing a film on the larger substrate. The higher operating powers can heat-up chamber components to temperatures that may hit or exceed high-temperature thresholds for those components. Thus, there is a need for components that can operate in the high power environments used with larger substrates that also have the low particle generation characteristics needed for efficient production of today's densely packed ICs.

HDP-CVD seasoning process for high power HDP-CVD gapfil to improve particle performance


The present invention provides a method of delivering immunogenic or therapeutic proteins to bone marrow cells using alphavirus vectors. The alphavirus vectors disclosed herein target specifically to bone marrow tissue, and viral genomes persist in bone marrow for at least three months post-infection. No or very low levels of virus were detected in quadricep, brain, and sera of treated animals. The sequence of a consensus Sindbis cDNA clone, pTR339, and infectious RNA transcripts, infectious virus particles, and pharmaceutical formulations derived therefrom are also disclosed. The sequence of the genomic RNA of the Girdwood S.A. virus, and cDNA clones, infectious RNA transcripts, infectious virus particles, and pharmaceutical formulations derived therefrom are also disclosed.

The Alphavirus genus includes a variety of viruses all of which are members of the Togaviridae family. The alphaviruses include Eastern Equine Encephalitis virus (EEE), Venezuelan Equine Encephalitis virus (VEE), Everglades virus, Mucambo virus, Pixuna virus, Western Equine Encephalitis virus (WEE), Sindbis virus, South African Arbovirus No. 86 (S.A.AR 86), Girdwood S.A. virus, Ockelbo virus, Semliki Forest virus, Middelburg virus, Chikungunya virus, O'Nyong-Nyong virus, Ross River virus, Barmah Forest virus, Getah virus, Sagiyama virus, Bebaru virus, Mayaro virus, Una virus, Aura virus, Whataroa virus, Babanki virus, Kyzylagach virus, Highlands J virus, Fort Morgan virus, Ndumu virus, and Buggy Creek virus.
The alphavirus genome is a single-stranded, messenger-sense RNA, modified at the 5'-end with a methylated cap, and at the 3'-end with a variable-length poly (A) tract. The viral genome is divided into two regions: the first encodes the nonstructural or replicase proteins (nsP1-nsP4) and the second encodes the viral structural proteins. Strauss and Strauss, Microbiological Rev. 58, 491-562, 494 (1994). Structural subunits consisting of a single viral protein, C, associate with themselves and with the RNA genome in an icosahedral nucleocapsid. In the virion, the capsid is surrounded by a lipid envelope covered with a regular array of transmembranal protein spikes, each of which consists of a heterodimeric complex of two glycoproteins, E1 and E2. See Paredes et al., Proc. Natl. Acad. Sci. USA 90, 9095-99 (1993); Paredes et al., Virology 187, 324-32 (1993); Pedersen et al., J. Virol. 14:40 (1974).
Sindbis virus, the prototype member of the alphavirus genus of the family Togaviridae, and viruses related to Sindbis are broadly distributed throughout Africa, Europe, Asia, the Indian subcontinent, and Australia, based on serological surveys of humans, domestic animals and wild birds. Kokemot et al., Trans. R. Soc. Trop Med. Hyg. 59, 553-62 (1965); Redaksie, S. Afr. Med. J. 42, 197 (1968); Adekolu-John and Fagbami, Trans. R. Soc. Trop. Med. Hyg. 77, 149-51 (1983); Darwish et al., Trans. R. Soc. Trop. Med. Hyg. 77, 442-45 (1983); Lundstrom et al., Epidemiol. Infect. 106, 567-74 (1991); Morrill et al., J. Trop. Med. Hyg. 94, 166-68 (1991). The first isolate of Sindbis virus (strain AR339) was recovered from a pool of Culex sp. mosquitoes collected in Sindbis, Egypt in 1953 (Taylor et al., Am. J. Trop. Med. Hyg. 4, 844-62 (1955)), and is the most extensively studied representative of this group. Other members of the Sindbis group of alphaviruses include South African Arbovirus No. 86, Ockelbo82, and Girdwood S.A. These viruses are not strains of the Sindbis virus; they are related to Sindbis AR339, but they are more closely related to each other based on nucleotide sequence and serological comparisons. Lundstrom et al., J. Wildl. Dis. 29, 189-95 (1993); Simpson et al., Virology 222, 464-69 (1996). Ockelbo82, S.A.AR86 and Girdwood S.A. are all associated with human disease, whereas Sindbis is not. The clinical symptoms of human infection with Ockelbo82, S.A.AR86, or Girdwood S.A. are a febrile illness, general malaise, macropapular rash, and joint pain that occasionally progresses to a polyarthralgia sometimes lasting from a few months to a few years.
The study of these viruses has led to the development of beneficial techniques for vaccinating against the alphavirus diseases, and other diseases through the use of alphavirus vectors for the introduction of foreign DNA. See U.S. Pat. No. 5,185,440 to Davis et al., and PCT Publication WO 92/10578. It is intended that all United States patent references be incorporated in their entirety by reference.
It is well known that live, attenuated viral vaccines are among the most successful means of controlling viral disease. However, for some virus pathogens, immunization with a live virus strain may be either impractical or unsafe. One alternative strategy is the insertion of sequences encoding immunizing antigens of such agents into a vaccine strain of another virus. One such system utilizing a live VEE vector is described in U.S. Pat. No. 5,505,947 to Johnston et al.
Sindbis virus vaccines have been employed as viral carriers in virus constructs which express genes encoding immunizing antigens for other viruses. See U.S. Pat. No. 5,217,879 to Huang et al. Huang et al. describes Sindbis infectious viral vectors. However, the reference does not describe the cDNA sequence of Girdwood S.A. and TR339, nor clones or viral vectors produced therefrom.
Another such system is described by Hahn et al., Proc. Natl. Acad. Sci. USA 89:2679 (1992), wherein Sindbis virus constructs which express a truncated form of the influenza hemagglutinin protein are described. The constructs are used to study antigen processing and presentation in vitro and in mice. Although no infectious challenge dose is tested, it is also suggested that such constructs might be used to produce protective B- and T-cell mediated immunity.
London et al., Proc. Natl. Acad. Sci, USA 89, 207-11 (1992), disclose a method of producing an immune response in mice against a lethal Rift Valley Fever (RVF) virus by infecting the mice with an infectious Sindbis virus containing an RVF epitope. London does not disclose using Girdwood S.A. or TR339 to induce an immune response in animals.
Viral carriers can also be used to introduce and express foreign DNA in eukaryotic cells. One goal of such techniques is to employ vectors that target expression to particular cells and/or tissues. A current approach has been to remove target cells from the body, culture them ex vivo, infect them with an expression vector, and then reintroduce them into the patient.
PCT Publication No. WO 92/10578 to Garoff and Liljestrom provide a system for introducing and expressing foreign proteins in animal cells using alphaviruses. This reference discloses the use of Semliki Forest virus to introduce and express foreign proteins in animal cells. The use of Girdwood S.A. or TR339 is not discussed. Furthermore, this reference does not provide a method of targeting and introducing foreign DNA into specific cell or tissue types.
Accordingly, there remains a need in the art for full-length cDNA clones of positive-strand RNA viruses, such as Girdwood S.A and TR339. In addition, there is an ongoing need in the art for improved vaccination strategies. Finally, there remains a need in the art for improved methods and nucleic acid sequences for delivering foreign DNA to target cells.

System for the in vivo delivery and expression of heterologous genes in the bone marrow

The University of North Carolina at Chapel Hill


This invention relates to an isolated nucleic acid fragment encoding a squalene metabolic enzyme. The invention also relates to the construction of a chimeric gene encoding all or a portion of the squalene metabolic enzyme, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the squalene metabolic enzyme in a transformed host cell.

The terpenoids constitute the largest family of natural products with over 22,000 individual compounds of this class having been described. The terpenoids play diverse functional roles in plants as hormones, photosynthetic pigments, electron carriers, mediators of polysccharide assembly, and structural components of membranes. Farnesyl pyrophosphate is converted to squalene in the first dedicated step towards sterol biosynthesis. Squalene is then converted to squalene-2,3-epoxide which, in photosynthetic organisms, is converted cycloarterenol.
Squalene monooxidase (EC 126.96.36.199), also referred to as squalene epoxidase, is an oxidoreductase which acts on paired donors with incorporation of molecular oxygen. This enzyme is located at the endoplasmic reticulum, and catalyzes the conversion of squalene to squalene 2,3-epoxide in the pathway to produce sterol. Squalene monooxygenase may be the rate limiting step in sterol biosynthesis. Oxygen, NADPH, FAD, and a cytosolic protein are required for squalene monooxygenase function. Squalene monooxygenase together with lanosterol synthase was formerly known as squalene oxydocyclase.
Whereas vertebrates and fungi synthesize sterols from epoxysqualene through the intermediate lanosterol, plants cyclize epoxysqualene to cycloartenol as the initial sterol. This reaction is catalyzed by cycloartenol synthase (EC 22.214.171.124), also called 2,3-epoxysqualene-cycloartenol cyclase.
Sequences encoding peptides with similarities to cycloartenol synthase and squalene monooxygenase are found in the NCBI database having General Identifier Nos. 5566676, 5468642, 5706248, 5608422, 5055959, 2309779, 2310417, 3107671, 702331, and 4874404.


A thin film solar cell thermal radiator configuration uses an embedded power bus to collect vertically migrating waste heat from electronic devices mounted on a thin film multilayer printed circuit board. The waste heat is vertically collected, horizontally spread, and vertically rejected through the a top coverglass glass layer around the side of a top silver contact of conventional thin film solar cells. The thermal radiator configuration enables the collection and rejection of the electronic component waste heat for maintaining thin film solar cells within desired temperature ranges.

The aerospace industry has developed designs for microsatellites and nanosatellites. This development of small space systems is likely to increase with new technologies that can be used to incorporate multiple functions in a small satellite. In U.S. Pat. No. 6,300,158 titled Integrated Solar Power Module, a method is described for producing thin film solar cells that are integrated with a multilayer printed wiring board and power processing electronics. However, the integrated solar power module does not address the problem of rejecting excessive heat produced by the active electronic components that are mounted on the back of the thin film solar cell substrate.
Active electronic components convert some of the electrical energy passing through the electronic components into heat. Most electronic components have maximum junction operating temperatures in the 80° to 125° C. range. The operating temperature of the thin film solar cells is in the 70° to 80° C. range. These thin film solar cells must reject excessive heat produced by the mounted electronic devices to maintain an operating temperature below the maximum operating temperature for the thin film solar cells. Further refinements of the integrated solar power module may have some portions of the thin film solar cell coated with a material that would have a low solar radiation absorptance and high thermal emissivity for maintaining the solar cells within a desirable temperature range. The entire solar cell assembly and current collectors are covered with a transparent polymer coverglass, which provides a front surface that has a high thermal emissivity. The existing design of the thin film solar cells presently has large silver current collectors located on the front of the solar cells. The silver collectors are silver bus bars that are highly reflective and have a low solar absorptance. The bus bars and solar cell structure obstruct thermal transmission through the solar cell. The amount of additional thermal energy that can be rejected from a thin film solar cell that has mounted active heat producing components is limited. These and other disadvantages are solved or reduced using the invention.